Trends in Food Science & Technology 21 (2010) 229e246

Review

Analytical strategies to evaluate antioxidants in food: a review

C. Andrea, I. Castanheiraa, J.M. Cruzb, P. Paseirob and A. Sanches-Silvaa,*
National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1600 Lisbon, Portugal (National Institute of Health Dr. Ricardo Jorge, Food and Nutrition, Av. Padre Cruz, 1600 Lisbon, Portugal. Tel./fax: D351 938380493; e-mail: ana.silva@insa.min-saude.pt) b Department of Analytical Chemistry, Nutrition and Food Science, Pharmacy Faculty, University of Santiago de Compostela, Campus Sur, 15782 Santiago de Compostela, Spain
Antioxidants are an important group of food additives that have the ability to protect against detrimental change of oxidizable nutrients and consequently they extend shelf-life of foods. The present paper is an updated review on the analysis of ten antioxidants (both synthetic and natural). Three types of gallates, propyl (PG), octyl (OG) and dodecyl (DG), tert-Butylhydroquinone (TBHQ), Butylated Hydroxytoluene (BHT), tertButylhydroxyanisole (BHA), a-tocopherol, ascorbic acid, erythorbic acid and sodium D-isoascorbate were the antioxidants selected. The physico-chemical properties, sample preparation procedures and analytical methods for their determination are compared and discussed.
a

Introduction Nowadays, antioxidants receive remarkable attention in the literature, due to the ability to preserve foodstuffs by retarding deterioration, rancidity and/or discoloration
* Corresponding author.
0924-2244/$ - see front matter 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.tifs.2009.12.003

caused by oxidation. Detrimental effects include undesirable chemical compounds formation like aldehydes, ketones and organic acids than yield off-avours (Saad et al., 2007). In biological systems, antioxidants protect against oxidative damage and help to prevent cardiovascular, neurological and/or carcinogenic diseases (Lanina, Toledo, Sampels, Kamal-Eldin, & Jastrebova, 2007). Following the recommendations of international organizations, the use of synthetic antioxidants like PG, OG, BHA, BHT and TBHQ is regulated by legal authorities in a limited number of foods, with maximum limits in each case or association. There are several documents legislating food additives such as European Union Directives and Regulations (Directive 95/2/EC, 1995; EC Regulation No 1333/ 2008, 2008), FDA in USA (FDA, 2001a, 2001b), Food Standards Australia New Zealand for Australia and New Zealand (The Australia New Zealand Food Standards Code, 2009), The Joint FAO/WHO Expert Committee on Food Additives (JECFA, 2003) and Codex Alimentarius (CGSFA, 1995). The use of antioxidants (AO) is not restricted to foodstuffs. They can also be used to prevent the degradation of food packaging during processing and storage, improving the end-use application. Nevertheless, they or their degradation products can migrate into foodstuffs. EU legislation imposes migration limits of substances that can be used in the manufacture of food packaging. Antioxidants can be classied as primary or long-term antioxidants and as secondary or processing antioxidants. Primary antioxidants include hindered phenols and secondary aryl amines and secondary antioxidants include orga nophosphites and thioesters (Dopico-Garca, Lopez Vilarino, & Gonzalez-Rodrguez, 2003). The primary are active radical scavengers or hydrogen donors or chain reaction breakers while secondary are peroxide decomposers. Most of the methods described in the literature for the quantitative analysis of antioxidants or antioxidant mixtures present interference problems, low resolution and are timeconsuming. The establishment of analytical methods to monitor the use of prohibited antioxidants and/or the excessive use of permitted AO and their degenerative products is a real need. In 2000, Karoviova & Simko (2000) have carc ried out a review regarding the determination of synthetic phenolic antioxidants in food by high-performance liquid

230

Table 1. Chemical and physical information of the antioxidants. Migrants CAS No Formula/ Elemental analysis C10H12O5 MW/Exact Mp ( C) mass 212.20a 145e146 Ca CAS Bp/Fp ( C) Density (g/cm3) Logkow Enthalpy of vap. Solubility

Propyl gallate, Benzoic acid, 3,4,5-trihydroxy-, propyl ester (9CI)

121-79-9

Fp: 181.320.8 C, bp: 448.640.0 C, Press: 760Torra

1.3630.06g/cm3, Temp: 20 C, Press: 760Torra

2.6000.333, Temp: 25 Ca

73.433.0kJ/mol, Press: 760Torra

Slightly soluble in water; freely soluble in ethanol, ether and propane1,2-diolc Insoluble in water, freely soluble in ethanol, ether and propane-1,2-diolc Insoluble in water; freely soluble in ethanol and etherc Freely soluble in water, soluble in ethanolc

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

Octyl gallate, Benzoic acid, 3,4,5-trihydroxy-, octyl ester (9CI)

1034-01-1

C15H22O5

282.33a

101e104 Ca 91e92 Ca

bp: 482.940.0 C, Press: 760Torr, Fp:177.120.8 Ca bp:521.745.0 C, Press: 760Torr, Fp:180.322.2 Ca bp: 552.750.0 C, Press: 760Torr, Fp: 238.223.6 Ca

1.1850.06g/cm3, Temp: 20 C, Press: 760Torra 1.1120.06g/cm3, Temp: 20 C, Press: 760Torra 1.9540.06g/cm3, Temp: 20 C, Press: 760Torr (calc), 1.654g/cm3 (exp)a

5.2560.333, Temp: 25 Ca

77.633.0kJ/mol, Press: 760Torra

Lauryl gallate, Benzoic acid, 3,4,5-trihydroxy-, dodecyl ester (9CI)

1166-52-5

C19H30O5

338.44

94e96 Ca

7.3820.333, Temp: 25 Ca

82.503.0kJ/mol, Press: 760Torra

89-65-6 Erythorbic acid, D-erythro-Hex-2-enonic acid, g-lactone (8CI, 9CI)

C6H8O6

176.12

167e174 Ca

2.4100.450, 95.786.0kJ/mol, Temp: 25 Ca Press: 760Torra

Sodium, D-isoascorbate monohydrate D-erythroHex-2-enonic acid, g-lactone, monosodium salt, monohydrate (9CI)

63524-04-9 C6H8O6, H2O, Na

216.12b

165 C (dec.) (lit.)b

50-81-7 Ascorbic acid, L-Ascorbic acid (8CI, 9CI)

C6H8O6

176.12

190e194 Cb

Fp: 238.223.6 C, bp: 1.9540.06g/cm3, 552.750.0 C, Press: Temp: 20 C (calc)a 760Torr (calc)a bp:485.90.0 C, Press: 760Torr (calc), Fp: 210.224.4 C (calc)a 0.9300.06g/cm3, Temp: 20 C, Press: 760Torr (calc),a 0.950g/mL at 20 C(lit.)b

2.4100.450, 95.786.0kJ/mol, Temp: 25 C Press: 760Torr(calc)a (calc)a

Freely soluble in water; sparingly soluble in ethanol; insoluble in etherc

a-Tocopherol, 2H-1Benzopyran-6-ol, 3,4-dihydro-2,5,7,8tetramethyl-2-(4,8,12trimethyltridecyl)-(9CI)

10191-410e

C29H50O2

430.71

11.9020.267, 78.003.0kJ/mol, Insoluble in water, Temp: 25 C Press: 760Torr (calc)a freely soluble in (calc)a ethanol, miscible with etherc

tert-Butylhydroquinone, 1948-33-0 1,4-Benzenediol, 2-(1,1dimethylethyl)-(9CI), TBHQ

C10H14O2

166.22

126.5e129 C (exp)a

bp: 291.320.0 C, Press: 760Torr, Fp: 138.716.4 Ca

1.0860.06g/cm3, Temp: 2.3330.225, 20 C, Press: 760Torra Temp: 25 Ca

55.203.0kJ/mol, Press: 760Torra

Practically insoluble in water; soluble in ethanolc

 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

231

Scinder, 2008. Available on: www.Sigma Aldrich.com. c Available on: www.fao.com. d Garca, Silva, Cooper, Franz, & Losada, 2006. e According to JEFCA (1986), there is no single denite number for this substance. CAS 59-02-9 is for vitamin E, 1406-18-4 is for alfa-tocopherol, 2074-53-5 is for all-rac-alpha-tocopherol and 10191-40-0 is for racemic-alpha-tocopherol synthesized from natural phytol or its derivative.

chromatography. Robbins (2003) has also carried out a review concerning the analytical methodologies to determine phenolic acids in foods. This review aims to carry out an updated evaluation of a wide range of analytical methods used to separate, identify and quantify antioxidants, phenolic and other, in foodstuffs and food packaging. Chemical and physical information of antioxidants Currently, food industry requires simple and rapid analytical assays for the control of synthetic phenolic antioxi dants added to food (Morales, Gonzalez, Reviejo, & n, 2005; Morales, Gonzalez, Serra, Reviejo, & Pin Pingarro garron, 2005). Many studies have been dedicated to the effects of antioxidants (EFSA, 2004a, 2004b, 2008; Fidler, Davidsson, Zeder, & Hurrell, 2004; Georgantelis, Blekas, Katikou, Ambrosiadis, & Fletouris, 2007; Gharavi & El-Kadi, 2005; Hirose et al., 1999; Kuwabara, Watanabe, Adachi, Nakanishi, & Matsuno, 2003; Sauberlich, Tamura, Craig, Freeberg, & Liu, 1996; Zurita et al., 2007). In fact, recently, the safety of AO has been questioned (Yang, Lin, & Choong, 2002). Studies demonstrate that excess use of PG, BHA, BHT and TBHQ, synthetic antioxidants added to foodstuff, may cause a loss of sustenance and even produce toxic substances to harm peoples health (Guan, Chu, Fu, Wu, & Ye, 2006). The use of high concentrations of BHA and BHT in rats was associated with cancer development. Therefore, although they unequivocal advantages, they have been subject of controversy (Saad et al., 2007). Limits are established according to the toxicity of substances. The Acceptable Daily Intake (ADI) recommended by the Joint FAO/WHO Expert Committee on Food Additives (JECFA, 2003) is 0e1.4mg of additive/kg body weight for Octyl gallate, Dodecyl gallate, BHA, BHT and TBHQ, while the Scientic Committee for Food (SCF) established a 0.5mg/kg value. Erythorbic acid is allocated a numerical ADI of 6mg/kg bw by the SCF. In the particular case of BHT, the ADI was established at 0e0.3mg/kg body weight but did not include BHT in the drinking water contaminants list (JECFA, 2003). In 2003, SCF set a tolerable upper intake of vitamin E, as a-tocopherol, for adults of 300mg of a-tocopherol equivalents per day while JECFA established an ADI of 0.15e 2mg/kg of body weight per day calculated as a-tocopherol (JECFA, 2003). Table 1 is summarizes chemical and physical information of 10 antioxidants and Table 2 resumes the methods used to determine antioxidants in foodstuffs (according to the type of food), packaging and pharmaceutical or cosmetic products. Sample preparation The present review has focused the analysis of antioxidants in foodstuffs. However, a few papers that have been found also extract antioxidants from food packaging

0.89

5.10

52.173.0kJ/mol, Press: 760Torra

71 C

220.36

Butylated hydroxytoluene, 128-37-0 2,6-di-tert-butyl-p-cresol, BHT

tert-Butylhydroxyanisole Phenol, (1,1dimethylethyl)-4methoxy-(9CI), BHA

25013-16-5 C11H16O2

C15H24O

180.25

51 C

Bp: 268 C

265 C

Insol. in water alkali; sol.: EtOH, H2O C6H6, acetone, petroleum ether, sol.: 0,6mg/Ld Insol. in water; freely sol. in ethanol and propane-1,2-diolc

Table 2. Methods used to determine antioxidants in foodstuffs (according to the type of food), packaging and pharmaceutical or cosmetic products. Type of food Antioxidants Analytical methods HPLCePDA GCeFID TLC RPeHPLCeUVevis RPeHPLCeUVeVis RPeHPLCeUV HPLCeUV LCeMSeAPCI LCeMSeESI LCeMS & GCeMS GCeFID GCeFID MECCeED MECC-UV VT HPLCeFl NPeHPLCeUVevis LCeUV References Sin et al., 2006

Fats, oils and their BHA, BHT, PG, OG, DG products BHA, BHT, TBHQ, PG, OG BHA, BHT, PG, OG, DG BHA, BHT, TBHQ, PG BHA, BHT, TBHQ Tocopherols BHA, BHT, TBHQ, PG BHA, BHT, TBHQ BHA, BHT, TBHQ TBHQ, BHA, BHT, PG BHA, BHT, DG BHA, BHT, TBHQ Tocopherols Cereals and cereals Tocopherols products (e.g. BHA, BHT, PG, OG, DG Bakery products) Vegetables and vegetables products Total AA AA Tocopherol TBHQ, BHA, BHT, PG LeAA Fruit and fruit products AA Total AA LeAA AA & EA AA & EA AA LeAA Milk and milk Total AA products (e.g. Tocopherol cheese and cheese products) Tocopherol BHA, TBHQ, PG, OG, DG BHA, BHT, TBHQ, PG BHA, BHT, TBHQ Meat and meat products BHA, BHT, TBHQ, PG, DG, OG

Perrin & Meyer, 2002 Aparicio et al., 2000 Saad et al., 2007 Maziero et al., 2001 Lanina et al., 2007 Tsuji, Nakanoi, Terada, Tamura, & Tonogai, 2005 Yang et al., 2002 Suh et al., 2005 Guan et al., 2006 Delgado-Zamarreno, Gonzalez-Maza, Sanchez-Perez, & Martnez, 2007 Raymundo, Paula, Franco, & Fett, 2007 Gliszczynska-Swig1o, & Sikorska, 2004 Devi, Jayalekshmy, & Arumughan, 2007 Rafecas et al., 1998

HPLCeUV & MECC Enzymatic/Spectrophotometric, RPeHPLCeFlu HPLCeDAD & HPLCeFlu MECCeED RPeHPLCeFlu RPeHPLCeFlu HPLCeUV & MECC HPLCePDA & HPLCeDADeESIeMS LCeHILICeUV CEeUVeVIS CE e ED RPeHPLCeFlu RPeHPLCeFlu RPeHPLCeUV & RPeHPLCeMSeAPCI & RPeHPLCeMSeESI NPeHPLCeFl HPLCeDAD RPeHPLCeUV GCeFID HPLCeUVePAD HPLCeDAD MECCeED HPLCeUV and HPLCeFl LCeMS & GCeMS CE-Microchip MEC Amperometric determination LCeMSeGCeMS RPeHPLCePDA LCeMSeGCeMS

Thompson & Trenerry, 1995 Raghu, Platel, & Srinivasan, 2007 Gomez-Coronado, Ibanez, Ruperez, & Barbas, 2004 Guan et al., 2006 Burini, 2007 Raghu et al., 2007 Thompson & Trenerry, 1995 Shui & Leong, 2004 Tai & Gohda, 2007 Versari, Mattioli, Parpinello, & Galassi, 2004 Wu, Guan, & Ye, 2007 Burini, 2007 Burini, 2007 Lanina et al., 2007 Rodrigo, Alegra, Barbera, & Farre, 2002 Noguera-Ort et al., 2000 Saad et al., 2007 Yang et al., 2002 Pinho, Ferreira, Oliveira, & Ferreira, 2000 Lopez-Cervantes et al., 2006 Guan et al., 2006 Quiros, Lopez-Hernandez, & SimalLozano, 2001 Tsuji et al., 2005 Ding et al., 2006 Tsuji et al., 2005 Perrin & Meyer, 2002 Tsuji et al., 2005

Fish, sh products Tocopherol and sea food TBHQ, BHA, BHT, PG AA Mixed foods BHA, BHT, TBHQ, PG PG, OG, DG BHA, BHT, TBHQ, PG BHA, BHT, TBHQ, PG, OG BHA, BHT, TBHQ, PG

Spice and other ingredients Dry foods (e.g. Sugar, honey and products thereof)

 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246 Table 2 (continued ) Food for special nutrition use Total AA Tocopherol BHA, TBHQ, PG, OG, DG AA & EA BHA, BHT, TBHQ BHA, PG (fatty foods) BHA, BHT BHA, BHT BHA, BHT Plastic products Beverages a-Tocopherol BHT AA & EA BHT AA BHT, BHA, PG, TBHQ AA AA Tocopherols RPeHPLCeUV NPeHPLCeFlu MLCeDAD LCeHILICeUV GCeFID Flow injection solid-phase spectrometry HPLCePDA RPeHPLCeDAD RPeHPLCeDAD HPLCeFlu & DRIFT-IR GCeMS HPLCeUV GCeMS CE conventional microchip HPLCeDAD HPLCeUVevis Iodometric titration/NIR RPeHPTLC NPeHPLCeUV RPeHPLCeUV GC Fontannaz, Kilinc, & Heudi, 2006 Rodrigo et al., 2002 Noguera-Ort et al., 2000 Tai & Gohda, 2007 Suh et al., 2005 Capitan-Vallvey, Valencia, & Nicolas, 2003

233

Other foods

Food simulants

Dopico-Garca, Lopez-Vilarino, & Gonzalez-Rodrguez, 2005 Dopico-Garca et al., 2003 Dopico-Garca, Vilarino, & Gonzalez Rodriguez, 2007 Graciano-Verdugo et al., 2006 Hill et al., 2003 Tai & Gohda, 2007 Tombesi & Freije, 2002 Law, Kuban, Zhao, Li, & Hauser, 2005 Ruiz, Garca-Moreno, Barbas, & Pingarron, 1999 Maia et al., 2007 Blanco & Alcala, 2006 Pyka & Sliwiok, 2001

Chewing gum Pharmaceutical &/or Cosmetic products

(Graciano-Verdugo, Peralta, Gonzalez-Ros, & Soto-Valdez, 2006; Hill, Shaw, & Wu, 2003) or pharmaceutical &/or cosmetic products (Blanco & Alcala, 2006; Maia et al., 2007; Pyka & Sliwiok, 2001). Most of the methods found in the literature, to determine AO in foodstuffs, were tested in fats, oils and their products, as it can be seen in Table 4. HPLC with UV was the most used technique (Aparicio, Andres, & Vera, 2000; Maziero, Baunwart, & Toledo, 2001; Perrin & Meyer, 2002; Sin, Wong, Mak, Sze, & Yao, 2006; Saad et al., 2007). The quantication of antioxidants in foodstuffs presents several difculties in what regards to incomplete extraction of antioxidants and co-extraction with interfering substances (Perrin & Meyer, 2002). Table 3 resumes some of the extraction techniques found in the literature to determine antioxidants from foodstuffs. The literature reports several extraction solvents due to the wide range of polarity of antioxidants. Some studies include comparison of several solvents for the extraction of antioxidants (Perrin & Meyer, 2002) and it has concluded that methanol was the most suitable for the antioxidants extraction. The most common method to extract these antioxidants is Solvent extraction (SE), which can be either LiquideLiquid Extraction (LLE) or liquidesolid extraction (LSE). In some matrices are also described Solid-Phase Extraction (SPE), SolideLiquid Extraction, SolideGas Extraction

and Solid-Phase Micro Extraction (SPME) (Tombesi & Freije, 2002). SolideLiquid Extraction and SolideGas Extraction are described in order to analyse a-tocopherol and BHT in plastics (Graciano-Verdugo et al., 2006; Hill et al., 2003). Direct sample injection in a chromatograph is not a com mon practise, although it has been used (Noguera-Ort, Villanueva-Camanas, & Ramis-Ramos, 2000). Solid-Phase Extraction is more rapid and environmentally friendly (due to the solvents save) than LLE. A lot of different solvents were used to extract BHA, BHT, TBHQ, PG, OG and DG from different matrices. For instance, to extract BHA, BHT, PG, OG and DG in oil samples Sin et al. (2006) used n-hexane, AcN and a methanolic ascorbic acid solution while Aparicio et al. (2000) used only hexane and AcN for BHT and petroleum ether and ethanol for the others. The gallates were extracted with methanol and AcN with ammonium acetate solution from food samples by Ding, Mora, and Garcia (2006). Regarding the analysis of vitamin C, the ofcial method involves the titration of vitamin C (AA and iso AA) by a mild oxidizing agent (2,6-dichlorophenol-indophenol) (AOAC, 1995). For the analysis of a-tocopherol, saponication is often required to obtain full extraction of lipids (Lanina et al., 2007; Lopez-Cervantes, Sanchez-Machado, & Rios-Vazquez, 2006).

234

Table 3. Extraction techniques used to determine antioxidants. Extraction technique LiquideLiquid Analyte determineda Phenolic antioxidants BHA, BHT, PG, OG, DG BHA, BHT, PG, OG, DG Sample Oil Oil sample Solvent used in the extraction procedure n-Hex, ACN For BHT: Hex, ACN For others: petroleum ether, EtOH (72%, v/v) MeOH References Sin et al., 2006 Aparicio et al., 2000

PG, TBHQ, OG, DG, BHA, BHT

Oils and fats

BHA, BHT, PG, OG, DG BHA, BHT, TBHQ BHA, BHT, TBHQ BHA BHT TBHQ PG BHA BHT TBHQ PG OG BHA BHT TBHQ PG DG OG BHA BHT BHA BHT DG PG OG DG Total AA

Bakery products Cooking oil, sh oil, butter, margarine, cheese, mayonnaise, salad dressing Mayonnaise Vegetable oil, butter, margarine, cheese Gravies, dehydrated soups, bouillons, dehydrated meat, dry pet food Liver pates LDPE Sunower oil, olive oil Food sample (gravy mix) Fortied food products

ACN-isopropanol (1:1, v/v) Diethyl ether MeOH MeOH/ACN (1:1, v/v) MeOH; Hex/2-propanol n-Hex saturated with ACN, ACN saturated with n-Hex n-Hex MeOH, ACN MeOH TCEP$HCl, trichloroacetic acid. For starch containing samples, taka-diastase was also used. For (1) and (2): ACNe66.7mM ammonium acetate solution (85:15, v/v) For (3): acetonitrileewater (30:70, v/v) Dilution 1:1, v/v with 10% Metaphosphoric acid 3% Metaphosphoric acid, 50% aqueous acetone 2% m-phosphoric acid

IUPAC, 1992 AOAC, 1995 AOCS, 1997 Rafecas et al., 1998 Yang et al., 2002 Raymundo et al., 2007 Saad et al., 2007 Perrin & Meyer, 2002 Pinho et al., 2000 Dopico-Garca et al., 2003 Delgado-Zamarreno et al., 2007 Ding et al., 2006 Fontannaz et al., 2006

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

Vitamin C

AA & EA

(1) Tea drinks, (2) Nutritional supplement food, (3)dried fruit

Tai & Gohda, 2007

AA & EA Total AA AA AA

Apricot Juices Fruit vegetables Star fruit Emblica ofcinalis, guava fruits, chilli fruits and amaranth leaves Grapefruit Shrimp waste Olive, grapeseed, corn rapeseed, peanut, sunower, soybean oils (1) Sunower oil, (2) Milk

Versari et al., 2004 Thompson & Trenerry, 1995 Shui & Leong, 2004 Raghu et al., 2007

Vitamin E

AA a-tocopherol a-tocopherol

EtOH 99.7% n-Hex 2-propanol

Wu et al., 2007 Lopez-Cervantes et al., 2006 Gliszczynska-Swig1o, & Sikorska, 2004 Lanina et al., 2007

tocopherol

For (1):Saponication with KOH in ethanol. Extraction with Hex For (2): Saponication with aqueous KOH. Extraction with Hex

 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

235

Delgado-Zamarreno et al., 2007 Tsuji et al., 2005

Graciano-Verdugo et al., 2006

Gomez-Coronado et al., 2004

Dopico-Garca et al., 2005 Shui & Leong, 2004 Thompson & Trenerry, 1995

Hill et al., 2003

Tombesi & Freije, 2002

Rodrigo et al., 2002 Devi et al., 2007

Analytical methods to determine antioxidants A large variety of methods were found in the literature. Antioxidants analysis of food and plastic samples are mainly performed by chromatographic methods as it can be conrmed by Table 4 which resumes the separation techniques and conditions used for the determination of the antioxidants.

Method performance Antioxidants have a wide polarity range. Therefore, when analysing them simultaneously a low recovery can be achieved for some of these compounds (Rafecas, Guardiola, Illera, Codony, & Boatella, 1998). As shown in Table 5 the difference methods have different behaviour concerning the property and kind of food. These antioxidants are most use in fats, oils and their products. TBHQ was determined in many samples but only found in fats and sh and sh products. All recoveries are good, being in the range 88.7e108.3%. There are no limits of quantication in the references and the higher limit of detection was achieved in fats, oils and their products and in other foods. BHA was found in more foods than TBHQ and also in plastics. In vegetables, sh, milk, food for special nutritional use and other foods BHA was not found. The limits of detection found were lower than TBHQ. The worst recovery was in oil and butter (81.80%). BHT is very similar to BHA but it was also found in beverages. The recoveries are not so good, the lowest was found in cereals (57.8%). The most used gallate is PG. It was determined in fats, oils, vegetables and mixed foods. OG and DG were not found in any kind of foods. The limit of detection (LD) is very similar being the higher in other foods. The worst recovery found was for DG in cereals and cereals products from 94.5 to 95.8%. From the selected food, tocopherol was not found in cereals and cereals products and presents the worst recovery in this kind of food. The LD was higher in the plastic products. Total vitamin C, was just checked in food for special nutritional use. The recoveries were around 100%. Ascorbic acid is used in different kind of food and also in cosmetic and pharmaceutical products. The only value not found was for food for special nutritional use. The higher limit of detection determined was for beverages. Recoveries were tested with 2 different solvents which present signicant different values. Erythorbic acid was just found in beverages with a limit of detection lower than the one found in fruit. Recovery was tested with 2 different methods of extraction, giving a good recovery for the rst method and worst for the second. For sodium D-isoascorbate there are no values in the references.

ChloroformeMeOH (2:1 v/v) Soxhlet extraction and extracted with MeOH, acetone, Hex

MeOH or ACN ACN/2-propanol/EtOH (2:1:1)

MeOH, THF Water and MeOH 3% Metaphosphoric acid

Acetone

THF

Infant formulas Deffated rice bran

Sunower oil Nikuman, olive oil, peanut butter, pasta sauce, chewing gum Aqueous food simulants Star fruit Fruit and vegetables

Air PVC tubes BHT

BHA, BHT, DG BHA BHT TBHQ PG

Fresh leaves

tocopherol tocopherol

BHA, BHT AA Total AA

a-Tocopherol

Tocopherol

BHT SPME
a

Drinking water From those studied in this review.

Phenolic antioxidants

Vitamin C

SolideLiquid

Plastic LDPE

DDTD

SPE

SFE

Fiber assembly with polydimethylsiloxane lm

236

Table 4. Separation techniques and conditions used for the determination of selected antioxidants. Technique HPLC Detector DAD (l280nm) Matrix Oil Analytes BHA, BHT, PG, OG, DG PG, OG BHA, BHT, TBHQ Conditions Column: C18 Hypersil ODS (150mm4.6mm, 5mm) Mobile phase solvents: A e acetic acid 5%; B e methanol/AcN 1:1 (Gradient elution) Column: Supelcosil LC-18 30 C (150mm4.6mm, 5mm) Mobile phase solvents: A e Water pH3.0 acidied with phosphoric acid 1% (v/v) B e MeOH-AcN 1:1 (v/v) (gradient elution) Column: Nova Pack C18 (150mm3.9mm, 4mm) Mobile phase solvents: MeOH/Water Column: Symmetry (150mm3.0mm, 3.5mm) Mobile phase solvents: MeOH/Water (gradient elution) Column: Symmetry (150mm3.0mm, 3.5mm) Mobile phase solvents: MeOH/Acn/THF/Water (gradient elution) Column: Shim-Pack VPeODS (250mm4.6mm) Column temperature: 27 C Mobile phase solvents: A e 0.1% formic acid in water; B e MeOH Column: SS Exil ODS (250m4.6mm, 5mm) Column temperature: 36 C Mobile phase solvents: MeOH:ACN:water (68:28:4, v/v/v) Column: Nova-Pak C18 (150mm3.9mm, 4mm) Column temperature: 30 C Mobile phase solvents: A e MeOH; B e Water (gradient elution) Column: Discovery C18 (250mm4.6mm, 5mm) Column temperature: 35 C Mobile phase solvents: A e Puried-for-HPLC water; B e AcNemethanol (70:30, v/v) with 0.1% (v/v) of acetic acid added (gradient elution) Column: Spherisorb C18 (250mm4.6mm, 5mm) Mobile phase solvents: A e 5% acetic acid in water; B e AcN with 5% acetic acid Column: C18 NOVAPAK Radial-PAK (100mm80mm,4mm) Mobile phase solvent: 0.2% (v/v) aqueous orthophosphoric acid Column: C18 (ODS) Vydac 201-TP 54 (250mm4.6mm, 5mm) Mobile phase solvents: AcN: methanol:5% acetic acid TBHQ and BHA: (25:25:5 0 v/v/v) BHT:(42.5:42. 5:15 v/v/v) Column: LiChrospher RP (250mm4.0mm, 5mm) Mobile phase solvents: A e AcN; B e water with 1% acetic acid (gradient elution) Column: Lichrosorb RP-18 (150mm3.9mm, 10mm) Mobile phase solvents: SDS/H3PO4/n-propanol 20e50% v/v References Sin et al., 2006

DAD (l280nm; 220e320nm)

Dry food

Perrin & Meyer, 2002

DAD (l276nm) DAD DAD

Aqueous food simulants Aqueous food simulants Fatty food simulants Star fruit

BHA, BHT BHA, BHT BHA, BHT

Dopico-Garca et al., 2005 Dopico-Garca et al., 2007 Dopico-Garca et al., 2007 Shui & Leong, 2004

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

DAD (l280nm) HPLCeDADeESIe MS-ESIeMS 250 C 4.5kV DAD (l208nm)

AA

Shrimp waste

a-Tocopherol

Lopez-Cervantes et al., 2006 Dopico-Garca et al., 2003

DAD (l276nm)

Food simulant

BHA, BHT

DAD (l295nm) Fl (lex295nm, lem350nm)

Fresh leaves

Tocopherol

Gomez-Coronado et al., 2004

UV DAD (l280nm) UV (l254nm)

Liver pates

TBHQ, BHA, BHT, PG, DG, OG AA, EA

Pinho et al., 2000

Fruits and vegetables Vegetable oil, hydrogenated vegetable fat, margarine, vegetable cream, halvarina Vegetable oil, butter, cheese margarine, Oil sample

Thompson & Trenerry, 1995 Maziero et al., 2001

UV (l280nm)

TBHQ, BHA, BHT

UV (l280nm)

PG, TBHQ, BHA, BHT BHA, BHT, PG, OG, DG

Saad et al., 2007

UVeVis (l290nm)

Aparicio et al., 2000

UV (l265nm)

Fortied food products

Total vitamin C

UV (l280nm)

Bakery products

PG, OG, DG, BHT, BHA AA, EA

UV (l260nm)

UVeVis (l 297nm) NPeUV (l290nm) RPeHPTLCeUV (l254nm) RPeUV (l290nm) UV (l292nm) MS-APCI()() MS-ESI()() UVevis (l254nm)

Tea drinks, dried fruit, nutritional supplement Deffated rice bran Mix of tocopherols in ethanol Mix of tocopherols in chloroform Mix of tocopherols in ethanol Sunower oil milk

Tocopherol Tocopherol Tocopherol Tocopherol Tocopherol

UV (l245nm)

Semisolid pharmaceutical/ cosmetic ormulations Sea urchin

AA

AA

Fl (lex350nm, lem430nm) Fl (lex295nm, lem330nm) Fl (lex290nm, lem330nm) DRIFT-IR Fl (lex295nm, lem325nm)

Sea urchin

AA

Infant formulas

tocopherol

Plastic LDPE

a-tocopherol

Fl (lex355nm, lem350nm) 425nm emission Fl (lex350nm,

Olive, peanut, grapeseed, rapeseed, sunower,corn, soybean oils Various food, milk

tocopherols

CTAB/H3PO4/n-propanol 20e50% v/v Column: LiChrospher RP-C18 (250mm x4.6mm, 5mm) Mobile phase solvents: decylamine, AcN, sodium acetate solution; distilled water (pH5.4 phosphoric acid 85%); TCEP.HCl (isocratic elution) Column: C18 Extrasil ODS2 (250mm4.6mm, 5mm) Mobile phase solvents: A e acetic acid-MeOH (5:95,v/v); B e acetic acid-water (5:95 v/v) (gradient elution) Column: GL Sciences (250mm x4.6mm, 5mm) Mobile phase solvents: AcN:water with 66.7mM ammonium acetate (85:15,v/v) Column: Shim-pack (250mm4.6mm) Mobile phase solvents: n-Hex/isopropanol (96:4); v/v Column: Hypersil Silica (250mm4.6mm, 5mm) Mobile phase solvents: Hex:amyl alcohol (99.5:0.5, v/v) Column: RPeHPTLC (10cm 10cm) Solvents: EtOH:water volume proportions 10:0; 9.5:0.5; 9:1 Column: RP C18 (150mm4mm, 5mm) Mobile phase solvents: MeOH-water (9:1, v/v) Column: Fluophase PFP (200mm4.6mm, 5mm) Column temperature: 23 C Mobile phase solvents: MeOH:water 95:5,v/v or AcN:water 78:22, v/v Column: Phenomenex LiChrospher 100-RP18 (250mm4.6mm, 5mm) Mobile phase solvents: 0.2% metaphosphoric acid/MeOH/ AcN (90:8:2, v/v/v) Column: Kromasil 100 (250mm4mm, 5mm), Teknokroma Mobile phase solvents: Milli-Q Water adjusted to pH 2.2 with metaphosphoric acid Column: Kromasil 100 (250mm4mm, 5mm), Teknokroma Mobile phase solvents: Methanol/0.1% metaphosphoric acid (7:1) ColumnNova-Pak: (150mm3.9mm, 5mm) Column temperature: room temp Mobile phase solvents: Hex-ethyl acetate (98:2, v/v) Column: Omnispher C18 column (150mm4.6mm, 5mm); Microsorb Mobile phase solvents: MeOH:water (98:2) (isocratic elution) Column: Symmetry C18 (150mm3.9mm, 5mm) Mobile phase solvents: AcN/MeOH (1:1) (elution gradient)

Fontannaz et al., 2006

Rafecas et al., 1998 Tai & Gohda, 2007

Devi et al., 2007 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246 Pyka & Sliwiok, 2001 Pyka & Sliwiok, 2001 Pyka & Sliwiok, 2001 Lanina et al., 2007

Maia et al., 2007

Quiros et al., 2001

Quiros et al., 2001

Rodrigo et al., 2002 Graciano-Verdugo et al., 2006

Gliszczynska Swig1o, & Sikorska, 2004

AA

Emblica fruits,

AA

Column: Nova-Pak C18 (150mm3.9mm, 4mm) Mobile phase solvents:phosphate pH7.8 containing16%MeOH (v/v) (isocratic elution) Column: SS Excil C18 (250mm4.6mm, 10mm)

Burini, 2007

Raghu et al., 2007 237 (continued on next page)

Table 4 (continued). 238 Technique Detector lem430nm) Amperometric detection GC MS m/z 35e500 V70eV MS m/z 10e550 70eV Matrix guava, chilli fruits, amaranth leaves Chewing gum Drinking water PVC tubes Analytes Conditions Mobile phase solvents: MeOH:water (55:45) (isocratic elution) Nickel phthalocyanine polymer-coated glassy carbon electrodes Column: HP5 column (30m0.25mm) Oven ramp: 40 C 5min; to 250 C 20 C/min 5min Column: Supelcowax 10(30m0.25mm, thickeness 0.25mm) Injector temperature: 250 C Oven ramp: 35 C held 2min, increased at 15 C/min to 250 C Column: ChromPack CP-Sil 8CB (30m0.53mm, 1.5mm) Injector temperature: 250 C; Detector temperature: 290 C Oven ramp:150 C held 1min, increased to 200 C at 7 C/ min, increased to 300 C at 40 C/min, held 5min at 300 C Carrier gas: H2 (4ml/min) References

AA BHT BHT

Ruiz et al., 1999 Tombesi & Freije, 2002 Hill et al., 2003

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

FID

FID

FID

Cooking oil, sh oil, butter, margarine, cheese, mayonnaise, salad dressing Soybean oil, corn oil, other vegetable oil, shortening, margarine, seasoned dried sh, dried sh, salted shery product and frozen shery product (combined), chewing gum, mayonnaise and breakfast cereal Oil

BHA, BHT, TBHQ

Yang et al., 2002

BHA, BHT, TBHQ

Suh et al., 2005

BHA, BHT, PG, OG, DG

Column: AT-5 (30m0.53mm) Injector temperature: and detector temperature: 250 C Column oven: 125 C held 20min; increased to 250 C at 20 C/min Carrier gas. H2 30mL/min Air 300mL/min Column: Fused-silica capillary (75cm25mm i.d. and 360mm o.d.) Electrochemical cell (WE: 300mm diameter carbon disk platimium electrode SCE) in combination with BAS LC4C AD Uncoated fused-silica capillary (total length50cm; effective length to the detector42.4cm; i.d.75mm) conditioned with 0.1M NaOH (3min); CE-grade water (5min) rinsed with running buffer 50mM tricine at pH 8.8 (5min) sample injection carried out at the anode for 5s. T30 C; V11kV across the capillary applied for 15min.

Sin et al., 2006

Electroforesis

CE Electrochemical detection

Grapefruit

AA

Wu et al., 2007

CE Capillary zone mode (CZE) UVe Vis

Fruit juice

AA and EA

Versari et al., 2004

MECC l254nm

Fruit and vegetables

AA and EA

MECC-UV l200, 214, 254, 280nm

Edible oils

BHA, BHT, DG

MECC electrochemical detection

Stock solutions

BHA, BHT, PG, TBHQ

Microchip MEC TLC

Food sample Edible oil

PG, OG, DG BHA, BHT, PG, OG, DG

Fused-silica capillary columns (65cm7.5mm i.d., effective length to the detector40cm for fruit; 75cm75mm id. effective length to the detector50cm for vegetables) Buffer 0.05M sodium deoxycholate, 0.01M sodium borate and 0.01M potassium dihydrogen orthophosphate. V25kV; T28 C; Buffer for 2min between runs. Column: Fused-silica capillary (57cm75mm i.d.) Separation solution: 20% AcN, 20mM boric-borate buffer (pH9.2) and 20mM bis-(2-ethylhexyl))sodium sulfosuccinate surfactant Column: fused-silica capillary (75cm25mm i.d. and 360mm o.d.) Injector: V18kV for 6s; T25 C Electrochemical cell (WE: 300mm diameter carbon discplatinium electrodeSCE in combination with BAS LC-4C AD Detection electrode: 25mm diameter, 99.95% gold wire T222 C TLC plate: silica gel 60, 2020cm2, Merck Mobile phases: Petroleum ether(40:60):toluene:acetic acid:AcN (4:4:1:0.4 by volume) Spraying 0.5% Gibbs reagent; dried at 105 C for 15min Exposed to ammonia vapour 1100e2498nm, 32 scans/spectrum 4000e400cm1 WE: glassy carbon disk (GC, 0.950cm2) and Pt disk (Pt, 0.502cm2). Reference: Hg/Hg2Cl2/KCl (sat.) electrodes and home made plate auxiliary electrode Cyclic voltammetry: yb50mVs1 Differential pulse voltammetry: DE25mV; yb50mVs1 Square wave voltammetry: DE25mV; f60Hz

Thompson & Trenerry, 1995

DelgadoZamarreno et al., 2007 Guan et al., 2006 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

Ding et al., 2006 Sin et al., 2006

NIR DRIFT-IR Voltametric techniques

Pharmaceutical products Plastic LDPE Mayonnaise

AA A-Tocopherol BHA, TBHQ, BHT

Blanco & Alcala, 2006 Graciano-Verdugo et al., 2006 Raymundo et al., 2007

239

240

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

Table 5. Method performance of analytical methods for determination of antioxidants. Compound TBHQ Food group Meat and meat products Fish and sh products Cheese and cheese products Milk and cheese products Fats, oils and their products Range n.d. 7.35103 mg/kg n.d. n.d. e n.d.e 20.2 mg/kg LOD e 0.80 106mol/l 0.1mg/ml 0.5mg/l 0.12 ng 0.5mg/l Recovery References

93.6% at 30.3mg/mLPinho et al., 2000 95.40% Guan et al., 2006

BHA

Other foods (mushroom cream) Several foods e 5mg/kg (shortening, e 2mg/kg seasoned dried sh, dried sh, salted shery product and frozen shery product, chewing gum, mayonnaise, breakfast cereal, etc) Food for special e 0.12 ng nutritional use Meat and meat 92.4 mg/kg e products Fish and sh n.d. 1.0106mol/l products Cheese and cheese n.d. 0.5mg/l products 5.4e12.4 mg/kg 0.1mg/ml Milk and cheese e 0.3 ng products Bakery (doughnuts, Sample: e biscuits, various 3.9e10.7 mg/kg types of cakes) Fat fraction: 17.5e55.6 mg/kg Other foods n.d. 1.0106mol/l (mushroom cream) Fats, oils and their n.d.e103.9 mg/kg 0.5mg/l products e n.d.-135.4 mg/kg n.d. 0.41-0.67pg 0.1mg/ml 0.45ppm

n.d.e70.3mg/kg n.d.e 143 mg/kg e 56.7e73.8 mg/kg n.d. n.d.

0.1mg/ml 3.0mg/kg 5mg/kg e 0.80 106mol/l 0.80 106mol/l

97.3e108.4% Yang et al., 2002 Cheese 85.3e93.1%Saad et al., 2007 98e105% Noguera-Ort et al., 2000 Oil: 103.9e104.0% Saad et al., 2007 Bread Spread: 95.0e 108.3% 97.3e108.4% Yang et al., 2002 103e108% Maziero et al., 2001 95.40% Suh et al., 2005 e Raymundo et al., 2007 95.80% Guan et al., 2006 99.70% Guan et al., 2006 88.70% e Suh et al., 2005 Perrin & Meyer, 2002

94.8e100.8% 97.80% 102.30% 97.4e101.2% 92e102% 94.5e95.8%

Noguera-Ort et al., 2000 Pinho et al., 2000 Guan et al., 2006 Saad et al., 2007 Yang et al., 2002 Noguera-Ort et al., 2000 Rafecas et al., 1998

97.40% Oil-96.7e97.6% Bread spread-97.5e 97.7% 96.2e105.7% 101.2e103.8% Found 0.75g/Kg adding 0.8g/Kg (MeOH) 0.79g/kg adding 0.8g/Kg (sunower oil) 0.70g/Kg adding 0.8g/Kg e 97.60% 81.80% 87e98% 96.50% e 93.4e99.7%

Guan et al., 2006 Saad et al., 2007

Aparicio et al., 2000 Yang et al., 2002 Delgado-Zamarreno et al., 2007

Food for special nutritional use

21.3e58.1mg/kg 164mg/kg e n.d. n.d. e e

e 2e4mg/g 3mg/kg 0.6mg/kg 1.0106mol/l 0.6mg/kg 0.3 ng

Raymundo et al., 2007 Sin et al., 2006 Suh et al., 2005 Maziero et al., 2001 Guan et al., 2006 Maziero et al., 2001 Noguera-Ort et al., 2000

 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246 Table 5 (continued ) Several foods e (shortening, seasoned dried sh, dried sh, salted shery product and frozen shery product, chewing gum, mayonnaise, breakfast cereal, etc) Dry foods e Chewing gum n.d. Food simulant e 4.2103mgdm2 (aqueous simulant) 4.4103mgdm2 (simulant D-olive oil) 16103e30103g/l 3mg/kg 89.20% Suh et al., 2005

241

5.5ng 0.052mgL1

95e104% e 104%

Perrin & Meyer, 2002 Ruiz et al., 1999 Dopico-Garca et al., 2005 Dopico-Garca et al., 2005

Plastic products BHT Meat and meat products Fish and sh products Cheese and cheese products

6.5mgl1

88% 99.90% 95.90%

Dopico-Garca et al., 2003 Pinho et al., 2000 Guan et al., 2006 Yang et al., 2002 Saad et al., 2007 Guan et al., 2006 Rafecas et al., 1998

e n.d. 56.7e87.1mg/kg n.d.

e 2.7106mol/l 0.1mg/ml 0.5mgl1 2.7106mol/l e

Other foods n.d. (mushroom cream) Bakery products Sample: 3.0e17.0mg/kg Fat fraction: 14.9e90.0mg/ kg Fats, oils and their e products 17.0e319mg/kg n.d. e 154.2mg/kg

Cheese e 89.5e 91.4% 94.30% 85.7e89.4%

3mg/kg 0.1mg/ml 0.5mgl1

46.5mg/kg e 17.6e26.7mg/kg n.d.

2e4mg/g/10mg/g 1.42e5.12pg e 1.47ppm

Other foods Beverages Chewing gum Food simulant

n.d. e 197mg/kg n.d. e e 0e38mg/L 513mgg1 e n.d.

2.7mg/kg 2.7106mol/l 2mg/kg 3mg/kg 4.2ml1 30ng 0.045mgl1

94.50% 92.5e97.6% Oil e 88.2e94.6% Bread spread e 73.9e80.2% 75.80% 83.0e85.5% e Adding 0.8g/kg found 0.064g/kg (MeOH) 0.99g/kg (Sunower oil) 0.64g/kg (olive oil) 78e98% 92.10% e 92.60% 84%e119% 974% 82%

Suh et al., 2005 Yang et al., 2002 Saad et al., 2007

Sin et al., 2006 Aparicio et al., 2000 Raymundo et al., 2007 Delgado-Zamarreno et al., 2007

Plastic products

5.1103 mgdm2 (aqueous simulant) 7.7103 mgdm2 (simulant D) 10 0.13nmol e 5.1e5.610 mol 70% 16e30mg/l/100mL simulant 5.6mgl1 n.d. n.d. 78.00% 98.70%

Maziero et al., 2001 Guan et al., 2006 Perrin & Meyer 2002 Suh et al., 2005 Tombesi & Freije, 2002 Ruiz et al., 1999 Dopico-Garca et al., 2005 Dopico-Garca et al., 2007 Hill et al., 2003 Dopico-Garca et al., 2003 Pinho et al., 2000 Guan et al., 2006 (continued on next page)

PG

Meat and meat products Fish and sh

e 0.29106mol/l

242 Table 5 (continued). Compound Food group

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

Range n.d. n.d.

LOD 0.3mgl1

Recovery Oil e 103.5e 108.3% Bread spread e 103.6e107.3% Spike 0.01 e 7.6% 95.3e100.9% 99.10% 93.3e99.5% 96e103% 101.80% 88.9e89.5% 100.6e101.3% 956% e 85e106% 82.10% 77.9e79.7% 99e100.1% 92.5e105.0% 95.80% 956% 100e103% 83e85%

References Saad et al., 2007

products Fats, oils and their products

e e 103mg/kg Cheese and cheese n.d. products Milk and cheese e products Other foods 217mg/kg (mushroom cream) Bakery products n.d. Food for special e nutritional use Mixed foods 45mg/kg Chewing gum n.d. Other foods e OG Meat and meat products Bakery products Milk and cheese products Fats, oils and their products Mixed foods Food for special nutritional use Other foods DG Meat and meat products Bakery products Fats, oils and their products n.d. n.d. e e e e e e n.d. n.d. e e n.d.

2e4mg/g 0.14e0.23pg 0.29106mol/l 0.3mgl1 0.05ng 0.29106mol/l e 0.05ng 2.2mM 7.5ng

Sin et al., 2006 Aparicio et al., 2000 Guan et al., 2006 Saad et al., 2007 Noguera-Ort et al., 2000 Guan et al., 2006 Rafecas et al., 1998 Noguera-Ort et al., 2000 Ding et al., 2006 Ruiz et al., 1999 Perrin & Meyer, 2002 Pinho et al., 2000 Rafecas et al., 1998 Noguera-Ort, et al., 2000 Aparicio et al., 2000 Sin et al., 2006 Ding et al., 2006 Noguera-Ort et al., 2000 Perrin & Meyer, 2002 Pinho et al., 2000 Rafecas et al., 1998 Sin et al., 2006 Aparicio et al., 2000 Delgado-Zamarreno et al., 2007

e e 0.2ng 0.31e0.40pg 2e4mg/g 2.3mM 0.2ng

e e 2e4mg/g 0.30e0.33pg 0.27ppm

94.30% 39.8e40.7% 97.60% 95.7e105.7% Adding 0.8g/kg, found 0.77g/kg (MeOH), 0.75g/kg (sunower oil, 0.82g/kg (olive oil) 956% 100.80%

Mixed foods Tocopherol Fish and sh products Fats, oils and their products

e 351mg/kg 100.16e203.78mg/kg

4.1mM 80ng/mL 28ng/mL (a-tocopherol) 8ng/mL (g and d tocopherol) APCI(-): 2.5e3ng/mL ESI(-): 5e20ng/mL 0.05mg/100g e

Ding et al., 2006 Lopez-Cervantes et al., 2006 Gliszczynska-Swig1o, & Sikorska, 2004 Lanina et al., 2007 Gomez-Coronado et al., 2004 Devi et al., 2007

393mg/kg Vegetables and 13e1322mg/kg vegetable products Cereals and cereal 51e347ppm (all products tocopherols)

102e112% a e 87% g e 57e60% 19.9%e47.7% based on crude extraction 104e109%

Milk and milk 0.939mg/kg APCI(): 3.0ng/mL Lanina et al., 2007 products ESI(): 20ng/mL Food for special 275mg/kg 1.60ng/50mL inj volume e Rodrigo et al., 2002 nutritional use Plastic product (LPPE0.26e1.59% (HPLC)/0.21e 0.25mg/mL (HPLC)/1300mg/ 71.3e86.0% (HPLC) Graciano-Verdugo et al., lm) 1.42% (DRIFT-IR) g (DRIFT-IR) 2006

 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

243

Table 5 (continued). Fruits 979.17106g/mL (grapefruit juice)/500 106g/g grapefruit peel 6.2e208.5mg/100g 30mg/L 31.9e267.5mg/l 1106g/mL 95% Wu et al., 2007

Total vitamin C Ascorbic acid

Food for special nutritional use Beverages

0.1mg/100g 3mg/L (conventional CE) 10mg/L (microship E) 0.3mg/ml

93e105%

Fontannaz et al., 2006 Law, Kuban et al., 2005

Fruit and fruit products

500106g/g (grapefruit peel)/979.17106g/g (grapefruit juice) 45e243mg/l 58.6e236.2mg/100g (HPLC) 52.4e213.5 (Enzymatic) 71.1e206.8 (indophenolxylene) 55.0e468.8 (2,4-DNPH) 2e38mg/100g 2e101mg/100g n.d.

1.0106g/mL

AcN/water 30/70 Tai & Gohda, 2007 using dithiothreitol: 92% AcN/water 30/70: Wu et al., 2007 38% 95%

1.6mg/l e

e e

Versari et al., 2004 Raghu et al., 2007

0.27mg/mL e 0.3mg/ml

e e

Vegetables and 41.1mg/100g (HPLC) e vegetable products 36.8mg/100g (Enzymatic) 60.4mg/100g (indophenolxylene) 62.3mg/100g (2,4-DNPH) 2e200mg/100g e 2.8e7.0mg/100g 1.15e1.44mg/kg 0.27mg/mL 0.27mg/mL

AcN/water 30/70 using dithiothreitol: 92% AcN/water 30/70: 38% e Raghu et al., 2007

Burini, 2007 Thompson & Trenerry, 1995 Tai & Gohda, 2007

e e e

Milk and milk products Sea Food

Thompson & Trenerry, 1995 Burini, 2007 Burini, 2007

Pharmaceutical/ cosmetic products Erythorbic acid Beverages

HPLCeFl: Fresh 26.57mg/ HPLCeFl: 0.19mg/ml, 100g 0.082mg/ml Canned 14.25mg/100g 0.05mg/mL1 1.909e10.154mg/ml 0.17mg/mL1 2.5% wt. e 23.4mg/ml 0.3mg/ml

98.12% (HPLCeFl) Quiros et al., 2001

95.46e101.54% e

Maia et al., 2007 Blanco & Alcala 2006

Food for special nutrition use Fruit and fruit products Fruit and fruit products

n.d. n.d. e 1.1mg/l

AcN/water 30/70 Tai & Gohda, 2007 using dithiothreitol: 92% AcN/water 30/70: 38%

Versari et al., 2004

244

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246

Conclusions The use of antioxidants has been growing in the past few years due to the advantages offered in the preservation of food. The most common method to analyse these antioxidants is reversed phase HPLC with UV detection. This UV detection can be a simple UVevis detector or DAD. Although tocopherol can be detected by RPeHPLC there are methods in normal phase. Various solvents can be used, but usually is selected an organic phase, AcN or methanol and an aqueous phase, water acidied or not. The present legislation is being very strict in the concentration limits of antioxidants that can be use in various foods. The analytical methods reviewed for this article presents limits of detection below the maximum permitted by legislation. In this way the methods used are suitable for detect and quantify the antioxidants in study. Acknowledgements Authors are grateful to the Fundac~o para a Ciencia e a Tecnologia, Portugal, for the Postdoctoral contract of Ana Sanches Silva in the frame of the Program Science 2007 and for the Bilateral Cooperation between Portugal and Italy. The authors are also grateful to Instituto Nacio nal de Saude Doutor Ricardo Jorge for the PhD scholar ship of Catarina Andre (BID 04/2006-I).

PDA: photodiode array RP: reverse phase SCE: saturated calomel electrode SFE: supercritical uid extraction SPME: solid-phase microextraction TCEP$HCl: tris-(2-carboxyethyl)-phosphine chloride THF: tetrahydrofuran TLC: thin layer chromatography UVevis: ultraviolet and visible detector VT: voltametric techniques WE: working electrode Wt.: weight References

hydro-

Abbreviations AA: ascorbic acid AcN: acetonitrile AD: amperometric detection APCI: atmosphere pressure chemical ionization CE: capillary electrophoresis DAD: diode array detector DDTD: dynamic thermal desorption DGLG: dodecyl gallate or lauryl gallate DRIFT: diffuse reectance Fourier transform infrared EA: erythorbic acid (D-isoascorbic) ED: electrochemical detection ESI: electrospray ionization EtOH: ethanol FID: ame ionization detector Flu: uorescence detector GC: gas chromatography Hex: hexane HILIC: hydrophilic interaction chromatography HPLC: high-performance liquid chromatography IR: infrared LC: liquid chromatography MECC: micellar electrokinetic capillary chromatography MeOH: methanol MLC: micellar liquid chromatography MS: mass spectrometry NIRS: near infrared spectroscopy NP: normal phase

AOAC Ofcial Method 983.15 AOAC (1995). Phenolic antioxidants in oils, fats and butter oil, liquid chromatographic method. AOCS, Ofcial Method Ce 6e86 AOAC (1997). Antioxidants. Liquid chromatographic method. Aparicio, A., Andres, M. P. S., & Vera, S. (2000). Separation and determination of phenolic antioxidants by HPLC with surfactant/npropanol mobile phases. Journal of High Resolution Chromatography, 23(4), 324e328. Blanco, M., & Alcala, M. (2006). Simultaneous quantitation of ve active principles in a pharmaceutical preparation: development and validation of a near infrared spectroscopic method. European Journal of Pharmaceutical Sciences, 27, 280e286. Burini, G. (2007). Development of a quantitative method for the analysis of total L-ascorbic acid in foods by high-performance liquid chromatography. Journal of Chromatography A, 1154, 97e102. Capitan-Vallvey, L. F., Valencia, M. C., & Nicolas, E. A. (2003). Simple resolution of butylated hydroxyanisole and n-propyl gallate in fatty foods and cosmetics samples by ow-injection solid-phase spectrophotometry. Journal of Food Science, 68(5), 1595e1599. CGSFA (1995). Codex general standards for food additivesdpreamble, Codex STAN 192-1995. (Revision 1997, 1999, 2001, 2003, 2004, 2005, 2006, 2007, 2008). Available on-line. http://www. codexalimentarius.net/gsfaonline/CXS_192e.pdf Delgado-Zamarreno, M. M., Gonzalez-Maza, I., Sanchez-Perez, A., & Martnez, R. C. (2007). Analysis of synthetic phenolic antioxidants in edible oils by micellar electrokinetic capillary chromatography. Food Chemistry, 100, 1722e1727. Devi, R. R., Jayalekshmy, A., & Arumughan, A. (2007). Antioxidant efcacy of phytochemical extracts from defatted rice bran in the bulk oil system. Food Chemistry, 104, 658e664. Ding, Y., Mora, M. F., & Garcia, C. D. (2006). Analysis of alkyl gallates and nordihydroguaiaretic acid using plastic capillary electrophoresis-microchips. Analytica Chimica Acta, 561, 126e132. Dopico-Garca, M. S., Lopez-Vilarino, J. M., & Gonzalez Rodrguez, M. V. (2003). Determination of antioxidant migration level from low-density polyethylene lms into food simulants. Journal of Chromatography A, 1018, 53e62. pez-Vilarin J. M., & Gonzalez-Rodrguez, M. V. Dopico-Garca, M. S., Lo o, (2005). Determination of antioxidants by solid-phase extraction method in aqueous food simulants. Talanta, 66, 1103e1107. Dopico-Garca, M. S., Vilarino, J. M., & Gonzalez Rodrguez, M. V. (2007). Antioxidant content of and migration from commercial polyethylene, polypropylene and polyvinyl chloride packages. Journal of Agricultural and Food Chemistry, 55, 3225e3231. EFSA (2004a). Opinion of the scientic panel on dietetic products, nutrition and allergies on a request from the commission related to the tolerable upper intake of vitamin C (L-ascorbic, its calcium, potassium and sodium salts and L-ascorbyl-6-palmitate). The EFSA Journal, 59, 1e21.

 C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246 EFSA (2004b). Opinion of the scientic panel on food additives, avourings, processing aids and materials in contact with foods on a request from the commission related to tertiary-butylhydroquinone (TBHQ). The EFSA Journal, 84, 1e50. EFSA (2008). Opinion on mixed tocopherols, tocotrienol tocopherol and tocotrienols as sources for vitamin E added as a nutritional substance in food supplements. Scientic opinion of the panel on food additives, avourings, processing aids and materials in contact with. The EFSA Journal, 640, 1e34. European Parliament and Council Directive N 95/2/EC of 20 February 1995 on food additives other than colours and sweeteners. Brussels. Ofcial Journal of the European Communities18 March 1995. 15.08.2006. 1a FDA. 2001a. Code of Federal Regulations (CFR), 2001a. CFR (Code of Federal Regulations), Title 21: Food and Drugs. Chapter I e Food and Drug Administration, Department of Health and Human Services, Part 170 e Food Additives, Ofce of the Federal Register, Washington, DC (Revised April, 2008). 1b FDA. 2001b. Code of Federal Regulations (CFR), 2001b. CFR (Code of Federal Regulations), Title 21: Food and Drugs. Chapter I e Food and Drug Administration, Department of Health and Human Services, Part 184 e Direct Food Substances afrmed as generally recognized as safe (GRAS), Ofce of the Federal Register, Washington, DC (Revised April, 2008). Fidler, M. C., Davidsson, L., Zeder, C., & Hurrell, R. F. (2004). Erythorbic acid is a potent enhancer of nonheme-iron absorption. The American Journal of Clinical Nutrition, 79, 99e102. Fontannaz, P., Kilinc, T., & Heudi, O. (2006). HPLCeUV determina tion of total vitamin C in a wide range of fortied food products. Food Chemistry, 94, 626e631. Garca, R. S., Silva, A. S., Cooper, I., Franz, R., & Losada, P. P. (2006). Revision of analytical strategies to evaluate different migrants from food packaging materials. Trends in Food Science and Technology, 17, 354e366. Georgantelis, D., Blekas, G., Katikou, P., Ambrosiadis, I., & Fletouris, D. J. (2007). Effect of rosemary, chitosan and a-tocopherol on lipid oxidation and colour stability during frozen storage of beef burgers. Meat Science, 75, 256e264. Gharavi, N., & El-Kadi, A. O. S. (2005). tert-Butylhydroquinone is a novel aryl hydrocarbon receptor ligand. Drug Metabolism and Disposition, 33(3), 365e372. Gliszczynska-Swig1o, A., & Sikorska, E. (2004). Simple reversed-phase liquid chromatography method for determination of tocopherols in edible plant oils. Journal of Chromatography A, 1048, 195e198. Gomez-Coronado, D. J. M., Ibanez, E., Ruperez, F. J., & Barbas, C. (2004). Tocopherol measurement in edible products of vegetable origin. Journal of Chromatography A, 1054, 227e233. Graciano-Verdugo, A. Z., Peralta, E., Gonzalez-Ros, H., & SotoValdez, H. (2006). Determination of a-tocopherol in low-density polyethylene (LDPE) lms by diffuse reectance Fourier transform infrared (DRIFT-IR) spectroscopy compared to high performance liquid chromatography (HPLC). Analytica Chimica Acta, 557, 367e372. Guan, Y., Chu, Q., Fu, L., Wu, T., & Ye, J. (2006). Determination of phenolic antioxidants by micellar electrokinetics capillary chromatography with electrochemical detection. Food Chemistry, 94, 157e162. Hill, S. S., Shaw, B. R., & Wu, A. H. B. (2003). Plasticizers, antioxidants, and other contaminants found in air delivered by PVC tubing used in respiratory therapy. Biomedical Chromatography, 17, 250e262. Hirose, M., Takahashi, S., Ogawa, K., Futakuchi, M., Shirai, T., Shibutani, M., et al. (1999). Chemoprevention of heterocyclic amine-induced carcinogenesis by phenolic compounds in rats. Cancer Letters, 143, 173e178. IUPAC Ofcial Method 2.642 IUPAC (1992). Determination of antioxidants by HPLC. Joint FAO/WHO Expert Committee on Food Additives (JEFCA), & World Health Organization (WHO) (2003). Summary of evaluations performed by the Joint FAO/WHO Expert Committee on Food

245

Additives (JECFA 1956e2007). Available on-line. http://jecfa.ilsi. org/index.htm. www.efsa.europa.eu/EFSA. www.fao.org Karoviova, J., & Simko, P. (2000). Determination of synthetic phec nolic antioxidants in food by high-performance liquid chromatography. Journal of Chromatography A, 882, 271e281. Kuwabara, K., Watanabe, Y., Adachi, S., Nakanishi, K., & Matsuno, R. (2003). Emulsier properties of saturated acyl L-ascorbates for preparation of O/W emulsions. Food Chemistry, 82, 191e194. Lanina, S. A., Toledo, P., Sampels, S., Kamal-Eldin, A., & Jastrebova, J. A. (2007). Comparison of reversed-phase liquid chromatographyemass spectrometry with electrospray and atmospheric pressure chemical ionization for analysis of dietary tocopherols. Journal of Chromatography A, 1157, 159e170. Law, W. S., Kuban, P., Zhao, J. H., Li, S. F. Y., & Hauser, P. C. (2005). Determination of vitamin C and preservatives in beverages by conventional capillary electrophoresis and microchip electrophoresis with capacitively coupled contactless conductivity detection. Electrophoresis, 25, 4648e4655. pez-Cervantes, J., Sanchez-Machado, D. I., & Rios-Vazquez, N. J. (2006). Lo High-performance liquid chromatography method for the simultaneous quantication of retinol, a-tocopherol, and cholesterol in shrimp waste hydrolysate. Journal of Chromatography A, 1105, 135e139. Maia, A. M., Baby, A. R., Yasaka, W. J., Suenaga, E., Kaneko, T. M., & Velasco, M. V. R. (2007). Validation of HPLC stability-indicating method for vitamin C in semisolid pharmaceutical/cosmetic preparations with glutathione and sodium metabisulte, as antioxidants. Talanta, 71, 639e643. Maziero, G. C., Baunwart, C., & Toledo, M. C. (2001). Estimated of the theoretical maximum daily intake of phenolic antioxidants BHA, BHT and TBHQ in Brazil. Food Additives and Contaminates, 18(5), 365e373. Morales, M. D., Gonzalez, M. C., Reviejo, A. J., & Pingarron, J. M. (2005). A composite amperometric tyrosinase biosensor for the determination of the additive propyl gallate in foodstuffs. Microchemical Journal, 80, 71e78. Morales, M. D., Gonzalez, M. C., Serra, B., Reviejo, A. J., & Pingarron, J. M. (2005). Composite amperometric tyrosinase biosensors for the determination of the additive propyl gallate in a reversed micellar medium. Sensors and Actuators B, 106, 572e579. Noguera-Ort, J. F., Villanueva-Camanas, R. M., & Ramis-Ramos, G. (2000). Determination of synthetic antioxidants in dairy products and dietetic supplements by micellar liquid chromatography with direct sample injection. Chromatographia, 51(1/2), 53e60. Perrin, C., & Meyer, M. (2002). Quantication of synthetic phenolic antioxidants in dry foods by reversed-phase HPLC with photodiode array detection. Food Chemistry, 77, 93e100. Pinho, O., Ferreira, I. M. P. L. V. O., Oliveira, M. B. P. P., & Ferreira, M. A. (2000). Quantication of synthetic phenolic anti oxidants in liver pates. Food Chemistry, 68, 353e357. Pyka, A., & Sliwiok, J. (2001). Chromatographic separation of tocopherols. Journal of Chromatography A, 935, 71e76. Quiros, A. R., Lopez-Hernandez, J., & Simal-Lozano, J. (2001). Determination of vitamin C in Sea urchin: comparison of two HPLC methods. Chromatographia, 53(Suppl.), S-246eS-249. Rafecas, M., Guardiola, F., Illera, M., Codony, R., & Boatella, J. (1998). Liquid chromatographic determination of phenolic antioxidants in bakery products. Journal of Chromatography A, 822, 305e309. Raghu, V., Platel, K., & Srinivasan, K. (2007). Comparison of ascorbic acid content of Emblica ofcinalis fruits determined by different analytical methods. Journal of Food Composition and Analysis, 20, 529e533. Raymundo, M. S., Paula, M. M. S., Franco, C., & Fett, R. (2007). Quantitative determination of the phenolic antioxidants using voltammetric techniques. LWT e Food Science and Technology, 40, 1133e1139. Regulation (EC) No 1333/2008 of the European parliament and the Council of 16 December 2008 on food additives. Ofcial Journal of the European Union, 31 December 2008, L 354. 16.

246

C. Andre et al. / Trends in Food Science & Technology 21 (2010) 229e246 hydroquinone (TBHQ) antioxidants in Korea. Food Additives and Contaminates, 22(12), 1176e1188. Tai, A., & Gohda, E. (2007). Determination of ascorbic acid and its related compounds in foods and beverages by hydrophilic interaction liquid chromatography. Journal of Chromatography B, 853, 214e220. The Australia New Zealand Food Standards Code (2009). Standard 1.3.1, food additives. Available on-line. http://www.foodstandards. gov.au/thecode/foodstandardscode/ Thompson, C. O., & Trenerry, V. C. (1995). A rapid method for the determination of total L-ascorbic acid in fruits and vegetables by micellar electrokinetic capillary chromatography. Food Chemistry, 53, 43e50. Tombesi, N., & Freije, H. (2002). Application of solid-phase microextraction combined with gas chromatography mass spectrometry to the determination of butylated hydroxytoluene in bottled drinking water. Journal of Chromatography A, 963, 179e183. Tsuji, S., Nakanoi, M., Terada, H., Tamura, Y., & Tonogai, Y. (2005). Determination and conrmation of ve phenolic antioxidants in foods by LC/MS and GC/MS. Shokuhin Eiseigaku Zasshi, 46(3), 63e71. Versari, A., Mattioli, A., Parpinello, G. P., & Galassi, S. (2004). Rapid analysis of ascorbic acid and isoascorbic acids in fruit juice by capillary electrophoresis. Food Control, 15, 355e358. Wu, T., Guan, Y., & Ye, J. (2007). Determination of avonoids and ascorbic acid in grapefruit peel and juice by capillary electrophoresis with electrochemical detection. Food Chemistry, 100, 1573e1579. Yang, M., Lin, H., & Choong, Y. (2002). A rapid gas chromatographic method for direct determination of BHA, BHT and TBHQ in edible oils and fats. Food Research International, 35, 627e633. Zurita, J. L., Jos, A., Peso, A., Salguero, M., Lopez-Artguez, M., & Repetto, G. (2007). Ecotoxicological effects of the antioxidants additive propyl gallate in ve aquatic systems. Water Research, 41, 2599e2611.

Robbins, R. J. (2003). Phenolic acids in foods: an overview of analytical methodology. Journal of Agricultural and Food Chemistry, 51(10), 2866e2887. Rodrigo, N., Alegra, A., Barbera, R., & Farre, R. (2002). High performance liquid chromatography determination of tocopherols in infant formulas. Journal of Chromatography A, 847, 97e102. Ruiz, M. A., Garca-Moreno, E., Barbas, C., & Pingarron, J. M. (1999). Determination of phenolic antioxidants by HPLC with amperometric detection at a nickel phthalocyanine polymer modied electrode. Electroanalysis, 11(7), 470e474. Saad, B., Sing, Y. Y., Nawi, M. A., Hashim, N. H., Ali, A. S. M., Saleh, M. I., et al. (2007). Determination of synthetic phenolic antioxidants in food items using reversed-phase HPLC. Food Chemistry, 105, 389e394. Sauberlich, H. E., Tamura, T., Craig, C. B., Freeberg, L. E., & Liu, T. (1996). Effects of erythorbic acid of vitamin C metabolism in young women. The American Journal of Clinical Nutrition, 64, 336e346. SciFinder. (2008). American Chemical Society, SciFinder database online (subscription database). Shui, G., & Leong, L. P. (2004). Analysis of polyphenolic antioxidants in star fruit using liquid chromatography and mass spectrometry. Journal of Chromatography A, 1022, 67e75. Sin, D. W. M., Wong, Y. C., Mak, C. Y., Sze, S. T., & Yao, W. Y. (2006). Determination of ve phenolic antioxidants in edible oils: method validation and estimation of measurement uncertainty. Journal of Food Composition and Analysis, 19, 784e791. Suh, H. J., Chung, M. S., Cho, Y. H., Kim, J. W., Kim, D. H., Han, K. W., et al. (2005). Estimated daily intakes of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl