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PALMITIC ACID
OCCURRENCE, BIOCHEMISTRY
AND HEALTH EFFECTS
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PALMITIC ACID
OCCURRENCE, BIOCHEMISTRY
AND HEALTH EFFECTS
LUCAS F. PORTO
EDITOR
New York
CONTENTS
Preface
vii
Chapter 1
Chapter 2
17
45
Chapter 3
Chapter 4
Chapter 5
105
Chapter 6
125
Chapter 7
145
Chapter 8
Chapter 9
Index
63
159
211
219
PREFACE
This book discusses the occurrence, biochemistry, and health effects of palmitic acid.
Chapter 1 - Fatty acids have traditionally been described as artery clogging species that is
detrimental to overall health. The most prevalent fatty acid is palmitic acid (PA), a sixteen
carbon chain fatty acid that is ubiquitous in biological systems. PA is prevalent in most
eukaryotic cell membranes and in the mitochondria derived from the Krebs cycle utilizing
acetyl-coenzyme A as its precursor. PA is found in a variety of plants with a high amounts in
coconut oil. Many cosmetics, shampoos, and commercialized beauty products contain PA
providing structure and substance to the gel or reagent.
An emerging field of study is the esterified form of PA or methyl palmitate, as it is
involved in biological signaling in the central nervous system. More specifically, methyl
palmitate or palmitic acid methyl ester can cause arterial vasodilation and is thought to be
involved in neurotransmission, as well as modulate vascular tonicity in cerebral circulation.
Methyl palmitate has also been implicated as a neuroprotective agent in both models of focal
and global cerebral ischemia; however, the exact mechanism(s) are still unknown. The
authors will focus on the known pharmacology, biochemistry, and clinical implications of PA
and other related fatty acids (i.e. Non-esterified v. esterified fatty acids) commonly found in
daily diets. Additionally, cellular target(s) of PA will be discussed as it relates to
improvement of disease states, synthesis, and possible health implications/benefits of methyl
palmitate in biological systems.
Chapter 2 - Palmitic acid (PA), one of the most abundant saturated fatty acid (SFAs)
within plants, humans, animals, microbial (bacteria), fungal, and marine organisms,
constitutes ~16 to 45 % of the lipid profile. The impressive abundance of PA throughout
nature could be attributed partly to its critical role in membrane lipid structural functionality,
formation of subcellular cysteine residue linkages and RNA posttranslational modifications in
eukaryotic and prokaryotic cells. PA has been demonstrated to undergo -oxidation to
produce short and medium chain fatty acids to maintain homeostasis in response to
endogenous and exogenous cues. Further, non-esterified PA is known to mediate numerous
biochemical and antimicrobial pathways towards the betterment of human health. Although
the importance of PA in human health and nutrition are established, critics attest that
excessive dietary PA may be unhealthy and even detrimental. Current microbiological and
epidemiological studies suggest that PA produces specific health benefits that are not
common to all other SFAs. For example, studies suggest that PA may activate nitric oxide
and superoxide, thus functioning as an antimicrobial agent against some strains of bacteria,
viii
Lucas F. Porto
algae, and helminthes and certain foodborne pathogens. This chapter reviews the occurrence,
biochemistry, and subsequent health implications of PA.
Chapter 3 - In the last few decades, disagreement between opinions and findings
concerning the ability of palmitic acid (PA) and other saturated fatty acids (SFAs) to raise
cholesterolaemia has led to discussions on whether PA, which has been positively related to
high serum cholesterol levels, could increase the risk of cardiovascular diseases. This study
aims to review the PA content of meat, dairy products, fish, and other food of animal origin in
the human diet and discusses nutritional issues related to the occurrence of this fatty acid
(FA) in these foods due to different diet supplementation. Meat and dairy products are
considerable dietary sources of SFAs, such as PA. In most industrialized countries, a high
meat or dairy intake contributes to a higher than recommended SFA intake. Palmitic and
myristic acids are common FAs in meat and dairy products, making up about 30-40% of total
FA intake and are the main factors responsible for raising cholesterol levels; indeed, strong
evidence indicates that these two SFAs increase serum cholesterol concentrations in humans.
Stearic acid is partially converted to oleic acid in vivo and has not been shown to elevate
blood cholesterol, while lauric acid is not as potent as PA at raising concentrations of total
cholesterol and LDL cholesterol in humans. The occurrence of PA in animal origin food is
influenced by both genetic and environmental factors, such as the composition of the animals
diet, its digestive system and its biosynthetic processes. The FA profile in food of animal
origin mainly reflects dietary lipid sources and has the potential to play a valuable role in
human nutrition by manipulating the composition of animal fat through diet. In order to
explain the variability in FA composition in food of animal origin, this review examines
different nutrition trials that have studied the effects of PA supplementation on the lipid
profile of animal origin food.
Chapter 4 - Palmitic acid or hexadecanoic acids is the most abundant saturated fatty acid
in human nutrition and represents about 17.6g per day in the United Kington diet. It is the
first fatty acid produced during the lipogenesis. During this process, glucose is converted to
fatty acids, which then react with glycerol to produce triacylglycerols. Palmitic acid mainly
occurs as its ester in triglycerides, especially in palm oil (40-44 %) but also in lard (20-30 %),
dairy products (25-40 %) and cocoa butter (25-27 %). One of the main applications of
palmitic acid in the food industry has been the formulation of interesterified fats, used as a
replacement of trans fats. In breast milk, native lard, enzyme-directed and randomly
chemically interesterified plant fats, palmitic acid is predominantly esterified to
triacylglycerol, center or -position, in native palm oil and cows milk, it is mainly at the
external or -positions. A higher palmitic acid absorption is obtained with formulas rich in
palmitic acid esterified in triacylglycerol sn-2 position, than with those containing palmitic
acid predominantly esterified in the sn-1,3 positions. These specific fatty acids distributions in
triacylglycerol, determine the physical properties of the fat, which affects its absorption,
metabolism and distribution into tissues. Many authors claim that a palmitic acid intake may
promote increased risk of hypercholesterolemia, liver disease, type 2 diabetes, insulin
resistance and toxicity. However, more recent investigations on the topic seem to have
reconsidered the negative role of the dietary saturated fatty acids as a risk factor for
cardiovascular diseases and show that not only the type of fat, but also that the triglyceride
structure plays a role in these diseases.
Chapter 5 - Current dietary recommendations are based on a reduced saturated fatty acid
(SFA) consumption to prevent cardiovascular disease (CVD). The role of individual SFA in
Preface
ix
metabolic disease is not fully understandable. One type of SFA present in many common
foods (dairy, meat, palm and coconut oil) is palmitic acid (16:0). A number of
epidemiological studies have shown that the populations who consume large amounts of
atherogenic SFA (especially palmitic, myristic, lauric) have elevated levels of LDL and HDLcholesterol. Saturated fatty acid exert their atherogenic and thrombogenic effect through
increased production of LDL, very-low-density lipoproteins particles and apolipoproteins A1,
with a decrease of LDL- receptors specific activity, and an increase in platelet aggregation.
The total cholesterol/ HDL-cholesterol ratio, the best overall indication of potential effects on
coronary heart disease (CHD) risk is nonsignificantly affected by consumption of palmitic
acid (PA). Compared with lipid effects, the influence of SFA intake on inflammation markers
is less well explored. The associations between circulating and tissue PA and dietary intake of
PA are diverse and most likely reflecting endogenous metabolism. Status of PA is not in
intakeresponse relationship biomarker, probably partly due to conversion of 16:0 to 16:1 by
steaoryl-CoA-desaturase (SCD-1). Increased SFA intake has been associated with increased
SCD-1 activity in which may predict mortality. Palmitoylation is the process involved in
proteinmembrane interactions and signal transduction. Increases in dietary intake of PA
decrease fat oxidation and daily energy expenditure with slight increases in adiposity.
Evidence for the effects of SFA, particularly PA consumption on insulin resistance, vascular
function, type 2 diabetes, and stroke is various. It is considered that circulating PA, as
nonesterified fatty acids stimulate insulin resistance by decreasing phosphorylation of the
insulin receptor and insulin receptor substrate-1. In muscle cells, PA decrease oxidation of
fatty acids and glucose which elevates fatty acid and glucose levels in tissues and blood, and
decreases adiponectin production, which may both promote insulin resistance. It was shown
that 16:0 and 14:0 stimulate -cells and endothelial dysfunction. The incidence of type 2
diabetes was associated with total SFA levels of plasma cholesterol esters (also demonstrated
for 16:0 levels independently) and phospholipids (also for 16:0 and 18:0). In skeletal muscle
phospholipids, PA has been negatively associated with insulin sensitivity and diabetes type 2.
Systematic reviews on prospective cohort studies indicated that CHD risk has not been
directly associated with SFA intake, although is associated with a dietary habits, high in SFArich foods. Taken together, there is collective convincing evidence for decreased CHD risk
when replacing SFA with polyunsaturated fats. Differences in cardiometabolic risk appear
greater between food groups and overall dietary patterns rather than between separate SFA.
Chapter 6 - Palmitic acid (C16:0) is one of the major fatty acids (FAs) forming virtually all
natural lipids. Both in eu-, and prokaryotes, C16:0 forms various lipid classes, which serve
either as the lipid background of storage fats and oils, or the hydrophobic matrix of cell
membranes, or the components of cuticle waxes and polymers. Non-esterified 16:0 does not
occur in living cells, and it is present there only as an acyl residue in various lipid classes,
such as mono-, di-, and triacylglycerols, glyco-, phospho-, and sphingolipids, wax and steryl
esters etc., where it esterifies the hydroxy groups of glycerol backbone or other alcohols
(sphingosine, higher and lower aliphatic alcohols etc.). Palmitic acid is known to be a primary
higher FA synthesized in the cell, while nearly all other FAs of natural lipids are the products
of its further modification caused by elongation, desaturation, insertion of various functional
groups, such as methyl, hydroxy, oxo, epoxy, etc. As a saturated FA, C16:0 is used by the cell
for regulating its functional state by shifting the membrane fluidity under adverse
environmental conditions and thus providing a necessary molecular species composition of
the membrane polar lipids. Among the latter, such classes as phosphatidylinositols,
Lucas F. Porto
phosphatidylserines, and other highly polar lipids are particularly rich in palmitic acid. In
accordance, its content in plant lipids rises as they became less TLC-mobile, more difficultly
extractable, or tightly bound. It is evident that further screening of plant lipids as regards this
index is of considerable interest.
Chapter 7 - Human breast milk provides the optimum nutrition for infants. Designed to
provide balanced nutrition, human breast milk naturally meets the needs of growing infants in
the first months after birth. In human breast milk, and in most infant formulas, approximately
50% of the energy is supplied to newborns as fat, of which more than 98% is in the form of
triglycerides. Triglyceride synthesis occurs in the mammary gland. The fatty acids are
specifically positioned to sn1, sn2 or sn3 positions on the glycerol backbone to yield the
structure-specific triglycerides that are found in human milk. Palmitic acid (C16:0) is the
predominant saturated fatty acid, comprising 17-25% of the fatty acids in mature human milk.
Surprisingly, the positioning of palmitic acid is highly conserved across populations, and
approximately 70-75% of palmitic fatty acids are esterified to the sn2 position of the
triglyceride (sn2 palmitate).
Clinical and pre-clinical studies over the last three decades have provided increasing
evidence that this specific positioning of palmitic acid on the triglycerides in human milk has
a significant holistic effect on optimal infant development and well-being that is related to the
increased absorption of both palmitic acid and calcium: softer stools, increased bone strength,
increased beneficial gut flora, controlled intestinal health, and reduced infant crying. All of
these contribute to the benefits of infant wellbeing.
The overall aim of the current review is to expand the understanding of the role of
palmitic acid and its specific sn2 position in infant nutrition.
Chapter 8 - Palmitic acid, either in its triglyceride form or hydrolysed as a free fatty acid
or an ester, needs to be extracted from its source, processed and isolated to obtain useful
products. The objective of this work is to consider the use of SCF (supercritical fluid)
processing as a method to extract and process palmitic acid and/or its derivatives. A phase
behaviour analysis, in supercritical CO2, ethane and propane, at temperatures close to the
critical point of the solvent show moderate solubility of palmitic acid and tripalmitin at
pressures below 50 MPa and total solubility of methyl and ethyl palmitate at pressures below
25 MPa. Analysis of the phase behaviour considered and studies presented in the literature
have shown that SCFs can be widely applied to the processing of palmitic acid containing
compounds. In particular SCFs can fractionate a mixture of acids or their derivatives
according to the chain length, it can de-acidify an edible oil and it is able to fractionate a
mixture containing palmitic acid and other compounds. Additionally, SCFs can also be used
to extract palmitic acid containing triglycerides from plant material. SCFs, in particular CO2,
are thus excellent alternative solvent to traditional organic solvents and should be considered
when processing palmitic acid containing products.
Chapter 9 - In this review the major saturated fatty acid, palmitic acid, of Virgin Olive
Oil (VOO) was studied. This oil is one of the oldest known vegetable oils and it plays a
fundamental role in human nutrition around the Mediterranean basin. This nature juice is the
only edible oil of great production obtained by physical methods from the fruit Olea europaea
L. Consideration of VOO as a natural functional fat is related to the presence of palmitic acid.
Updating of its levels in Virgin olive oils throughout the Tunisian olive oil as well as
information on expecting levels in other products from olive tree establish the authors view
point. Studies on levels palmitic acid upon maturity stage in the oil are also discussed.
Preface
xi
Major analytical practices are given in brief. Palmitic acid (C16:0) is the principal
saturated fatty acid in olive oil, responsible for its figeability at low temperature.
Few are the exceptions as palmitic acid content depends heavily on the genetic factor.
Palmitic fatty acids, important for the nutritional properties of an olive oil, showed a crucial
rule in the characterization of olive oils.
In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 1
ABSTRACT
Fatty acids have traditionally been described as artery clogging species that is
detrimental to overall health. The most prevalent fatty acid is palmitic acid (PA), a
sixteen carbon chain fatty acid that is ubiquitous in biological systems. PA is prevalent in
most eukaryotic cell membranes and in the mitochondria derived from the Krebs cycle
utilizing acetyl-coenzyme A as its precursor. PA is found in a variety of plants with a
high amounts in coconut oil. Many cosmetics, shampoos, and commercialized beauty
products contain PA providing structure and substance to the gel or reagent.
An emerging field of study is the esterified form of PA or methyl palmitate, as it is
involved in biological signaling in the central nervous system. More specifically, methyl
palmitate or palmitic acid methyl ester can cause arterial vasodilation and is thought to be
involved in neurotransmission, as well as modulate vascular tonicity in cerebral
circulation. Methyl palmitate has also been implicated as a neuroprotective agent in both
models of focal and global cerebral ischemia; however, the exact mechanism(s) are still
unknown. We will focus on the known pharmacology, biochemistry, and clinical
implications of PA and other related fatty acids (i.e. Non-esterified v. esterified fatty
acids) commonly found in daily diets. Additionally, cellular target(s) of PA will be
discussed as it relates to improvement of disease states, synthesis, and possible health
implications/benefits of methyl palmitate in biological systems.
Corresponding author: Hung Wen Lin, Ph.D. University of Miami, Miller School of Medicine, Department of
Neurology, Cerebral Vascular Disease Research Center, Two Story Laboratory (TSL), Medical Campus,
Locator: D4-5, 1420 N.W. 9th Avenue, Miami, FL 33136. E-mail: hwlin@med.miami.edu.
Keywords: Palmitic acid methyl ester, methyl palmitate, stearic acid, stearic acid methyl
ester, cerebral ischemia, stroke, vasodilation
INTRODUCTION
High fatty acid content has been commonly associated with an increased risk of
cardiovascular diseases in humans [1]. The most prevalent of fatty acids are palmitic (16
carbon) and stearic (18 carbon) acids, which are fatty acids ubiquitously present in biological
systems abundant in eukaryotic cell membranes. Palmitic acid (PA) is the most common
saturated fatty acid found in various organisms. Synthesis of PA is well-known and occurs
naturally in mammalian cells via acetyl-coenzyme A (CoA) and malonyl-CoA precursors.
Alternatively, the synthesis and function of the esterified form of palmitic and stearic acid,
palmitic acid methyl ester (PAME) and stearic acid methyl ester (SAME), has not been welldocumented (Figure 1) [2]. Only recently has PAME and SAME been introduced to the
forefront of fatty acid research in biology and disease. Specifically, PAME results in aortic
vasodilation and neuroprotection, while SAME causes neuroprotection all in the context of
cerebral ischemia. Thus far, PAME and SAME are thought to act as a neurotransmitter/
modulator released from a neuronal source (SCG, superior cervical ganglion SCG) [3].
-3 FATTY ACIDS
Eicosapentaenoic acid (EPA, 20:5n-3), docosahexaenoic acid (DHA, 22:6n-3) (Figure 4),
and -linolenic acid (ALA, 18:3n-3, rich in plant oils) are three major -3 essential fatty
acids involved in cardiovascular function and general circulation. Administration of EPA can
1) induce endothelium-independent aortic and mesenteric vasodilation via activation of K+ATP
channels on VSMCs via EPA-derived prostanoids [27], 2) Activation of largeconductance/Ca2+-mediated K+ channels (BK) on VSMCs by EPA metabolite, 17,18-EET
[28].
In addition, clinical research studies on the administration of EPA can inhibit the onset
and progression of atherosclerosis and decrease the prevalence of myocardial infarction [29].
[35]. Furthermore, dietary intake of fish oil has also shown to enhance the coronary
vasomotor activity in patients with coronary artery disease [36] and reduce the number of
deaths derived from chronic cardiac failure [29]. Altogether, these data suggest that dietary
intake of -3 fatty acid supplements are beneficial to counteract against cardiovascularrelated diseases such as hyperlipidemia, hypertension, atherosclerosis, and myocardial
infarction.
Modified from Ratledge C. 2004; Catal A, 2013; Condary R and Yao JK, 2011.
Figure 6. Synthesis of unsaturated fatty acids.
10
Other fatty acids such as alpha-linolenic acid (ALA), have been shown to increase CBF
and vasodilation in vivo and in vitro [33]. ALA, PA, or saline was administered onto rat
carotid or basilar arteries resulting in ALA-induced vasodilation in basilar arteries. This was
also confirmed by laser-Doppler flowmetry suggesting an increase in CBF further supporting
ALA as a vasodilator [33] thought to be activated by the TREK-1 K+ channel. Additionally,
ALA has been shown to provide some degree of neuroprotection in animal models following
transient global ischemia [72].
Interestingly, Wang et al., 2006 and 2007 suggested that treatment with SA alone has
been shown to mitigate neuronal cell death due to ischemia-induced oxidative stress [73, 74].
More research with SAME, the esterified form of SA, is necessary to prove therapeutic
efficacy under ischemic conditions. Moreover, since PAME and SAME were co-released
from a neuronal source, it would be interesting to co-administer PAME/SAME and observe
the possible effects of CBF and neuroprotection [5]. Taken together, it is important to note
that the esterified from of PA produces consistent vasodilation while, PA alone does not
possess any vasodilatory properties.These results illustrate the need for further investigation
into PAME regarding its potential receptor/binding site, synthesis, and localization of stored
PAME. Assuming that further studies of PAME, SAME, and other fatty acid signaling
molecules reinforce data already available, this class of endogenous molecules should be
highly considered for interventional therapy against stroke and general ischemia.
ACKNOWLEDGMENTS
This work was supported by National Institutes of Health grant NS073779-03, American
Heart Association-Philips grant AHA-13SDG13950014 and ASA-14BFSC17690007, and
Miami Evelyn F. McKnight Brain Institute.
Disclosure/Conflict of Interest: The authors have no conflict of interest in this
manuscript.
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In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 2
ABSTRACT
Palmitic acid (PA), one of the most abundant saturated fatty acid (SFAs) within
plants, humans, animals, microbial (bacteria), fungal, and marine organisms, constitutes
~16 to 45 % of the lipid profile. The impressive abundance of PA throughout nature
could be attributed partly to its critical role in membrane lipid structural functionality,
formation of subcellular cysteine residue linkages and RNA posttranslational
modifications in eukaryotic and prokaryotic cells. PA has been demonstrated to undergo
-oxidation to produce short and medium chain fatty acids to maintain homeostasis in
response to endogenous and exogenous cues. Further, non-esterified PA is known to
mediate numerous biochemical and antimicrobial pathways towards the betterment of
human health. Although the importance of PA in human health and nutrition are
established, critics attest that excessive dietary PA may be unhealthy and even
detrimental. Current microbiological and epidemiological studies suggest that PA
produces specific health benefits that are not common to all other SFAs. For example,
studies suggest that PA may activate nitric oxide and superoxide, thus functioning as an
antimicrobial agent against some strains of bacteria, algae, and helminthes and certain
foodborne pathogens. This chapter reviews the occurrence, biochemistry, and subsequent
health implications of PA.
mjonson@mytu.tuskegee.edu;
dabugri@mytu.tuskegee.edu; abugrigh@yahoo.com.
18
1.1. INTRODUCTION
The most common and abundant saturated fatty acid (SFA) in nature, as well as within
plants, animals and humans, palmitic acid (PA) has many critical functions (Figure 1).
Serving as the primary storage form of excess dietary carbohydrates and the precursor for
longer chain fatty acids, palmitic acid occupies a critical role in long-term energy storage, the
synthesis of other biomolecules (e.g. glycolipids, phospholipids, vitamins, prostaglandins,
prostacyclins, thromboxanes, etc.) and cellular membrane structural constituents. Palmitic
acid has also been evaluated based on its antimicrobial, antifungal, antibacterial contributions,
when bound to specific proteins. Applications within the health and beauty (e.g. cosmetics,
soap production), food (e.g. additive, texturing agent), pharmaceutical (e.g. palmate ester as a
carrier medium and releasing agent) and biotechnology industries further add credence to the
virtual diversity of this fatty acid to function as a multipurpose entity and challenges one to
consider this fatty acid beyond the conventional understanding as a fatty acid whose primarily
purpose for existence is to serve as a fuel source.
Palmitic acid and its bioactive metabolites and successors exhibit unique absorption,
transport, metabolic, site-specific distribution(s), mechanisms of action and implications
(Figure 1). These characteristics contribute to localized and generalized observed and
subsequent potential repercussions- both acute and chronic. The ability of palmitic acid to
influence protein-protein interactions, hydrophobicity, membrane association, and subcellular
trafficking within and between membrane constituents, exemplifies the capability of palmitic
acid to purposefully engage in covalent binding to specific protein residues. The occurrence,
biochemistry and health effects of palmitic acid within the micro and macro bio-arenas of
plants, animals, humans and other microorganisms accentuate the transformative nature of
this fatty acid and its influence on the lipidome, metabolome, proteome, transcriptome and
genome. This chapter seeks to integratively explore the multi-faceted nature of palmitic acid
and provide the reader with an interactive learning experience, with appropriate visual aids to
abridge concepts and aid in comprehension, which allows for the enhanced appreciation of
one of the most abundant fatty acids in nature.
*Excess carbohydrates (CHOs) are converted to palmitic acid; negative feedback (inhibition) by acetyl-CoA carboxylase (Benoit et al. 2009) the enzyme
catalyzing the conversion of acetyl-CoA to malonyl-CoA; positive feedback (synthesis) catalyzing the synthesis of palmitic acid; CVD- cardiovascular
disease.
Figure 1. Summary of the occurrence, biochemical courses of action and implications of palmitic acid.
20
21
Figure 2. Functions and contributions of palmitic acid. (Image Sources: en.wikipedia.org; Microsoft office
clipart).
22
lipid levels increased in both visceral and subcutaneous adipocytes, in comparison to primary
accumulation in visceral adipocytes when in triglyceride form.
Liver (SHR)
Liver (mice)
Fungal
Agaricus bisporus
Cortinarius glaucopus
Hygrophoropsis
aurantiaca
Hypholoma capnoides
Laccaria laccata
Lactarius salmonicolor
Lespista inversa
Turkey tail
% PA
References
20-25%
17.6-30.1%
36.0%
23.4-23.8%
19.5-19.6%
19.8-21.5%
69% (Phosphoglycerides)
3% (Monoacylglycerides)
8% (Cholesterol)
4% (Free fatty acids)
6% (1,2-Diacylglyceride)
3% (1,3-Diacylglyceride)
2%(Cholesterol ester)
6% (Triglyceride)
16-21%
21.0-21.5%
(Rule, 1997)
(Shirai, Suzuki, & Wada, 2005)
(Shirai, Suzuki, & Wada, 2005)
13-14%
12%
10%
16%
12%
7.4%
16.4%
23%
% PA
14%-15.1%
Artis Conk
17.3% -17.4%
Algae
Phaeodactylum
tricornutum
Thalassiosira weissflogii
14.7-26.8%
28.8-36.6%
Dunaliella primolecta
21.8-26.0%
Nannochloris sp.
15.1-17.8%
Parietochloris incisa
10.0-19.8%
Nostoc commune
25.3-43.5%
Synechocystis sp.
18.8-26.5%
Pavlova lutheri
Emiliana huxleyi
11.1-23.6%
10.3-17.7%
Heterosigma akashiwo
40.0-46.3%
Yeast
Nadsonia fulvescens and
N. commutata
R. bisporidii and R.
dibovatum, R.Toruloides
Bacteria
Lactobacillus arabinosus
References
(Abugri, McElhenney, & Willian,
2012)
(Abugri, McElhenney, & Willian,
2012)
(Lang et al., 2011; Tonon et al., 2002;
Viso & Marty, 1993)
(Lang et al., 2011; Viso & Marty,
1993)
(Lang et al., 2011; Viso & Marty,
1993)
(Lang et al., 2011; Viso & Marty,
1993)
(Bigogno et al., 2002; Lang et al.,
2011)
(Lang et al., 2011; Temina et al.,
2007)
(Lang et al., 2011; Viso & Marty,
1993)
(Lang et al., 2011; Tonon et al., 2002)
(Lang et al., 2011; Viso & Marty,
1993)
(Lang et al., 2011; Viso & Marty,
1993)
14-15%
14-19.0%
18.7%
Lactobacillus casei
24.3%
Lactobacillus
delbrueckii
Streptococcus sp
27.3%
Clostridium butyricum
Escherichia coli
Argrobacterium
tumefaciens
Plant
Malva sylvestris
(leaves,flowers,immature
fruits, leafy flowered
stems)
Sweet potatoes (Ipomoea
batatasi) leaves
Capsicum chinnese
49.0%
25-38.6%
8.2%
26.6%
10% (leaves)
17.2% (flowers)
20%(immature fruits)
13% (leafy flowers stems)
10-21%%
16-21%
23
24
% PA
16.2%
References
(Abugri et al., 2013)
16 %-29.20%
Source: http://en.wikipedia.org/wiki/File:Palmitic-acid-3D-balls.png.
Figure 3a. Ball-and-stick 3D structure of palmitic acid molecule.
Source: http://en.wikipedia.org/wiki/File:Palmitic_acid.svg.
Figure 3b. Linear structure of palmitic acid molecule.
25
Source: http://ercfre86.wordpress.com/2012/03/19/applications-of-palmitic-acid/
Figure 3c. Physical appearance of palmitic acid.
26
cell will experience disruption in the cell membranes which could lead to greater
conformational changes within membrane proteins and their functionality (Liu et al. 2008;
Avis & Belanger, 2001; Altieri et al., 2007; Pohl et al. 2011). Other effects meditated by PA
at high fluidity could be the release of intracellular components, cytoplasmic disorder and
eventually cell disintegration and apoptosis (Liu et al. 2008; Avis & Belanger, 2001; Altieri et
al., 2007; Pohl et al. 2011).
Fatty acids are known in nature to function as anionic surface agents and these roles
make them nonfunctional at certain physiological pH conditions (Armstrong, 1957; Kabara et
al., 1972; Scharff & Beck, 1959). Palmitic acid acts as antimicrobial agents against bacteria
(Kabara et al., 1972), fungal and other pathogens. For instance, palmitic acid has been
demonstrated to greatly inhibit Alernaria solani (Liu et al., 2008), Aspergillus niger (Altieri
et al., 2007), Aspergillus terreus (Altieri et al., 2007), Cucumerinum lagenarium (Liu et al.,
2008), Emericella nidulans (Altieri et al., 2007) and Fusarium oxysporum (Liu et al., 2008).
With this knowledge about palmitic acid, its biochemical pathways of inhibition of the
enzymatic secretory pathways used by such microorganisms are speculated to be either
disruption of mitochondrial machinery resulting in much electron transport influx or
disruption of the cell membrane integrity (Kabara et al., 1972; Desbois & Smith, 2010; Pohl
et al., 2011).
27
A. Cell Lysis
Fluidity of fatty acid depends on the chain length and the degree of unsaturation,
therefore because of these unique property, fatty acids have the ability to insert themselves
into the inner membranes of pathogens resulting in more fluidity and permeability
(Chamberlain et al., 1991; Greenway & Dyke, 1979). PA mechanism of cell lysising may be
due to high permeability of membrane due to the insertion of PA, and has inner consequence
such as allowing of internal contents to leak from cells which can result in growth inhibition
and death (Galbraith & Miller, 1973; Shin et al., 2007; Wang & Johnson, 1992; Boyaval et al.
1995; Liu et al. 2008; Avis & Belanger, 2001; Altieri et al., 2007). The higher the fluidity in
the cell the greater the cell tendency to lysising or bursting because of the issue of unbalance
membrane fluidity of the cell (Desbois & Smith, 2010).
B. Inhibition of Enzymatic Activity and Disruption of Electron Transport Chains
Minor and major biochemical processes in the body depend upon enzymes for catalysis
to bring about homeostasis. However, these enzymes can be hampered by the physiological
conditions. Palmitic acid just like any other fatty acid might have the capability to penetrate
through the cell wall, which could cause irreversible deformation of the cell membranes (Pohl
et al., 2011; Liu et al. 2008; Avis & Belanger, 2001; Altieri et al., 2007). Since these are
protected by membrane proteins, the conformational forms of these proteins could be
disrupted depending on the levels require in bring such effect within the cell. When the
enzymes are inhibited it results in the disruption of the ATP synthesis due to decoupling
effect on the energy chain which affects the ATP synthase responsible for production and
regulation of ATPs in the cell (Desbois & Smith, 2010; Harold, 1972). Palmitic acid might
cause a direct binding to the electron carriers, insertion between carriers preventing
interaction, complete displacement of carriers from membranes and prevention of carriers
interaction by reducing fluidity of the membranes (Pohl et al., 2011; Liu et al. 2008; Avis &
Belanger, 2001; Altieri et al., 2007). However, further studies will be needed to actually
conclude its mechanism of action.
C. Impartment of Nutrient Uptake and Release
Every microorganism has well defined coordination systems that allow nutrient uptake
into the cellular level for the cell growth and bioenergetics purposes. However, if the cell
membranes and its associated proteins that is performing this role is disrupted then the
coordination becomes poor for the cell to do its selectivity in terms of nutrient uptake. It has
been proposed by (Desbois & Smith, 2010) that the presence of holes created by free fatty
acids causes leakages which cause impairment of active nutrient uptake by either indirectly
or direct on transport proteins. In most nutrient uptake the effect of this fatty acid is directly
influence by the inability for the transport protein to carry their function properly due to
conformational changes during cell membrane and its associated machinery disruption. On
the part of the indirect route used by free fatty acids to destabilized proper nutrient uptake into
vital parts of the cell could be attributed to the not favorable condition received by the cell
due to leaking, unregulated exchange of palmitic acid between inner and out membranes
machinery which results in the cells inability to produce ATP. The interesting biochemistry of
these effects is that in larger animals the cell organization is totaling different and that helps
prevent leakages in humans and associated mammalian species.
28
29
improperly disruption of cell energy dispensation (Lehninger & Remmert, 1959; Wojtczak &
Lehninger, 1961; Lehninger, 1962).
The possible causes for these swelling in the mitochondrial depended on the chain length
and degree of unsaturation. This potential is similar to the features used to disrupt the energy
coupling processes in cellular (Pressman & Lardy, 1956).
30
There have been strong correlations between fatty acids and stimulation of mitochondrial
respiration and their potentials as protonophoric properties (Cunarro & Weiner, 1975).
Furthermore, the longer the chain of the fatty acid, the more the proton conductance will
occur in the phospholipid bilayer membranes (Gutknecht, 1988). This implies that multiple
studies are in agreement with the observation (Gutknecht, 1988). Palmitic acid may be using a
similar mechanism as any other fatty acids which are able to facilitate transmembrane flux of
some other cations, namely monovalent alkali metal cations ( Liu et al. 2008; Avis &
Belanger, 2001; Altieri et al., 2007; Zeng, Han, & Gross, 1998). However, research is needed
to draw a conclusive mechanism of PA under the membrane flux potential. Most of the
monovalent elements that palmitic can help in exchange between membranes are potassium,
sodium and lithium (Zborowski & Wojtczak, 1963). The mechanism is made possible by
(Pressman & Lardy, 1956; Zborowski & Wojtczak, 1963; Schonfeld, Schild & Kunz, 1989).
The capacity of palmitic acid in its undissociated form to perform flip-flop in the inner
mitochondrial membrane (Zborows; ki & Wojtczak, 1963) makes it potent for interfering
with the pathogens monovalent machinarys. The second possible way palmitic acid is
utilizing in inhibiting or killing pathogens could be due to its ability to exist in the anionic
form. This form is transferred by the adenine nucleotide translocator as well as other proteins.
It is important to note that the process mentioned above is possible based on size and the
structural nature of the fatty acid molecule in question. Many authors have found out that
fatty acid has highest potency to uncouple oxidative phosphorylation (Pressman & Lardy,
1956; Schnfeld, Schild, & Kunz, 1989), to induce mitochondrial swelling (Zborowski &
Wojtczak, 1963), and to promote energy-dependent accumulation of monovalent cations in
mitochondria (Wojtczak, 1974). This could possibly be the same as what has been observed
in palmitic acid in fungus inhibition and killing (Wojtczak, 1974; Choi et al. 2010).
31
hepatocytes (Bruce & Salter, 1996). Others found palmitic acid to be metabolized at a much
lower rate than myristic acid in rat hepatocytes; myristic acid exhibited a significantly greater
rate of -oxidation and elongation (Rioux, Lemarchal, & Legrand, 2000). In the presence of
other fatty acids, the rates of gluconeogenesis, palmitic acid metabolism and the metabolism
of long-chain fatty acids is adapted accordingly in both humans and animals (Emken, 1994;
Mashek, Bertics, & Grummer, 2002). Palmitic acid also influences the metabolism of lipids,
lipoproteins, total and HDL cholesterol (Mensink et al., 2003; Ramamoorthy, Gupta, &
Khosla, 2000; Sanders et al., 2011; Snook et al., 1999). However, others found diets high in
palmitic acid to alter neither fasting nor postprandial levels of homocysteine or other
inflammatory biomarkers (e.g. TNF-, IL-1, IL-6 and IL-8, high-sensitivity C-reactive
protein and interferon-) (Voon et al., 2011).
Although researchers found palmitic acid supplementation to facilitate an increase in
plasma cholesterol concentrations in a metabolic-diet study in comparison to lauric acid
(Denke & Grundy, 1992), conflicting findings were presented elsewhere, with dietary
palmitic acid resulting in lower serum cholesterol concentrations than a lauric-myristic acid
combination (Sundram, Hayes, & Siru, 1994). In a study examining the effect of palmitic acid
intake (high vs low) on the endogenous synthesis of cholesterol and plasma lipoprotein
cholesterol levels, palmitic acid was unable to illicit a significant effect on lipoprotein
profiles, when recommended intakes of dietary linoleic acid (C18:2n6) were achieved
(Clandinin et al., 2000). Researchers have found increasing dietary palmitic acid to decrease
fatty acid oxidation and daily energy expenditure (Kien, Bunn, & Ugrasbul, 2005).
32
Figure 8. Simplified illustration outlining the catabolism of palmitic acid as indicated by: 1) -oxidation
of palmitic acid into acetyl CoA; 2) acetyl CoA entry into the Citric Acid Cycle; and 3) electron
transfer via the Electron Transport Chain.
Palmitic acid serves as the major metabolic fuel, particularly in the brain, which utilizes
fatty acids as a major fuel source during fatty acid oxidation and active transport into cerebral
microvessels (Williams et al., 1997). Up on transport across the palmitic is incorporated into
brain phospholipids, contributing to ~29% of phospholipids (mol/g) in rat brain (Rapoport,
2001). The uptake of palmitic acid is enhanced with the expression of the fatty acid transport
protein, which facilitates the transcellular transversion of fatty acids across the blood-brain
33
barrier (Mitchell & Hatch, 2011). During metabolic conditions of starvation and diet-induced
obesity, palmitic acid in triglyceride form but not free fatty acid form, when administered
intravenously was able to inhibit the transport of leptin across the blood-brain barrier (Banks
et al., 2004). This suggests the role of increased triglyceride levels and subsequent
hypertriglyceridemia during starvation and conditions such as diabetes in mediating the
homeostasis of hormones such as leptin, which regulates fat storage and energy expenditure
and in certain cases inducing the resistance of certain hormones to enter the brain.
34
palmitoleic acid content have been observed in metabolic disorders such as anorexia nervosa,
obesity, metabolic syndrome (Kremmyda et al., 2011).
Recently it has been suggested that not all saturated fats increase the risk for
cardiovascular and other diseases, but that the triglyceride structure of the fatty acid is of
more importance when assessing disease risk (Fattore & Fanelli, 2013). Studies investigating
the influence of palm oil, which is comprised of ~50% palmitic acid, 40% oleic acid and 10%
linoleic acid, have yielded conflicting evidence in the relationship between palm oil and
cardiovascular disease risk. However, the replacement of trans fatty acids with palm oil more
favorably influenced biomarkers for disease (Fattore et al., 2014). The inclusion of palm oil
into the diets of moderately hyperlipidemic individuals adversely altered plasma lipid
profiles, more noticeable increases LDL-cholesterol concentrations (Vega-Lpez et al., 2006).
Insulin sensitivity was reduced, HDL- and total cholesterol significantly increased and fatty
acid oxidation was moderately increased in individuals consuming diets enriched in saturated
fatty acids (9% palmitic acid) compared to those consuming diets enriched in
monounsaturated fatty acids (9% oleic acid) (Lovejoy et al., 2002). Dietary monounsaturated
fatty acids have been proposed to provide protection from the risks associated with metabolic
syndrome and cardiovascular diseases (Gillingham, Harris-Janz, & Jones, 2011). Palmitoleic
acid (C16:1n7), the monounsaturated complement of palmitic acid (C16:0), is believed to
provide such protection.
CONCLUSION
This chapter emphasized palmitic acid, its physical, chemical and structural features as
pertaining to its functional characteristics, occurrence and percent distribution in plants, algae,
fungus, human and animal tissues. The antimicrobial mechanisms of action, metabolism,
general biochemistry and health implications of palmitic acid provide insight into the versatile
35
nature of this fatty acid. Palmitic acid, one of the most abundant saturated fatty acids in nature
and within living systems, offers more than meets the eye. Palmitic acid is a master and
universal saturated fatty acid that has several benefits and effects when in high concentration.
Both de novo and salvage pathways ultilze palmitic acid for the biochemical synthesis of
short, medium, and long(er) chain fatty acids as well as other phospholipids, which are
required for cell mebrane structure and function, brain functionality and neuronal stability.
Palmitic acid is the only fatty acid posttranslation modification is reversible and because of
this unique feature cell signalling and trafficking is effect in the cell. Although, palmitic acid
has unique benefits, its biochemistry is still not explored into in both prokaryotes and
eukaryotes. Also because of the mechanisms exerted by palmitic acid for instance cell
signalling and trafficking, there is a possibility of it been used in theraupeutics and as
nutraceuticals for human microbial infection prevention. Although the evidence is conflicting
regarding the role of palmitic acid in health and its implications for health promotion and
disease prevention, there exists the potential for palmitic acid to serve as a functional and
bioactive endogenous and exogenous constituent of homeostasis.
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Chapter 3
ABSTRACT
In the last few decades, disagreement between opinions and findings concerning the
ability of palmitic acid (PA) and other saturated fatty acids (SFAs) to raise
cholesterolaemia has led to discussions on whether PA, which has been positively related
to high serum cholesterol levels, could increase the risk of cardiovascular diseases. This
study aims to review the PA content of meat, dairy products, fish, and other food of
animal origin in the human diet and discusses nutritional issues related to the occurrence
of this fatty acid (FA) in these foods due to different diet supplementation. Meat and
dairy products are considerable dietary sources of SFAs, such as PA. In most
industrialized countries, a high meat or dairy intake contributes to a higher than
recommended SFA intake. Palmitic and myristic acids are common FAs in meat and
dairy products, making up about 30-40% of total FA intake and are the main factors
responsible for raising cholesterol levels; indeed, strong evidence indicates that these two
SFAs increase serum cholesterol concentrations in humans. Stearic acid is partially
converted to oleic acid in vivo and has not been shown to elevate blood cholesterol, while
lauric acid is not as potent as PA at raising concentrations of total cholesterol and LDL
cholesterol in humans. The occurrence of PA in animal origin food is influenced by both
genetic and environmental factors, such as the composition of the animals diet, its
digestive system and its biosynthetic processes. The FA profile in food of animal origin
mainly reflects dietary lipid sources and has the potential to play a valuable role in human
nutrition by manipulating the composition of animal fat through diet. In order to explain
the variability in FA composition in food of animal origin, this review examines different
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46
P. G. Peiretti
nutrition trials that have studied the effects of PA supplementation on the lipid profile of
animal origin food.
INTRODUCTION
Palmitic acid (PA) or n-hexadecanoic acid is the most common saturated fatty acid (SFA)
found in plants, animals, and microorganisms (Gunstone et al., 2012). As the name suggests,
PA is characteristic of palm oil and its saponified form was discovered and isolated for the
first time by Frmy (1840). The major difference between palm oil and other vegetable oils is
the higher proportion of PA (Table 1).
The impact of excess SFA in the diet on cardiovascular diseases has been studied and
discussed both in animal and human studies (Crawford, 1968; Keys, 1970; Temme et al.,
1996). In the past, because of its high PA content, palm oil has been attacked as highly
saturated oil and accused of raising blood cholesterol and increasing the risk of
cardiovascular disease (Mukherjee and Mitra, 2009).
Several studies attack SFAs with regard to their hypercholesterolaemic and atherogenic
effects, which adversely affect cardiovascular risk (Kromhout et al., 1989; Menotti et al.,
1989; Verschuren et al., 1995). Lauric and myristic acid are the main cholesterol-raising
SFAs, whereas PA and stearic acid have much weaker cholesterol-raising potential (Sundram,
1994).
Table 1. Palmitic acid content of oils and fats of vegetable sources (expressed as
percentage mass-fraction of total fatty acids)
Palm oil
Cottonseed oil
Cocoa butter
Illipe fat
Olive oil
Oat bran oil
Avocado oil
Rice brain oil
Wheat germ oil
Corn oil
Tomato seed oil
Peanut oil
Soya bean oil
Pistachio nut oil
Grapeseed oil
Babassu fat
Poppyseed oil
Sesame oil
40.1-47.5
21.4-26.4
25.4
18.0-22.0
7.5-20.0
17.4
17.2
16.9
16.6
8.6-16.5
12.0-15.5
8.3-14.0
8.0-13.3
11.6
5.5-11.0
5.2-11.0
10.6
7.9-10.2
47
6.5-10.0
8.9
8.2
3.9-7.2
4.0-7.0
6.5
6.4
4.8-6.2
6.0
4.0-6.0
1.5-6.0
5.4-5.9
5.2
4.0
The negative effect attributed to lauric and myristic acid explains why foods rich in SFA
should be consumed in moderation and there is convincing evidence that PA increases the
risk of cardiovascular disease (World Health Organization, 2003). Palm oil raises plasma
cholesterol only when the diet contains excess dietary cholesterol, in which case the risk of
coronary heart disease may rise (Jones, 1989). Temme et al. (1996) reported that both PA and
lauric acid are hypercholesterolaemic compared with oleic acid.
Lauric acid raises total cholesterol concentrations more than PA, which is partly due to a
greater rise in HDL cholesterol. Enig (1993) reported that PA increased the level of blood
cholesterol more than other SFAs, including lauric acid and myristic acid, which are more
abundant in palm kernel oil and coconut oil than in palm oil. Clarke et al. (1997) concluded
that, compared to carbohydrates, PA raises blood cholesterol levels. However, some reviews
do not seem to support these conclusions (Edem, 2002; Ong and Goh, 2002; Sundram et al.,
2003; Oguntibeju et al., 2009; McNamara, 2010) and indicate that the effect of PA (found
mainly in palm oil) on blood cholesterol is relatively neutral when compared to other fats and
oils. We must therefore realize, although it may seem simplistic, that what matters is the
dietary context, rather than the individual nutrient. In a balanced diet, in fact, PA is usually
harmless (it is even synthesized by the body), but can become dangerous when consumed as
part of frequent caloric excesses or is consumed in particularly large quantities.
Hayes and Khosla (1992) suggested that PA may be neutral in normocholesterolaemic
subjects if the diet contains little cholesterol and linoleic acid intake is adequate. Fattore and
Fanelli (2013) reviewed the scientific literature on the evidence of the relationship between
palm oil and adverse effects on human health and concluded that there is no clear evidence of
a negative role of PA on health and much less of native palm oil, which is a complex
alimentary matrix, in which PA is only one of its components. However, more recent lipid
research on the topic seems to have reconsidered the negative role of dietary SFAs as a risk
factor for cardiovascular diseases. For instance, lauric acid and myristic acid have a greater
total cholesterol-raising effect than PA, whereas stearic acid has a neutral effect on the
concentration of total serum cholesterol, including no apparent impact on either LDL or HDL
(Daley et al., 2010).
48
P. G. Peiretti
Fattore and Fanelli (2013) showed that not only the type of fat, but also its triglyceride
structure, play a role in cholesterolaemia. PA is located at the Sn-1 position of the three
principal triglyceride species in palm oil (Small, 1991) and a high fraction of PA in palm oil
is bound at the Sn-1 or Sn-3 position of the glycerol molecule (Mu and Hy, 2004). This
location confers the non-hypercholesterolaemic property to the oil (Ng, 1997). This is in
contrast with the major triglyceride species in animal fats such as butter, which contains PA
in the Sn-2 position with resultant hypercholesterolaemic and atherogenic effects (Ng, 1997).
Evidence is now growing that the molecular structures of dietary triacylglycerols play an
important role in the development of atherosclerosis (Patsch, 1994), because triacylglycerols,
enriched with SFA at the Sn-2 position, exhibit different metabolic behaviour than
triacylglycerols with SFA at the Sn-1/3 position (Redgrave et al., 1988; Tuten et al., 1993;
Carnielli et al., 1995).
The enzymatic hydrolysis of dietary triglycerides by pancreatic and lipoprotein lipases
preferentially targets fatty acids (FAs) in the Sn-1/3 position rather than those esterified to the
Sn-2 position, which are substantially preserved in chylomicrons (Karupaiah and Sundram,
2007). These authors showed that the positioning of unsaturated versus SFAs in the Sn-2
position may explain the modulatory effects on atherogenicity and thrombogenicity.
Kritchevsky (2000) reported a higher degree of absorption of PA at the Sn-2 position in rabbit
models and this could be related to the increased atherogenicity of interesterified palm oil, in
comparison with the native one.
The predominant SFAs that occur naturally in animal fats and the main products of
cytosolic FA synthetase multienzyme complex in lipogenesis are stearic acid and PA, which
can be biosynthesized de novo by all known organisms, including fish (Sargent et al., 1989).
Some authors have determined enzyme activities in subcutaneous adipose tissue. StearoylCoA or 9 desaturase is the terminal step in the desaturation and conversion of PA, myristic
and stearic acid into the 9 monounsaturated FAs palmitoleic, myristoleic, and oleic acid,
respectively (De Smet et al., 2004). Kazala et al. (1999) suggested that FA elongation was
unable to keep pace with the de novo production of PA in animals that deposited greater
amounts of intramuscular fat. However, based on tissue incubations, it has been suggested
that 9 desaturation and not elongation is the rate-limiting step for the conversion of PA to
oleic acid in bovine subcutaneous adipose tissue (St. John et al., 1991).
This review examines the occurrence and variation of PA content in food of animal origin
in relation to different diets and different dietary regimens.
49
24.02
21.07
5.73-7.55
7.41
5.67-5.99
5.70
3.93-4.93
4.03
3.71
2.89
2.19
2.06
0.58-1.99
1.95
1.90
1.63-1.77
1.66
1.65
50
P. G. Peiretti
Whole turkey with skin (raw)
Whole rabbit
Beef meat
Sheep meat
Goat meat
Ostrich meat
Deer meat
1.54
1.22
0.31-1.14
0.58
0.40
0.16
0.12
Adapted from Italian food composition tables - National Research Institute for Food and Nutrition
(INRAN).
A survey conducted by Enser et al. (1996) illustrated the differences in fat content and
FA profile of muscle between lamb, pork and beef; in particular, PA content, expressed as mg
per 100 g muscle, decreased in the order lamb>beef>pork, while PA, expressed as percentage
of total FAs, decreased in the order beef>pork>lamb.
According to Banskalieva et al. (2000), FA composition of fat depots in goats appears to
be in the range typical for ruminants, with average percentages of PA in goat muscles being
similar to those for other ruminant species. The PA concentration in kidney fat is similar in
goats and lambs, but lower for goat meat than in beef. Hilditch and Williams (1964) noted
that land animals tend to have a relatively constant amount of PA in fat depots.
20.86
8.04
7.31
5.72
4.10
3.49
2.85
1.97
1.58
1.34
0.92
0.92
0.45
0.05
Adapted from Italian food composition tables - National Research Institute for Food and Nutrition
(INRAN).
51
The FA composition of milk varies in relation to the influence of several factors related
to the animal and its environment (Palmquist et al., 1993; Perdrix et al., 1996). These include
numerous highly significant factors such as the nature of the food (Grummer, 1991; Coulon et
al., 1994), the type of ration, mode of administration, energy concentration and energy level
of the ration, the physical state of the food and the entire ration, quantity, quality and length
of the fiber, the type, shape and physical treatment of cereals, etc.
9.96
5.98
3.00
2.98
2.72
1.90
Adapted from Italian food composition tables - National Research Institute for Food and Nutrition
(INRAN).
52
P. G. Peiretti
Scollan et al. (2001) examined the effects of different sources of dietary n-3 PUFA on the
FA composition of muscle and adipose tissue in beef cattle. These authors found that the
proportion of PA was decreased by the linseed diet in both neutral lipids and phospholipids of
muscle. However, fish oil significantly increased the proportion of PA in neutral lipids, while
Mandell et al. (1997) also found that fish meal feeding increased PA content, contrary to the
lack of effect observed by Mills et al. (1992) on feeding fish meal and Ashes et al. (1992)
who fed ruminally-protected fish oil.
Furthermore, there was a lower proportion of PA in phospholipids as a result of feeding
the linseed diet compared with the control diet. However, compared to the control diet, fish
oil did not alter the proportion of PA, but produced lower proportions of stearic and oleic acid
(Scollan et al., 2001).
Nuernberg et al. (2005) examined the effects of feeding system and breed on the content
of the beneficial n-3 PUFA and conjugated linoleic acids in beef muscle, finding that the
percentage of PA decreased significantly with the grass-based system, while there was no
influence of breed on the percentages of PA. Daley et al. (2010) report that grass-finished
cattle are typically lower in total fat as compared to grain-fed contemporaries. Interestingly,
there is no consistent difference in total SFA content between these two feeding regimens.
Those SFAs considered to be more detrimental to serum cholesterol levels, i.e., PA and
myristic acid, were higher in grain-fed beef as compared to grass-fed contemporaries in 60%
of the studies reviewed. Grass-finished meat contains elevated concentrations of stearic acid,
the only SFA with a net neutral impact on serum cholesterol. Thus, grass-finished beef tends
to produce a more favorable SFA composition, although little is known of how grass-finished
beef would ultimately impact serum cholesterol levels in hyper-cholesterolaemic patients as
compared to grain-fed beef (Daley et al., 2010).
Diet is known to affect the FA composition of pig adipose tissue, particularly perirenal
and back fat (Fontanillas et al., 1997; Larick et al., 1992) and to improve the FA profile of
carcass fat in pigs (Morgan et al., 1992; Van Oeckel and Boucqu, 1992). In particular,
dietary oils affected total FA content in pig longissimus muscle (Corino et al., 2002; Kouba
and Mourot, 1999; Van Oeckel et al., 1996). Teye et al. (2006) evaluated the effects of palm
oil, palm kernel oil and soyabean oil, in combination with high and low protein levels, on the
FA composition of the longissimus dorsi muscle in pigs; they found that palm kernel oil
increased the concentrations of PA and other SFAs, while it decreased linoleic acid levels
(Figure 1).
Wood et al. (2004) examined the effects of diet, breed and muscle on FA composition in
pigs and concluded that PA and myristic acid in neutral lipids were higher in Berkshire and
Tamworth than in Duroc and Large White, while the dietary effects on these FAs were small.
Dietary FA modification is considered a viable method of adding value to poultry
products for the health-conscious consumer (Hargis and Vanelswyk, 1993). The profile of
dietary FAs is of importance because it influences the quality of the fat deposited on the
broiler carcass (Figure 2). PA was the predominant SFA in the adipose tissue of birds (Kang
et al., 2001). These authors found that dietary supplementation of palm oil resulted in a
significant increase in PA in the liver and adipose tissue of broilers. According to Rodriguez
et al. (2002), palm oil or mixtures of palm oil, and FAs distilled from palm and calcic soap
are sources of vegetal oils with an FA profile that might replace animal fats without any kind
of negative impact on carcass quality. Consistent parallels between dietary fat and abdominal
fat in broilers have been described by Scaife et al. (1994). The FA pattern of abdominal fat
53
represents the FA pattern in the dietary fat sources very well and the key FAs which occur at
a somewhat constant rate in broiler adipose tissue are PA, stearic acid, and to some extent,
oleic acid (Zollitsch et al., 1997). Smink et al. (2008) found that the feeding of randomized
instead of native palm oil significantly raised the PA content of breast meat and abdominal fat
and lowered the ratio of unsaturated fatty acids to SFA.
Figure 1. Relationship between dietary palmitic acid percentage and muscle palmitic acid percentage in
pig nutritional trials: Peiretti et al. (2013; ), Teye et al. (2006; ), and Corino et al. (2002; ).
Figure 2. Relationship between dietary palmitic acid percentage and muscle palmitic acid percentage in
poultry nutritional trials: Zollitsch et al. (1997; ), Kang et al. (2001; ), and Smink et al. (2010; ).
Rabbits, like other monogastric animals, are able to directly incorporate dietary FAs into
adipose and intramuscular tissue lipids, thus making it possible to modify the FA profile of
54
P. G. Peiretti
rabbits through the strategic use of unsaturated dietary fat sources (Dalle Zotte, 2002). FA
levels vary a great deal on the basis of the nature of the rabbit diets. The influence of the FA
profile in the diet seems to be more pronounced on the FA composition of adipose tissue than
intramuscular fat (Xiccato, 1999). Peiretti (2012) showed that FA profile was clearly
influenced by diet composition and it was possible to linearly characterize the incorporation
of certain FAs, while PA failed to show a good correlation, both in muscle fat and in perirenal
fat, with its percentage in feed (Figures 3 and 4).
Figure 3. Regressions of palmitic acid percentage in rabbit muscle fat according to its contents in feed.
Lactation responses to dietary fat supplementation have been variable and have been
dependent on fat source, stage of lactation, and dry matter intake (Coppock and Wilks, 1991).
Several studies have examined the effects of PA supplementation on milk FA profile.
Grummer (1991) demonstrated that de novo FA synthesis decreased linearly as
supplementation of dietary fat increased, and that the changes in stearic acid and PA were
dependent on the ratio in the added fat. Steele and Moore (1968) reported reductions in yield
and concentration of short and medium-chain FAs (from butyric to myristic acids) and
dramatic increases in PA with increased dietary intake of PA; the concentration of PA in milk
increased from 38.7% of total FA in controls to 60.7% of total FA in cows supplemented with
PA. Noble et al. (1969) reported similar changes in milk FAs when diet was supplemented
with PA at 10%, they found that short- and medium-chain FAs decreased when compared
with a no-fat control, while milk PA increased from 36.4% of total FA in controls to 49.8% of
total FA in PA-treated cows. Banks et al. (1976) also observed decreases in short- and
medium-chain FAs in milk, with increases observed in concentrations of PA, palmitoleic, and
oleic acids. Using duodenal infusions of 500 g of PA, Enjalbert et al. (2000) reported that
concentrations of PA in milk increased 30% compared with controls. Mosley et al. (2007)
determined the optimum feeding level of a by-product rich in PA (86.6%) on dry matter
intake, milk yield, milk components, and milk FA profile in dairy cattle. They found that milk
FA concentrations were affected by the addition of this by-product. As the intake of PA
increased with the supplemented diets, milk PA concentrations increased. When 1.5 kg/d of
this by-product was consumed, milk PA concentration increased by 50% compared with the
55
control. This increase was countered by a general decrease in the weight percentage of many
other FAs, with the major difference being an increase in SFAs driven by the linear increase
in PA.
Figure 4. Regressions of palmitic acid percentage in rabbit perirenal fat according to its contents in
feed.
Chouinard et al. (1998) found that dietary Ca salts had no effect on the proportion of PA
in milk fat, but that dietary Ca salts of FA from palm oil typically increased PA (Atwal et al.,
1990; Klusmeyer and Clark, 1991; Elmeddah et al., 1994). Chouinard et al. (1998) reported
two possible explanations for the higher concentration of PA observed for dietary Ca salts of
FA from palm oil. First, palm oil contains a high proportion of PA (>40%) that may be
directly incorporated in milk fat. Second, exogenous PA stimulated synthesis and
incorporation of PA into triacylglycerols by dispersed mammary gland epithelial cells in
vitro, as found by Hansen and Knudsen (1987). Chouinard et al. (1998) reported that PA of
dietary origin might not have been sufficient to compensate for the depression in the de novo
synthesis that occurs, as observed by Grummer (1991), when a fat supplement containing
long-chain FA is fed.
Several researchers (Yu et al., 1977; Mugrditchian et al., 1981; Greene and Selivonchick,
1990; Ng et al., 2000) have reported that fish maintained a constant level of total SFA
regardless of the amount in their diet. Ng et al. (2001; 2004) have shown that palm oil has
some advantages compared to other vegetable oils when used in feeds for warm-water fish
species such as tilapia and catfish. Ng et al. (2007) found that FA composition of Atlantic
salmon fillet total lipid showed close correlations with dietary palm oil inclusion, such that
the concentrations of PA, oleic acid, linoleic acid, SFAs and monounsaturated FAs increased
linearly with increasing dietary oil supplementation.
Bell et al. (2002), studying the FA composition of muscle total lipid in Atlantic salmon
post-smolts fed diets containing increasing levels of crude palm oil, found that the
concentration of PA and other FAs increased linearly with increasing dietary palm oil (Figure
5).
56
P. G. Peiretti
Figure 5. Relationship between dietary palmitic acid percentage and muscle palmitic acid percentage in
Atlantic salmon (Bell et al., 2002; ) and in African catfish (Ng et al., 2003;).
Similarly, Ng et al. (2003) found that FA composition in the muscle lipids of African
catfish were strongly influenced by dietary treatments. In general, the FAs found in high
concentrations in the diet were also the most abundant in muscle and the converse was true of
the least abundant FAs. In particular, the concentration of PA was generally high in muscle
lipid irrespective of diet. PA was present in low concentrations in the sunflower oil and crude
palm kernel oil diets (7.2% and 7.7% of total FA, respectively) and at high concentrations in
the refined, bleached, deodorized palm olein and crude palm oil diets (44.3% and 49.5% of
total FA, respectively), but its concentration in muscle remained somewhat constant at a mean
of about 26%. Nevertheless, relatively higher concentrations of PA were still found in the
muscle of fish fed with diets having higher PA levels. PA was found in muscle lipids at a
relatively uniform concentration of 19.430.2% of total FA despite being fed with diets
containing varied levels of PA at concentrations of 7.249.5% of total FA. The conservation
of PA levels might be because this FA is the major FA in phosphatidylcholine, found in the
Sn-1 position.
The amounts of SFA and unsaturated FAs in egg yolk could be altered by dietary
manipulation (Milinsk et al., 2003). These authors found that there is a decrease in the
percentage of PA and stearic acid in egg yolk produced by hens fed diets enriched with
canola, flaxseed, soybean or sunflower oils in comparison with those of hens fed a control
diet. Kang et al. (2001) found that PA composition of egg yolk was not influenced by hen
diets containing different levels of palm oils and their PA percentage ranged from 21.8 to
24.0 % of total FA.
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In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 4
ABSTRACT
Palmitic acid or hexadecanoic acids is the most abundant saturated fatty acid in
human nutrition and represents about 17.6g per day in the United Kington diet. It is the
first fatty acid produced during the lipogenesis. During this process, glucose is converted
to fatty acids, which then react with glycerol to produce triacylglycerols. Palmitic acid
mainly occurs as its ester in triglycerides, especially in palm oil (40-44 %) but also in lard
(20-30 %), dairy products (25-40 %) and cocoa butter (25-27 %). One of the main
applications of palmitic acid in the food industry has been the formulation of
interesterified fats, used as a replacement of trans fats. In breast milk, native lard,
enzyme-directed and randomly chemically interesterified plant fats, palmitic acid
is predominantly esterified to triacylglycerol, center or -position, in native palm oil and
cows milk, it is mainly at the external or -positions. A higher palmitic acid absorption
is obtained with formulas rich in palmitic acid esterified in triacylglycerol sn-2
position, than with those containing palmitic acid predominantly esterified in the sn-1,3
positions. These specific fatty acids distributions in triacylglycerol, determine the
physical properties of the fat, which affects its absorption, metabolism and distribution
into tissues. Many authors claim that a palmitic acid intake may promote increased risk of
hypercholesterolemia, liver disease, type 2 diabetes, insulin resistance and toxicity.
However, more recent investigations on the topic seem to have reconsidered the negative
role of the dietary saturated fatty acids as a risk factor for cardiovascular diseases and
show that not only the type of fat, but also that the triglyceride structure plays a role in
these diseases.
Keywords: Palmitic acid, palm oil, triacylglycerol, saturated fatty acid, dietetic fatty acid
64
LIST OF ABBREVIATIONS
3HB 3-Hydroxybutyrate;
AcAc Cetoacetate;
ACC Acetyl-Coenzyme-A Carboxylase;
ACP Acyl Carrier Protein;
ACSL Long-Chain Acyl-CoA Synthetase;
AMP Adenosine Monophosphate;
apo Apolipoproteins;
ARA Arachadonic Acid;
ATP Adenosine Triphosphate;
BC Biotin Carboxylase;
CACT Carnitine/Acylcarnitine Translocase;
CB Cocoa Butter;
CBE Cocoa Butter Equivalents;
CBR Cocoa Butter Replacers;
CBS Cocoa Butter Substitutes;
CoA Coenzyme-A;
CPT Carnitine Palmitoyltransferase;
CT Carboxyltransferase;
DHA Docosahexaenoic Acid;
DPA Docosapentaenoic Acid;
ELOVL Elongation of Very Long-Chain Fatty Acids;
EPA Eicosapentaenoic Acid;
ETC Electron Transport Chain;
FA Fatty Acids;
FAD Flavin Adenine Dinucleotide;
FADH Reduced Flavin Adenine Dinucleotide;
FAS Fatty Acid Synthase;
FABP Fatty Acid-Binding Protein;
FABPpm Plasma Membrane Fatty Acid-Binding Protein;
FAT/CD36 Fatty Acid Translocase/Cluster of Differentiation;
FAO Fatty Acid Oxidation;
FATP Fatty Acid Transport Protein;
GPx Glutathione Peroxidase;
GTP Guanosine-5'-Triphosphate;
HADC 3-Hydroxyacyl-CoA Dehydratase
HDL High Density Lipoprotein;
HMF Human Milk Fat;
HMG-CoA 3-Hydroxy-3-Methyl-Glutaryl Coenzyme A;
IL Interleukins;
JNK Jun Amino Terminal Kinase;
KAR 3-Ketoacyl-CoA Reductase;
LCAD Long-Chain Acyl Coenzyme A Dehydrogenase;
LCFA Long-Chain Fatty Acids;
65
66
INTRODUCTION
Fatty acids (FAs) have attracted the attention of the scientific community owing to their
striking fundamental properties which are interesting for science and technology. Fatty acids
are required not only for membrane synthesis, modifications of proteins and carbohydrates,
construction of various structural elements in cells and tissues, production of signaling
compounds, and energy storage, but also for solubilizing a variety of nonpolar and poorly
soluble cellular and extracellular constituents [1, 2].
All fats and oils are esters of glycerol and fatty acids. It is commonly referred to as
triglyceride or triacylglycerol (TAG) because of the glycerol molecule has three hydroxyl
groups where a fatty acid can be attached. All TAGs have the same glycerol units, and then
fatty acids contribute to the different properties. Fatty acids are often categorized into short
chain (up to 6 carbons), medium chain (8 to 12 carbons), or long chain (>12 carbons).
Although hydrocarbon chain length is an important determinant of function, fatty acids are
often classified based on whether or not the fatty acid carbon chain contains no double bonds
(saturated fatty acid SFA), one double bond (monounsaturated fatty acid MUFA), or more
than one double bond (polyunsaturated fatty acid PUFA), as well as the configuration of the
double bonds (cis or trans). In addition, PUFAs are often further classified based on the
position of the first double bond from the fatty acid methyl terminus, creating n-3 and n-6
fatty acids [3, 4].
It is the differences in chain length and saturation status that dictate their performance in
food and cooking, as well as their role in the body and impact on human health and disease
risk [5]. The major types of FAs in the circulation and in the tissues of mammals are the longchain and very-long-chain FAs with many degrees of saturation. These include palmitic acid
(C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), oleic acid (C18:1n-9), linoleic acid
(C18:2n-6) and, particularly in smaller mammals, arachidonic acid (20:4n-6) and
docosahexaenoic acid (22:6n-3) [6].
The variety of fatty acid in common fats and oils is provided in Table 1.
Table 1. Fatty acid profiles (%)a of select animal and vegetable fats and oils
LIPID
SFA
8:0
10:0
12:0
14:0
16:0
18:0
MUFA
18:1
PUFA
18:2
18:3
Avocado oil
Beef tallow
Butterj
Canola oil
Coconut oil
Corn oil
Flaxseed oil
Grapeseed oil
Lard
Olive oil
Palm oil
Palm kernel oil
Rice bran
Salmon oil
Soybean oil
11.9
46.8
53.6
7.6
11.8
12.9
9.0
9.6
36.9
13.7
49.3
81.5
19.7
19.9
15.7
0.0
0.0
5.1
0.0
1.0
0.0
0.0
0.0
0.0
0.0
0.0
3.3
0.0
0.0
0.0
0.0
2.6
0.0
0.8
0.0
0.0
0.0
0.1
0.0
0.0
3.7
0.0
0.0
0.0
0.9
2.7
0.0
6.1
0.0
0.0
0.0
0.2
0.0
0.1
47.1
0.0
0.0
0.0
3.5
7.8
0.0
2.3
0.0
0.1
0.1
1.3
0.0
1.0
16.4
0.7
3.3
0.0
11.3
23.5
22.6
4.4
1.1
10.6
5.1
6.7
22.4
11.2
43.5
8.1
16.9
9.9
10.4
0.7
17.8
10.4
6.8
0.4
1.8
3.4
2.7
12.7
1.9
4.3
2.8
1.6
4.3
4.4
72.6
39.3
22.0
65.1
0.8
27.6
18.5
16.1
42.5
72.4
37.0
11.4
39.3
29.0
22.8
69.9
33.9
20.8
63.5
0.8
27.4
18.3
15.8
38.8
70.7
36.6
11.4
39.1
17.0
22.6
13.9
3.8
3.2
29.0
0.2
54.7
67.9
69.9
10.5
10.4
9.3
1.6
35.0
40.3
57.7
12.9
2.9
2.9
19.6
0.2
53.5
14.3
69.6
9.6
9.7
9.1
1.6
33.4
1.5
51.0
1.0
0.6
0.4
9.4
0.0
1.2
53.4
0.1
1.0
0.7
0.2
0.0
1.6
1.0
7.1
EPAe
DPAf
DHAg
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
31.3
0.0
ARAh
TFAi
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.7
0.0
0.0
0.0
0.0
0.4
0.0
0.3
0.1
0.0
0.5
Listed as percent of total fatty acid content, based on 13.6 g fatty acids/tablespoon. Cells without numbers did not have data in United
States Department of Agriculture Nutrient Database; bSFA saturated fatty acid; cMUFA monounsaturated fatty acid; dPUFA
polyunsaturated fatty acid; eEPA eicosapentaenoic acid; fDPA docosapentaenoic acid; gDHA docosahexaenoic acid; hARA
arachadonic acid; iTFA trans-fatty acid; jButter contains 16 % water and therefore the percentages are unable to be directly
compared with percentages of the other fats/oils [5].
68
PALMITIC ACID
The word 'palmitic' is French in origin, derived from the word palmitique which refers to
the pith of the palm tree. The palmitic acid (PA) (C16:0, hexadecanoic acid) is one of the
most common saturated fatty acids found in animals and plants with chemical formula
CH3(CH2)14COOH. It is a white solid that melts at 63-64 C, have a boiling point of 351352 C, a mass of 256.42 gmol and a density of 0.853 gcm3 at 62 C. PA is water-insoluble
(7.2 x 10-4 g/100 g) at 20 C and slightly soluble in ethanol and iced acetone, but it is highly
soluble in alcohol, heated acetone and chloroform with 4.76 0.02 pKa value [7]. It is
constituted of carbon atoms linked to each other through single bonds as shown in Figure 1.
The molecules are arranged as dimers through OHO hydrogen bonds. These dimers are
packed in bilayers with terminal methyl groups at both external faces and these layers are
parallel to the crystallographic (100) plane [8].
PA mainly occurs as its ester in TAG, especially in palm oil but also in lard, dairy
products, avocado oil, butter and beef tallow (Table 1). Palmitate is a term for the salts and
esters of palmitic acid. The palmitate anion is the observed form of palmitic acid at basic pH.
Aluminum salts of palmitic acid and naphthenic acid were combined during World War II to
produce napalm. The word "napalm" is derived from the words naphthenic acid and palmitic
acid [9].
Beyond PA presence in feed, it has utilities ranging from inks antioxidants application,
waterproofing in textile industry, candle manufacture together with paraffin and liquid
crystal, widely used in electronic industry [7].
Palmitic acid can participates in several chemical reactions as other acids of this same
class, which include:
- Neutralization reactions: salts of palmitic acid are formed through this reaction.
The palmitic acid reacts with a hard base, forming a salt of palmitic acid and water. Using
69
sodium hydroxide as a base, for example, sodium palmitate is obtained as shown in Equation
1 (Figure 2) [7].
Those salts, especially lithium, sodium and potassium, have vast application as ink
constituent, lubricants and insulations that protect against water corrosive action [7].
- Esterification reaction: directly reaction of palmitic acid and a monohydric alcohol
obtaining an ester (Figure 3. Equation 2), or by a reaction between a salt of palmitic acid and
a haloalkane (Figure 3. Equation 3) [7].
(2)
(3)
Figure 3. Equation 2 and 3. Esterification reactions.
RETYNIL PALMITATE
The two most abundant retinoid forms that are present in the diet are retinol and retinyl
esters - a fatty acyl group is esterified to the hydroxyl terminus of retinol [10]. Dietary retinol
is taken up directly by mucosal cells. Nevertheless, dietary retinyl esters are cleaved in the
intestine by the pancreatic triglyceride lipase and intestinal brush border enzyme,
phospholipase B [11]. The free retinol taken up by the enterocyte is complexed with cellular
retinol-binding protein type 2 and the complex serves as a substrate for reesterification of the
retinol by the enzyme lecithin retinol acyltransferase (LRAT). The resulted retynyl ester are
incorporated with other neutral lipid esters (i.e., triacylglycerols and cholesteryl esters) into
chylomicrons and absorbed via the lymphatics [11, 12]. In the vascular compartment, much of
the chylomicron triacylglycerol is hydrolyzed by lipoprotein lipase in extrahepatic tissues,
resulting in the production of a chylomicron remnant that contains most of the newly
absorbed retinyl esters [13]. Under conditions of adequate vitamin A nutrition, the liver is the
main site of vitamin A storage, with more than 95 % of the total neutral retinoid being present
70
as retinyl esters, predominately retinyl palmitate and stearate [12, 14, 15]. According to Ihara
et al. [12] the explanation for the use of those fatty acids would be that the retinyl esters
together cholesterol present in its particle would be release into the liver cells via LDL
receptor wherein saturated fatty acid regulate it pathway largely mRNA level. Once the cells
has met it requirements for retinol, saturated fatty acids inhibit receptor activity, so that the
receptor is no longer able to internalize retynil esters. In the response to cellular requirements
the liver release retinol in forms of a retinol-binding protein (RPB4), target cells, a cell
surface receptor for retinol RBP4 remove retinol from RPB4.
Esterification techniques using palmitic acid have yielded more stable esters in the form
of retinyl palmitate which has been used successfully as a supplement as well as a way to
fortify numerous foods, including vegetable oil, rice, monosodium glutamate, cereal flours
and sugar [review 16, 17]. This application is due to its high stability in relation to vitamin A
and its low cost [16]. The oil matrix protects against the oxidation of vitamin A during
storage, improves stability of the retinol and facilitates the vitamins absorption by the body
[18].
The advantages of oil fortified with retinyl palmitate have historically been utilized by
food aid programs, where a daily intake of 16 g of oil provided approximately 50 % of the
recommended daily intake (RDI) of an adult male [19]. Surman et al. [20] reports that retinyl
palmitate it also can be associated with toxicities at high doses. The precise human dose
required to ensure efficacy without toxicity remains a point of controversy. Also, retinyl
palmitate (RP) is widely used in pharmaceutical and cosmetics products to improve the skin
elasticity [21, 22].
71
72
Fatty acid
12:0
14:0
16:0
16:1
18:0
18:1
18:2
18:3
20:0
Saturates
Monounsaturates
Polyunsaturates
[23].
Palm olein
0.10.2
0.91.0
39.540.8
Trace0.2
3.94.4
42.743.9
10.611.4
00.4
0.10.3
45.8
42.5
11.6
Super olein
0.4
1.4
31.5
3.2
49.2
13.7
0.3
0.4
36.6
49.2
14.0
Super stearin
0.10.2
1.01.3
46.568.9
Trace0.2
4.45.5
19.938.4
4.19.3
0.10.2
0.10.3
52.176.2
19.938.6
4.29.5
Butter
Butter or buttermilk is an important edible fat in northern Europe, North America and
Brazil, being about the third product of worlds milk production. It is a yellow-to-white solid
and an emulsion of fat globules, water and inorganic salts produced by churning the cream
from cows milk. Butter has high energy (~715 Kcal/100 g), cholesterol (~215 mg/100 g) and
a major content of FAs, respectively: palmitic (25-32 %, C16:0), oleic (22-29 %, C18:1),
myristic (C14:0) and stearic (C18:0) about same proportions (8-13 %), linoleic (C18:2) and
lauric (C12:0) with less than 4.5 %. In despite of high SFAs contents, MUFAs are higher than
PUFAs in buttermilks [39, 40]. Verardo et al. [40] determined the fatty acid composition of
different samples of butter and the samples manufactured by a traditional method showed
higher levels of MUFAs and PUFAs compared with industrial samples. Palmitic acid
presented the higher fatty acids contents, 29.8 to 31.1 % and 30.3 to 33.6 % from traditional
and industrial process, respectively.
Cocoa Butter
The generic name of cocoa is Theobroma belonging to the family of Sterculiaceae, also
called Food of God. It contains about 3050 beans, covered with pulp. About 500 years
ago, cocoa beans were originated from Latin America and within a few years it was spread to
Europe [41, 42].
Cocoa butter (CB) is a highly valued ingredient primarily used in the confectionery
industry due to its specific physical and chemical properties. CB is solid at room temperature
(below 25 C), and liquid at body temperature (~37 C) [43]. Furthermore, the predominant
presence of symmetrical TAG, about 90 % of the TAG species in CB, is mainly responsible
for the functionality of this fat [44]. The major FAs of cocoa butter are palmitic acid (C16)
2533.7 %, stearic acid (C18:0) 33.740.2 %, oleic acid (C18:1) 26.335 % and linoleic acid
73
(C18:2) 1.73 % which contribute about 98 % of the total fatty acid [45]. Regarding the
palmitic acid composition of natural cocoa butter produced from various countries ranged
from 24.1 to 27.9 % [43]. CB fat contains significantly higher amount of saturated acid
leadings to triglycerides of glycerol-1,3-dipalmitate-2-oleate (POP), glycerol-1-palmitate-2oleate-3-stearate (POS) and glycerol-1,3-distearate-2-oleate (SOS). Among these three TAGs,
POS is the major leading triacylglycerol component present in cocoa butter with range 42.5
46.4 % yield followed by SOS (27.833.0 %) and POP (18.922.6 %). Therefore, palmitic
acid occupies mostly the sn-1 position in cocoa butter [45].
Avocado Oil
Avocado (Persea americana Mill) is an important tropical fruit and a good source of
lipophilic phytochemicals such as monounsaturated fatty acids, carotenoids, vitamin E and
sterols [46]. It has several cultivars that present great variation on time of fruit production and
oil content in the pulp. Studies have indicated that the avocado oil is similar to olive oil and
can be used in cosmetics and also for human consumption [47, 48]. New Zealand, Mexico,
Chile United States and South America are among the main avocado oil producers. Avocado
oil has the advantage that can be obtained from the fruit by means of a cold extraction
methods, which is an easy and low technology that allow maintain in the oil significant
amounts of the bioactive phytochemicals present in the fruit [47].
In comparison to other vegetable sources, avocado oil is characterized by its contents of
palmitic (C16:0), linoleic (C18:2), palmitoleic (16:1) and alpha-linolenic (18:3) FAs that are
13.5, 12.6, 3.26 and 1.0 % of total fatty acids, respectively. Stearic (18:0), tridecanoic (13:0),
tetradecanoic (14:0), cis-10-heptadecenoic (17:1) and cis-13-16-eicosenoic (20:2) FAs are
present in trace amounts [49]. Ozdemir and Topuz [50] showed in Fuerte and Hass avocado
varieties present a reduction of palmitc acid according to fruit ripening, with variations of
22.4-12 % to Fuerte variety and 23.3-16.8 % to Hass variety. Yanty et al. [51] studying three
Malaysia avocados varieties, found oleic acid as the major fatty acid (43.6551.22 %)
followed by palmitic (26.4130.37 %) and linoleic (12.7517.45 %) FAs. Oils of avocado
fruits are generally found to have extremely low amounts of stearic acid (0.271.56 %).
Beef Tallow
Beef tallow is one residual material from slaughterhouses which main destination is the
soap industry, however because of its high melting point (45 C) and low level of
polyunsaturated fatty acids (< 3 %) [52, 53] beef tallow is considered as a less valuable fat
not suitable for direct human consumption [54]. From the nutritional point of view, vegetable
oils are preferred over animal fats, because contain a high proportion of saturated fatty acids
and low proportion of polyunsaturated fatty acids. Regarding fatty acids composition, tallow
has about 29 % palmitic acid, 25-37 % stearic acid and 23-31 % oleic acid. Thus, saturated
fatty acid content is responsible for over 50 % of total fatty acids in the beef tallow [52, 53].
The higher stearic and palmitic acid content of beef tallow are accounted for the unique
properties of high melting point and high viscosity [55]. In relation to TAG, beef tallow and
other bovine adipose tissues have nearly 50% of the fatty acids in the sn-2 position, which are
74
oleic acid (unsaturated fatty acid), palmitic and stearic acids. Nevertheless, oleic acids are the
most fatty acids in the sn-1,3 positions [52, 56].
Lard
Animal fats such as lard and tallow have long been recognized as raw material for food
and industrial applications. Lard has exceptional properties compared to other vegetable oils
such as firmness and special flavor values. Nevertheless, lard has lost its significance to
numerous substitutes such as hydrogenated cottonseed and soybean oil due to its negative
nutritional values such as low digestibility, high calories and high content of saturated fatty
acids. Nonetheless, lard remains a major ally in the meat product industry due to its positive
contributions in flavor and texture [57, 58].
Lard contains about 28.4 % palmitic acid (C16:0), 21 6 % stearic acid (C18:0), 31.4 %
oleic acid (C18: 1) 11.1 % linoleic acid (C18: 2). It is unique among animal depot fats,
because it has a strong predominance of saturated fatty acids in the sn-2 position [52]. In lard,
C16:0 is located exclusively at the sn-2 position, with an unsaturated fatty acid at sn-3 but the
fatty acid occupying the sn-1 position is highly variable, as in SPO, OPL and OPO TAGs
(species dominants in lard) [56].
Milk
Bovine milk and dairy products have long traditions in human nutrition. Its fat fractions
are widely used in a variety of food products such as liquid milk, cream, butter, cheese and
ice cream due to many favorable physical, chemical and nutritional properties of milk fat
[59].
Milk contains, in average, about 33 g total lipid/L, being 95-98 % of triacylglycerols
composed of fatty acids of different length and saturation, less than 0.5 % of cholesterol,
about 1 % of phospholipids and less than 0.5 % of free fatty acids. SFA represent more than
half of total milk fat, about 19 g/L especially lauric (C12:0), myristic (C14:0) and palmitic
(C16:0). Furthermore it is rich in oleic acid (about 25 %), however, a relatively poor source of
polyunsaturated fatty acids as linoleic (C18:2) and alpha linolenic (C18:3), with contents in
the order of 3 % and 2 %, respectively [59; 60; 61].
In relation to TAG molecules, the most probable sn-position of the main fatty acids in
milk fat are: sn-position 1 C16:0, C18:0 and C18:1 with 44.1 %, 54.0 % and 37.3%
respectively; sn-position 2 C12:0 (62.9 %), C14:0 (65.6 %) and C16:0 (45.4 %); and snposition 3 98.1 % of C4:0, 93.0 % of C6:0, 34.5 % of C8:0 and 41.5 % of C18:1. Therefore,
the most probable sn-position of palmitic acid in TAG molecules of milk are in sn-1 and sn-2
[61].
Human Milk
The human mammary gland has evolved with unusual pathways for acylation of fatty
acids into triglycerides for secretion in milk, and major portion of milk fat is comprised by
75
these molecules that represent 98 % of total fat [61, 62]. The average fat content of human
milk is about 3.83.9 g/100 ml, but it varies widely [61]. The predominant fraction are
saturated fatty acids, followed by a relatively high proportion of monounsaturated fatty acids
such as oleic acid (18:1n 9) [59]. Palmitic acid is the qualitatively and quantitatively major
SFA in human milk and constitutes approximately one fourth of this with a concentration
highly conserved, regardless of ethnic origin or the nutritional status of the woman [62]. PA
comprises 17 % to 25 % of the total FAs [63] and it is an important source of energy thereby
contributing 1012 % of breast-fed infants' dietary energy intake [64].
The stereospecific numbering (sn) designates the location of fatty acids within the
triglyceride molecule. If the glycerol is drawn with the first and third hydroxyl groups to the
right and the second to the left, the first carbon is termed sn-1; the second, sn-2, and the third
sn-3 [61]. Most of the 16:0 in human milk is located in the sn-2 position of the triacylglycerol
molecules (70 % to 75 %) [63], in contrast to cows milk and vegetable oils which have 40 %
and 5 % to 20 %, respectively, of the 16:0 in the sn-2 position [62]. The major unsaturated
fatty acid in human milk is oleic acid (18:1n-9) and this is mostly esterified at the triglyceride
sn-1,3 (outer) positions, with the result that triglycerides with the structure 18:1n-916:0
18:1n-9 (1,3-dioleoyl-2-palmitoylglycerol, OPO) are a major triglyceride species in human
milk and represent an estimated 11.8 % of the total triglyceride species [65]. Since lipolytic
enzymes will cleave the FA in sn-1 and sn-3-positions, human milk palmitic acid will appear
primarily in the remaining monoglyceride which has a higher polar than free palmitic acid
[61]. Hence, most of the 16:0 is absorbed as the sn-2 and this structure is preserved through
and beyond the intestinal wall.
SPECIAL OILS
Pequi Tree
Pequi tree (Caryocar brasiliensis Camb.) is a member of the Central and South American
family Caryocaraceae [66, 67]. It stands out by high occurrence in Brazilian Cerrado and
extensive period of fruit production, which can be collected from September to February in
Cerrado of Goias (Brazil) [68]. It is a Brazilian oleaginous fruit, rich in A, E, C and B2
vitamins in both edible parts: pulp and kernel [69, 70].
The oil is extracted by rendering the mesocarp and kernel of pequi fruit and the oil is
generally used to cook rice with the objective of adding specic avors and a light-yellow
color to the nal product [71]. The fruit is rich in -carotene and selenium. It is even used to
produce fermented liquor. Pequi has a considerable economic importance in some parts of
Brazil and has a substantial ecological impact on the country. In relation to fatty acids
composition, kernel and pulp present high content of palmitic acid (35.17 % and 43.76 %)
and oleic (43.59 % and 55.87 %), respectively. The TAG composition of pequi oil is also
relatively simple with trioleoyl glycerol (OOO, 56 gkg1), palmitoyl dioleoyl glycerol (POO,
466 gkg1) and dipalmitoyl oleoyl glycerol (POP, 452 gkg1) comprising 974 gkg1 of the
total. Dioleoyl stearoyl glycerol (OOS) was found in small amounts (5.2 gkg1). The
relatively simple composition of pequi oil may be of interest for selected applications.
76
It appears that is has some promise as a less expensive chocolate substitute upon fractionation
[23, 38].
77
78
at position 2 and in other fatty acids at positions 1 and 3 (oleic acid, caprylic acid, etc.). These
fat compositions can be obtained by subjecting fatty mixtures comprising glycerides
consisting substantially of more saturated 2-palmitoyl glycerides to a rearrangement catalyst,
such as a lipase, which is regiospecific in activity in the 1- and 3-positions of the glycerides.
Under the influence of the catalyst, unsaturated fatty acid residues may be introduced into the
1- and 3-positions of the 2-palmitoyl glycerides by exchange with unsaturated free fatty acids
or their alkyl esters.
Unfortunately, such formulas have resulted in poor absorption of fats and minerals,
particularly when studied in infants during the first few weeks of life [62, 91, 92]. This is
because PA is present in the sn-2 position (human breast milk, lard native, enzyme-directed
and randomly chemically interesterified fats plant) compared with the sn-1 and sn-3 positions
(bovine milk, randomly chemically interesterified lard or crude palm oil). It was found the
relative absorption of palmitic acid and full fat was linearly related to the proportion of
palmitic acid in the sn-2 position of the TAG in human infants. Pancreatic lipase selectively
hydrolyses the fatty acids at the sn-1 and sn-3 positions, yielding free fatty acids and
monoacylglycerols (MGs). The TAG sn-2 position is absorbed more efficiently than free
palmitic acid and it is conserved through the digestion, absorption and chylomicron TAG
synthesis [63, 91].
Nonetheless, after digestion, the free PA solidify in the intestine because of their high
melting temperature, creating insoluble and indigestible complexes with dietary minerals (eg,
calcium), and causing hard stools [87]. Quinlan et al. [93] were able to relate stool hardness to
stool composition. They concluded that differences in the triacylglycerol palmitate content of
formula and breast milk resulted in more calcium soap formation in formula- fed infants and
thus in harder stools. HMF containing palmitic acid at the sn-2 position yields 2-MG during
digestion which does not lead to the formation of calcium soaps. Consequently, both calcium
and 2-MG become bioavailable for the infant [94]. Yaron et al. [63] studying two infant
formulas demonstrates that -palmitate may affect the intestinal microbiota composition
during the first weeks of life by increasing Lactobacillus and Bifidobacteria abundance in the
stool, and thus may provide beneficial effects for the health and well-being of formula-fed
infants. They concluded that the effects of the infant formulas of the gut microflora are due to
lipid estructure. It is therefore important to synthesize TAGs with a composition and
distribution of fatty acids similar to those of human milk. In particular there is considerable
interest in the synthesis of 1,3-diolein-2-palmitin (OPO), which is the most abundant TAG in
human milk.
79
80
Figure 5. Acetyl-coenzyme-A carboxylase (ACC) has critical roles in fatty acid metabolism. The ACCcatalyzed biotin carboxylase (BC) and carboxyltransferase (CT) reactions [103].
Basically, this reaction consists of elongating the acetyl group by C2 units derived from
malonyl-CoA, so that each step takes place by condensation, reduction, dehydration and
further reduction [99] (Figura 6):
1. Transfer of the acetyl group of acetyl-CoA to ACP-catalyzed by acetyl-CoA-ACP
transacylase;
81
2. Next, this two-carbon fragment is transferred to a temporary holding site, the thiol
group of a cysteine residue on the enzyme;
3. The now-vacant ACP accepts a three-carbon malonate unit from malonyl CoA
catalyzed by malonyl CoA-ACP transacylase;
4. The acetyl group on the cysteine residue condenses with the malonyl group on ACP
as the CO2 originally added by acetyl CoA carboxylase is released. The result is a
four-carbon unit attached to the ACP domain. The loss of free energy from the
decarboxylation drives the reaction catalyzed by 3-Ketoacyl-ACP synthase;
5. Reduction of the Beta-keto group to a Beta-hydroxyl group with NADPH catalyzed
by Beta-keto-ACP reductase;
6. Dehydration between the alpha and Beta by Beta-hydroxyacyl-ACP dehydrase. A
molecule of water is removed to introduce a double bond between carbons 2 and 3
(the - and -carbons);
7. Reduction of the trans double bond by NADPH catalyzed by enoyl-ACP reductase).
Figure 6. Reaction sequence for biosynthesis of fatty acids de novo by the animal FAS.
The result of these seven steps is production of a four-carbon compound (butyryl) whose
three terminal carbons are fully saturated, and which remains attached to the ACP. These
seven steps are repeated, beginning with the transfer of the butyryl chain from the ACP to the
Cys residue, the attachment of a molecule of malonate to the ACP (3), and the condensation
of the two molecules liberating CO2 (4). The carbonyl group at the -carbon is then reduced
(5), dehydrated (6), and reduced (7), generating hexanoyl-ACP. This cycle of reactions is
repeated five more times, each time incorporating a two-carbon unit (derived from malonyl
CoA) into the growing fatty acid chain at the carboxyl end. When the fatty acid reaches a
length of 16 carbons, the synthetic process is terminated with palmitoyl-S-ACP. All the
carbons in PA have passed through malonyl CoA except the two donated by the original
acetyl CoA, which are found at the methyl-group () end of the fatty acid [98, 99, 105, 106].
82
Palmitic acid can be further elongated by the addition of two-carbon units in the smooth
endoplasmic reticulum (SER). Elongation involves the addition of two-carbon units to a fatty
acyl-CoA, employing malonyl-CoA as the donor and NADPH as the reducing agent. In
mammals, the initial and rate-controlling condensation reaction is catalysed by the elongase
enzymes referred to as elongation of very long-chain fatty acids (ELOVLs) [108]. To date,
seven ELOVL proteins (ELOVL1-7) have been identified, with ELOVL1, ELOVL3,
ELOVL6 and ELOVL7 preferring saturated and monounsaturated fatty acids as substrate; and
ELOVL2, ELOVL4 and ELOVL5 being selective for PUFAs [109, 110]. The process of
elongation requires four separate enzymatic reactions: condensation between the fatty acylCoA and malonyl-CoA to yield 3-ketoacyl-CoA; reduction of 3-ketoacyl-CoA to generate 3hydroxyacyl-CoA; dehydration of 3-hydroxyacyl-CoA to produce trans-2-enoyl-CoA, and;
reduction of trans-2-enoyl-CoA to form the two-carbon elongated acyl-CoA (Figure 7). [108,
109, 111].
Figure 7. Enzymatic steps in long-chain fatty acid elongation. Enzymatic steps of microsomal fatty acyl
chain elongation. ELOVL, elongation of very-long-chain fatty acids; KAR, 3-ketoacyl-CoA reductase;
HADC, 3-hydroxyacyl-CoA dehydratase; TER, trans-2,3-enoyl-CoA reductase [108].
83
Fatty acids are also formed in endoplasmic reticulum and the reaction is catalyzed by
enzymatic systems, generically designated as Acyl-CoA desaturases [112]. Acyl-coenzymeA
(CoA) desaturases introduce a double bond at a specific position on the acyl chain of longchain fatty acids, thereby influencing several of the key biological properties of the fatty acid
itself and of more complex lipids containing this acyl chain. Mammalian cells express 9, 6
and 5-desaturase activities [108]. The desaturation process involves an oxidoreductase chain
(including cytochrome b5) that O2 works as last oxidant of Acyl-CoA and NADPH (or
NADH). The PA product of FAS and its metabolite produced by Stearoyl-Coa Desaturase
(SCD-1), C16:1n-7, can both be further elongated by ELOVL6 to yield stearic acid (C18:0) and
vaccenic acid (C18:1, n7), respectively [108; 109].
Lipolisis
In the fasted state, most tissues, except the brain and red blood cells, rely heavily on the
direct utilization of FA to generate energy. The prime pathway for the degradation of fatty
acids is mitochondrial fatty acid -oxidation (FAO), a key metabolic pathway for energy
homoeostasis in organs [113, 114]. Long-chain fatty acids (LCFA) (C16-18) in tissues exist
as components of TAG or phospholipids. Adipose tissue TAG storages are the primary source
of fatty acids used for FAO during fasting conditions [115].
TAGs are first hydrolyzed by the action of endothelium-bound lipoprotein lipase release
free FAs, which are transported to tissues via the bloodstream. The uptake of FAs seems to be
largely mediated by membrane proteins, although passive uptake probably also occurs. This
implies that several transport steps are necessary before fatty acids are oxidized [115, 116,
117].
The solubility of LCFA in aqueous solutions is extremely low, so the fatty acids must
cross the cell membrane via a protein-mediated mechanism. Membrane-associated fatty acidbinding proteins (fatty acid transporters) are small (15kDa) cytosolic proteins that enhance
the uptake of long chain and very long chain fatty acids into cells. Through their control of
fatty acid transport, metabolism and storage, FABPs are proposed to be central regulators of
lipid metabolism, inflammation and energy homeostasis. In humans, FATPs comprise a
family of six highly homologous proteins, FATP1FATP 6, which are found in all FAs
utilizing tissues of the body [118, 119]. Besides FATPs, FAT/CD36 (fatty acid translocase
/Cluster of Differentiation 36) has been shown to function as a plasma membrane LCFA
transporter in various tissues, including skeletal muscle, heart, liver, adipose tissue and the
small intestine [120, 121]. Other mechanism is fatty acid uptake plasma membrane fatty acidbinding protein (FABPpm) which is associated with the plasma membrane in many tissues
including liver, adipose tissue, cardiac muscle and vascular [122, 123, 124, 125].
After transport across the plasma membrane, FAs must be esterified to coenzyme A, on
the outer mitochodrial membrane by long chain acyl-CoA synthetase activity (ACSL; C12 to
C20) before they can undergo oxidative degradation. This reaction is coupled with two ATP
hydrolysis to AMP and 2Pi. The mitochondrial membrane is not permeable to long chain
acyl-CoA (i.e., C16-C18), therefore requires the initial conversion of acyl-CoA to an ester
acylcarnitine, followed by transport of the acylcarnitine across the inner mitochondrial
membrane into the mitochondrial matrix and subsequent delivery of acyl-CoA [126]. This
process is referred to as carnitine shuttle and requires the concerted action of 3 proteins 6:
84
The complete oxidation PA is achieved in the following three steps (Figure 9): betaoxidation of fatty acid chain yielding acyl-CoA; the oxidation of acetyl CoA to CO2 and
production FADH2 and NADH2 in citric acid cycle; the transfer of electron from reduced
electrons carries FADH2 and NADH2 to mitochondrial respiratory chain resulting into ATPs
[130]. This process involves a variety of enzymes: long-chain acyl coenzyme dehydrogenase
(LCAD), enoyl-CoA hydratase, hydroxyacyl-CoA dehydrogenase and ketoacyl-CoA thiolase
[117, 131];
1. In the first step, LCAD, catalyzes oxidation of the fatty acid moiety of acyl-CoA to
produce a double bond is introduced into a carboxylic acid between the and
carbons, FAD is the electron acceptor, and electrons from the reaction ultimately
enter the respiratory chain and are carried to O2 with the concomitant synthesis of
two ATP molecules per electron pair;
2. In the second step of the fatty acid oxidation cycle, water is added to the double bond
of the trans-2-enoyl-CoA to form the L stereoisomer of -hydroxyacyl-CoA. This
reaction, catalyzed by enoyl-CoA hydratase;
3. In the third step, the L--hydroxyacyl-CoA is dehydrogenated to form -ketoacylCoA by the action of -hydroxyacyl-CoA dehydrogenase; NAD+ is the electron
acceptor. The NADH formed in this reaction donates its electrons to NADH
85
dehydrogenase an electron carrier of the respiratory chain. Three ATP molecules are
generated from ADP per pair of electrons passing from NADH to O2 via the
respiratory chain;
4. Finally hydroxy-acyl-CoA is dehydrogenated to 3-keto-acyl-CoA. Then, thiolytic
cleavage of the 3-keto-acyl-CoA produces a two-carbon chain-shortened acyl-CoA
plus acetyl-CoA. Each cycle yields an acyl-CoA shortened by two carbon atoms, an
acetyl-CoA, and one nicotinamide adenine dinucleotide (NADH) and one flavin
adenine dinucleotide (FADH2) as electron carriers (or reducing equivalents).
The PA undergoes seven passes through this oxidative sequence, in each pass losing two
carbons as acetyl-CoA. At the end of seven cycles the last two carbons of palmitate
(originally C-15 and C-16) are left as acetyl-CoA. Generally, the total ATP yield due to the
complete oxidation of palmitic acid in the following equation [99, 130]:
Palmitic acid + 8 Coenzyme A + 7 FAD+ + 7NAD+ + ATP 8 CH3CO-SCoA (AcetylCoA) + 7FADH2 + 7 (NADH + H+) + AMP + PPi.
If one acetyl CoA involved in TCA cycle gives = 10 ATPs; ATPs due 8 acetyl-CoA = 8
x 10 = 80; ATPs due to 7 FADH2 = 1.5 x 7= 10.5; ATPS due 7 (NADH + H+) = 2.5 x 7.5 =
17.5. The total of number ATPs produced 108. During the initiation of the - oxidation
pathway a 2 ATPs converts into a 2 AMP and 2 Pi for the activation of fatty acid. So, net
ATPs produced by palmitic acid are 106. These calculations assume that mitochondrial
oxidative phosphorlation produces 1.5 ATPs/FADH2 oxidized and 2.5 ATP/NADH2 oxidized.
The Guanosine-5'-triphosphate (GTP) produced directly in the acid citric cycle yields ATP in
the reaction catalyzed by nucleoside diphosphate kinase [99, 130].
Figure 9. The -oxidation of saturated fatty acids involves a cycle of four enzyme-catalyzed reactions.
Each cycle produces single molecules of FADH2, NADH, and acetyl-CoA and yields a fatty acid
shortened by two carbons.
86
87
88
substitution of palm oil or its liquid fractions (palm olein, super olein) for habitual fats in the
diet does not result in an elevation of total serum cholesterol. In another review, Edem [23]
concluded that, in animal experiment and human studies, palm oil administration with
approximately 50 % of saturated fatty acid, does not behave as saturated oil, reducing the
blood levels of total cholesterol, LDL cholesterol. A review by Sundram [29] also concluded
that high levels of palmitic acid in the diet do not significantly affect serum total and LDL
cholesterol levels.
Khosla and Hayes [141] conclusions about cholesterolaemic effects of the saturated fatty
acid of palm oil suggest that not all SFAs are cholesterol-raising. According to authors, when
fatty acids contents are similar, the palmitic acid appears to have no impact on the plasma
cholesterol in normocholesterolaemic subjects. Above 400 mg of dietary cholesterol intakeed
per day, PA might be cholesterol increasing, even more than myristic acid and quite neutral
underneath this value. Nevertheless, if cholesterol consumption exceeds the critical value or
when hypercholesterolaemic subjects are studied, the PA appears to increase the plasma
cholesterol. Furthermore, authors linked the different PA actions to the differences in LDLreceptor status. It seems that more studies are needed to explain these inconclusive results.
Free FAs absorption rate depends on the type of fatty acid and intestine emulsifier
environment. An important explanation of why palm oil does not "follow the Keys [138] and
Hegsted [139] model is due to unsaturated fatty acids are in sn-2 position (> 58.25 % of oleic
acid and > 18.41 % of linoleic acid) and a high proportion of PA is in sn-1 and 3 positions
(17-23 %) [23]. Therefore, as already mentioned in this text, fatty acids in sn-2 position are
preferentially absorbed at bowel wall and, thereby, more bioavailable than the fatty acids in
sn-1 and 3 positions [87]. On the assumption that all SFA localized at sn-1 and 3 positions in
palm oil are preferentially absorbed, whilst saturated are faecal excreted as salts, therefore
only 8 % of SFA localized at sn-2 position would be absorbed as consequence there is a less
caloric intake and a lower serum TGA content.
Another explanation to palm oil hypocholesterolemic action would be the presence of
tocotrienols. They are admittedly considered hypocholesterolemic once they regulate
cholesterol synthesis through 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA)
reductase inactivation enzyme that primarily synthesizes the cholesterol [147].
In a review of Hayes and Khosla [148], the authors concluded that cholesterolemic
effects of PA are large extent determined by the concomitant level of linoleic FAs. It is why
C18:2 regulate numerous lipogenic genes involved in fatty acid synthesis. PUFA maximally
inhibit hepatic gene transcription when they provide 20 % of the dietary calories.
Nonetheless, as little as 5% of calories as PUFA will inhibit gene expression 50 %. In this
way, once 18:2 intake is above threshold (57 %) the detrimental effects of the SFA on
LDL are no longer observed in part because LDL receptors are maximally up-regulated and
sterol regulatory element-binding protein (SREBP1c; that group of proteins uptake
cholesterol and FAs biosynthesis) is inhibited, resulting in decreased FAs synthesis and
decreased VLDL secretion. As a consequence, PA appears neutral above the threshold
requirement for linoleic FAs (Figure 10). Simply stated it implies that a certain level of 18:2
is required by an individual to prevent certain SFA-rich fats from raising the serum LDL
level. If you drop below your 18:2 threshold, LDL will increase, with the increase being most
severe during consumption of 12:0 1 14:0-rich fats. Also note that if 18:2 intake is high
enough (610 % en depending on the LP setpoint), SFA no longer have a significant
cholesterol-raising effect. Simply stated it implies that a certain level of 18:2 is required by an
89
individual to prevent certain SFA-rich fats from raising the serum LDL level. If you drop
below your 18:2 threshold, LDL will increase, with the increase being most severe during
consumption of 12:0 1 14:0-rich fats. Also note that if 18:2 intake is high enough (610 %
depending on the LP setpoint), SFA no longer have a significant cholesterol-raising effect
[148].
Figure 10. The scheme depicts the 18:2/SFA ratio hypothesis, which can be applied to data in Simply
stated it implies that a certain level of 18:2 is required by an individual to prevent certain SFA-rich fats
from raising the serum LDL level. If you drop below your 18:2 threshold, LDL will increase, with the
increase being most severe during consumption of 12:0 1 14:0-rich fats. Also note that if 18:2 intake is
high enough (610 % depending on the lipids setpoint), SFA no longer have a significant cholesterolraising effect [148].
90
development of hepatic steatosis to NASH. The increase in FA oxidation in the steatotic state
could potentially induce an increased electron flux through the electron transport chain
(ETC), which may lead a major production of reactive oxygen/nitrogen specie (ROS/RNS).
These reactive lipid derivatives have the potential to amplify intracellular damage by
mediating the diffusion of ROS/RNS into the extracellular space thus causing tissue damage
[160].
Enzymatic and no enzymatic systems are also capable to avoid hepatic damage, inducing
an inflammatory process initiation. Damage and lipid peroxidation products induce an
inflammatory response with up-regulation of pro-inflammatory cytokines including alpha
tumoral necrosis factor (TNF-), interleukins 6 and 1 (IL-6 and IL-1). These cytokines are of
major importance for directing polymorphonuclear and mononuclear leukocytes into inflamed
tissues [158]. The greater cytokines pro-inflammatory production, especially TNF- and IL-6
and IL-1, can contribute to peripheral and hepatic of insulin, which induce to an infiltration in
the hepatic parenchyma, in a vicious cycle that promotes more tissue injury. This mechanism
is described as the two hit model with the first hit being steatosis and insulin resistance,
and the second hit needed to initiate NASH requiring other factor(s) that promote lipid
peroxidation, inflammatory cascade, oxidative stress, tissue injury and inflammatory process
[161]. Moreover a multiple parallel hits model has been suggested to promote progression
of steatosis to NASH because of failure of the antilipotoxic protection systems of the liver
and multiple hits from the gut and/or adipose tissue [162].
Palmitic acid roles in NAFLD installation and development have been discussed. PA
overloading is known to induce apoptotic cell death and a large number of molecular
mechanisms have been implicated in this action: nitric oxide (NO) synthesis, suppression of
antiapoptotic factors such as Bcl-2 [163, 164] reactive oxygen species generation,
endoplasmic reticulum stress [165], nuclear factor-kB activation [166].
Apoptosis can be triggered by mitochondrial damage, which is followed by the release of
cytochrome c and the caspase cascade [167]. The BAX protein activates mitochondrial
release of cytochrome c [168], while Bcl-2 is a mitochondrial protein inhibits the apoptotic
process and promotes cell survival [169]. Ji et al. [163] research showed that occur a decrease
in mitochondrial Bcl-2 and a markedly increase in mitochondrial level of Bax in the HepG2
cells treated with PA from 200 to 400 mM concentrations. The authors suggest that this
mechanism can contribute to NASH and NAFLD installation, and especially may play an
important role in the transition from steatosis to steatohepatitis in human. Other studies [170]
on the effect of FAs-induced steatosis on cellular apoptosis have demonstrated that palmitic
and oleic FA mixtures-induced steatosis is associated with apoptosis in hepatocyte cell
cultures. Joshi-Barves et al. [171] studies also showed that exposure to excess palmitic acid
induces apoptosis and IL-8 production in hepatocytes in a relation of dose-dependent and
time dependent manner, via activation of c-Jun amino terminal kinase (JNK/AP-1), and
nuclear factor kappa B (NF-B) transcription factors for IL-8 expression.
91
diabetes fall into two broad etiopathogenetic categories. In one category, type 1 diabetes, the
cause is an absolute deficiency of insulin secretion. Individuals at increased risk of
developing this type of diabetes can often be identified by serological evidence of an
autoimmune pathologic process occurring in the pancreatic islets and by genetic markers. In
the other, much more prevalent category, type 2 diabetes, is associated with a combination of
pancreatic -cell dysfunction and insulin resistance [174]. The chronic hyperglycemia
produces glucotoxicity characterized by -cell function gradual deterioration and insulin
resistance aggravation. It is similar to the paradoxically deleterious effects of chronic
hyperglycemia, if lipotoxicity is produced, once the free FAs which are essential fuels in
the normal state, become toxic when they are chronically present in excessive levels
[164, 175].
Under diabetic conditions, oxidative stress and endoplasmic reticulum stress are induced
in various tissues [173, 176, 177, 178, 179]. Moreover, the -cells have very low levels of
antioxidative enzymes, becoming them more susceptible to the stress [172]. ROS can function
as signaling molecules to activate a number of cellular stress-sensitive pathways that cause
cellular damage, and are ultimately responsible for the late complications of diabetes.
Evidence suggests that common stress-activated signaling pathways such as nuclear factor
nuclear factor-B (NF-B) [180, 181, 182], p38 mitogen-activated protein kinase (MAPK)
[183], protein kinase C (PKC) [184], toll-like receptors (TLRs) [185, 186], and c-Jun Nterminal kinase (JNK) [187; 188] underlie the development of these diabetic complications.
Nuclear factor kappa-B (NFkB), a redox-sensitive transcription factor regulating a battery
of inflammatory genes, has been indicated to play a role in the development of numerous
pathological states [189]. Activation of NFkB induces gene programs leading to transcription
of factors that promote inflammation, such as leukocyte adhesion molecules, cytokines and
chemokines [181].
Mitogen-activated protein kinases (MAPKs) are a family of serine/threonine kinases and
consist, among others, Jun N-terminal protein kinase (JNK), p38s MAP kinase, cyclic AMP
dependent protein kinase (PKA), protein kinase B (PKB) and protein kinase C (PKC) [190].
It is involved in the regulation of a wide range of cellular responses, including cell
proliferation, differentiation, and survival [191]. Its also well established that p38 and JNK
play important roles in mediating apoptosis caused by various stimuli [190]. There are three
isozymes of JNK: JNK1, JNK2 and JNK3, and that only JNK1 has been shown to be
implicated in type 2 diabetes [192] probably to reduce insulin gene expression [187]. The
TLR family is known to consist of 10 members (TLR1-TLR10) that are pattern recognition
receptors which initiate innate immune responses upon recognition of a wide range of
pathogen-associated molecular patterns [185, 193].
In vitro studies have shown that -cells are vulnerable to palmitate, in the presence of
high glucose concentration [194, 195]. Many authors report that the palmitate induces -cell
dysfunction in vivo by activating inflammatory processes within islets, for example:
activation of nuclear factor-kB, resulted in increased expression of several proinflammatory
cytokines (TNF-, IL-1, IL-6, MCP1) in rat liver as well as an increase in circulating MCP1
levels. The rise in plasma MCP1 is particularly interesting because MCP1 is well established
to regulate macrophage recruitment to sites of inflammation [196].
Additionally, palmitate increased the expression and secretion of inflammatory cytokines
(e.g. IL-6 and TNF-) and impaired insulin sensitivity via an NFkB/PKC pathway in muscle
cells [197, 198]. Too increased intracellular concentration of PA producing diacylglycerol
92
have been shown to activate and cause cellular redistribution of protein kinase C isoforms
[199], which result in the induction of inflammatory pathways via NF-B activation [200].
Jiang [201] studies with PA showed to induce endothelial progenitor cells apoptosis via p38
and JNK mitogen activated protein kinases MAPK pathways in a time-and-dose-dependent
manner. Eguchi et al. [186], through a combination of in vivo and in vitro studies, reported
that SFA palmitate induces -cell dysfunction. According to authors this cell responds to
palmitate via the TLR4/MyD88 pathway and produce chemokines that recruit M1-type
proinflammatory monocytes/macrophages to the islets. Depletion of M1-type cells protected
mice from palmitate-induced -cell dysfunction.
After insulin binds to insulin receptor on cell surface, insulin receptor and its substrates
are phosphorylated, which leads to activation of various insulin signaling pathways [202].
Reynoso et al. [203] evaluated several aspects of the insulin resistance induced by palmitic
acid in rats and found that after treatment with 0.09 g/kg of palmitic acid there is a delay in
the curve of tolerance to glucose. The authors concluded that occur an increase in the
phosphorylations in serine of the insulin receptor after the treatment with palmitate,
suggesting that PKC has a role as negative regulator of the insulin receptors activation in the
insulin resistance induced by palmitic acid.
CONCLUSION
Palmitic acid is the most abundant saturated fatty acid in human nutrition. It is a major
component of palm oil, but can also be found in beef tallow, lard, cocoa butter, human, cows
milk and interesterification food. PA is the first fatty acid produced during fatty acid synthesis
and the precursor to longer fatty acid. Through this bioprocesses, glucose is converted to fatty
acids, which then react with glycerol to produce triacylglycerols. Furthermore, palmitic acid
can participates in several chemical reactions as other acids of this same class, being attached
to the alcohol form of vitamin A which has been used successfully as a supplement due to its
high stability in relation to vitamin A and low cost. Several studies have documented that
palmitic acid position in TGC molecule has a great importance in fatty acids action in several
human metabolic bioprocesses. In addition, a higher palmitic acid absorption is obtained with
formulas rich in palmitic acid esterified in triacylglycerols sn-2 position, than those
predominantly esterified in sn-1,3 positions. Some authors suggest that palm oil does not
behave as saturated oil, reducing the blood levels of total cholesterol, LDL cholesterol due to
TGC sn position. In the other hand, if cholesterol consumption exceeds the critical value or
when hypercholesterolaemic subjects are studied, the palmitic acid appears to increase those
cholesterol levels. There are still other researches that observed adverse healthy effects by the
use of palmitic acid. Moreover, also several studies have documented roles in NAFLD
installation and insulin resistance higher levels development in diets rich in palmitic acid.
Nonetheless, those studies have still much divergent results, being necessary more researches
to clarify the real participation of PA in these processes.
93
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104
In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 5
PALMITIC ACID AS
A CARDIOMETABOLIC RISK FACTOR
Danijela Risti-Medi* and Vesna Vui
Centre of Research Excellence in Nutrition and Metabolism, Institute for Medical
Research, University of Belgrade, Belgrade, Serbia
ABSTRACT
Current dietary recommendations are based on a reduced saturated fatty acid (SFA)
consumption to prevent cardiovascular disease (CVD). The role of individual SFA in
metabolic disease is not fully understandable. One type of SFA present in many common
foods (dairy, meat, palm and coconut oil) is palmitic acid (16:0). A number of
epidemiological studies have shown that the populations who consume large amounts of
atherogenic SFA (especially palmitic, myristic, lauric) have elevated levels of LDL and
HDL-cholesterol. Saturated fatty acid exert their atherogenic and thrombogenic effect
through increased production of LDL, very-low-density lipoproteins particles and
apolipoproteins A1, with a decrease of LDL- receptors specific activity, and an increase
in platelet aggregation. The total cholesterol/ HDL-cholesterol ratio, the best overall
indication of potential effects on coronary heart disease (CHD) risk is nonsignificantly
affected by consumption of palmitic acid (PA). Compared with lipid effects, the influence
of SFA intake on inflammation markers is less well explored. The associations between
circulating and tissue PA and dietary intake of PA are diverse and most likely reflecting
endogenous metabolism. Status of PA is not in intakeresponse relationship biomarker,
probably partly due to conversion of 16:0 to 16:1 by steaoryl-CoA-desaturase (SCD-1).
Increased SFA intake has been associated with increased SCD-1 activity in which may
predict mortality. Palmitoylation is the process involved in proteinmembrane
interactions and signal transduction. Increases in dietary intake of PA decrease fat
oxidation and daily energy expenditure with slight increases in adiposity. Evidence for
the effects of SFA, particularly PA consumption on insulin resistance, vascular function,
type 2 diabetes, and stroke is various. It is considered that circulating PA, as nonesterified
*
Corresponding author: Danijela Risti-Medi, MD. PhD. Institute for Medical Research, Centre of Research
Excellence in Nutrition and Metabolism, University of Belgrade, Tadeusa Koscuska 1, 11129 Belgrade,
Serbia. Tel:+38111303-1997; Fax: +381 11 2030-169; e-mail: dristicmedic@gmail.com.
106
INTRODUCTION
Previously, low fat intakes were traditionally recommended in the prevention of
cardiovascular disease (CVD) as a component of a health promoting diet, without much
attention to the quality of fat. However, current dietary guidelines generally put more
emphasis on the quality of fat [1-4]. Imbalances in the amounts of individual fatty acids in the
diet may have an impact on the occurrence of dyslipidemia, atherosclerosis, thrombosis,
hypertension and obesity. Saturated fatty acids (SFA) have shown to be particularly important
for development of the above mentioned diseases. However, in spite of an increasing body of
new data, the role of individual dietary SFA in metabolic diseases is not fully clarified (Micha
2010). The reachest dietary sources of SFA include fast foods, processed foods, high-fat dairy
products, red meats, and pork [1,5].
One of the most abundant SFA in many common foods (dairy, meat, palm and coconut
oil) is palmitic acid (PA, 16:0). The amount of PA is the highest in palm oil (around 50%),
but significant amounts of PA (25-26%) can also be found in butter, chicken fat, lard, beef
and lamb fat, as well as in cocoa butter. Even olive oil, which is one of basic components of
the healthy Mediterranean diet contains around 16% of PA [6]. Furthermore, PA is present in
human milk with 20-25% of total fats. Overall, PA and stearic acid (18:0) are the most
common dietary SFA and therefore they are also the major SFA in human plasma and tissues.
Their concentration in serum/plasma phospholipids and cholesterol esters reflect dietary high
fat intake.
Dietary saturated fats are of particular scientific interest because of their association with
CVD. In some countries, e.g. in Finland, there has been a decline in coronary heart disease
(CHD) mortality along with the decreased intake of saturated and total fats [7]. Some
epidemiological studies showed that total dietary fats intake is positively associated with
metabolic syndrome [8-11]. De Oliveira et al [11] have recently reported that saturated fat
intake greater than 10% of total caloric value represented a double risk for metabolic
syndrome diagnosis, with odds ratio (OR) 2.0 (1.04-3.84). This association is mostly
attributed to palmitic acid, due to the fact that excessive intake of PA increases the visceral
adipose tissue in greater proportion than other fat types [12]. Metabolic syndrome or
cardiometabolic risk refers to a cluster of metabolic abnormalities including disturbances in
107
glucose and insulin metabolism, central obesity, dyslipidemia (high triglyceride levels, low
HDL-cholesterol and high levels of small dense LDL-particles) and hypertension [13,14].
Central to the etiology of metabolic syndrome is an interrelated triad comprising
inflammation, obesity (particularly abdominal), and aberrations in fatty acid metabolism
[15,16].
108
major component in the diet, palmitate strongly increased the expression and secretion of
TNF and IL-10 in murine 3T3L1 adipocytes, compared with monounsaturated oleic acid
and polyunsaturated docosahexanoic acid (DHA) [31]. It has been considered that in
adipocytes, palmitic acid modulated intracellular signaling and induced endoplasmic
reticulum stress by increasing C/EBP homologous protein and glucose regulatory protein 78.
Furthermore, palmitic acid alters phosphorylation of eIF2 and increases phosphorylation of
JNK and extracellular receptor kinase [32].
Experimental studies [33,34] have shown that high level of palmitic acid in the diet
impaired insulin sensitivity by reducing adiponectin secretion and impairing insulin signaling
pathways required for glucose uptake. Adiponectin is an insulin-sensitizing protein produced
by adipocytes. Reducing the levels of adiponectin appears to be the mechanism by which
palmitate caused insulin resistance in isolated rat adipocytes [35]. In accordance, experiments
in mice fed a high-fat diet showed that overexpression of adiponectin decreased insulin
resistance [36]. Furthermore, PA leads to insulin resistance due to changes of phosphorylation
level of the insulin receptor and insulin receptor substrate.
Saturated fatty acid induced inflammatory response in the interaction between adipocytes
and macrophages by the TLR4/NFkB signaling patway [26]. Supplementation with palmitic
acid induced IP-10 inflammatory gene expression in human macrophages (U937) by an
NFkB-dependent mechanism [37]. It was shown that adipocytes containing SFA have the
capability to activate macrophages to a greater extent than smaller adipocytes. It is even more
pronounced when compared adipocytes containing SFA with adipocytes enriched in
unsaturated fatty acid. For instance, SFA increased TNF mRNA levels in cultures of
adipocytes and murine macrophages, whereas unsaturated fatty acid had no effect. Factors
secreted from macrophages increase adipocyte inflammation and insulin resistance [38,39]
and high-fat feeding in mice increased TLR4 signaling in macrophages and adipocytes and
impaired insulin signaling effects [29].
109
110
pancreatic lipase to hydrolyze triglycerides at the sn-1 and sn-3positions leads to the
production of free fatty acids and 2-monoglyceride [69]. In this way, the unique position of
SFA in milk fat may affect postprandial metabolism, leading to prevention of
hypercholesterolemia and hypertriglyceridemia that would otherwise be associated with
consumption of saturated fat [69,68,70]. The beneficial effect of milk fat on serum lipids may
partially explain why milk fat, despite its contribution of SFA to the diet, has not been
consistently associated with higher incidence of CVD [69,72] or risk factors for cardiometabolic syndrome [72,73]. In a recent cohort study, butter and dairy intake did not predict
all-cause and ischemic heart disease mortality in men, and slightly increased risk in women,
whereas fermented full-fat milk was inversely associated with mortality in both men and
women [74] Moreover, similar findings are reported for palm oil, which also does not raise
blood cholesterol as expected based on the content of PA. It may be explained by the position
of palmitic acid in palm oil triacilglycol (10% of total PA is in the middle position). Because
triacilglycols with SFA in the sn-2 position may be absorbed more efficiently and cleared
from circulation more slowly than triacilglycols with SFA in the sn-1 and sn-3 positions,
intake of these dietary triacilglycols often leads to a more pronounced postprandial lipemia,
which is an independent risk factor for CHD [75]. Therefore, it can be assumed that PA
esterified to the sn-2 position is more atherogenic than when esterified to the sn-1 and sn-3
positions. Studies in animals and in human infants have supported this assumption since they
reported higher plasma triglycerides levels after diets with PA in the sn-2 position than after
diets containing PA in the sn-1/3 positions [76]. However, no significant differences were
found in one adult trial [77], whereas another study [78] reported larger LDL-cholesterol
concentrations caused by diets including palmitic acid in the sn-2 position in men but not in
women. Thus further research is needed to elucidate these relationships.
111
112
lowest category of SFA intake has been shown in meta-analyses of eight prospective cohort
studies [98]. Results from Women Health initiative trial [99], indicated that reduction in SFA
consumption does not appear to increase risk of stroke over 8 years. Some links between
dietary PA and cardiometabolic risk factors are presented in Figure 1.
Palmitic acid
Dietary sources:
Palm oil, dairy,
meat
Elevated LDL
Elevated HDLcholesterol
Tissue inflammation
Weight gain
Figure 1. Relationship between dietary intake of palmitic acid and cardiometabolic risk.
113
experimental data suggesting that palmitic acid has unique effects on several cellular
functions, such as apoptosis [127], endoplasmic reticulum stress [128], and up-regulation of
SCD-1 [129]. The lipogenic enzyme SCD catalyzes the synthesis of MUFAs, eg, oleic and
palmitoleic acids. Estimated SCD activity (16:1/16:0 ratio), together with palmitoleic acid,
has been considered as a strong predictor of mortality [126]. It may be associated with
increased lipogenesis [129], ectopic fat deposition and thereby insulin resistance
[130,131,132]. Accordingly, the estimated SCD ratio was established as an independent
predictor of directly measured insulin sensitivity over 20 years [81]. In contrast, our data on
patients with non-Hodgkin lymphoma showed very low proportion of 16:1 and activity of
SCD, especially in patients with progression of disease, but the role of SDC in cancer should
be further investigated [133].
Table 1. Palmitic acid status in patients with cardiometabolic risk
Cardiometabolic risk patients
1.Hyperlipidemic patients (n=29) [113]
2. Hyperlipidemic patients (n=39) [114]
Obesity women (n=30) [15]
I-NGT group (n=12)
II-IR group (n=18)
DM type 2 with hyperlipidemia (n=28) [115]
I-IHTG (n=14)
II-CHL (n=14)
1. Hemodialysis patients (n=35) [116]
2. Hemodialysis patients (n=37) [117]
PA status
(source of FA, mol%)
1.serum PL 30.35 5.94
Er 25.58 4.15
2.serum PL 30.30 1.39
Er 23.64 0.90
Er 22.631.39
I-Er 22.491.67
II-Er 22.731.21
serum PL 30.01 2.70
I-serum PL 29.05 1.43
II-serum PL 30.71 3.67
1.serum PL 29.93 3.52
2. serum PL 28.09 3.34
controls 26.46 2.44
Er 21.63 1.85
control 22.42 2.59
serum PL 30.76 4.75
control 26.53 2.44
I. serum CE 10.07 0.77
II. serum CE 9.91 0.77
p< 0.001
III. serum CE 10.21 0.83
IV. serum CE 9.94 0.77
p< 0.001
114
Table 1. (Continued)
Cardiometabolic risk patients
PA status
(source of FA, mol%)
I. Patients with incident CHD (n=282) [101]
I- serum PL 25.5 1.5
II-Patients with no incident CHD (n=3309)
II-serumCE 10.02 0.8
I- serum PL 25.4 1.7
II-Serum CE 10.0 0.8 p < 0.01
Patients with no MS (n=640) [121]
Er 22.7 1.2
Patients with MS (n=396)
Er 23.1 1.22 p<0.001
I- Normal weight patient (n=60) [112]
I-Serum PL 30.18 0.29
I-Serum CE 12.67 0.18
II-Overweight with no MS (n=45)
II-Serum PL 30.80 0.32
II-Serum CE 13.35 0.20
III-Overweight with MS (n=15)
III-Serum PL 31.52 0.55
III-Serum CE 13.97 0.32 p<0.001
The values are means SD; CHD, coronary heart disease; DM, diabetes mellitus; MS, metabolic
syndrome; IHTG, isolated hypertriglyceridemia; CHL, combined hyperlipidemia; CE, cholesterol
esters; PL, phospholipids; FA, fatty acid; Er, erythrocytes; IR,insulin resistence; NGT, normal
glucoso tolerance.
MF/24 3
(n=16) [122]
26.21 2.16
21.56 0.96
MF/57 (19-74)
(n=29) [65]
26.4 2.5
n.a.
F/ 23.671.56
(n=14) [123]
27.72 1.60
23.39 0.40
MF/ 54.412
(n=37) [117] )
26.46 2.44
22.42 2.59
The values are means SD; PL. phospholipids; Er, erythrocytes; M, male; F, female;
115
cohort study from Australia, dietary intake of 16:0 and 18:0 assessed by food frequency
questioners (FFQ) at baseline nonsignificantly predicted diabetes incidence, whereas dietary
15:0 was inversely associated with diabetes [100]. In baseline plasma phospholipids, total
SFA and 18:0 were positively associated with diabetes risk, 16:0 was nonsignificantly
associated, whereas 15:0 was negatively associated [102]. Since dietary intake correlates with
serum lipids fatty acid composition, changes in dietary habits and intake of proper fats can
prevent development of metabolic and cardiovascular diseases.
CONCLUSION
Patients with cardiometabolic risk factors represent a group at high lifetime risk for CVD.
Among other factors, nutrition markedly contributes to the development of metabolic
diseases. Although cardiometabolic risk is associated with SFA-rich foods, systematic
reviews on prospective cohort studies indicated that CHD risk has not been directly
associated with SFA intake. Nevertheless, there is convincing evidence for decreased CHD
risk when replacing SFA with polyunsaturated fats. Differences in cardiometabolic risk
appear greater between food groups and overall dietary patterns rather than between separate
SFA, even though it has been documented that palmitic acid increased the risk. Based on the
current data, it is not possible to give dietary recommendations solely based on the content of
individual SFA. Strictly controlled short-term and longer-term intervention studies are needed
to establish convincing link between palmitic acid and various alterations reported in
observational studies.
ACKNOWLEDGMENT
This work was supported by the Project III41030 financed by the Ministry of Education,
Science and Technological Development of the Republic of Serbia.
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In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 6
ABSTRACT
Palmitic acid (C16:0) is one of the major fatty acids (FAs) forming virtually all natural
lipids. Both in eu-, and prokaryotes, C16:0 forms various lipid classes, which serve either
as the lipid background of storage fats and oils, or the hydrophobic matrix of cell
membranes, or the components of cuticle waxes and polymers. Non-esterified 16:0 does
not occur in living cells, and it is present there only as an acyl residue in various lipid
classes, such as mono-, di-, and triacylglycerols, glyco-, phospho-, and sphingolipids,
wax and steryl esters etc., where it esterifies the hydroxy groups of glycerol backbone or
other alcohols (sphingosine, higher and lower aliphatic alcohols etc.). Palmitic acid is
known to be a primary higher FA synthesized in the cell, while nearly all other FAs of
natural lipids are the products of its further modification caused by elongation,
desaturation, insertion of various functional groups, such as methyl, hydroxy, oxo, epoxy,
etc. As a saturated FA, C16:0 is used by the cell for regulating its functional state by
shifting the membrane fluidity under adverse environmental conditions and thus
providing a necessary molecular species composition of the membrane polar lipids.
Among the latter, such classes as phosphatidylinositols, phosphatidylserines, and other
highly polar lipids are particularly rich in palmitic acid. In accordance, its content in plant
lipids rises as they became less TLC-mobile, more difficultly extractable, or tightly
bound. It is evident that further screening of plant lipids as regards this index is of
considerable interest.
126
INTRODUCTION
Lipids are an essential constituent of all plant cells. Fatty acids (FAs) are the mandatory
structural components of almost all lipids both storage triacylglycerols (TAGs), the seed oil
base, and membrane lipids forming every cell membrane bilayer. To date, more than 1000
different FAs are described. Such a FA diversity were found mainly in plant seed oils, and it
has been estimated that thousands more could be present throughout the plant kingdom. [1,
2]. All FA diversity is subdivided into major, minor and unusual ones [3]. Structural
membrane glycerolipids of all plant cells contain almost exclusively C16- and C18-FAs, with
none or up to three methylene-interrupted double bonds. Seven FAs with even number of
carbon atoms in the aliphatic chain lauric (C12:0), myristic (C14:0), palmitic (C16:0), stearic
(C18:0), oleic (C18:19), linoleic (C18:29,12) -linolenic (C18:39,12,15) acids, belongs to major
FAs. These FAs are widespread in nature and are the main lipid components, as a rule. The
acids homologous to major FAs with the greater or fewer number of carbon atoms or
differing in the position or configuration of double bonds and present in lower amounts in
lipids (no more than 5% of total FAs) of limited number of plant species are known as minor
FAs. Major and minor FAs are present in the lipids of most plant species; therefore, they are
called as common or usual FAs. The rest FAs deviate significantly from the common
ones in double bond configuration or position or in the presence of coupled double or triple
bonds or presence of different functional groups and also met in the plant lipids belong to
unusual FAs [3, 4]. Meanwhile, the attribution of a FA to the group of major, minor, or
unusual FAs is rather arbitrary and may be used only in a comparative study of the total lipids
in a large number of objects.
Palmitic acid occupies a distinct place among the all FA diversity. This FA in one or
another quantity present in every lipid class in all plant objects. Moreover virtually all FAs
are derived from palmitic acid by its modification, namely desaturation, elongation,
hydroxylation, oxidation, etc. Although fatty acids are major constituents of every membrane
in a cell and are also found outside cells in the cuticular lipids, their major site of synthesis is
within the plastid. In this regard, the process of lipid biosynthesis in plants is fundamentally
different from that in animals and fungi, which produce fatty acids primarily in the cytosol
[5].
127
FatA and FatB acyl-ACP thioesterases hydrolyzing predominantly oleoyl-ACP and diverse saturated
and unsaturated acyl ACP, respectively; ENR enoyl ACP reductase; KAR ketoacyl ACP
reductase; KAS ketoacyl synthase; MCMT malonyl-CoA : ACP malonyl transferase; HADH
hydroxyacyl ACP dehydratase.
Figure 1. FA biosynthesis in higher plants.
The synthesis of FA from acetyl-CoA starts in plastids (Figure 1) and occurs in three
stages. At the first stage the irreversible carboxylation of acetyl-CoA catalyzed by acetyl-CoA
carboxylase with the formation of malonyl-CoA occurs. Then the malonyl group is
transferred to the acyl-carrier protein (ACP) and, due to the operation of plastidial acetyl-CoA
carboxylase complex the primary substrate of FA synthetase, malonyl-ACP, is formed. [6, 7].
During the second stage, successive condensation of newly formed malonyl-CoA with
growing, bound to ACP acyl chain occurs. In such a way, by the successive addition of twocarbon fragments to the growing chain, palmitoyl-ACP is finally produced [6, 7]. Each cycle
of condensation includes four reactions. Firstly, due to the condensation of acetyl-CoA and
malonyl-CoA, 3-ketobutyl-ACP is formed catalyzed by ketoacyl synthase III (KAS III); then
3-ketobutyl-ACP is reduced to 3-oxyacyl-ACP, dehydrated to enoyl-ACP, and further
reduced to butyryl-ACP. Subsequent cycles of condensation of acyl-ACP with malonyl-CoA
are catalyzed by KAS I and proceed to the formation of myristoyl-ACP and palmitoyl-ACP
[4, 6-8]. The elongation of palmitoyl-ACP to stearoyl-ACP and its desaturation by 9desaturase to oleoyl-ACP are catalyzed by KAS II and occur also in plastids. Then, during the
third stage of FA biosynthesis, palmitoyl-ACP, stearoyl-ACP, and oleoyl-ACP are exported
into the cytosol and included into acyl-CoA and acyl-lipid pools [6, 9]. In most plants, the
process of FA elongation can continue, and due to functioning of the microsomal fatty acid
elongation (FAE) system so-called very long chain FAs (VLCFAs) including 20 and more
carbon atoms are formed [3, 6, 10].
The termination of elongation is catalyzed by ACP-thioesterases (enzymes belonging to
the class of acyl-ACP hydrolases). These enzymes hydrolyze acyl-ACP with the formation of
free FA, which can cross the plastid membrane to be reactivated outside the organelle [11].
128
Two main types of plant thioesterases are described: FatA preferably cleaving oleate from
ACP and FatB hydrolyzing diverse saturated and unsaturated acyl-ACP. In some plant
species, thioesterases, which are specific to acyl-ACP with short-chain acyls were found [12,
13]. The interaction between FA synthetase, 9-desaturase, and the two types of thioesterases
determines the ratio between FA produced in plastids, which will be further used by the cell
for the synthesis of glycerolipids [2].
129
FAE fatty acids elongase; FAS fatty acid synthetase; KASII ketoacyl synthase II.
Figure 2. Fatty acid desaturation.
130
AD I & AD III aldehyde decarbonilases, encoded by A. thaliana CER1 and CER3 genes resp.; FAE
fatty acids elongase; FAR fatty acids reductase; FAS fatty acid synthetase; S and SO
substrate and oxidized substrate respectively.
Figure 3. Some other modifications of palmitate.
In plant tissues C16:0 can undergo some other modifications (Figure 3), namely
elongation, hydroxylation, oxidation, epoxydation, reduction, oxidative decarboxylation, etc.
As a result of these modifications many different lipophilic substances are produced. Among
these substances very long-chain FAs (VLCFAs, C20), different unusual FAs (hydroxy-,
epoxy-, acetylenic, dicarboxylic), fatty aldehydes and alcohols, hydrocarbons, oxilipins, etc.
are formed. Some of them are present in plants in free form (are embedded in the complex
cuticular lipid matrix or as a components of epicuticular waxes), the others are used as a
substrates for more complex lipids and lipid polymers biosynthesis (see below).
Once synthesized, the palmitoyl moiety is transported out of the plastid to the
endoplasmic reticulum where it serves as an initial substrate for further modifications. For
instance, it can be elongated to very long chain fatty acids which contain up to 34 carbon
atoms. This process requires the sequential use of 4 reactions to add each C2 unit [25]. These
reactions are analogous to de novo synthesis (see above) but elongases, involved in synthesis
of very long chain fatty acids, are membrane-bound enzymes which use acyl-CoA substrates
and malonyl-CoA directly as the source of the C2 unit. Once elongation is complete, VLCFAs
may be reduced to fatty primary alcohols by an acyl-CoA reductase (FAR3/CER4) of the
acyl-reduction pathway, which appears to be associated with the ER. The primary fatty
alcohols generated and C16:0 acylCoA are condensed into wax esters by the bi-functional
wax synthase/acylCoA:diacylglycerol acyltransferase (WS/DGAT) enzyme, WSD1. Wax
synthase, catalyzing this reaction, is an integral membrane protein, but its site of action is not
known [26].
VLCFAs, besides, may be converted to the alkanes another component of epicuticular
waxes through fatty aldehydes by the action of membrane-associated enzymatic complex
131
comprised of two subunits of aldehyde decarbonylases [27, 28]. It was proposed that
VLCFAs activated by the long chain acyl CoA synthases in very long-chain acyl-CoAs would
be used as precursors of very long-chain alkane synthesis. A mandatory CER1/CER3
heterodimer would efficiently catalyze a two-step reaction starting with the reduction of acylCoA to a potential intermediate aldehyde subsequently decarbonylated to alkane with the loss
of one carbon potentially in carbon monoxide or formate as reported in cyanobacteria [29].
CYTB5s would interact with the di-iron catalytic core of CER1, providing electron(s)
required for the decarbonylation reaction. Additionally, palmitoyl-CoA and VLCFAs may be
converted into 2-hydroxy fatty acids, aldehydes and odd-numbered fatty acids through the
action of peroxigenases on the 2-hydroperoxy FAs which arise by the action of dehydrogenases [30].
VLCFAs form components of epidermal lipids, sphingolipids or storage lipids and are
synthesized by the acyl-CoA elongase complex [31]. The elongase is a multienzyme complex
which comprises four dissociable subunits, each possessing a unique enzymatic activity: 3ketoacyl-CoA synthase (KCS), 3-ketoacyl-CoA reductase (KCR), 3-hydroxyacyl-CoA
dehydratase and trans-2,3-enoyl-CoA-reductase (ECR); this elongase complex is membranebound which use acyl-CoA substrates and malonyl-CoA directly as the source of the C2-unit
[32]. VLCFAs are further modified to make different kinds of epicuticular wax precursors
(e.g. alkanes, fatty alcohols, aldehydes, etc., Figure 3). On the other hand, C16 and C18 FAs
are also modified to produce major monomers of lipid polymers, cutin and suberin (e.g. C16
and C18 hydroxy- and dicarboxylic FAs). The cuticle monomers are synthesized inside the
epidermal cells and deposited outside the epidermis [3]. This fact suggests the existence of
transporter(s) involved in the transportation of cuticle lipids across the plasma membrane.
Cutin is a polyester polymer composed of complex mixture of inter-esterified long chain
-hydroxy FAs and is considered as the major constituent of the plant cuticle. Cutin
monomers build up a complex, three-dimensional network by cross-links through the primary
and secondary hydroxyl and carboxyl groups [33]. C16- and C18- -hydroxy-FAs, with midchain functional groups such as epoxy- and hydroxy-, were predominant in all cutins; in some
plant cutins a small quantities of -hydroxy-C20 and C22 FAs may also occur. The other
monomers have been found in plant cutins are FAs, dicarboxylic FAs, alkanols, some
aromatic acids, and glycerol. The usual and in many cases the major constituents of most
plant cutins are different dihydroxy- derivatives of palmitic acid (C16:0di-OH). Several C16:0di-OH
isomers have been found in many cutin hydrolysates, the major ones being 10,16-C16:0di-OH
and 9,16-C16:0di-OH acids; smaller concentrations of 8,16-C16:0di-OH and 7,16-C16:0di-OH isomers
also often occur [34]. In some plant cutins a substantial quantities of 16-hydroxy-10oxohexadecanoic acid and related positional isomers were found [35].
While the cuticle lies on the outer face of the primary cell wall, suberin is located on the
inside of the primary cell wall, usually close to the plasma membrane. Plants synthesize
suberin to create a hydrophobic barrier to water and solute diffusion through cell walls during
normal development, or to provide a barrier in response to environmental stresses. A familiar
example of suberized tissue is cork, constituted by the outer bark cells (periderm) of cork oak
(Quercus suber). Suberin is also deposited in periderm walls of underground organs (i.e.
tubers, roots), and in the root endodermis. A material with composition intermediate between
suberin and cutin is often deposited surrounding the bundle sheaths of monocot leaves,
perhaps to prevent CO2 released during decarboxylation reactions from diffusing out of the
bundle sheath cells. In mature seeds, suberin has been found in seed coat layers, and sealing
132
off the chalazal region of the inner seed coat after disconnection from vascular tissue.
Exposure to cold, mineral stress and fungal infection are some examples where suberization
occurs as a response to external factors. It is also deposited as a wound response by injured
plant cells, even in those cells that normally synthesize cutin. [36].
The other lipid polymer, suberin, is a heteropolymer, consisting of an aliphatic polyester
associated with cross-linked polyaromatics and embedded waxes. Upon transesterification of
suberin, the monomers released include C16-C28 -hydroxy fatty acids and C16-C26 ,dicarboxylic acids, the latter of which are diagnostic monomers, unsubstituted very-longchain fatty acids (VLCFAs; C>18) and alcohols, glycerol and ferulate. Usually the major
components of suberin are -hydroxy derivatives of palmitic and/or oleic acids, but in some
cases -hydroxy C22:0 also is a dominant component [37]. Dicarboxylic FAs derived from
further oxidation of the -hydroxy-FAs are also found in suberin.
The synthesis of cutin and suberin aliphatic polyesters usually are described as distinct
pathways, with chain elongation and conversion of -hydroxy acids to dicarboxylic acids
being specific for suberin monomers. However, data from Arabidopsis has shown that
dicarboxylates are not exclusively found in suberin, and that the same gene families seem to
be involved in the synthesis of both types of polyesters. Hence, monomers with chain lengths
beyond C20, higher levels of aromatics, and primary alcohol synthesis can be considered as
specifically associated with suberin. Hydroxylated fatty acids are major constituents of plant
lipid polyesters. In plants, hydroxylation of the terminal methyl of aliphatic chain (-position)
is catalyzed by cytochrome-P450-dependent (CYP) enzymes, most of them belonging to the
CYP86 and CYP94 subfamilies. Although Arabidopsis leaf and stem cutin has an unusual
composition rich in unsaturated dicarboxylic acids, its flowers present a classical cutin
composition with high content of 10,16-dihydroxy fatty acid. CYP86A4 is involved in hydroxylation forming 16-hydroxy fatty acid, and CYP77A6 catalyzes the in-chain
hydroxylation of this monomer to produce 10,16-dihydroxy fatty acid [36]. At least two other
members of the CYP86 subfamily participate in suberin synthesis. CYP86A1 catalyzes root
suberin saturated and unsaturated C<20 monomer oxidation and is expressed in tissues where
suberization takes place; the potato CYP86A1-homologous gene plays a similar role in potato
periderm. CYP86B1 is involved in C22 and C24 fatty acid -oxidation in Arabidopsis root and
seed coat suberin [30]. Two different pathways for the synthesis of dicarboxylic acids have
been proposed. One postulates that CYP multifunctional enzymes are able to catalyze the
complete set of reactions leading to the oxidation of a terminal methyl group to the carboxyl
function, similar to tobacco CYP94A5 or Arabidopsis CYP94C1. The second route involves
two subsequent dehydrogenases that oxidize -hydroxy fatty acids to -oxo fatty acids and
then to ,-dicarboxylic acids. This was first supported by in vitro labeling experiments, and
later by genetic evidence shown in Arabidopsis cutin [36].
133
134
aliphatic alcohols are rarely present in plant lipids. These fatty acid short-chain-alkyl esters
(FASCAE) were found in the fruits of 12 plant species of Celastraceae family [57], in the
neutral lipids of corn pollen [58], walnut fruit oil [59], callus culture from mint leaves [60],
dry rhizomes of ginseng [61], dry matter of liverwort [62]; small amounts of FASCAE were
also found in the volatile compounds of some other plant sources [57]. The concentrations of
FASCAE were low in almost all instances (<1% of total lipids). In all cases, the FASCAE
found in plants did not represent artifacts of the experiment and were represented mostly by
FA methyl esters which predominated in the FASCAE of the investigated plants, but FA
esters of other short-chain alcohols as a rule were also present. As a FA part of FASCAE
several C12-24 saturated and unsaturated fatty acyls were identified. In most cases palmitic acid
is one of the major FAs of FASCAE, and in dry rhizomes of ginseng the only FASCAE found
was methylpalmitate [61]. The qualitative and quantitative composition of various FASCAE
fractions varies considerably depending on the taxonomic position of a plant species, plant
tissue and/or organ, the extent of fruit maturity, etc.
135
136
137
138
PE accumulated in soybean seeds at concentrations much higher than those observed in wildtype plants.
139
individual contributions in tissues that accumulate oil in different plants may be unequal.
However the similar transcript amounts detected in the study of Tranbarger et al. suggest that
the contribution of these three enzymes could be of similar importance in the mesocarp of oil
palm [82].
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In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 7
ABSTRACT
Human breast milk provides the optimum nutrition for infants. Designed to provide
balanced nutrition, human breast milk naturally meets the needs of growing infants in the
first months after birth. In human breast milk, and in most infant formulas, approximately
50% of the energy is supplied to newborns as fat, of which more than 98% is in the form
of triglycerides. Triglyceride synthesis occurs in the mammary gland. The fatty acids are
specifically positioned to sn1, sn2 or sn3 positions on the glycerol backbone to yield the
structure-specific triglycerides that are found in human milk. Palmitic acid (C16:0) is the
predominant saturated fatty acid, comprising 17-25% of the fatty acids in mature human
milk. Surprisingly, the positioning of palmitic acid is highly conserved across
populations, and approximately 70-75% of palmitic fatty acids are esterified to the sn2
position of the triglyceride (sn2 palmitate).
Clinical and pre-clinical studies over the last three decades have provided increasing
evidence that this specific positioning of palmitic acid on the triglycerides in human milk
has a significant holistic effect on optimal infant development and well-being that is
related to the increased absorption of both palmitic acid and calcium: softer stools,
increased bone strength, increased beneficial gut flora, controlled intestinal health, and
reduced infant crying. All of these contribute to the benefits of infant wellbeing.
The overall aim of the current review is to expand the understanding of the role of
palmitic acid and its specific sn2 position in infant nutrition.
146
INTRODUCTION
Palmitic acid (16:0) (PA) is the most abundant and widespread natural saturated acid; it is
present in plants, animals, and microorganisms. Twenty to thirty percent of the mammalian
body lipids and 10 to 40% of vegetable oils are palmitic acid. Palm oil is a rich commodity oil
source, and over 40% of it is palmitic acid (Scrimgeour and Harwood 2007). Palmitic acid
was discovered in 1840 by Edmond Fremy (Fremy 1842) in saponified palm oil. The word
palmitic is derived from the French word palmitique, for the pith of the palm tree. As its
name indicates, it is a major component of the oil derived from palm trees (palm oil and palm
kernel oil). Recent evidence suggests that PA plays an important role beyond serving as an
energy source.
Palmitic acid is the predominant saturated fatty acid in mature human milk, comprising
17-25% of the total fatty acids, of which 70-75% is esterified to the sn2 () position of the
triglyceride (Breckenridge et al. 1969). The significance of this fatty acid and its position is
evident from the fact that this positioning is conserved in all women, regardless of their ethnic
origin or nutrition, unlike the general fatty acid profile of human milk (Jensen et al. 1978,
Jensen 1999). In contrast, the palmitic acid in vegetable oils that are commonly used in the
manufacturing of infant formulas is esterified primarily at the sn1 and sn3 positions, while the
sn2 position is predominantly occupied by unsaturated fatty acids (Jensen 1995).
NUTRIENT ABSORPTION
Infants require a bio-available source of energy and nutrients to meet the requirements of
their rapid growth, development and expanding skeletal mass. An infants nutritional
147
environment involves a high fat diet with frequent feedings; therefore, efficient fat absorption
is required. In breastfed infants, this is achieved using complex fat globule lipids and TG
structures that enable the efficient absorption and processing of a high-fat diet with highly
saturated fatty acids without requiring a high metabolic effort from the infant. For formulafed infants, the availability of nutrients and energy depends on the composition of the formula
(Giovannini et al. 1995).
Gastric lipase, reponible for intial TG digestion, hydrolyzes fatty acids from the sn3
position of dietary TG to release sn1,2 diacylglycerols and accounts for 10% or more of
dietary TG hydrolysis, depending on the composition of the milk or formula (Mu and Hoy
2004). Fat digestion continues with pancreatic colipase-dependent lipase (pancreatic lipase),
which has specificity for TG sn1 and sn3 ester linkages and completes TG hydrolysis to give
sn2 monoacylglycerol (2-MAG) and unesterified fatty acids, each of which are absorbed
separately. The absorption of unesterified fatty acids depends on their physical and chemical
properties (Innis 2011). The palmitic acid and stearic acid have melting points above the
intestine temperature (63C and 70C, respectively), in contrast to unsaturated fatty acids
such as oleic acid, linoleic acid and linolenic acid, which have melting points below the
intestinal temperature. Therefore, together with the pH of the intestine, unesterified longchain saturated fatty acids have an increased tendency to form insoluble fatty acid soaps that
are subsequently lost in the stools (Innis 2011). Fat absorption is influenced by the position of
the individual fatty acids on the triglyceride molecule and the importance of the positional
distribution of the fatty acids in human milk or infant formula fat applies particularly to
palmitic acid because it is one of the major constituents (Jensen et al. 1986). Palmitic acid is
absorbed from human milk as sn2 monoacylglycerol (Innis et al. 1994), and it is conserved
through digestion and absorption (Nelson and Innis 1999).
Figure 1. Soaped fatty acids in 24 hr stool collection at 6 weeks postnatal. The significance was
calculated for the two groups by the Mann Whitney test. Different letters indicate statistical significance
(p<0.05) between groups.
148
Clinical studies in term (Carnielli et al. 1996, Kennedy et al. 1999, Lopez-Lopez et al.
2001) and preterm infants (Carnielli et al. 1995, Lucas et al. 1997) have demonstrated the
correlation between the level of palmitic in the sn2 position on fatty acids and calcium
absorption. The reduction in calcium and fatty acid absorption is accompanied by increased
calcium soaps and, consequently, hard stools (Quinlan et al. 1995). Infants receiving the high
sn2 formula had softer stools and fewer formed and hard stools than did control regular
vegetable oil formula group (Sidnell and Greenstreet 2011, Kennedy et al. 1999, Litmanovitz
et al. 2014, and Yao et al. 2014).
Prebiotics are routinely added to infant formulas. A prebiotic is a non-digestible food
ingredient that has several potential beneficial effects on neonatal intestinal development,
including protection against infection and facilitation of nutrient absorption (Calder et al.
2006). Sn2 palmitate was recently shown to enhance fat absorption and reduce calcium soap
formation in infant formulas containing prebiotics (GOS) (Bar-Yoseph et al. 2014). In a
multicenter clinical study on Chinese term infants, sn2 palmitate formula consumption
resulted in benefits to the infant in terms of the nutrient absorption, specifically fat absorption.
Comparable with breastfeeding, the formula with sn2 palmitate reduced fat excretion,
primarily in form of calcium soap (Figure 1 (Bar-Yoseph et al. 2014) and Yao et al. 2014).
149
docosahexanoic acid in plasma unesterified fatty acids. This finding suggests the need for
paying further attention to the plasma unesterified fatty acids in young infants as a possible
source of fatty acids for membrane phospholipid synthesis and turnover (Innis and Nelson
2013). It is therefore possible to have the same average fatty acid composition in plasma TGs
with very different TG structures (Innis 2011).
BONE HEALTH
During the last decade, substantial efforts were made to determine the factors that
influence bone mineral accretion in healthy children. This may be attributed to the suggestion
that osteoporosis originates in childhood (Janz 2002), thereby providing optimal nutrition for
reaching the highest possible peak bone mass. The absorption of nutritional factors, such as
minerals, fats, carbohydrates, and proteins, is significantly important for normal infant growth
and development and may contribute to early bone mineral accretion (Specker 2004).
Kennedy et al., by using dual-energy X-ray absorptiometry (DEXA) to assess bone
mineralization showed in a randomized, controlled, double-blind study higher body bone
mass in infants after 12 weeks of feeding with sn2 palmitate formula (Kennedy et al. 1999),
Litmanovitz et al., showed that bone speed of sound (SOS) at 12 weeks in term newborns fed
with sn2 palmitate formula was significantly higher than that of newborns fed with regular
formula and comparable to that of breastfed newborns (Litmanovitz et al. 2013). In this study,
the SOS measurements showed a decrease in SOS in the first 12 weeks for all infants,
regardless of the type of feeding, but the decline was slighter in the sn2 palmitate and
breastfed groups than in the control group (figure 2), indicating higher bone strength in both
sn2 palmitate and breastfed groups.
Figure 2. Bone speed of sound (SOS) during the first 12 postnatal weeks for infants fed with control
formula, sn2 palmitate formula or breast milk.
150
This finding is in agreement with studies in both preterm infants () and term infants that
demonstrated a decrease in SOS (Eliakim et al. 2002, Litmanovitz et al. 2003, Liao et al.
2005) and DEXA (Rauch and Schoenau 2001). The reason for these phenomena is not clear.
It was suggested that the decline in bone mineral density (BMD) in healthy newborns is
associated with a relative physiological decrease in the cortical area and a redistribution of
bone tissue from the endocortical to the periosteal surface rather than with bone loss (Liao et
al. 2005). It is also possible that this decrease represents a delay between rapid linear bone
growth and mineralization.
QUALITY OF LIFE
One of the major concerns of parents is their baby's comfort and wellbeing. Various
parameters may provide comfort to the newborn infants. Those parameters include stool
characteristics, such as consistency, frequency and volume, as well as sleeping and crying
duration and their frequency during the day. Thus, comfort parameters can be the result of
neurological or gastrointestinal pathways and involve much more complex physiological
mechanisms.
Clinical studies have revealed beneficial effects of sn2 palmitate on the comfort and wellbeing of healthy term infants through observations of softer stools (Carnielli et al. 1996,
Kennedy et al. 1999). Recently, in a randomized controlled study, formula containing sn2
palmitate was shown to reduce crying in infants compared to a control formula (Bar-Yoseph
et al. 2013). Additionally, the study showed that the consumption of formula with sn2
palmitate improved infant crying patterns during the day. The crying duration and patterns in
the sn2 palmitate group were similar to those of breastfed infants, and the percentage of
crying infants was significantly lower in the sn2 palmitate group compared to the control
group during the afternoon hours (Litmanovitz et al. 2014).
Crying is a basic, instinctive response that is governed by basic neuro-chemical
mechanisms similar to those that control feeding (Brazelton 1962). Although crying is a
common spontaneous behavior, it can induce parental concern, which often results in requests
for assistance from health services (Brazelton 1962). Most infants follow a universal crying
pattern during the first few months of life, in which crying peaks at 6 weeks of age and then
declines leading up to 3 months of age (Brazelton 1962, Barr et al. 1989, St James-Roberts
and Halil 1991, St James-Roberts et al. 1993, Baildam et al. 1995, Evanoo 2007). Crying also
has a typical diurnal pattern: 40% of crying episodes occur between 1600h and 2200h, and
after the third month of life, crying episodes are distributed throughout the day. Infant crying
is usually believed to be related to abdominal discomfort (Rasquin et al. 2006). Due to the
mechanisms involved in crying, it is conceivable that diet, by modifying neurochemistry,
could alter crying that is either endogenous for no apparent reason or associated with
physiological stimuli, such as hunger/separation. Dietary approaches to reducing crying
episodes in formula-fed infants have already been introduced. Savino et al., (2006) evaluated
the efficacy of a formula containing partially hydrolyzed whey proteins, prebiotic
oligosaccharides (OS), and a high sn2-palmitic acid content (41% in the sn2 position), and the
authors observed a significant difference in crying, which most likely resulted from the
significant decrease in colic episodes (Savino, Palumeri et al. 2006). More recently, the effect
151
of sn2 palmitate on infant crying was re-assessed in a double blind study using infant
formulas that already contained prebiotics (GOS). This study demonstrated an additional
beneficial effect of sn2 palmitate beyond the effect of prebiotics in crying reduction (BarYoseph et al. 2014).
The authors concluded that if the reduction of crying in infants fed formula with sn2plamitate is simply due to the looser stools, one would not expect an effect using formulas
with prebiotics that result in softer stools, and there would definitely not be a diurnal pattern
of altered crying (random during the day and likely associated with time of passing stool).
Therefore, this effect could not be attributed to softer stools alone, and the mechanism had not
yet been revealed.
GASTROINTESTINAL HEALTH
Over the past two decades, easy digestion and improved nutrient absorption following
sn2 palmitate-based infant formulas have been shown in preclinical and clinical studies in
preterm (Carnielli et al. 1995, Lucas et al. 1997) and term infants (Carnielli et al. 1996,
Kennedy et al. 1999, Lopez-Lopez et al. 2001); recently, this effect was shown even in infant
formulas containing prebiotics (Bar-Yoseph et al. 2014). The benefits of sn2 palmitate on
better digestion, promote a comfortable environment for the gut, thereby indicating a possible
health benefit to the gut. The potential mechanism of sn2 palmitate in improving gut function
is not clear; however, in view of its significant contribution to the digestion process, it is
reasonable to believe that it has further effects on the gastrointestinal surroundings.
The human gut is the natural habitat for a large and dynamic microorganisms' community
(Guarner and Malagelada 2003). The structure and composition of the gut flora reflects
natural selection at both the microbial and host levels, which promotes mutual cooperation
within and functional stability of this complex ecosystem (O'Hara and Shanahan 2006). The
fetal gut is sterile colonization begins immediately after birth and is influenced by the mode
of delivery, the infant diet, hygiene levels and medication (Niers et al. 2007). Throughout life,
the gastrointestinal tract becomes heavily inhabited by a complex community of
microorganisms, which are far in excess of the eukaryotic cells of the human body
(Dethlefsen et al. 2006). The intestinal microflora is an essential organ that serves
numerous important functions for the human host, including protection against pathogens and
provision of enhanced metabolic capabilities. It acts as important source of energy and
regulates intestinal epithelial proliferation, gut maturation and modulation of the
inflammatory immune response (Lodinova et al. 1967, Dethlefsen et al. 2006, Kau et al.).
Moreover, the characteristics of the intestinal bacterial community have been associated with
allergies, late-onset autism, inflammatory bowel disease and cancer (Dethlefsen et al. 2006).
It has been shown that bifidobacteria and lactobacillus protect infants from pathogenic
intestinal microorganisms and therby decrease the incidence of infantile diarrhea (Yoshioka et
al. 1983; Huang et al. 2002). They are associated with a higher production of short-chain fatty
acids (acetic and lactic acids), which are a source of energy for colonocytes. Furthermore,
strains of bifidobacteria and lactobacillus influence gut maturation processes in infants and
have anti-inflammatory effects (Hedin et al. 2007). Specific components of the intestinal
microflora, including Lactobacilli and Bifidobacteria, are beneficial for the host, such as
152
promoting gut maturation and integrity, acting against pathogens and participating in immune
modulation (Calder et al. 2006, Pai and Kang 2008).
The human diet plays a major role in the gut microbiota composition and development
(Fanaro et al. 2003,Kau et al. 2011, Moore et al. 2011). Following the first days after birth,
the flora composition rapidly changes under the influence of the infant's diet; consequently,
the composition is different between breastfed and formula-fed babies (Harmsen et al. 2000,
Fanaro et al. 2003). Within 1 week after birth, bifidobacteria becomes the predominant
species in the intestine of breastfed infants, whereas formula-fed infants have a more diverse
flora without any prevalent microorganisms (Harmsen et al. 2000, Fanaro et al. 2003, Morelli
2008). Yaron et al., showed that after 6 weeks of feeding the infants fed the sn2 palmitate
formula have a profile of intestinal microorganisms similar to that found in breastfed infants,
with high Lactobacilli and Bifidobacteria compared with infants fed control formula (Yaron
et al. 2013). A similar observation was reported in a recently published study by Yao et al.,
(Yao et al. 2014). The effect may be attributed to direct or indirect mechanisms. The 2-MAG
may directly induce the adhesion and proliferation of bifidobacteria and lactobacillus and/or
inhibit other competitive bacterial species in the gut. Alternatively, the indirect mechanism
might be from the non-absorbed vegetable oil products (i.e., calcium-palmitate insoluble
complexes) that reduce the growth of the beneficial bacteria in the intestine or from fat
degradation products that activate metabolic pathways in the intestine, affecting bacterial
growth. Although the mechanism by which sn2 palmitate affects the microbiota composition
is not well understood, these data indicate the beneficial long-term health effects for infants
fed with a sn2 palmitate-containing formula.
Furthermore, the direct interaction of microflora with the intestinal mucosa might
stimulate inflammatory activity in the gut lesions in IBD (Monteleone et al. 2014). Therefore,
there might be a relationship between the beneficial effect of sn2 palmitate on the gut flora
and its beneficial effect of protection from inflammation.
Proper development of immune tolerance is necessary for the maintenance of gut
homeostasis and an efficient response against pathogens. Dysregulation of the involved
mechanisms can lead to inappropriate intestinal inflammation against microbiota, such as
inflammatory bowel disease. The immaturity of the neonatal immune system explains the
age-dependent differences of the immune responses against pathogens and the susceptibility
to different types of infection (Tourneur and Chassin 2013).
Recently, it was shown in an animal model of Mucin2 deficient mice (Muc2-/-) with
colitis (Lu et al. 2013) that sn2 palmitate controls the intestinal inflammation. Mucins are the
principal components of the intestinal mucus layer (Velcich et al. 1997), which forms a
physical barrier that protects the underlying epithelium against luminal substances and
microbes (Hecht 1999, Hollingsworth and Swanson 2004, Dharmani et al. 2009). A
deficiency in Muc2 affects the protective capacities of the mucus layer (Johansson et al.
2011), leading to the development of spontaneous colitis in Muc2-/- mice, a well-described
animal model for enterocolitis (Velcich et al. 2002, Van der Sluis et al. 2006, Lu et al. 2011).
In this study, low sn2 palmitate diet increased the incidence of erosion and mucosal
damage in the distal colon of Muc2-/- mice compared with the AIN-93G reference diet, while
the sn2 palmitate diet controled the damage by enhancing the immunosuppressive Treg
response in Muc2-/- mice, suggesting beneficial effects of sn2-palmitic acid in limiting
intestinal inflammatory diseases (Figure 3, (Lu et al. 2013)).
153
Moreover, the sn2 palmitate diet induced an immunosuppressive Treg response (Lu et al.
2013). Increasing evidence has shown that Treg cells have a protective role in inflammatory
diseases (Sakaguchi 2005), and patients with inflammatory bowel diseases (IBD) have
reduced Treg cell numbers compared with patients who have non-IBD inflammatory diseases
(Boden and Snapper 2008).
These data suggest that replacing the control fat with sn2 palmitate (i.e., fat containing
high levels of palmitic acid esterified to the sn2 position) may enhance the
immunosuppressive Treg response in the intestine, thereby preventing or limit intestinal
inflammation. Furthermore, sn2 palmitate might have significant beneficial effects on
necrotizing enterocolitis (NEC), which is the leading cause of morbidity, mortality and longterm complications in preterm infants (Neu and Walker 2011).
Figure 3. Morphology of the distal colon of Muc2-/- mice fed AIN93G, CF (control diet, low sn2
palmitate), or sn2 palmitate (INFAT). Distal colonic sections of mice fed with the different diets were
stained with hematoxylin and eosin. Shown are representative sections for each diet group.
Data from other fatty acids, such as LC-PUFA, with similar effects merit further
investigation. Some postulated mechanisms of this observed effect are presumably attributed
to alterations in signal transduction pathways, membrane fluidity, intraluminal bacteria, and
gene expression (Teitelbaum and Allan Walker 2001).
CONCLUSION
The sn2 palmitate research area began with a focus on increased calcium and fat
absorption and its role in bone mineralization. The increasingly broad research efforts on sn2
palmitate gradually revealed additional, and significant clinical effects, such as intestinal flora
composition, intestinal inflammation, and the infant's comfort in the form of crying patterns.
The efficient absorption of fat by breast-fed infants emphasizes the importance of the human
milk TG structures for digestion. The structure of sn2sn2 palmitate is also conserved
following digestion, entering the blood stream to reach target peripheral tissues. This
observation strongly suggests that the significance of human milk TG structures goes beyond
maintaining efficient palmitic acid absorption. As demonstrated in this review, the effect of
sn2sn2 palmitate extends to different biological benefits beyond fat and calcium absorption,
while the mechanism and the active pathways involved in those effects remain to be
elucidated.
154
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In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 8
ABSTRACT
Palmitic acid, either in its triglyceride form or hydrolysed as a free fatty acid or an
ester, needs to be extracted from its source, processed and isolated to obtain useful
products. The objective of this work is to consider the use of SCF (supercritical fluid)
processing as a method to extract and process palmitic acid and/or its derivatives. A
phase behaviour analysis, in supercritical CO2, ethane and propane, at temperatures close
to the critical point of the solvent show moderate solubility of palmitic acid and
tripalmitin at pressures below 50 MPa and total solubility of methyl and ethyl palmitate at
pressures below 25 MPa. Analysis of the phase behaviour considered and studies
presented in the literature have shown that SCFs can be widely applied to the processing
of palmitic acid containing compounds. In particular SCFs can fractionate a mixture of
acids or their derivatives according to the chain length, it can de-acidify an edible oil and
it is able to fractionate a mixture containing palmitic acid and other compounds.
Additionally, SCFs can also be used to extract palmitic acid containing triglycerides from
plant material. SCFs, in particular CO2, are thus excellent alternative solvent to
traditional organic solvents and should be considered when processing palmitic acid
containing products.
160
C. E. Schwarz
FFA
SC
SCF
SCFE
SCFF
SVE
Tr
VLE
wi
INTRODUCTION
Palmitic acid is a major constituent of many naturally derived oils and extracts with
applications in, amongst others, the cosmetic, pharmaceutical and surfactant industries.
Palmitic acid is a linear saturated carboxylic acid with 16 carbon atoms, hence its IUPAC
name, hexadecanoic acid.
As its common name indicates, palmitic acid is a major constituent of the oil from palm
trees. However, palmitic acid does not only occur in the products of palm trees but occurs
widely in naturally derived oils and extracts from both plant and animal sources. Plant
material sources include soybean oil [1], cocoa butter [2], palm kernel oil [3], wheat bran oil
[4], pumpkin seed oil [5], to name but a few. Additionally by-products from oil production
also contain palmitic acid, for example soybean oil and palm oil deodoriser distillates [6,7].
Fish oils also contain a significant amount of palmitic acid, as shown in the review project of
Gruger et al. [8].
Table 1. Physical properties of palmitic acid and its derivatives
Component
Palmitic acid
Methyl palmitate
Ethyl palmitate
Tripalmitin
Melting point
336 to 338 K [9]
303 to 304 K [9]
297 to 298 K [9]
337 to 338 K [10]
Boiling point
544 K [9]
469 K at 2 kPa [9]
464 K at 1.33 kPa [9]
(Not readily available)
Molecular mass
256.42 g/mol
270.45 g/mol
284.48 g/mol
807.32 g/mol
In nature, palmitic and other high molecular mass acids generally do not occur as free
fatty acids (FFAs) but rather primarily as triglycerides. In general, these triglycerides are
usually mixed, in other words the triglycerides contains different acids e.g. one palmitic acid
and two oleic acid groups. While extraction from the source material results in triglycerides,
should palmitic acid be isolated or concentrated, the triglycerides need to be hydrolysed to
acids or esters. Therefore, palmitic acid and its derivatives, while present in many sources,
also occur in combination with many other components. In order to obtain pure palmitic acid
(or its derivative), or even a product concentrated in palmitic acid, some type of separation
process is required.
As palmitic acid may occur as a triglyceride, or may be hydrolysed or esterified, this
study will thus concern itself with not only the triglyceride form but also the FFA (palmitic
acid) and the methyl and ethyl esters (methyl and ethyl palmitate) form of palmitic acid. The
161
molecular mass and melting and boiling temperatures (where available) of palmitic acid and
its derivatives are presented in Table 1.
The physical properties of these compounds indicate that while they are liquids or solids
near their melting temperature at room temperature, they have very high boiling points, even
at vacuum. Additionally, their high molecular mass and hydrocarbon backbone nature results
in significant thermal degradation at elevated temperatures, in particular above 420 K. These
properties, together with the surfactant nature of these compounds points to difficulties in
separating these compounds from others using traditional separation methods, such as
distillation and liquid-liquid extraction.
Supercritical fluid (SCF) processing is an attractive alternative separation method that
can readily be used to separate high molecular mass thermally sensitive compounds. This
separation method is analogous to separation with a liquid organic solvent except instead of
the use of an organic solvent a high pressure gas near its critical point is used. Previously SCF
processing has been used to fractionate waxes according to their chain length [1113],
separate detergent range alkanes and alcohols [14,15], and extract components from plant and
animals sources [16], to name but a few.
The aim of this chapter is to investigate the supercritical fluid extraction (SCFE) and
supercritical fluid (SCFF) of solid matrices and liquid mixtures containing palmitic acid and
its derivatives (tripalmitin, methyl palmitate and ethyl palmitate). SC (supercritical) CO2,
ethane and propane will be considered as SC solvents with ethane, propane and other
compounds as possible co-solvents to CO2.
In particular, this chapter will focus on:
(1) The phase behaviour of palmitic acid and its derivatives in SCFs and
(2) Using the phase behaviour information, how SCFF and SCFE can be used to obtain a
product enriched in palmitic acid and/or its derivatives.
This chapter will start with a brief overview of SCF processing. Thereafter, a phase
behaviour analysis of palmitic acid, methyl palmitate, ethyl palmitate and tripalmitin in CO2,
ethane and propane, as well as CO2 together with a co-solvent will be considered. The
analysis will investigate phase equilibrium data present in the literature, trends observed the
data, a comparison between the phase behaviour of the various derivatives as well as the
various solvents, and the effect of a co-solvent. The phase behaviour data will then be used to
analyse how SCFF and SCFE can be used to obtain extracts enriched in palmitic acids or its
derivatives. In particular this study will consider how SCFF can be used to distinguish
between various fatty acids and their derivatives and also between palmitic acid and/or its
derivatives and other components present in the sources. Finally, SCFE of palmitic acid
containing triglycerides from solid matrices will be considered. The overarching aim of this
chapter is thus to show how SCFs can be used to process palmitic acid and its derivatives.
162
C. E. Schwarz
change. In general, SCF processes are operated close to the critical temperature and
preferably the critical pressure of the solvent. At these conditions a SCF is neither a liquid nor
a gas but has physical and transport properties are intermediate of that of a liquid and a gas.
Additionally, in the vicinity of the critical point a small change in pressure and/or temperature
results in a large change in density, and thus solubility of components in these solvents.
Therefore, SCFs have excellent mass transfer properties and are often used as extraction or
fractionation media. Processing using SCFs are analogous to using a liquid organic solvent
with the exception that instead of a liquid solvent a high pressure gas is used as the mass
transfer medium. CO2 is the most popular SCF but low molecular mass alkanes are also
sometimes used.
While this separation method does require high pressure equipment and is generally not
the first method of choice to achieve a separation, it does hold a number of advantages.
Typically a SCF process operates a few degrees above the critical temperature of the solvent
(See Table 2) and as such thermal degradation of temperature sensitive compounds is
avoided. The nature of the solvents are such that they are generally non-toxic and in some
cases also non-flammable. In fact, the most popular SC solvent, CO2, is classified as generally
regarded as safe (GRAS). Additionally, at atmospheric temperatures and pressures these
solvents generally have a very low solubility in high molecular mass solutes resulting in a
negligible solvent residue. Lastly, due to the pressure-volume-temperature behaviour of the
solvent in the operating range, solvent-solute separation can easily be achieved through a
change in pressure and/or temperature. A complicated solvent removal process is thus not
generally required.
SCF separation processes can be divided into two main groups namely SCFF of a liquid
feed and SCFE of compounds from a solid matrix. Basic schematic representations of these
two processes are shown in Figure 1 and Figure 2 for SCFF and SCFE, respectively.
Table 2. Critical properties of selected SC solvents [17]
SC solvent
Carbon dioxide
Methane
Ethane
Propane
n-Butane
Water
Critical Temperature
304.1 K
190.4 K
305.1 K
369.8 K
425.3 K
647.1 K
Critical pressure
7.38 MPa
4.60 MPa
4.88 MPa
4.24 MPa
3.80 MPa
22.06 MPa
As pointed out by Peter and Brunner [18], SCF processing can extract and separate
compounds containing low/non-volatile and heat sensitive substances at temperatures far
lower than that of vacuum distillation. Both SCFF and SCFE are techniques that can be
applied to sources containing palmitic acid. For example, palm kernel oil can be extracted
from the kernels using SCFE and then fractionated by SCFF. In order to design such
processes one requires knowledge of the solubility of palmitic acid (or its derivate) as well as
the other components present in the SC solvent. A difference in solubility between
compounds is a necessary requirement for separation to occur.
163
Carbon dioxide (CO2) is by far the most studied SCFs. This is mainly due to its suitable
critical temperature, its abundance and its non-toxicity. The popularity of CO2 can be seen in
number of studies considering the phase behaviour of compounds in CO2 [1922] and
fractionation and extraction studies using SC carbon dioxide [23]. However, CO2 is not
always a good solvent for high molecular mass compounds. In particular, acids and
triglycerides often require very high pressures for significant solubility in SC CO2 [24,25].
164
C. E. Schwarz
Although methyl and ethyl esters of acids have a significantly higher solubility in SC CO2
and total solubility can be attained [26], relatively high pressures (approximately 20 MPa) are
still required.
Low molecular mass alkanes (in particular ethane and propane) are also popular SCFs
and have shown improved solubility of acids in SC CO2 [27,28] with total solubility below
30 MPa at moderate temperatures. Additionally, for methyl and ethyl esters ethane and
propane also show a marked increase in solubility [29,30]. These alkanes can be used either
as solvents on their own or may be used as a co-solvent together with CO2. The critical
temperature of ethane is very close to that of CO2, hence its suitability at an alternative
solvent to CO2. On the other hand, propane has a significantly higher critical temperature
(369.8 K). However, this temperature is quite close to the melting points of palmitic acid and
tripalmitin and, as pointed out by Coorens et al. [31], may therefore be a suitable SC solvent
to process palmitic acid and its derivative. Additionally, propane in general has a higher
solubility of high molecular mass compounds compared to ethane [30], thus requiring lower
pressures and/or lower solvent to feed ratios.
In addition, besides propane and ethane, a number of studies have considered other low
molecular weight organic compounds such as ethanol and n-octane as possible co-solvents for
SCF processes. Ideally, the addition of the co-solvent generally decreases the operating
pressure significantly while hopefully not influencing the selectivity significantly.
This study will thus consider the SCFF and SCFE of sources containing palmitic acid or
its derivatives using predominantly CO2 as extraction medium. However, ethane and propane
as possible alternative solvents will also be considered, as well as the use of co-solvents.
165
CO2/Palmitic Acid
The systems CO2/palmitic acid has been extensive studied. Outlined in Table 3 is a
summary of the available phase equilibrium data.
Palmitic acid has a normal melting point of 336 to 338 K [9]. However, in the presence of
a SC CO2 palmitic acid undergoes a melting point depression. Bertakis et al. [46] studied the
vapour-liquid-solid phase equilibria of, amongst others, the CO2/palmitic acid system. Their
results showed that the melting point decreased significantly as the pressure increased to a
minimum of approximately 319.5 K at about 10.6 MPa, after which the melting temperature
increased slightly as the pressure increased. This information is significant at it indicates
which of the data listed in Table 3 is of vapour-solid nature and which data is of vapour-liquid
nature. As most of the data is above 10 MPa, and the melting point does not significantly
change at pressure above 10 MPa, it is assumed that all data above 320 K is of vapour-liquid
in nature, the remainder being of vapour-solid in nature.
Figure 3 shows the solubility pressure of solid palmitic acid in CO2 as a function
composition at 308.5 to 318.15 K. The data of Brunetti et al. [35] does not agree with that of
the other authors and was therefore omitted from Figure 3. Maheshwari et al. [42] also
questioned the data of Brunetti et al. [35] as they found that the data of Brunetti et al. was
consistently higher than their data as well as that of Ohgaki et al. [43]. They ascribed the
difference due to the presence of impurities in the materials. They suggested that impurities
may act as entrainers and could therefore either increase or decrease the solubility of the
molecules in CO2, depending on the impurities present.
Table 3. Literature data for the CO2 (1)/palmitic acid (2) system
Reference
Purity
Temperature
range
Pressure range
Composition range
(w2) a
(1) 99.92 %
(2) 99-100 %
313 K
(1) 99.9 %
(2) 90 %
353.15 and
373.15 K
13.60 to
30.52 MPa
Vapour: 0.000582 to
0.0155 (5)
Liquid: 0.580 to 0.810
(7)
(1) 99.995 %
(2) 99 %
10.14 to
23.35 MPa
8.361E-4 to 6.44E-3
(11)
166
C. E. Schwarz
Table 3. (Continued)
Reference
Purity
Temperature
range
Pressure range
Composition range
(w2) a
(2) 96 % min.
308.15 to
323.15 K
20 and 30 MPa
Garlapati and
Madras [36]
(1) 99.9 %
(2) 98 %
12.8 to 22.6
MPa
1.50E-3 to 6.06E-3
(10)
Garlapati and
Madras [37]
(1) 99.9 %
(2) 98 %
328 K
12.8 to 22.6
MPa
308.15 K to
328.15 K
10.0 to 35.0
MPa
3.79E-3 to 9.13E-3
(18)
(1) 99.9 %
(2) 99 %
308.15 K
(1) 99.9 %
(2) 99 %
313.2 K
15.0 MPa
3.24E-3 (1)
(1) 99.8 %
(2) 95 %
318 to 338 K
14.21 to
57.48 MPa
Maheshwari et al.
[42]
(1) 99.9 %
(2) 99 %
308 to 328 K
13.8 to
41.4 MPa
(1) 99.99 %
(2) 99 %
298.15 and
313.15 K
7.89 to 18.7
MPa
4.08E-4 to 3.34E-3
(22)
(1) 99.99 %
(2) 99.99 %
313.15 to
353.15 K
6 to 12 MPa
Vapour: 0.0087 to
0.0345 (9)
Liquid: 0.2107 to
0.3372 (9)
Schwarz and
Knoetze [24]
(1) 99.95 %
(2) 99 %
337.1 to 358.2
K
10.53 to
25.93 MPa
Vapour: 0.0142 to
0.0224 (23)
Liquid: 0.585 to 0.756
(18)
(1) 99.5 %
(2) 98 %
373.15 to
473.15 K
1.01 to 5.07
MPa
Vapour: 0.0000 to
0.0031 (15)
Liquid: 0.931 to 0.993
(15)
As seen, a reasonable solubility (greater than 0.2 mass %) can be attained at moderate
pressures (less than 20 MPa), indicating that SC CO2 is able to extract palmitic acid from a
solid matrix. The data also shows, as pointed out by Bamberger et al. [32], that an increase in
temperature at constant pressure leads to an increase in solubility, and, as stated by Garlapati
and Madras [37], that an increase in pressure at constant temperature also leads to an increase
in solubility.
The VLE phase behaviour of the CO2/palmitic acid system using the data of Schwarz and
Knoetze [24] is shown in Figure 4. Schwarz and Knoetze found their data for
CO2/hexadecanoic acid to be in agreement with that of Kramer and Thodos [41] and Bharath
et al. [33]. However, the data of Penedo et al. [44] appeared to be at too low pressures, most
probably due to the fact that they used an extraction type set-up instead of a phase equilibrium
167
type set-up. Penedo et al. [44] claim that their results agree well with that of Chen et al. [47]
for the system CO2/linoleic acid and thus regarded their results as verified. However,
inspection of the graphs showing the comparison points towards a significant difference. For
all further analyses the data of Schwarz and Knoetze will be used as these authors have
regularly verified their method, they use a synthetic method which is highly accurate for
binary phase equilibrium data, and the palmitic acid used by Schwarz and Knoetze is of high
purity.
40
35
Pressure (MPa)
30
318.15 K
25
313.15 K
20
308.15 K
15
10
5
0
0.000
0.002
0.004
w2
0.006
0.008
Figure 3. Pressure composition (w2) plot for the CO2 (1)/palmitic acid (2) system at 308.15 to 318.15
K [34,36,3843].
32
28
Pressure (MPa)
24
20
358.15 K
16
348.15 K
12
338.15 K
8
4
0
0.0
Figure 4. (Continued).
0.2
0.4
(a) w2
0.6
0.8
1.0
168
C. E. Schwarz
32
28
Pressure (MPa)
24
20
358.15 K
16
348.15 K
12
338.15 K
8
4
0
0.012
0.015
0.018
0.021
0.024
(b) w2
Figure 4. Pressure composition (w2) plot for the CO2 (1)/palmitic acid (2) system at 338.15 to 358.15
K [24] (a) entire composition range and (b) detail of low molecular mass composition range.
Schwarz and Knoetze [24] found that for their VLE data an approximately linear
relationship exists between temperature and the phase transition pressure at constant
composition. This relationship has a positive gradient and indicates a higher solubility at
lower temperatures, converse to that of the solid-vapour equilibrium (SVE) phase behaviour.
This positive gradient was also found through the entire mass fraction range studied and the
authors did not find any indications of temperature inversions in this system.
Comparing Figure 3 and Figure 4(b) it is clear that the solubility of liquid phase palmitic
acid in SC CO2 is of an order of magnitude higher than that of the solid phase palmitic acid
(compare the x-axes). Thus, SCFE of palmitic acid as a solid from a matrix will, in all
likelihood, occur at much higher pressures than SCFF processes involving palmitic acid.
CO2/Methyl Palmitate
Outlined in Table 4 is a summary of data for the system CO2/methyl palmitate. Due to the
low melting point of the data (See Table 1), all data is of a vapour-liquid equilibrium (VLE)
nature.
Figure 5 shows the phase equilibrium data of the system CO2/methyl palmitate of both
Inomata et al. [48] and Lockemann [49]. Figure 5 hints towards significant differences
between the data sets, yet insufficient information is available to determine which data set us
superior. However, the data clearly shows that an increase in temperature results in an
increase in phase transition pressure. Due to the scatter in the data and slight inconsistencies
between the two sources the exact nature of the relationship between temperature and the
phase transition pressure can not be determined. Additional measurements would be required
therefore. Both sets of data do, however, indicate that total solubility can be achieved at
moderate pressures (less than 25 MPa at temperatures below 343 K).
169
Table 4. Literature data for the CO2 (1)/methyl palmitate (2) system
Reference
Purity
Inomata et al.
[48]
Lockemann
[49]
a
(1) 99.9 %
(2) 95 %
Temperatur
e range
313.15 to
343.15 K
Pressure
range
1.01 to
18.28 MPa
(1) 99.95 %
(2) 95 %
313.15 to
333.15 K
0.98 to
13.0 MPa
Composition range
(w2) a
Vapour: 0.00165 to
0.0676 (24)
Liquid: 0.333 to
0.960 (16)
Vapour: 0.0006 to
0.1028 (16)
Liquid: 0.3041 to
0.9614 (38)
16
Pressure (MPa)
12
343.15 K
8
333.15 K
323.15 K
313.15 K
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 5. Pressure composition (w2) plot for the CO2 (1)/methyl palmitate (2) system at 313.15 to
343.15 K [48,49].
CO2/Ethyl Palmitate
Table 5 summarises data available for the system CO2/ethyl palmitate and the data is
illustrated in Figure 6. Liang and Yeh [50] also studied the solubility of ethyl palmitate in
CO2. However, they presented their data only in figures and as a correlation (Chrastil
correlation). As such their data has not been included in this study. The data of Crampon et al.
[26] and that of Gaschi et al. [51] appear to be in agreement.
The data also indicates, as pointed out by Crampon et al., that an increase in temperature
leads to an increase in phase transition pressure and suggests that a linear relationship
between temperature and pressure exists. Gaschi et al. observed three phase behaviour at
303.15 K but not at 313.15 K and higher temperatures. Three phase regions are interesting
from a thermodynamic point of view yet in practice should be avoided due to the
complications associated therewith. Importantly, as for the CO2/methyl palmitate system,
total solubilities can be attained at moderate pressures (< 25 MPa) in the temperature range
under investigation. This affords lower operating pressures and as such lower capital and
operating costs.
170
C. E. Schwarz
CO2/Tripalmitin
A significant amount of data has been published on the VSE and VLE for the system
CO2/tripalmitin, as outlined in Table 6. To date, no detailed study has been conducted on the
melting point depression of tripalmitin in SC CO2. Due to the similar melting point of
tripalmitin and palmitic acid [9], a cut-off temperature of 320 K similar to that of palmitic
acid in CO2 is assumed. The VSE and VLE of the system CO2/tripalmitin is shown in Figure
7 and Figure 8, respectively.
Table 5. Literature data for the CO2 (1)/ethyl palmitate (2) system
Reference
Purity
Temperatur
e range
Pressure range
Composition
range (w 2) a
Crampon et al.
[26]
(1) 99.9 %
(2) 99 %
313.15 to
333.15 K
1.98 to
15.89 MPa
0.0376 to 0.931
(77)
Gashi et al.
[51]
(1) 99.9 %
(2) 95 %
303.15 to
353.15 K
3.10 to
20.86 MPa
0.05368 to 0.8645
(60)
21
Pressure (MPa)
18
15
353.15 K
12
343.15 K
333.15 K
323.15 K
313.15 K
303.15 K
3
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 6. Pressure composition (w2) plot for the CO2 (1)/ethyl palmitate (2) system at 303.15 to
353.15 K [26,51].
As for the VSE of the CO2/palmitic acid system, the VSE of the CO2/tripalmitin system
shows that an increase in temperature leads to an increase in solubility. However, there
appears to be significant discrepancies between the various sources, as seen for the 313.15 K
data. Bamberger et al. [32] found that the data of Chrastil [52] is an order of magnitude
higher than theirs and speculate that it may be due to purity. Bamberger et al. conducted
additional tests (data not given) using a lower purity tripalmitin (90%) and found that the
solubilities were much higher. The purity of the tripalmitin used by Chrastil was not given.
171
It may therefore be that the discrepancies between the data sets are due to the purity of the
components used.
Table 6. Literature data for the CO2 (1)/tripalmitin (2) system
Reference
Purity
Bamberger
et al. [32]
Bharath et
al. [33]
(1) 99.92 %
(2) 99 %
(1) 99.9 %
(2) 90 %
Chrastil
[52]
Mnkl et
al. [53]
Nilsson and
Hudson
[54]
Ohgaki et
al. [43]
Weber et
al. [25]
(1) 99.98 %
313 to 353 K
(1) 99.995 %
(2) 99 %
Not given
336 to 363 K
(1) 99.99 %
(2) 96.5 %
(1) 99.5 %
(2) 95 %
298.15 to
313.15 K
333 and 353
K
Temperature
range
313 K
353.15 K
313.15 and
333.15 K
Pressure
range
12.2 to 29.7
MPa
5.55 to
24.31 MPa
8.11 to
25.33 MPa
3.24 to
24.75 MPa
17.2 to 31.0
MPa
8.60 to 18.2
MPa
10 to 50 MPa
Composition
range (w 2)a
5.56E-5 to 4.97E-4
(7)
Vapour: 0.00183
to 0.0286 (5)
Liquid: 0.561 to
0.787 (5)
1.02E-4 to 1.96E-3
(18)
0.700 to 0.950 (20)
2.8E-4 to 1.08E-4
(9)
8.36E-5 to 1.42E-4
(16)
Vapour: 0.00018
to 0.03340 (9)
Liquid: 0.5873 to
0.9183 (9)
40
35
Pressure (MPa)
30
313.15 K
25
20
298.15 K
15
10
5
0
0.00E+00
5.00E-04
w2
1.00E-03
1.50E-03
Figure 7. Pressure composition (w2) plot for the CO2 (1)/tripalmitin (2) system at 292.15 and 313.15
K [32,43,52,54].
172
C. E. Schwarz
50
45
Pressure (MPa)
40
35
30
363.15 K
25
353.15 K
20
343.15 K
15
10
5
0
0.0
0.2
0.4
(a) w2
0.6
0.8
1.0
50
45
40
Pressure (MPa)
35
30
25
353.15 K
20
15
10
5
0
0.000
0.006
0.012
(b) w2
0.018
0.024
Figure 8. Pressure composition (w2) plot for the CO2 (1)/tripalmitin (2) system at 343.15 to 363.15 K
[25,33,52,53] (a) entire composition range and (b) detail of low molecular mass composition range.
173
Pressure (MPa)
28
24
Tripalmitin
20
Palmitic acid
16
Methyl
palmitate
12
8
4
0
0.0
0.2
0.4
(a) w2
0.6
0.8
1.0
32
28
Pressure (MPa)
24
20
Tripalmitin
16
Palmitic acid
12
8
4
0
0.000
0.002
(b) w2
0.004
0.006
Figure 9. Comparison of the pressure composition (w2) for the systems CO2 (1)/palmitic acid (2)
[34,43], CO2 (1)/methyl palmitate (2) [48,49], CO2 (1)/ethyl palmitate (2) [26,51] and CO2
(1)/tripalmitin (2) [32,43,54] systems at 313.15 K (a) entire composition range and (b) detail of low
molecular mass composition range.
174
C. E. Schwarz
The data shows that at both temperatures the esters have the highest solubility followed
by palmitic acid and that tripalmitin has the lowest solubility. It is also noted that the systems
CO2/methyl palmitate and CO2/ethyl palmitate have very similar phase behaviour and similar
phase transition pressures. This is expected due to the similarity of the molecules. However,
despite the fact that Bharath et al. [55] found that for C18 and higher esters the methyl ester
has a higher phase transition pressure than the ethyl ester, Figure 9 indicates that the phase
transition pressures are indeed very similar and from the data available in the present analysis
no outcome can be given in this regard.
32
28
Pressure (MPa)
24
20
Tripalmitin
16
Palmitic acid
12
8
4
0
0.0
0.2
0.4
0.6
0.8
1.0
(a) w2
32
28
Pressure (MPa)
24
20
Tripalmitin
16
Palmitic acid
12
8
4
0
0.000
0.005
0.010
0.015
0.020
0.025
(b) w2
Figure 10. Comparison of the pressure composition (w2) for the CO2 (1)/palmitic acid (2) [24], CO2
(1)/ethyl palmitate (2) [51]and CO2 (1)/tripalmitin (2) [25,33,52,53] systems at 353.15 K (a) entire
composition range and (b) detail of low molecular mass composition range.
175
Importantly, however, is the fact that the solubility of methyl palmitate and ethyl
palmitate in SC CO2 is significantly higher than that of palmitic acid and tripalmitin.
Significantly lower pressures would therefore be required for processes involving methyl
palmitate or ethyl palmitate compared to those involving palmitic acid or tripalmitin.
Purity
Schwarz and
Chobanov [27]
(1) 99.5 %
Temperature
range
332.4 to 354.4 K
Pressure
range
9.17 to
21.20 MPa
Composition
range (w 2) a
0.0164 to 0.671
(2) 99 %
Value in brackets indicates the number of data points published
Ethane/Methyl Palmitate
As for the ethane/palmitic acid system, only a single source contains data for the
ethane/methyl palmitate system. The data is outlined in Table 8 and the phase behaviour for
the system ethane/methyl palmitate is shown in Figure 12.
Schwarz et al. [29] found that total solubility can be attained for the system
ethane/methyl palmitate at low pressures (< 15 MPa) and that, as for the ethane/palmitic acid
system, a linear relationship exists between the temperature and the phase transition pressure
at constant composition in the temperature range studied.
176
C. E. Schwarz
24
Pressure (MPa)
20
16
353.15 K
12
343.15 K
333.15 K
8
4
0
0.0
0.1
0.2
0.3
w2
0.4
0.5
0.6
0.7
Figure 11. Pressure composition (w2) plot for the ethane (1)/palmitic acid (2) system at 333, 343 and
353 K [27].
Table 8. Literature data for the ethane (1)/methyl palmitate (2) system
Reference
Purity
Temperatu
re range
Pressure range
Composition range
(w2) a
Schwarz et
al. [29]
(1) 99.95 %
(2) 99 %
311.5 to
365.5 K
5.21 to 13.8
MPa
15
Pressure (MPa)
12
9
353.15 K
343.15 K
333.15 K
323.15 K
313.15 K
3
0
0.0
0.1
0.2
0.3
w2
0.4
0.5
0.6
0.7
Figure 12. Pressure composition (w2) plot for the ethane (1) /methyl palmitate (2) system at 313 to
353 K [29].
177
Ethane/Ethyl Palmitate
Once again, only a single data source is available for the ethane/ethyl palmitate system is
available. The data is summarised in Table 9 and the phase behaviour of the ethane/ethyl
palmitate data is presented in Figure 13.
The data for the ethane/ethyl palmititate system is very similar to that of the
ethane/methyl palmitate system and the same comments thus apply.
Table 9. Literature data for the ethane (1)/ethyl palmitate (2) system
Reference
Purity
Temperature
range
312.7 to 357.3 K
Pressure range
Schwarz et
(1) 99.5 %
5.38 to 13.52
al. [30]
(2) 99 %
MPa
a
Value in brackets indicates the number of data points published
Composition
range (w 2) a
0.0155 to
0.593 (48)
15
Pressure (MPa)
12
9
353.15 K
343.15 K
333.15 K
323.15 K
313.15 K
3
0
0.0
0.1
0.2
0.3
w2
0.4
0.5
0.6
0.7
Figure 13. Pressure composition (w2) plot for the ethane (1)/ethyl palmitate (2) system at 333, 343
and 353 K [30].
Ethane/Tripalmitin
No data is available for the system ethane/tripalmitin. In order to estimate the phase
behaviour of the system ethane/tripalmitin one can approximate the data to that of an alkane
with the same molecular mass. Peters previously suggested this analogy for the
propane/tripalmitin system [56]. The oxygen atoms are located centrally in the molecule and
thus well shielded. Tripalmitin has 57 carbon and oxygen atoms in the molecular backbone
and as such the ethane/tripalmitin system may be approximated by the ethane/nheptapentacosane system as a first order approximation. While the VLE for the ethane/nheptapentacosane system has not been measured, Schwarz et al. [57] published correlations
that link the phase transition pressure with the number of carbon atoms. These correlations
are used and the approximated phase behaviour of the system ethane/tripalmitin is presented
in Figure 14. It should be noted that these correlations are based on data generated with the
178
C. E. Schwarz
solute in the liquid phase and are thus only valid above the melting point of the solute.
Additionally, data for ethane/n-heptapentacosane system would be an extrapolation and while
extrapolation may be possible [58] the data generated would therefore be more of an
indication of the phase transition pressure rather than an accurate representation. Figure 14
has been generated at 340 and 360 K, and while n-heptapentacosane may be a solid at this
temperature, tripalmitin will be in the liquid state, hence its validity.
As expected, an increase in temperature leads to an increase in phase transition pressure.
Importantly, total solubility can be attained, albeit at quite high pressures (approximately 35
MPa at 360 K).
35
Pressure (MPa)
28
21
360 K
340 K
14
7
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 14. Approximation of the ethane (1)/tripalmitin (2) pressure composition (w2) plot at 340 and
360 K based on the ethane/n-alkane correlations for 57 carbon atoms, as proposed by Schwarz et al.
[57].
179
35
Tripalmitin
30
Palmitic acid
Pressure (MPa)
25
Methyl
palmitate
20
15
10
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 15.Comparison of the pressure composition (w2) for the systems ethane (1)/palmitic acid (2)
[27], ethane (1)/methyl palmitate (2) [29], ethane (1)/ethyl palmitate (2) [30] and ethane (1)/tripalmitin
(2) (generated as described above using the ethane/n-alkane correlation of Schwarz et al. [57]) systems
at 360 K.
180
C. E. Schwarz
their graphs indicates that the three phase region occurs between approximately 370 and 374
K and therefore the observation of Schwarz et al. that no three phase regions are present
between 378 and 408 K is in agreement with the work of Peters [56].
Table 10. Literature data for the propane (1)/palmitic acid (2) system
Reference
Purity
Temperature
range
Pressure
range
Composition range
(w2) a
Schwarz et
al. [28]
(1) 99.95 %
(2) 99 %
376.4 to 409.5
K
4.18 to 9.03
MPa
0.0184 to 0.655
10
Pressure (MPa)
8
6
408.15 K
393.15 K
378.15 K
2
0
0.0
0.1
0.2
0.3
w2
0.4
0.5
0.6
0.7
Figure 16. Pressure composition (w2) plot for the propane (1)/palmitic acid (2) system at 378 to 408 K
[28].
Propane/Methyl Palmitate
Rovetto et al. [59] published phase equilibrium data for the system propane/methyl
palmitate. The data is summarised in Table 11 and Figure 17 shows the phase behaviour.
Figure 17 indicates that total solubility can be attained at very low pressures (< 8 MPa).
The data also shows that as temperature increases, the phase transition pressure increases. In
fact, the data shown in Figure 17 was obtained by fitting a linear relationship between the
temperature and phase transition pressure at constant composition on the data published by
Rovetto et al. [59]. Data between approximately 370 and 410 K was used and a fit with a
Pearsons R2 value greater than 0.997 was attained in all cases for a linear relationship
between the temperature and the phase transition pressure.
181
Table 11. Literature data for the propane (1)/methyl palmitate (2) system
Reference
Purity
Temperature
range
Pressure
range
Composition range
(w2) a
Rovetto et
al. [59]
(1) 99.95 %
(2) 99 %
312.11 to
450.63 K
1.23 to 9.87
MPa
0.1798 to 0.7647
(84)
10
Pressure (MPa)
8
6
408.15 K
393.15 K
378.15 K
2
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 17. Pressure composition (w2) plot for the propane (1)/methyl palmitate (2) system at 378 to
408 K [59].
Propane/Ethyl Palmitate
Together with their study on the phase behaviour of the ethane/ethyl ester homologous
series, Schwarz et al. published data on the propane/ethyl ester homologous series. This is the
only known source of data for the propane ethyl/palmitate system, is summarised in Table 12
and graphically illustrated in Figure 18.
As for the propane/methyl palmitate system, total solubility can be attained at very low
pressures (< 8 MPa) for the propane/ethyl palmitate system. Similar to the observed linear
relationship between temperature and the phase transition pressure, Schwarz et al. [30] also
found a linear relationship between the phase transition pressure and temperature. They also
did not observe any three phase behaviour or indications thereof.
Table 12. Literature data for the propane (1)/ethyl palmitate (2) system
Reference
Purity
Temperature
range
377.2 to 409.6 K
Pressure
Composition range
range
(w2) a
Schwarz et
(1) 99.95 %
3.52 to 7.36 0.0201 to 0.642 (36)
al. [30]
(2) 99 %
MPa
a
Value in brackets indicates the number of data points published
182
C. E. Schwarz
8
Pressure (MPa)
4
408.15 K
2
393.15 K
378.15 K
0
0.0
0.1
0.2
0.3
w2
0.4
0.5
0.6
0.7
Figure 18. Pressure composition (w2) plot for the propane (1)/ethyl palmitate (2) system at 378 to 408
[30].
Propane/Tripalmitin
Coorens et al. [31] conducted a detailed study on the phase behaviour of the
propane/tripalmitin system. Their data is summarised in Table 13 and illustrated in Figure 19
for SVE and in Figure 20 for VLE phase behaviour.
The VSE of the propane/tripalmitin system shows that the solubility is very much
temperature and pressure dependent and that at constant pressure, significantly higher
solubility can be attained at higher temperatures. Similar observations were made for the
CO2/tripalmitin system.
The VLE phase behaviour of the propane/tripalmitin system is similar to that of the other
compounds in propane. Total solubility can be attained at moderate pressure (< 12 MPa).
Table 13. Literature data for the propane (1)/tripalmitin (2) system
Reference
Coorens et al. [31]
Purity
Temperatur
e range
317.59 to
413.77 K
Pressure
range
0.351 to
12.731 MPa
(1) 99.95 %
(2) GC
standard
Coorens et al. [31] (1) 99.95 %
314.36 to
1.051 to
(2) GC
339.37 K
13.051 MPa
standard
a
Value in brackets indicates the number of data points published
Composition
range (w 2) a
0.1258 to
0.9752 (139)
0.1258 to
0.9752 (51)
183
15
Pressure (MPa)
12
0.975
9
0.861
0.717
0.491
0.303
0.126
0
310
315
320
325
330
335
340
Temperature (K)
Figure 19. Pressure temperature plot for the propane (1)/tripalmitin (2) system at various tripalmitin
compositions (w2) K [30].
12
Pressure (MPa)
9
408.15 K
6
393.15 K
378.15 K
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 20. Pressure composition (w2) plot for the propane (1)/tripalmitin (2) system at 378, 393 and
408 K [30].
As for the propane/methyl palmitate data, the propane/tripalmitin data presented was
generated using a linear relationship fitted to the experimentally published data of Coorens et
al. [31]. Once again an excellent fit was obtained (R2 > 0.997 in all cases), therefore
illustrating the linear relationship between temperature and the phase transition pressure.
Coorens et al. [31] also published vapour-liquid-liquid equilibrium data showing that the
system has a three phase region between 349 and 370 K. However, compositions were not
included.
184
C. E. Schwarz
Comparing Figure 19 and Figure 20, it can be seen that the phase behaviour for the VSE
and VLE is significantly different. As for the CO2/palmitic acid and CO2/tripalmitin systems,
the VSE data shows an increase in solubility with temperature while the converse is true for
the VLE data. Additionally, the solubility of the solid in propane is again much lower than
that of the liquid in propane. Thus, while the absolute values of the pressure differ (a detailed
analysis is presented below) the same trends as for the CO2/tripalmitin system are present.
Pressure (MPa)
Tripalmitin
Palmitic acid
Methyl
palmitate
4
2
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 21. Comparison of the pressure composition (w2) for the systems propane (1)/palmitic acid (2)
[28], propane (1)/methyl palmitate (2) [59], propane (1)/ethyl palmitate (2) [30] and propane
(1)/tripalmitin (2) [31] systems at 393 K.
As for CO2 and ethane as SC solvents, the same trends are observed for propane as SC
solvent. Methyl palmitate and ethyl palmitate behave very similarly and are the most soluble,
followed by palmitic acid. Tripalmitin is the least soluble in propane, yet even for tripalmitin
only moderate pressures are required for total solubility.
Despite the high temperatures, relatively low pressures are required for total solubility,
therefore decreasing the capital cost investment. Propane should thus be regarded as a good
substitute or co-solvent to CO2 to reduce the processing pressures.
185
Pressure (MPa)
20
Propane
15
Ethane
10
Carbon
dioxide
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 22. Comparison, at similar reduced temperatures (Tr ~ 1.11), of the pressure composition (w2)
phase behaviour of the CO2 (1)/palmitic acid (2) at 338 K [24], ethane (1)/palmitic acid (2) at 338 K
[27] and propane (1)/palmitic acid (2) at 410 K [28] systems.
15
Pressure (MPa)
12
9
Propane
Ethane
Carbon
dioxide
3
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 23. Comparison, at similar reduced temperatures (Tr ~ 1.11), of the pressure composition (w2)
phase behaviour of the CO2 (1)/methyl palmitate (2) at 323 K [48,49], ethane (1)/methyl palmitate (2)
at 323 K [29] and propane (1)/methyl palmitate (2) at 393 K [59] systems.
186
C. E. Schwarz
15
Pressure (MPa)
12
9
Propane
Ethane
Carbon
dioxide
3
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 24. Comparison, at similar reduced temperatures (Tr ~ 1.06), of the pressure composition (w2)
phase behaviour of the CO2 (1)/ethyl palmitate (2) at 323 K [26,51], ethane (1)/ethyl palmitate (2) at
323 K [30] and propane (1)/ethyl palmitate (2) at 393 K [30] systems.
35
Pressure (MPa)
30
25
Ethane
20
Carbon dioxide
15
Propane 436 K
10
Propane 429 K
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 25. Comparison, at similar reduced temperatures of the pressure composition (w2) phase
behaviour of the CO2 (1)/tripalmitin (2) at 353 K (Tr = 1.16) [25,33,52,53], ethane (1)/tripalmitin (2) at
360 K (Tr = 1.18) (data generated using the ethane/n-alkane correlations for the system ethane/ntetrapentacontane, as proposed by Schwarz et al. [57]) and propane (1)/tripalmitin (2) at 429 (Tr = 1.16)
and 436 K (Tr = 1.18) [31] systems.
The results show that, as expected from the analysis above, the phase transition pressure
of propane is the lowest and as such its solubility the highest. Additionally, for palmitic acid,
methyl palmitate and ethyl palmitate the phase transition pressures are lower for propane than
for ethane, and CO2 has the highest phase transition pressure and thus the lowest solubility.
187
For tripalmitin, it should be noted that the CO2 and ethane data are not at the same
temperature not at the same reduced temperature. The CO2 and propane at 429 K data are at
similar reduced solvent temperatures and the ethane and propane at 436 K data are at similar
reduced solvent temperatures. Tripalmitin is thus considerably more soluble in propane than
in ethane or CO2. It should be recalled (See Figure 8) that significant scatter exists for the
CO2/tripalmitin data. Before an outcome regarding a comparison of the solubility of
tripalmitin in CO2 and ethane can be given, issues relating to this scatter need to be resolved.
The accuracy of the ethane/tripalmitin approximation also needs to be verified. In all
likelihood, additional CO2/tripalmitin as well as ethane/tripalmitin data need to be measured.
The low molecular mass alkanes are thus able to dissolve palmitic acid and its derivatives
at lower pressures than CO2. This is in agreement with Mnkl et al. [53] who compared the
solubility of CO2 and in propane in a hardened rape seed oil and found that propane is able to
dissolve in the oil a lot better than the CO2. While ethane and in propane are not toxic, they
are highly flammable and as such care is required if they are used as alternative solvents or
even in high concentrations as co-solvents. However, despite the flammability issues
associated with ethane and propane, these two solvents are non-toxic and excellent
alternatives to CO2. As they are both also SC solvents, the solvent residue would be similar
than when using CO2 as solvent and the solvent recycle systems are of similar complexity to
using CO2.
188
C. E. Schwarz
the phase transition pressure in the region of the critical point (as shown in Figure 27), at low
CO2 compositions this observation is reversed. Similar phase behaviour was observed for the
CO2/biodiesel/ethanol system [62].
Table 14. Literature data phase equilibria of palmitic acid or its derivatives (2) in CO2
(1) in the presence of a co-solvent (3)
Reference
System
Purity
Temperature
range
Pressure
range
Composition
range (w2, w3) a
Brandt et al.
[34]
(2) palmitic
acid
(3) ethanol
(1) 99.995 %
(2) 99 %
(3) 99.8 %
313 K
8.21 to
24.61 MPa
w2 = 0.010 to
0.1271;
w3 = 0.00525 to
0.0948 (11)
Brandt et al.
[34]
(2) palmitic
acid
(3) 2propanol
(1) 99.995 %
(2) 99 %
(3) 99.8 %
313 K
10.90 to
20.69 MPa
w2 = 0.0326 to
0.1272
w3 = 0.0110 to
0.118 (9)
Garlapati
and Madras
[36]
(2) palmitic
acid
(3) ethanol
(2) 98 %
(3) 99.9 %
308 an 318 K
12.8 to 22.6
MPa
w2 = 3.07E-3 to
0.0349;
w3 = 7.57E-3 to
0.0429 (20)
Garlapati
and Madras
[36]
(2) palmitic
acid
(3) 3methyl-1butanol
(2) 98 %
(3) 98 %
308 an 318 K
12.8 to 22.6
MPa
w2 = 2.39E-3 to
0.0145;
w3 = 0.0144 to
0.0387 (15)
Gaschi et al.
[51]
(2) ethyl
palmitate
(3) ethanol
(1) 99.9 %
(2) 95 %
(3) 99.5 %
303.15 to
353.15 K
3.52 to
19.05 MPa
w2 = 0.0530 to
0.700;
w3 = 0.0140 to
0.343 (108)
Iwai et al.
[40]
(2) palmitic
acid
(3) water
(1) 99.9 %
(2) 99 %
(3) Ultrapure
313.2 K
15.0 MPa
w2 = 0.00659
and 0.00568;
w3 = 0.00352
and 0.00391 (2)
Koga et al.
[60]
(2) palmitic
acid
(3) ethanol
(1) 99.9 %
(2) 99 %
(3) 99.5 %
308.2 K
9.9 and
19.7 MPa
w2 = 8.91E-3 to
0.0622;
w3 = 0 to 0.0879
(22)
Koga et al.
[60]
(2) palmitic
acid
(3) octane
(1) 99.9 %
(2) 99 %
(3) 98 %
308.2 K
9.9 and
19.7 MPa
w2 = 8.91E-4 to
0.0331;
w3 = 0 to 0.0184
(22)
Rosso
Comin et al.
[61]
(2) palmitic
acid
(3) ethanol
(1) 99.9 %
(2) 99 %
(3) 99.9 %
313 to 343 K
8.34 to
19.94 MPa
w2 = 0.105 to
0.705;
w3 = 0.0611 to
0.186 (34)
189
25
Pressure (bar)
20
With Entrainer
333 K
Without
Entrainer 333K
With Entrainer
343 K
Without
Entrainer 343 K
15
10
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 26. Pressure composition (w2) plot for the system CO2 (1)/palmitic acid (2)/ethanol (3) at 323
and 343 K without ethanol [24] and for an ethanol to CO 2 molar ratios of 0.25 [61].
18
15
Pressure (bar)
12
No
entrainer
1:1
1:3
6
3
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 27. Pressure composition (w2) plot for the system CO2 (1)/ethyl palmitate (2)/ethanol (3) at
333.15 K for various ethanol to ethyl palmitate molar ratios [51].
190
C. E. Schwarz
24
0.73 mol %
2Me3BuOH
1.98 mol %
2Me3BuOH
Pressure (bar)
18
0.73 mol %
EtOH
12
1.98 mol %
EtOH
4.16 mo l%
EtOH
No entrainer
0
0.000
0.010
0.020
0.030
w2
0.040
Figure 28. Pressure composition (w2) plot for the system CO2 (1)/palmitic acid (2)/co-solvent (3) at
318 K for 3-methyl-1-butanol and ethanol as co-solvents at a range of co-solvent concentrations [36].
0.07
0.06
0.05
Ethanol, 9.9 MPa
w2
0.04
Ethanol, 19.7
MPa
Octane, 9.9 MPa
0.03
0.02
0.01
0.00
0.00
0.05
0.10
w3
0.15
0.20
Figure 29. Palmitic acid (w2) co-solvent (w3) concentration plot at 308 K and constant pressure for the
system CO2 (1)/palmitic acid (2)/co-solvent(3) for ethanol and octane as co-solvent [60].
Garlapati and Madras [36] and Koga et al. [60] compared various co-solvents and found
the increase in solubility to be more significant using ethanol compared to 3-methyl-1-butanol
and octane, respectively. Iwai et al. [40] considered the effect of water and found that the
solubility of palmitic acid increased with increasing molarity of water, especially near the
saturation point of water. They also found that the solubility of palmitic acid in CO2 saturated
with water was 16 times higher than that of pure CO2. (It should be noted that only 2 data
points are tabulated; the remaining data is all presented in figures.)
191
Gl-stnda and Temelli [63] reviewed the effect of a co-solvent on the phase
behaviour of lipids in SC CO2. They found that physical interactions between the solutes and
co-solvent, such as dipole dipole, dipole induced dipole or induced dipole induced
dipole (dispersion) interactions and specific interactions such as H-bonding and charge
transfer complexes, are important contributors to the co-solvent effect. The use of a cosolvent may also lead to a change in selectivity. The magnitude of the effect of the co-solvent
is thus a combination of the solvent, the co-solvent, the solute and the operating conditions.
The use of a co-solvent can therefore significantly reduce the phase transition pressure
and thus increase the solubility of palmitic acid and its derivatives in a SC solvent. However,
while this reduction leads to a decrease in operating pressure, it comes at the cost of a more
complicated solvent recycling system, a more complicated control philosophy and increased
solvent residue in the products. The choice of the use of a co-solvent thus needs to take these
aspects, as well as the nature of the co-solvent itself, into account when the use of a cosolvent is evaluated.
192
C. E. Schwarz
phase behaviour of methyl and ethyl esters in SC ethane and propane and found similar
results.
30
Pressure (MPa)
25
20
Stearic acid
Palmitic acid
15
Myristic acid
Lauric acid
10
Decanoic acid
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 30. Pressure Composition (w2) of various CO2 (1)/linear saturated acid (2) systems at 353 K
[24].
30
Pressure (MPa)
25
Behenic acid
20
Stearic acid
15
Palmitic acid
Myristic acid
10
Lauric acid
Capric acid
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 31. Pressure Composition (w2) of various ethane (1)/linear saturated acid (2) systems at 353 K
[27].
193
30
Pressure (MPa)
25
20
Tristearin
Tripalmitin
15
Trilaurin
10
Tricaprylin
5
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 32. Pressure Composition (w2) of various CO2 (1)/triglyceride (2) systems at 353 K [25,33].
20
Pressure (MPa)
16
Ethyl
stearate
12
8
Ethyl
palmitate
Ethyl
myristate
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 33. Pressure Composition (w2) of various CO2 (1)/ethyl ester (2) systems at 333 K [26].
The phase behaviour clearly shows that SCFs are able to distinguish between molecules
based on the number of carbon atoms present. Bharath et al. [33] found, from their phase
equilibria study, that that fatty acid and triglycerides can be fractionated based on their carbon
number using SC CO2. This is in agreement with their previous study [64] that considered the
phase behaviour of palm kernel oil and sesame oil in SC CO2, where they found that SC CO2
is able to selectively dissolve triglycerides based on their carbon number. Numerous other
studies came to similar conclusions [48,6567]. Additionally, Soares et al. [68] compared
the solubility of various fats and oils in SC CO2 and they found, in agreement with the trends
noted above, that oils with more lower molecular mass acids have a higher solubility.
194
C. E. Schwarz
As temperature and pressure affect the solubility, these effects can also be varied to
optimise the difference in solubility. Liang and Yeh [50] studied the separation of ethyl
palmitate, ethyl oleate, eicosapentaenoic acid (EPA) ethyl ester and docosahexaenoic acid
(DHA) ethyl ester. They considered the phase behaviour and used the determined coefficients
for the Chrastil equation to estimate the separation efficiency that can be attained. In general
better separation efficiencies were achieved at higher temperatures and lower pressures. They
verified their qualitative prediction through extraction of esterified fish oil and found the
results correlated well.
While it is believed that solute-solute interactions are not as large as solute-solvent
interactions, there is clear evidence that some type of solute-solute interaction is present in
SCF/high molecular mass systems. Lockemann [49] studied the phase behaviour of the
ternary system CO2/methyl myristate/methyl palmitate and found that these two components
can be separated using SC CO2. However, the separation factor, which dictates the degree or
difficulty of separation, is dependent on the composition of the feed to be separated and the
operating pressure and temperature. They found that while the composition does not
significantly affect the separation factor, better separation can be achieved at lower
composition of the component to be extracted.
In addition to saturated fatty acids with varying chain length, palmitic acid and its
derivatives usually occur in the presence of unsaturated fatty acids and their derivatives, in
particular those with 18 carbon atoms (oleic acid, linoleic acid and linolenic acid). Figure 34
compares the phase behaviour of various methyl esters of C18 acids in SC CO2.
Phase behaviour shown in Figure 34 suggests that it is also possible to achieve separation
according to the degree of saturation, but comparing Figure 33 and Figure 34, it is noted that
the difference in phase behaviour due to unsaturation is less than due to variations in the chain
length. Separation according to unsaturation, while possible, will be more difficult than
according to the chain length. The higher the degree of saturation the lower the phase
transition pressure and thus the higher the solubility. Liong et al. [65] and Nilsson et al. [67]
came to similar conclusions. Normal counter-current fractionation may not be sufficient to
separate compounds only differing in hydrocarbon backbone length and an additional
stationary phase (such as the column in a SCF chromatograph) may be required to achieve
such separations.
Gl-stnda and Temelli [66] as well as Zou et al. [70] studied the ternary system
CO2/oleic acid/linoleic acid. Gl-stnda and Temelli found that depending on the initial
composition, SC CO2 can also distinguish between the two acids. The interactions are,
however, complex and there are indications that solute-solute interactions are present. These
interactions can both enhance and decrease the partition coefficients determined based on the
binary data, depending on the temperature, pressure and acid ratios. Interestingly Zou et al.
found that while the ratio of the components in the liquid phase remains essentially the same
due to the fact that the majority of the components are in the liquid phase, the ratio is the
vapour phase is significantly different, but the difference depends on the molar ratio of the
components.
195
15
Pressure (MPa)
12
9
Methyl
linoleate
Methyl
oleate
Methyl
stearate
0
0.0
0.2
0.4
w2
0.6
0.8
1.0
Figure 34. Pressure Composition (w2) of various CO2 (1)/C18 ethyl ester (2) systems at 313 K [48,69].
Therefore, provided the correct operating conditions are applied, SC CO2 and other
solvents are able to distinguish between acids of the same chain length but different degrees
of saturation.
The analysis presented above has focused primarity on CO2 as SC solvent, mainly due to
an abundance of information on this solvent and a lack of information on other SC solvents.
However, qualitatively similar trends are expected in other SC solvents and as such it is
expected that these solvents could also achieve the desired separation.
Separation Set-Up
The phase behaviour analysis presented above shows that SC CO2 can be used to
fractionate fatty acids and their derivatives primarily according to their chain length, but also
according to their degree of saturation. Most sources of palmitic acid contain other acids of
both longer and shorter chain length and as such a two-step separation process is required
where, in one step, components that are more soluble (generally those with less than 16
carbon atoms) are removed while, in the other step, components with that are less soluble
(generally those with more than 16 carbon atoms) are removed. Two process options, shown
in Figure 35 and Figure 36 are therefore possible. A decision as to the better process option
would depend on the feed stock composition and the associated technical and economic
analysis.
196
C. E. Schwarz
Figure 35. Possible set-up for fractionation of acids or esters according to their hydrocarbon backbone
where the light fraction is removed in the first column and heavy fraction removed in the second
column.
Figure 36. Possible set-up for fractionation of acids or esters according to their hydrocarbon backbone
where the heavy fraction is removed in the first column and the light fraction removed in the second
column.
197
In the setup shown in Figure 35 the light fraction (i.e. components with less than 16
carbon atoms) is first removed and the raffinate from the first column is fed to the second
column where the C16 component is extracted and the raffinate ideally contains components
with 18 and more carbon atoms. By and large this option would be preferred as the solvent
usage would be less.
However, there may be occasions where it is more viable to first remove the heavy
components (C18 and greater) after which the extract is fractionated into the light components
(C14 and less) and the resultant product (C16). Such a typical process is shown in Figure 36.
It should be noted that these setups were compiled where only palmitic acid or its
derivative is the only desired product. However, in many cases palmitic acid occurs in
combination with other valuable fatty acids (e.g. oleic acid, linoleic acid, stearic acid and
even DHA and EPA) and as such more complicated separation sequences are likely.
198
C. E. Schwarz
extract. They thus showed it is possible to separate high molecular mass acids based on the
number of carbon atoms on the hydrocarbon backbone.
Using the same concept, only applying it to triglycerides Asep et al. [2] fractionated
cocoa butter according to the molecular mass of the triglyceride. Their triglycerides contained
mainly palmitic, stearic and oleic acid and triglycerides containing more palmitic acid were
preferentially extracted. Acetone, ethanol and isopropanol were used as co-solvents to
increase the extraction yield. Interestingly, an increase in co-solvent concentration also
showed an increase in palmitic acid selectivity while it did not influence the selectivity of the
other acids significantly. Ethanol was found to be the best polar co-solvent.
199
low molecular mass esters (C14 to C18) from high molecular mass esters (C18 to C20). The high
molecular mass ethyl ester products were achieved at 95 % recovery and 95 % purity. They
thus showed that practically fatty acid ethyl esters can also be separated according to their
molecular mass using SC fluids.
Figure 37. Possible set-up for obtaining palmitic acid or the ester thereof from fish oil using SCFF
(Modification of process proposed by Staby and Mollerup [75]).
200
C. E. Schwarz
presence of acetone as an entrainer, is able to separate mono-, di- and triglycerides. Later
Peter and Ender [79] showed that monoglycerides can be separated from a mixture of
glycerides using CO2 with propane as an co-solvent. A product containing 99.5 %
monoglycerides was obtained from a glyceride mixture containing 55 % monoglycerides.
Outlined below is a brief discussion on how SCFs can be used to achieve fractionations
involving palmitic acid where the required separation is not (only) between fatty acids of
differing chain length, but rather obtaining palmitic acid from a mixture of a wide range of
compounds. In particular, this part of the chapter will focus on the reduction of FFA content
of oils and the separation of palmitic acid from tocopherols, sterols and other components.
201
the process operated at a lower pressure. They did not consider a detailed acid content of the
triglycerides, but did show fractionation is possible by separating them into fractions with 48,
50, 52 and 54 carbon atoms.
While literature studies on the removal of FFAs have focussed almost exclusively on CO2
as SC solvent, the work by Peter and co-workers [18,79] on the fractionation of mono-, diand triglycerides using propane or CO2 with propane as co-solvent, shows that it would also
be possible to remove FFAs from oils using other SC solvents.
Pressure (MPa)
25
20
Cholesterol
15
alpha-tocopherol
Palmitic acid
10
Squalene
5
0
0.00
0.01
w2
0.02
0.03
Figure 38. Comparison of the pressure composition (w2) phase behaviour of the CO2 (1)/squalene (2)
[84], CO2 (1)/palmitic acid (2) [24], CO2 (1)/alpha-tocopherol (2) [85] and CO2 (1)/cholesterol (2) [86]
systems at 333 K.
202
C. E. Schwarz
Published Studies
Araujo et al. [6] modelled the SCFF with SC CO2 of Soybean Oil distillates, containing
FFAs, squalene, tocopherols, sterols and triglycerides, using the Peng Robinson equation of
state. Their results show that squalene and the FFAs present themselves in the extract, the
remainder of the product in the raffinate. However, the squalene has a much higher partition
coefficient and therefore separation from the FFAs is possible in a second column. In their
study the FFAs were lumped together and they did not consider palmitic acid alone. However,
the concept should be applicable to palmitic acid, as suggested by Figure 38.
Stoldt and Brunner [87] considered the phase behaviour of various deacidified palm oils
and soybean oil deodoriser distillates. They focused their study in the compounds other than
the acids/triglycerides present and showed that the other components can all be concentrated
either in the extract or the raffinate of a SCF process. However, improved purity palmitic acid
fractions are obtained as by-products in their process, showing that palmitic acid can also be
recovered from these sources.
203
zkal and co-workers [89,90] studied the extraction of palmitic acid containing oil from
apricot kernels. They studied the effect of, amongst others, temperature, pressure and CO2
flow rate on the extraction yield and built a mass transfer model to describe the extraction.
Their results show that irrespective of temperature, pressure or solvent flow rate, at very long
extraction times the same yield is obtained. However, particle size is important as higher
yields at long extraction times were obtained for smaller particles. zkal and co-workers
concentrated their efforts on the extraction yields and, save the optimum point, did not
consider the composition of their extracts. The operating parameters may influence the
sequence in which the components are extracted and would thus warrant a separate study.
Use of Entrainers
As for SCFF entrainers can be used in SCFE to increase the solubility and/or decrease the
operating pressure. SCFE is by and large more concerned with extraction rather than
204
C. E. Schwarz
fractionation and thus the loss of selectivity sometimes encountered with the use of entrainers
is far out-weighed by the increase in solubility and/or decrease in operating pressure. A large
number of studies have been published on the SCFE using a co-solvent with a selection of
these focussing on products that contain palmitic or similar acids (or rather their
triglycerides).
Bimakr et al. [92] considered the extraction of seed oil from winter melon using SC CO2
and ethanol as a co-solvent with a 10:1 CO2 to ethanol mass ratio. While they did not
investigate the effect of the co-solvent per se, they did conclude that their optimised SCFE
process yields similar results to that of traditional solvent extraction using ethanol. Their
optimised the SCFE process did yield a slightly lower saturated fatty acid contents, with
palmitic acid being approximately 40 % less. However, only the yield was optimised and the
only sample for which analysis was presented was that of the optimised condition. The
process was not optimised for a specific component and it may thus be that higher palmitic
acid content, possibly even higher than that of the traditional ethanol extraction, may be
obtained.
Mendes et al. [93] used ethanol as co-solvent (10 mol %) to increase the lipid yield
during the SC extraction of Spirulina. The CO2 modified with ethanol gave lipid yields
comparable with those of traditional organic solvents. Mendes et al. focussed their research
on the extraction of -linoleinic acid but did also analyse the palmitic acid content of their
samples. However, no comments regarding palmitic acid were given and too little information
is presented to provide definitive conclusions.
zkal et al. [89] investigated the effect of parameters on the SCFE of apricot kernels.
Amongst others, they considered the effect of the use of and the amount of ethanol as cosolvent. They found that an increase in the amount of co-solvent lead to an increase in the
fatty acid yield. The fatty acid contents was not presented for all samples, only the maximum
yield sample and a composition similar to that using traditional organic solvent extraction was
obtained.
In another study on the SC CO2 of extraction of apricot kernels zkal et al. [90]
developed a mass transfer model. They used the model to predict, amongst others, the effect
of the co-solvent and found that while the co-solvent initially increases the extraction yield at
a very long times the extraction yield with or without the co-solvent was the same. The effect
of the co-solvent therefore appears to be to reduce the amount of solvent required and thus
also the extraction time rather than the absolute maximum extraction that can be attained.
CONCLUSION
This chapter has shown that SCF processing is a viable method to process products
containing palmitic acids and its derivatives (tripalmitin, ethyl palmitate and methyl
palmitate). This study was conducted by investigating the solubility of these compounds in
SCFs, in particular CO2, ethane and propane, followed by an investigation as to how SCFF
and SCFE can be applied based on the phase behaviour and verified using published studies.
This study concludes that SCF processing is able to (i) extract palmitic acid and/or its
derivatives from a solid matrix, (ii) fractionate a fatty acid mixture or a mixture of its
derivatives mainly according to the chain length and (iii) provided sufficient difference in the
205
phase behaviour is present, is able to separate palmitic acid from other components. The
fundamental basis for the possibility of the extraction and separation is that palmitic acid and
its derivatives have a reasonable solubility in SC solvent. While the solubility in CO2 may be
low at times, the solubility can be improved through the use of co-solvents, or ethane or
propane may be used as alternative solvents.
This study has shown that on technical level SCF processing of palmitic acid containing
products is a viable method. In particular, a more environmentally friendly and generally
regarded as safe solvent is used resulting in a product with minimal, if any, solvent residue.
This study was conducted by conceptually considering the separation of palmitic acid or
its derivatives. While examples were mentioned, a more global approach was taken to prove
the technical viability. In order to implement SCF processing technology, the separation
required needs to be investigated based on the raw materials. In addition, an economic and
energy analysis would be required to provide a final outcome as to the viability of the process
as a whole.
NOMENCLATURE
Details pertaining to the acids (and their derivatives) studied in this works are presented
in Table 15.
Table 15. Nomenclature and structure of acids mentioned in this work
Common
name
IUPAC ID
Number of
carbon atoms
Degree of unsaturation
Capric acid
Decanoic acid
10
None
Lauric acid
Dodecanoic acid
12
None
Myristic acid
Tetradecanoic acid
14
None
Palmitic acid
Hexadecanoic acid
16
None
Stearic acid
Octadecanoic acid
18
None
Oleic acid
Octadecenoic acid
18
Linoleic acid
Octadecadienoic
acid
18
Linolenic acid
Octadecatrienoic
acid
18
EPA
Eicosapentaenoic_a
cid
20
Behenic acid
Docosanoic acid
22
None
DHA
Docosahexaenoic
acid
22
206
C. E. Schwarz
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In: Palmitic Acid: Occurrence, Biochemistry and Health Effects ISBN: 978-1-63321-519-1
Editor: Lucas F. Porto
2014 Nova Science Publishers, Inc.
Chapter 9
ABSTRACT
In this review the major saturated fatty acid, palmitic acid, of Virgin Olive Oil
(VOO) was studied. This oil is one of the oldest known vegetable oils and it plays a
fundamental role in human nutrition around the Mediterranean basin. This nature juice is
the only edible oil of great production obtained by physical methods from the fruit Olea
europaea L. Consideration of VOO as a natural functional fat is related to the presence of
palmitic acid. Updating of its levels in Virgin olive oils throughout the Tunisian olive oil
as well as information on expecting levels in other products from olive tree establish our
view point. Studies on levels palmitic acid upon maturity stage in the oil are also
discussed.
Major analytical practices are given in brief. Palmitic acid (C16:0) is the principal
saturated fatty acid in olive oil, responsible for its figeability at low temperature.
Few are the exceptions as palmitic acid content depends heavily on the genetic
factor. Palmitic fatty acids, important for the nutritional properties of an olive oil, showed
a crucial rule in the characterization of olive oils.
Keywords: Palmitic acid, olive oil, saturated fatty acid composition, storage olive oil
Ghayth Rigane: Laboratoire de Chimie Organique-Physique UR11ES74, Facult des Sciences de Sfax,
Dpartement de Chimie, B.P 1171 3038, Sfax, Universit de Sfax, Tunisie.
Corresponding author: Ridha Ben Salem. Laboratoire de Chimie Organique-Physique UR11ES74, Facult des
Sciences de Sfax, B.P 1171 3038 Sfax, Universit de Sfax, Tunisie. E-mail : ridha.bensalem@voila.fr
212
1. INTRODUCTION
Olives (Olea europea L.) are the most widespread and valuable plant in Mediterranean
countries. Chehab et al. (2013) mentioned that world-wide production of olive oil during the
last 20 years increased by almost 70% (from 1.7 to 2.8 million tons). Olive oils makes up a
small proportion (<3.5%) of the volume in the world vegetable oil market. However, in terms
of product value, only olive oil has a 15% share of world trade (Luchetti, 2000). According to
the International Olive Council (IOC) statistics, there is an equilibrium exists between olive
oil production and consumption worldwide, which is interesting taking into account the
increase in production the last decade.
Olive oil production currently reaches the 3 Mt (Tsimidou, 2012).
Olives (Olea europaea L.) are a major part of the agriculture and gastronomy in many
countries in Europe, North Africa, and Asia Minor surrounding the Mediterranean Sea. They
are one of the main sources of fat in the Mediterranean diet. The oval-shaped olives can be
consumed as table olives or as (virgin) olive oil. The edible part of the olive is called pulp and
the hard central part, which usually is discarded as waste, is called the kernel (Rigane et al.,
2013 a; b; Moghaddam et al., 2012).
Fatty acids, which are made up by long chains of carbon atoms, are extremely useful in
the characterization of olive oils. The health-related benefits of olive oil have also partly been
attributed to the high amount of oleic acid present in this type of vegetable oil (Moghaddam et
al., 2012). The virgin olive oil composition depends on numerous factors such as the
interaction between the cultivar and the environment, cultivation techniques, fruit ripeness
and the oil extraction system.
The influence of the cultivar on oil quality is being considered with increasing interest
because its determination could be used to produce mono-varietal virgin olive oils, thus
increasing product value (Haddada et al., 2008).
213
Bouchouka, Gemri-Dhokar and Dhokar olive oils grown under the same growth conditions in
the south of Tunisia. In these works, they mentioned that Dhokar olive oil was characterized
by its high palmitic acid level which means this oil freezes at low temperatures (19.37 %). In
addition, Oueslati et al., (2009) have studied four olive oils from southern of Tunisia. They
found that Chemlali variety yields oils of rather high concentration of palmitic acids (13.8716.06 %). Compared to some secondary Tunisian VOOs (Baccouri et al., 2007; Mana et al.,
2007) Chemlali Tataouine, Fakhari Douirat and Zarrazi Douirat varieties produced oils with
excellent fatty acid (FA) composition, i.e., low palmitic acid content (minimum of 8.67%)
which was lower than the upper limit of 5% established for extra virgin olive oil (EVOO)
(EEC, 2003).
Some of these FA ratios may prove useful in chemometric studies to discriminate oils on
the basis of origin (Aparicio et al., 1999) because they showed great variability depending on
the crop seasons, maturity index and the genetic factor.
To our knowledge such data are limited or missing. Among the many autochthonous
varieties the major ones are Chemlali (central and southern Tunisia) and Chtoui (northern
Tunisia), the former being more resistant to lack of rainfall. Some interesting data for the
evolution of palmitic acid during maturation are given by Damak et al. (2008). In this report,
this research team mentioned that during maturation, as fruit ripened, the content of palmitic
acids slightly decreased from 16.230.28% to 9.350.65%. Furthermore, it is well-known
that, in addition to the maturity stage, fatty acid composition could be affected by
environmental factors such as rainfall and geographical origin (Ben Temime et al., 2006).
While, Issaoui et al. (2008) studied the effect of harvesting time on the FA composition of the
five cultivars including: Zarrazi Zarzis, Jeddaria Chaal, Chemchali Chouamekh, Chemlali
Zarzis and Chemchali Gafsa. They found that the decreases in the level of palmitic acid are
variety dependent (55.92% of variability), and there was a significant cultivar kind (P <
0.001) and harvest timing (42.65% of variability; p < 0.001) effect in all varieties studied.
Hence, Zarrazi Zarzis show a decline of about one-half at the last part of maturation (from
12.83 to 7.74%); in contrast, a slight decrease was shown for the cultivars Chemlali
Chouamekh and Chemlali Zarzis (from 19.02 to 13.81% and from 19.06 to 13.61%,
respectively). the decrease of the palmitic acid during maturation was explained by GomezRico et al. (2005): In fact, the increase in oleic acid content is a result of the active
biosynthesis of TAGs, which takes place through fruit ripening, involving a fall in the relative
percentage of the oils palmitic acid content.
On the other hand, Bedbabis et al. (2010) reported the effect of waste water irrigation on
the extra virgin olive oil quality from the Tunisian cultivar Chemlali. The objective of this
work was to verify if an irrigation with Treated Waste Water (TWW) over 4 years affects the
Extra Virgin olive oil quality from Tunisian cv. Chemlali. Moreover, the amounts of
palmitic (C16:0) varied from 16.28 to 19.92%. The variability of fatty acid composition of the
sampled oil was within the normal range expected for olive oils (Codex, 1989). Differences
between our data and those reported for other varieties indicated that the olive cultivar affects
oil fatty acid composition, confirming previous studies (Inglesse et al., 1996; Ranalli et al.,
1997; Patumi et al., 1999; Dhifi et al., 2004; Ben Temime et al., 2006; Baccouri et al., 2007).
In addition, Gharsallaoui et al., (2011) studied the effect of irrigation with TWW on
Chemlali olive oil quality. This study focuses also on the determination of the quality of
oils extracted from hand picked olives and from olives fallen under the trees irrigated with
reclaimed water. Regarding palmitic acid, the difference between the treatments was not
214
significant with values ranging between 19.88 and 20.23%. Moreover, it has been note that
the percentages of this acid in the samples under investigation were very close to the upper
limit set by the IOC (International Oleicol Council) (20%) and even exceed the limit for the
WW-Fl plot. These values are similar to those found for Chemlali variety grown in the Sfax
region and in the South of Tunisia (Dabbou et al., 2010; Issaoui et al., 2010).
Several studies have reported that irrigation can affect the fatty acid composition (Ranalli
et al., 1997; Aparicio and Luna, 2002). At the end of our experiment, palmitic, palmitoleic,
stearic and linoleic acids were found in high concentrations in oils coming from TWW
irrigated plot, respect to acid contents in oil from trees grown under rain-fed conditions. In
addition, these results indicated that irrigation with TWW seems to stimulate these fatty acid
enzymes synthetase.
CONCLUSION
As can be seen from this review, the level of palmitic acid in olive oil, as the major
saturated acid form, is the result of a complex interaction of various factors. From the existing
literature concerning the characterization of palmitic acid in the Tunisian olive oils, it is clear
that the main focus has been directed toward the two most important varieties: Chtoui and
Chemlali.
Currently, most cultivars may yield virgin olive oils with more than 19 %.
215
ACKNOWLEDGMENTS
The authors thank the Tunisian Ministry of Higher Education and Scientific Research for
financial support and are grateful to Professor Mohamed Rigane for useful discussions about
the English.
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INDEX
#
21st century, 94
A
accounting, 70
acetone, 68, 86, 200
acetylcholine, 5, 13
acidity, 25, 71
active site, 34
active transport, 32
acylation, 74, 134, 135, 138, 146
AD, 97, 100, 119, 130
adaptation, 157
adenine, 30, 85
adenosine, 6, 36, 79
adenosine triphosphate, 6
adhesion, 20, 91, 152
adipocyte, 34, 107, 108, 118
adiponectin, ix, 106, 107, 108, 117
adipose, 3, 11, 21, 35, 37, 38, 40, 48, 49, 52, 53, 58,
61, 73, 78, 79, 83, 86, 90, 99, 106, 107, 114, 116,
117, 119, 123
adipose tissue, 3, 11, 21, 37, 38, 40, 48, 49, 52, 54,
58, 61, 73, 78, 79, 83, 86, 90, 99, 106, 107, 114,
116, 117, 119, 123
adiposity, ix, 105, 119
adolescents, 112, 122
ADP, 85
adults, 33, 39, 42, 115, 116, 118, 121, 122, 148
adverse effects, 47, 108
Africa, 71
age, 21, 112, 114, 150, 152, 156
agriculture, 212
Agrobacterium, 38
AIDS, 39
alcohols, ix, 125, 130, 131, 132, 133, 161
220
Index
B
bacteria, vii, 17, 21, 25, 26, 28, 152, 153
bacterial cells, 37
Bangladesh, 38
base, 68, 126
basilar artery, 5
beauty products, vii, 1
beef, 22, 42, 49, 50, 52, 57, 58, 59, 61, 68, 73, 92,
106
beneficial effect, 78, 110, 148, 150, 151, 152, 153
benefits, vii, x, 1, 17, 35, 145, 148, 151, 153, 212
bile, 146
biochemical action, 24, 25
biochemical processes, 27
biochemistry, vii, viii, 1, 17, 18, 27, 34, 93, 94, 99,
100, 139, 156
biodegradation, 141
biodiesel, 20, 36, 188
biological processes, 9
biological signaling, vii, 1
biological systems, vii, 1, 2, 20
biomarkers, 31, 34, 122, 123
biomolecules, 18
biosynthesis, 13, 28, 37, 79, 81, 88, 126, 127, 130,
135, 136, 137, 138, 139, 140, 141, 142, 213
biosynthetic pathways, 29, 134
biotechnology, 18, 20, 38
biotin, 79, 80
birds, 52
birth weight, 156
Black Sea, 51
bleaching, 71, 94
blends, 77, 96, 97
blood, viii, ix, 2, 3, 9, 12, 32, 36, 37, 45, 46, 47, 62,
86, 87, 88, 92, 101, 106, 108, 109, 110, 112, 118,
120, 123, 153
blood pressure, 3, 12
blood stream, 86, 153
blood vessels, 3
blood-brain barrier, 33, 36
bloodstream, 83, 86
body fat, 57
body mass index, 111, 119
body weight, 148, 154
bonds, 24, 66, 68, 126, 128
bone, x, 93, 107, 145, 149, 150, 153, 155, 156, 157
bone growth, 150
bone marrow, 107
bone mass, 149
Botswana, 141
bowel, 88
bradykinin, 4
brain, 11, 28, 32, 35, 40, 41, 42, 46, 83, 86, 100
Brazil, 63, 71, 72, 75, 95, 97
breast milk, viii, x, 63, 78, 145, 146, 149
breastfeeding, 148
building blocks, 79
by-products, 160, 202
C
Ca2+, 4, 5
cacao, 133
caffeine, 202
calcium, x, 3, 4, 9, 12, 57, 58, 59, 78, 98, 145, 148,
152, 153, 156
calcium channel blocker, 4, 9, 12
caloric intake, 87, 88
cancer, 13, 107, 113, 151, 155, 157
capillary, 214
carbohydrate(s), 18, 19, 34, 39, 41, 47, 66, 109, 112,
118, 119, 149
carbohydrate metabolism, 41
carbon, vii, 1, 2, 3, 7, 8, 14, 20, 24, 26, 35, 66, 68,
75, 79, 81, 82, 85, 86, 99, 109, 126, 127, 129,
130, 131, 136, 139, 142, 146, 159, 160, 163, 177,
178, 193, 194, 195, 197, 198, 201, 205, 212
carbon atoms, 68, 85, 126, 127, 129, 130, 136, 159,
160, 177, 178, 193, 194, 195, 197, 198, 201, 205,
212
carbon dioxide, 35, 86, 163
carbon monoxide, 131
carboxyl, 7, 9, 18, 81, 131, 132
carboxylic acid, 84, 160
carcinogenicity, 37, 58
cardiac arrest, 9
cardiac muscle, 83
cardiomyopathy, 102
cardiovascular disease(d), viii, 2, 4, 6, 12, 19, 33, 34,
37, 41, 45, 46, 47, 63, 97, 105, 106, 109, 112,
115, 118, 119, 120, 121
cardiovascular function, 4
cardiovascular risk, 46, 102
carotene, 75, 94
carotenoids, 71, 73
cascades, 103
catabolism, 32
catalysis, 27
catalyst, 76, 78
catfish, 55, 56, 60
cattle, 52, 54, 59, 60, 61
CBS, 64
C-C, 18
cell culture, 90
cell death, 3, 9, 10, 14, 90, 102
Index
cell line, 102
cell membranes, ix, 20, 26, 27, 86, 125, 129
cell organelles, 25
cell organization, 27
cell signaling, 20, 28, 37
cell surface, 70, 92, 155
central nervous system, vii, 1
central obesity, 37, 107
ceramide, 39, 107, 108, 111, 117, 123
cerebral blood flow, 3
challenges, 18, 94
channel blocker, 3
cheese, 46, 50, 74, 95
chemical(s), 24, 34, 38, 39, 40, 68, 70, 71, 72, 74,
76, 77, 92, 97, 147, 150, 201
chemical characteristics, 97
chemical properties, 24, 70, 71, 72, 147
chemical reactions, 68, 92
chemokines, 91, 92
chicken, 49, 96, 106
childhood, 149, 155, 157
children, 40, 149
Chile, 73
chloroform, 68
chloroplast, 128, 139
cholesterol, viii, ix, 6, 31, 34, 36, 39, 41, 45, 46, 47,
49, 52, 59, 61, 70, 72, 74, 86, 87, 88, 89, 92, 100,
105, 106, 107, 109, 111, 112, 114, 115, 116, 119,
121, 122, 124, 136, 146, 201
cholesterolaemia, viii, 45, 48
choline, 136, 137, 138, 143
chromatography, 37, 133, 214
chronic diseases, 62
circulation, vii, 1, 2, 4, 5, 6, 66, 110
cirrhosis, 89, 113
classes, ix, 26, 57, 87, 125, 141
classification, 42
cleavage, 85
climates, 71
clinical trials, 118, 155
clusters, 134
CNS, 10
CO2, x, 79, 81, 84, 131, 159, 161, 162, 163, 164,
165, 166, 167, 168, 169, 170, 171, 172, 173, 174,
175, 178, 179, 182, 184, 185, 186, 187, 188, 189,
190, 191, 192, 193, 194, 195, 198, 200, 201, 202,
203, 204, 205
cocoa, viii, 63, 71, 72, 77, 92, 95, 106, 133, 160,
197, 198
cocoa butter, viii, 63, 71, 72, 77, 92, 95, 106, 160,
197, 198
coconut oil, vii, ix, 1, 47, 76, 77, 87, 105, 106
coenzyme, vii, 1, 2, 80, 83, 84, 88, 98, 140
221
coffee, 202
colic, 150
colitis, 152, 156, 157
collagen, 89
colon, 152, 153
colonization, 41, 151
color, 75, 94
combustion, 89
commercial, 202, 212, 216
commodity, 146
communication, 33
community, 66, 122, 151, 157
complement, 34
complexity, 187
complications, 12, 91, 102, 103, 153, 169
compounds, x, 7, 28, 35, 66, 71, 84, 134, 159, 161,
162, 163, 164, 182, 194, 200, 201, 202, 204, 215
comprehension, 18
condensation, 79, 80, 81, 82, 127
conductance, 4, 30
configuration, 66, 126
conflict, 10
conflict of interest, 10
Congress, 57, 102
conservation, 56
conserving, 49
constant rate, 53
constituents, 6, 18, 41, 66, 111, 126, 131, 132, 142,
147, 198
Constitution, 58, 141
construction, 66
consumers, 49
consumption, viii, 34, 48, 58, 59, 73, 86, 87, 88, 89,
92, 105, 109, 110, 111, 116, 120, 148, 150, 212
consumption patterns, 59
control group, 149, 150
controlled trials, 39, 119, 121
controversial, 87
convergence, 102
COOH, 18, 26, 68
cooking, 66, 71, 76
cooling, 96
cooperation, 128, 151
coordination, 27, 28
coronary arteries, 6, 11
coronary artery disease, 6, 10
coronary heart disease, ix, 42, 47, 49, 62, 87, 105,
106, 109, 114, 116, 119, 121
correlation(s), 30, 54, 55, 148, 169, 177, 178, 179,
186
cortical neurons, 15
cosmetic(s), vii, 1, 18, 20, 38, 70, 73, 76, 160
cost, 60, 70, 77, 92, 94, 184, 191
222
Index
cotton, 77
covalent bond, 28
CPT, 8, 64, 84, 136
crises, 102
critical value, 88, 92
crop, 213
crude oil, 197
crystalline, 24, 77
crystallization, 71, 96
CT, 64, 79, 80
cues, vii, 17
cultivars, 73, 95, 212, 213, 214, 215, 216, 217
cultivation, 212, 216
culture, 37, 39, 102, 134, 136, 215
culture conditions, 37
cuticle, ix, 125, 131
cutin, 131, 132, 141
CVD, viii, 19, 33, 105, 106, 107, 109, 111, 115, 116
cycles, 8, 85, 127
cysteine, vii, 17, 20, 81
cytochrome, 3, 11, 83, 90, 132
cytokines, 34, 90, 91, 102
cytoplasm, 8, 79, 134
D
dairy products, viii, 45, 48, 63, 68, 74, 106, 109
data set, 168, 171, 172
database, 95
deaths, 6, 9
decomposition, 71
decoupling, 27
Deer, 50
defects, 90
deficiency, 91, 94, 107, 117, 152
deformation, 27
degradation, 8, 30, 83, 142, 152
degumming, 71
dehydration, 80, 82
dendritic cell, 107
deoxyribonucleic acid, 116
Department of Agriculture, 67, 93
Department of Health and Human Services, 115
deposition, 61, 62, 113
depression, 55, 165, 170
derivatives, x, 38, 90, 131, 132, 138, 159, 160, 161,
164, 165, 173, 175, 178, 179, 184, 187, 188, 191,
194, 195, 197, 199, 200, 204, 205
detection, 116
detergents, 35, 70
detoxification, 89
developed countries, 49
developmental process, 15
diabetes, ix, 4, 33, 39, 91, 100, 102, 103, 106, 107,
110, 111, 114, 116, 120, 121, 124
diabetic kidney disease, 103
diabetic patients, 122
diacylglycerol, 91, 104, 107, 117, 130, 135, 136,
142, 143
diarrhea, 151
diet, viii, 31, 33, 45, 46, 47, 48, 49, 52, 54, 55, 56,
57, 59, 61, 62, 63, 69, 78, 86, 87, 88, 89, 100,
101, 106, 108, 110, 111, 116, 118, 119, 122, 146,
147, 150, 151, 152, 153, 212
diet composition, 54
dietary fat, 37, 39, 52, 54, 57, 59, 60, 61, 62, 87, 100,
106, 109, 110, 118, 119, 120, 121, 122
Dietary Guidelines, 115
Dietary Guidelines for Americans, 115
dietary habits, ix, 106, 115
dietary intake, ix, 5, 6, 28, 54, 93, 105, 110, 112,
114, 123
dietary lipid sources, viii, 45
dietary sources, viii, 42, 45, 106
dietary supplementation, 52
diffusion, 90, 131
digestibility, 60, 74
digestion, 57, 59, 61, 78, 93, 120, 147, 148, 151,
153, 156
digestive system, viii, 45
discomfort, 150
discrimination, 139
diseases, viii, 6, 20, 34, 49, 63, 90, 102, 106, 107,
115, 153
disorder, 26, 30
dispersion, 97, 191
displacement, 27
distillation, 71, 161, 162
distribution, viii, 18, 21, 22, 28, 34, 39, 57, 63, 78,
98, 119, 120, 123, 141, 142, 146, 147, 156, 201
diversification, 39
diversity, 18, 126, 215
DNA, 26
docosahexaenoic acid, 4, 5, 66, 67, 96, 194
DOI, 98, 101
donors, 3, 11, 136
double blind study, 151
double bonds, 66, 126, 128, 198, 205
down-regulation, 103, 108, 118
drugs, 40
dry matter, 54, 95, 134
duodenum, 59
dyslipidemia, 106, 107, 109
223
Index
E
ecosystem, 151
editors, 139, 140, 143
egg, 46, 51, 56, 57, 59
eicosapentaenoic acid, 12, 13, 41, 67, 194
electron(s), 9, 26, 27, 32, 84, 85, 90, 131
elongation, ix, 6, 8, 14, 31, 48, 61, 82, 99, 122, 125,
126, 127, 128, 130, 132
e-mail, 105, 125
employment, 212
emulsions, 61
encoding, 142
endogenous synthesis, 31
endosperm, 140
endothelial dysfunction, ix, 4, 106
endothelial NO synthase, 13
endothelium, 3, 4, 5, 6, 11, 12, 13, 83
energy, ix, x, 8, 18, 20, 26, 27, 28, 29, 30, 31, 33, 34,
37, 38, 42, 51, 57, 66, 72, 75, 77, 79, 83, 86, 87,
89, 98, 105, 109, 110, 111, 117, 118, 134, 138,
145, 146, 148, 149, 151, 205
energy expenditure, ix, 26, 31, 33, 38, 98, 105, 117
engineering, 93
environment, 21, 37, 51, 88, 147, 151, 212
environmental conditions, ix, 125
environmental factors, viii, 45, 213
environmental stress(s), 131
enzymatic activity, 131
enzyme(s), viii, 6, 3, 8, 11, 19, 20, 24, 26, 27, 34, 48,
49, 59, 61, 63, 69, 75, 78, 79, 81, 82, 84, 85, 86,
88, 91, 99, 100, 113, 127, 130, 132, 134, 136,
137, 138, 139, 142, 143, 146, 214
EPA, 4, 12, 41, 64, 159, 194, 197, 198, 205
epidemiologic, 111
epidermis, 131
epinephrine, 79
epithelial cells, 55
epithelium, 86, 152
equilibrium, 160, 161, 165, 166, 168, 175, 180, 183,
197, 212
equipment, 128, 162
erosion, 152
erythrocytes, 112, 114
ESI, 141
essential fatty acids, 4, 61
ester, vii, viii, x, 1, 2, 7, 14, 18, 22, 63, 68, 69, 83,
121, 140, 147, 159, 174, 181, 191, 193, 194, 195,
199
ethanol, 68, 164, 187, 188, 189, 190, 198, 200, 204
etiology, 101, 107
eukaryotic, vii, 1, 2, 17, 28, 128, 134, 151
eukaryotic cell, vii, 1, 2, 28, 151
F
FAD, 8, 64, 84, 85
families, 129, 132
family members, 102
FAS, 64, 79, 81, 83, 129, 130
fast food, 106
fasting, 31, 42, 83, 86, 114, 119, 120, 124
fasting glucose, 114
fat intake, 106, 110, 121
feedstocks, 139
fermentation, 59, 95
fetal growth, 33, 38
fiber, 51
fibrate, 122
fibrinolysis, 62
fibrosis, 89
financial, 215
financial support, 215
Finland, 106, 116
fish, viii, 4, 5, 6, 45, 46, 48, 51, 52, 55, 56, 57, 58,
59, 60, 61, 191, 194, 198, 199
fish oil, 4, 5, 6, 52, 56, 57, 60, 191, 194, 198, 199
flammability, 187
flavor, 74
flora, x, 145, 151, 152, 155, 157
flowers, 23, 36, 132
fluid, x, 159, 160, 161
fluid extract, 160, 161
food, viii, ix, 18, 20, 40, 41, 45, 48, 49, 50, 51, 57,
58, 60, 63, 66, 70, 71, 74, 76, 77, 92, 94, 95, 106,
111, 115, 116, 121, 148
food industry, viii, 63, 71, 76
food products, 74, 77, 95
food spoilage, 41
foodborne pathogens, viii, 17
football, 123
formation, vii, 6, 17, 20, 39, 78, 127, 128, 135, 136,
137, 138, 140, 142, 143, 148
224
Index
formula, 68, 78, 97, 98, 147, 148, 149, 150, 151,
152, 156, 157, 158
fragility, 60
fragments, 127
France, 57
free energy, 81
freshwater, 51, 58, 61
fruits, 23, 36, 70, 73, 97, 129, 133, 134, 138, 140,
141, 142, 212
FTICR, 141
fungal infection, 132
fungi, 26, 39, 126
fungus, 21, 25, 30, 34
fusion, 39, 40
G
ganglion, 2, 11
gastrointestinal tract, 151
gel, vii, 1
gene expression, 88, 91, 103, 108, 111, 119, 153,
156
genes, 88, 91, 107, 130, 142
genetic factors, 57
genetic marker, 91
genome, 18
geographical origin, 213
germination, 134
gestational age, 156
ginseng, 134, 142
gland, x, 55, 74, 145, 146
glucagon, 79
gluconeogenesis, 31, 39, 86
glucose, viii, ix, 33, 39, 63, 79, 86, 91, 92, 103, 106,
107, 108, 110, 118, 124
glucose tolerance, 124
glutathione, 89
glycerol, viii, ix, x, 48, 61, 63, 66, 70, 73, 75, 76, 77,
92, 125, 128, 131, 132, 135, 138, 145, 146
glycogen, 89
glycolysis, 79, 135
God, 72
Gori, 95
GRAS, 162
grass, 52, 57, 60
growth, 25, 27, 33, 37, 38, 41, 42, 57, 59, 60, 61, 77,
136, 146, 149, 152, 213, 216
growth temperature, 41
guidelines, 106, 119
Guinea, 35
H
habitat, 151
hair, 42
hardness, 71, 78, 98, 157
harvesting, 95, 213, 216
H-bonding, 191
HE, 95
health, vii, x, 1, 9, 17, 18, 33, 34, 35, 38, 39, 42, 47,
49, 52, 57, 58, 59, 78, 98, 99, 106, 116, 145, 150,
151, 152, 154, 155, 212
health care, 9
health care costs, 9
health effects, vii, 18, 42, 152
health promotion, 35
health services, 150
heart disease, 14, 59, 110
heart failure, 33, 39
helminthes, viii, 17
hematology, 58
heme, 11
hemodialysis, 122
hepatocellular carcinoma, 89
hepatocytes, 31, 36, 39, 40, 90, 102
hepatoma, 33, 43
hexane, 202
high fat, 86, 106, 147
hippocampus, 9
HIV, 34, 39, 40
HIV-1, 39, 40
HM, 95, 96, 98, 124, 142
homeostasis, vii, 17, 27, 33, 35, 83, 116, 118, 152
homocysteine, 31, 42
hormone(s), 33, 89
host, 21, 28, 36, 151, 155, 157
House, 59
housing, 58
human body, 151
human health, vii, 17, 38, 47, 66, 76
human milk, x, 38, 75, 77, 78, 96, 98, 106, 145, 146,
147, 148, 153, 154, 155, 156
human subjects, 36, 37
Hunter, 119
hybrid, 60
hydrocarbons, 41, 130
hydrogen, 24, 68
hydrogen bonds, 68
hydrogenation, 71, 97
hydrolysis, 34, 48, 83, 147
hydrophobicity, 18, 20
hydroxide, 69
hydroxyl, 7, 66, 69, 75, 81, 131
hydroxyl groups, 66, 75
225
Index
hygiene, 151
hypercholesterolemia, viii, 12, 63, 87, 110
hyperglycemia, 90
hyperlipidemia, 6, 113, 114, 122
hypertension, 4, 5, 6, 11, 106, 107, 113, 122
hypertriglyceridemia, 33, 110, 114
hypertrophy, 107, 112, 122
hypothalamus, 33
hypothesis, 89, 102, 138
hypoxia, 13
I
IBD, 152, 153
ID, 36, 205
ideal, 87
identification, 22
identity, 3, 38
IL-8, 31, 90
immune modulation, 152
immune reaction, 156
immune response, 33, 34, 91, 151, 152
immune system, 152, 155, 157
immunity, 154
immunomodulation, 156
improvements, 117
impurities, 71, 165
in vitro, 3, 6, 9, 10, 36, 55, 92, 107, 132
in vivo, viii, 6, 9, 10, 12, 13, 45, 91, 92, 99
incidence, ix, 87, 106, 110, 111, 114, 120, 121, 122,
151, 152
individuals, 34
Indonesia, 70
induction, 92, 107
industrialized countries, viii, 45, 49
industry(s), 18, 20, 38, 40, 68, 71, 72, 73, 74, 76, 77,
160
infancy, 154, 157
infants, x, 38, 57, 75, 77, 78, 96, 98, 110, 145, 146,
147, 148, 149, 150, 151, 152, 153, 154, 155, 156,
157, 158
infection, 34, 35, 39, 148, 152
inflammation, ix, 38, 83, 89, 91, 102, 105, 107, 108,
109, 111, 116, 117, 118, 121, 123, 152, 153, 157
inflammatory bowel disease, 151, 152, 153, 154,
155, 156
inflammatory disease, 102, 152, 153
ingestion, 93
inhibition, 5, 14, 19, 24, 26, 27, 30, 40, 42, 79
inhibitor, 2, 9, 11, 39
initiation, 34, 85, 90
injury, 3, 11, 89, 90, 100, 102
innate immunity, 117
INS, 102
insertion, ix, 25, 27, 125, 128
insulin, viii, ix, 4, 33, 34, 36, 37, 38, 39, 63, 79, 90,
91, 92, 98, 101, 103, 104, 105, 107, 108, 109,
110, 111, 113, 114, 116, 117, 118, 119, 120, 121,
122, 123, 124
insulin resistance, viii, ix, 33, 34, 36, 38, 63, 90, 91,
92, 98, 101, 103, 104, 105, 107, 108, 109, 110,
113, 116, 117, 118, 119, 120, 121, 122
insulin sensitivity, ix, 39, 91, 106, 108, 110, 113,
114, 116, 117, 118, 119, 120, 121, 123, 124
insulin signaling, 92, 107, 108, 116, 117
integrity, 25, 26, 28, 37, 152
interference, 110, 137
interferon, 31
interferon-, 31
intervention, 9, 37, 115, 122, 124, 156
intestinal flora, 153, 156, 158
intestine, 69, 78, 88, 147, 152, 153, 155
intracellular calcium, 5
intravenously, 33
investment, 184
ion channels, 20
ions, 26
IP-10, 108, 117
iron, 131
irrigation, 213, 214, 215, 216
ischemia, vii, 1, 2, 3, 5, 9, 10, 11
isolation, 43, 217
isomers, 131, 141
isozymes, 91
Israel, 145, 154
issues, viii, 45, 187
Italy, 45
K
K+, 3, 4, 5, 10
keratinocytes, 34
kidney, 50
kill, 26, 102
kinetics, 34, 118
KOH, 214
L
labeling, 132
lactation, 54, 58
lactic acid, 151
lactobacillus, 151, 152
Lactobacillus, 23, 38, 78
L-arginine, 2
226
Index
Latin America, 72
lauric acid, viii, 31, 36, 45, 47, 77, 203
LDL, viii, ix, 6, 13, 34, 45, 47, 59, 65, 70, 86, 87, 88,
89, 92, 105, 107, 109, 110, 111, 118, 119
lead, 26, 49, 78, 79, 89, 104, 108, 111, 117, 152,
161, 191, 204
learning, 3, 18
lecithin, 69
leptin, 33, 36, 154
lesions, 152
leukocytes, 90
life cycle, 142
lifetime, 115
light, 75, 196, 197
linoleic acid, 21, 31, 34, 37, 43, 47, 49, 52, 55, 59,
66, 71, 73, 74, 88, 110, 112, 120, 121, 123, 147,
148, 167, 194, 197, 198, 214
lipases, 38, 48, 76, 97
lipemia, 101, 110
lipid dysregulation, 107
lipid metabolism, 41, 61, 83, 119, 128, 139
lipid peroxidation, 90, 102
lipoproteins, 6, 31, 36, 60, 61, 86, 100, 101, 109,
118, 119
liquid chromatography, 37
liquid phase, 71, 168, 178, 191, 194
liquids, 161
Listeria monocytogenes, 42
lithium, 30, 69
liver, viii, 3, 6, 28, 30, 36, 38, 39, 40, 41, 43, 52, 61,
63, 69, 78, 79, 83, 86, 89, 90, 91, 93, 99, 101,
102, 122, 123, 199
liver cells, 70, 79, 123
liver cirrhosis, 122
liver disease, viii, 39, 63, 89, 101, 102
localization, 10, 20, 36, 93, 134
locus, 157
low temperatures, 200, 213
low-density lipoprotein, ix, 13, 105, 109
lower lip, 35, 123
low-grade inflammation, 112, 122
lubricants, 69
lumen, 134
Luo, 43, 117
lymphatic system, 86
lymphoma, 113, 119
lysis, 26
M
machinery, 26, 27
macrophages, 6, 92, 107, 108, 117
magnesium, 98, 156
227
Index
methyl group(s), 18, 68, 132
methyl palmitate, vii, 1, 2, 11, 161, 164, 168, 169,
173, 174, 175, 176, 177, 178, 179, 180, 181, 183,
184, 185, 186, 191, 194, 204
methylation, 41, 214
methylene chloride, 202
Mexico, 73
Miami, 1, 10
mice, 13, 22, 33, 92, 99, 102, 108, 117, 152, 153,
156, 157
microbiota, 78, 152, 154, 158
microorganism(s), 13, 18, 24, 26, 27, 41, 46, 146,
151, 152
Microsoft, 21
microsomes, 28, 61, 136
microstructures, 97
mineralization, 149, 150, 153, 156
Ministry of Education, 115
misuse, 59
mitochondria, vii, 1, 6, 8, 26, 28, 30, 36, 39, 41, 42,
43, 79, 102, 126
mitochondrial damage, 90
mitogen, 91, 92, 103, 104, 107, 118
models, vii, 1, 9, 10, 48, 89
modifications, vii, 17, 66, 130
molar ratios, 189
molecular mass, 24, 160, 161, 162, 163, 164, 168,
172, 173, 174, 177, 187, 191, 193, 194, 198, 199,
203
molecular structure, 48, 68
molecular weight, 164
molecules, 6, 8, 9, 10, 20, 68, 74, 75, 79, 81, 84, 85,
86, 91, 135, 138, 165, 174, 193, 199, 201
monolayer, 134
monomers, 131, 132, 141
monosodium glutamate, 70
monounsaturated fatty acids, 34, 37, 39, 73, 75, 82,
109, 115, 118
Moon, 14, 99
morbidity, 9, 153
morphology, 77
mortality, ix, 9, 59, 62, 105, 106, 110, 112, 120, 121,
122, 153
Moscow, 125
MR, 94, 139
mRNA, 20, 59, 65, 70, 87, 108
mucin, 156, 157
mucosa, 30, 152
mucus, 152, 155
muscles, 49, 50, 57, 107
mutagenesis, 155
mutation, 117
mycobacteria, 41
N
Na+, 5
NAD, 8, 65, 79, 84
NADH, 8, 9, 79, 83, 84, 85
National Institutes of Health, 10
National Research Council, 45
natural selection, 151
necrosis, 14, 90
Netherlands, 120
neurons, 9
neuroprotection, 2, 9, 10, 11
neuroprotective agent, vii, 1
neurotransmission, vii, 1
neurotransmitter, 2, 20
neutral, 47, 52, 69, 88, 134, 140, 142
neutral lipids, 52, 134, 142
neutrophils, 107
New Zealand, 10, 73
NH2, 107
nicotinamide, 85
Nigeria, 70
nitric oxide, vii, 2, 11, 12, 17, 90, 111
nitric oxide synthase, 2, 11
nitrogen, 89, 214
NMR, 141
non-esterified PA, vii, 17
norepinephrine, 5, 11
normal development, 131
North Africa, 212
North America, 72
nutraceutical, 36
nutrient(s), 21, 26, 27, 37, 47, 49, 59, 146, 148, 151,
212
nutrition, vii, viii, x, 17, 45, 57, 61, 62, 63, 69, 74,
92, 94, 96, 115, 119, 145, 146, 149, 154, 155,
156, 157, 211
nutritional status, 75
O
obesity, 4, 33, 34, 103, 106, 107, 108, 117, 118, 119,
123
obstruction, 9
octane, 164, 188, 190
OH, 131, 138
oil production, 160, 212
oilseed, 46, 129
228
Index
oleic acid, viii, 7, 13, 21, 30, 34, 35, 36, 42, 45, 47,
48, 52, 53, 54, 55, 58, 61, 66, 71, 72, 73, 74, 75,
76, 78, 88, 96, 108, 110, 120, 123, 132, 138, 147,
148, 160, 194, 197, 198, 203, 212, 213
olive oil, x, xi, 73, 95, 101, 106, 200, 211, 212, 213,
214, 215, 216, 217
omega-3, 12, 13, 61
operating costs, 169
operating range, 162
opportunities, 20
organ(s), 83, 89, 131, 133, 134, 151, 157
organelle(s), 26, 28, 127, 128, 134
organic compounds, 164
organic matter, 138
organic solvents, x, 159, 187, 200, 202, 204
organism, 78, 79
osteoporosis, 149, 155
overweight, 112, 122
ox, 6
oxidation, vii, ix, 8, 9, 13, 14, 17, 20, 30, 31, 32, 33,
34, 38, 39, 42, 70, 71, 83, 84, 85, 86, 90, 99, 100,
105, 107, 108, 111, 117, 123, 126, 130, 132, 134
oxidative stress, 10, 15, 90, 91, 101, 102, 103, 111,
116
oxygen, 7, 9, 24, 90, 101, 177
P
Pacific, 59, 61
palm oil, viii, 34, 46, 47, 48, 52, 55, 56, 57, 58, 59,
60, 61, 63, 68, 70, 71, 72, 77, 78, 87, 88, 92, 94,
95, 97, 100, 101, 106, 110, 120, 133, 146, 160,
197, 200, 202
palmate, 18
palmitic acid methyl ester, vii, 1, 2
Pamirs, 140
pancreas, 14, 111
parallel, 68, 90, 102
parasite, 26
parenchyma, 89, 90
parents, 150
participants, 111
partition, 194, 202
pasta, 97
pathogenesis, 11, 20, 103
pathogens, viii, 17, 25, 26, 27, 28, 30, 36, 40, 151,
152
pathology, 101
pathophysiology, 100
pathways, vii, 17, 20, 26, 30, 35, 74, 91, 92, 101,
103, 104, 108, 128, 132, 134, 135, 137, 150, 153
pattern recognition, 91
perfusion, 6, 13
229
Index
Portugal, 37
positive feedback, 19
positive relationship, 111
potassium, 3, 5, 12, 30, 69
potato, 132
poultry, 22, 48, 49, 52, 53, 58
precipitation, 71
premature infant, 148, 154
preparation, 77
preterm infants, 148, 150, 153, 156
prevention, 27, 35, 62, 106, 110, 120, 121, 155
probiotics, 155
producers, 70, 73
progenitor cells, 92, 104
pro-inflammatory, 90
project, 160
prokaryotes, ix, 28, 35, 125
prokaryotic cell, vii, 17, 28
proliferation, 33, 91, 151, 152
propane, x, 159, 161, 164, 177, 179, 180, 181, 182,
183, 184, 185, 186, 187, 191, 199, 200, 201, 204,
205
prostacyclins, 18
prostaglandins, 18
prostate cancer, 123
proteasome, 104
protection, 34, 90, 148, 151, 152, 155, 157
protective role, 153
protein kinase C, 6, 13, 91, 92, 103, 104, 107, 111,
118
protein kinases, 91, 92, 107
protein synthesis, 14, 26, 89
protein-protein interactions, 18
proteins, 18, 20, 26, 27, 28, 30, 40, 41, 66, 82, 83,
86, 88, 93, 99, 118, 134, 149, 150
proteome, 18
pulp, 72, 73, 75, 76, 212
purification, 164
purity, 167, 170, 199, 202
quality of life, 60
R
rainfall, 213
rape, 77, 187
rape seed, 77, 187
raw materials, 205
reactions, 6, 7, 68, 69, 77, 79, 80, 81, 82, 85, 127,
130, 131, 132, 134, 135
230
Index
seed, 46, 70, 76, 77, 126, 129, 131, 132, 133, 134,
137, 138, 139, 140, 141, 142, 143, 160, 197, 204
seedlings, 142
selectivity, 27, 109, 164, 191, 198, 204
selenium, 75
sensitivity, 4, 31, 34, 114, 122, 124
sequencing, 198
Serbia, 105, 115, 116
serine, 91, 92, 110
serum, viii, 28, 31, 37, 39, 41, 45, 47, 52, 56, 61, 86,
87, 88, 89, 100, 106, 107, 109, 110, 111, 112,
113, 114, 118, 119, 120, 121, 122, 123, 124
serum albumin, 28
serum cholesterol, viii, 31, 41, 45, 47, 52, 61, 86, 88,
100, 112, 114, 121, 122, 124
shape, 51
sheep, 50
shelf life, 71
shellfish, 51, 58
shock, 26
showing, 148, 167, 183, 202
side chain, 20
signal peptide, 155
signal transduction, ix, 40, 105, 153
signaling pathway, 40, 91
signalling, 35
signals, 79
silica, 141
silver, 141
skeletal muscle, ix, 14, 30, 35, 79, 83, 103, 106, 108,
110, 111, 114, 116, 118, 122, 123, 124
skin, 25, 36, 41, 49, 50, 70, 94, 95
small intestine, 83
smooth muscle, 3
smooth muscle cells, 3
sodium, 3, 30, 68, 69, 76, 214
sodium hydroxide, 69
solid matrix, 162, 163, 166, 202, 204
solid phase, 165, 168
solubility, x, 83, 159, 162, 163, 164, 165, 166, 168,
169, 170, 172, 173, 174, 175, 178, 179, 180, 181,
182, 184, 186, 187, 190, 191, 193, 194, 200, 203,
204, 205
solution, 214
solvents, 161, 162, 164, 184, 187, 190, 195, 198,
201, 202, 205
South Africa, 159
South America, 73, 75
soybeans, 57, 203
specialization, 143
species, vii, ix, 1, 3, 21, 22, 27, 38, 41, 48, 50, 51,
55, 58, 60, 72, 74, 75, 76, 89, 90, 97, 101, 107,
125, 126, 128, 129, 134, 138, 139, 140, 142, 152
specifications, 95
Spring, 117
SS, 99, 102, 121, 123, 141
SSS, 66, 70
stability, 35, 58, 60, 70, 76, 92, 151, 215
starvation, 33
state(s), vii, ix, 1, 39, 51, 83, 89, 91, 97, 125, 148,
178, 202
statistics, 14, 212
Stearic acid, viii, 15, 45, 109, 205
sterile, 151
sterols, 73, 200, 201, 202
stimulation, 2, 30, 39, 89
stock, 195
storage, ix, 18, 21, 33, 34, 66, 69, 70, 71, 78, 83, 89,
93, 125, 126, 128, 129, 131, 133, 134, 135, 139,
141, 142, 211
stress, 33, 43, 86, 89, 90, 91, 102, 107, 111, 113,
117, 123, 132
stroke, ix, 2, 5, 6, 9, 10, 14, 15, 105, 111, 112, 116,
122, 123
stroma, 135
structure, vii, viii, x, 1, 3, 9, 24, 34, 35, 48, 61, 63,
75, 77, 97, 100, 120, 129, 134, 145, 148, 151,
153, 154, 155, 178, 205, 217
subarachnoid hemorrhage, 11
suberin, 131, 132, 141
substitutes, 74, 77, 96
substitution, 88, 95
substrate(s), ix, 39, 69, 82, 84, 86, 92, 104, 106, 108,
117, 127, 128, 129, 130, 131, 134, 135, 139
sulfur, 20
Sun, 12, 41, 87, 101, 117, 121, 156
superoxide, vii, 17, 89
supplementation, viii, 12, 31, 45, 54, 55, 58, 122
suppression, 90, 137, 143
surfactant, 160, 161
surplus, 86
survival, 9, 28, 90, 91
susceptibility, 152
swelling, 28, 29, 30, 39, 42, 43
Switzerland, 62, 115
syndrome, 106, 112
synthesis, vii, x, 1, 2, 6, 7, 10, 14, 18, 19, 20, 27, 28,
30, 34, 35, 54, 55, 66, 78, 79, 84, 86, 88, 90, 92,
98, 111, 113, 126, 127, 128, 129, 130, 131, 132,
134, 138, 139, 140, 142, 145, 146, 149
T
T cell(s), 154, 157
target, vii, 1, 26, 70, 86, 153
target organs, 86
231
Index
techniques, 70, 162, 212
technological advances, 40
technology, 66, 73, 205
temperament, 154
temperature, xi, 26, 72, 138, 139, 146, 147, 160, 161,
162, 163, 164, 165, 166, 168, 169, 170, 172, 173,
175, 178, 179, 180, 181, 182, 183, 184, 185, 187,
191, 194, 198, 203, 211, 214
temperature dependence, 173
texture, 74, 77
TGA, 88
Thalassiosira, 23
therapeutic interventions, 9
therapy, 9, 10, 122
thermal analysis, 96
thermal degradation, 161, 162, 200, 202
thermograms, 96
threonine, 91
thrombin, 9
thrombosis, 106
thromboxanes, 18
thrombus, 9
tissue, ix, 22, 33, 39, 48, 52, 53, 56, 61, 78, 83, 90,
99, 105, 107, 112, 114, 123, 131, 133, 134, 138,
142, 150
TLR, 66, 91, 103, 107, 117
TLR4, 92, 108, 117
TNF, 31, 34, 66, 90, 91, 107, 117
TNF-alpha, 117
TNF-, 31, 34, 66, 90, 91
tobacco, 132, 140
tocopherols, 71, 200, 201, 202
total cholesterol, viii, ix, 34, 45, 47, 62, 87, 88, 92,
105, 109
total energy, 87
toxicity, viii, 9, 41, 63, 70, 101, 163
trade, 212
traditions, 74
trafficking, 18, 35
traits, 60
transcription, 14, 88, 90, 91, 117
transcription factors, 14, 90
transesterification, 40, 132
transformation, 37, 97, 137
transforming growth factor, 103
transient ischemic attack, 122
translocation, 34
transmission, 11
transport, 18, 26, 27, 32, 40, 42, 79, 83, 90, 99, 100,
118, 162
transportation, 131
treatment, 9, 10, 51, 92, 100, 101, 116
triacylglycerols, viii, ix, 41, 48, 55, 59, 63, 69, 74,
92, 111, 125, 126, 129, 139, 141, 142, 143, 146,
148, 156
trial, 110, 112, 148, 155, 156
triglycerides, viii, x, 4, 12, 18, 41, 48, 62, 63, 71, 73,
74, 75, 77, 98, 109, 110, 120, 123, 141, 142, 145,
155, 159, 160, 161, 163, 191, 193, 197, 198, 200,
201, 202, 203, 204
tumor, 14, 107, 117, 118
tumor necrosis factor, 107, 117, 118
Turkey, 22, 51
turnover, 33, 39, 149
Tuskegee University, 17
type 1 diabetes, 91
type 2 diabetes, viii, ix, 63, 91, 102, 105, 112, 114,
121, 124
U
U.S. Department of Agriculture, 115
UK, 58
ultrasound, 156
uniform, 56
United States (USA), 9, 67, 73, 93, 94, 98, 140, 143
updating, 217
USDA, 58, 93, 119
V
vacuum, 71, 161, 162
vagina, 39
variables, 123
variant angina, 13
variations, 73, 128, 194, 215
varieties, 73, 95, 212, 213, 214, 215, 216
vascular tonicity, vii, 1
vasculature, 9
vasoconstriction, 5, 6, 13
vasodilation, vii, 1, 2, 3, 4, 5, 6, 9, 10, 11, 12, 13
vasodilator, 2, 3, 10, 11, 12
vasomotor, 6
vasopressin, 5
vasospasm, 6, 13
VAT, 3
vegetable oil, 46, 55, 70, 73, 74, 75, 77, 87, 97, 116,
146, 148, 152, 212
vegetables, 35, 38, 133, 142
Vereshchagin, 133, 140, 141, 142, 143
victims, 9
virus infection, 37
viruses, 21
viscosity, 73
232
Index
W
Y
Washington, 94, 115, 141
waste, 212, 213, 215
waste water, 213, 215
wastewater, 216
water, 21, 55, 67, 68, 69, 72, 81, 84, 86, 123, 131,
188, 190, 213