J Package Technol Res (2017) 1:2531
DOI 10.1007/s41783-017-0004-3
RESEARCH ARTICLE
Modified Atmosphere Packaging (MAP) and the Effect
of Chemical Preservative to Enhance Shelf Life of Khoa
(Heat Desiccated Milk Product)
Tanima Chowdhury1 Sanjay Kr. Chattopadhyay1 Narayan Ch. Saha1
Received: 18 April 2016 / Accepted: 13 July 2016 / Published online: 16 March 2017
Indian Institute of Packaging 2017
Abstract Khoa is an indigenous heat desiccated milk
product having very poor shelf life. At ambient condition,
shelf-life of Khoa is normally only 2 days. The aim of
present study was to determine the effect of benzoic acid as
preservative as well as modified atmosphere packaging
(MAP) technology to enhance shelf life of Khoa at
27 2 C and 65% RH. During storage, analysis of
chemical, sensory as well as microbiological characteristics
were taken into consideration to mark distinguishable
changes between the package of modified atmosphere
technology (MAP) and ordinary packed Khoa (with and
without preservative) samples. The results indicated a
significant decrease of moisture content, pH and sensory
scores and increase in titratable acidity, standard plate
count and yeast and mould count during storage, irre-
spective of the type of packaging conditions. However, the
rate of changes in characteristics of product packed in
modified atmosphere was found to be slow. The storage
study indicated that the Khoa packed in ordinary packag-
ing, with and without preservative, was acceptable for 8
and 4 days, respectively, whereas for modified atmosphere
packed samples, it was consumable up to 12 and 8 days
respectively.
Keywords Khoa
Shelf life
Benzoic acid
Modified
atmosphere packaging
& Narayan Ch. Saha
director-iip@iip-in.com
1
Indian Institute of Packaging, Mumbai 400093, India
Introduction
India produces about 140.6 million tons of milk annually.
About 50% of milk out of total production is converted to
various traditional dairy products like heat desiccated milk
products viz., Khoa, Basundi, Lalpeda, Rabri, Kalakand
etc., coagulated milk products viz., Dahi, Shrikhand,
Paneer, Chhana and Chhana based products and heat
clarified products viz., butter oil, Ghee etc., which have a
strong social and cultural heritage in the Indian society [1].
Among the indigenous milk products, Khoa occupies a
prominent place as it forms a base for number of sweet
delicacies.
Khoa is a concentrated whole milk product obtained by
open pan condensing of milk under atmospheric pressure.
According to Food Safety and Standard Authority of India
(FSSAI) [2] Khoa, by whatever variety of names it is sold
such as Pindi, Danedar, Dhap, Mawa or Kava, means the
product obtained from cow or buffalo or goat or sheep milk
or milk solids or a combination thereof by rapid drying.
The milk fat content shall not be less than 30% on dry
weight basis of finished product. It may contain citric acid
not more than 0.1% by weight. It shall be free from added
starch, added sugar and added colouring matter. All of the
Khoa varieties are in demand which are required for
making value added Khoa based products like Burfi, Peda,
Gulabjamun, pantua, Kalakand, milk cake, Kunda, etc.
Khoa is also used for stuffing vegetables in many food
items.
Shelf-life is defined as the number of days the product
can be stored on shelf at a particular climatic condition and
can be consumed safely without deterioration of its
organoleptic properties. The shelf-life depends on the
microbiological status and chemical characteristics of the
product [3]. With innovations in packaging materials and
123
26
systems like vacuum packaging, modified atmosphere
packaging (MAP), smart packaging, etc., globally, it was
possible to retain the freshness of many perishable as well
as non-perishable foods. MAP can maintain the respiration
rate and reduce the microbial growth, oxidation reactions
and thus affects the shelf life of food products [4]. Gases
like CO2, O2 and N2 are normally used in MAP. These
gases are tasteless and have peculiar characteristics such as
CO2 effectively inhibits the growth of spoilage bacteria and
molds [5] and N2 is an inert, tasteless gas mostly used as a
filler gas. However, commercial application of modern
packaging techniques for traditional dairy foods is still
limited [6]. Reports have been published on the application
of MAP for preservation of Peda [7], Paneer [8, 9], Malai
peda [10], Brown peda [11] and Lal peda [6] which shows
that this packaging technique holds a promise for enhanc-
ing the shelf life of traditional dairy foods.
The shelf life of Khoa is influenced by the moisture
contents, which permit the growth of the molds within
2 days of storage at room temperature [12]. Various
attempts were taken by the researchers to prevent the
deterioration of Khoa by application of steam for
1520 min [13], ultraviolet radiation [14] and by the
addition of cane-sugar [15]. Shelf-life extension of Khoa
and Khoa based products with various approaches like
different packaging materials [1618], addition of preser-
vatives (potassium sorbate, nisin, herbal extracts) and
antioxidants [1924] were reported. A significant reduction
of yeast and mold count was reported by the use of solar
radiation [25]. However, no information is available on the
effect of MAP technology and addition of benzoic acid as
preservative for the enhancement of shelf life of Khoa. The
objective of the present study is to determine the effect of
benzoic acid as preservative and MAP technology to
extend the shelf life of Khoa.
Materials and Methods
Preparation of Khoa Samples
Khoa was prepared following traditional method i.e., open
pan evaporation process as reported by Kumar [26]. Fresh
buffalo milk (&25 litre) was procured from an organized
dairy of Mumbai. Milk was heated in an open pan over non
smoky fire and continuously stirred and scraped with the
help of a scraper to avoid the scorching of milk solids
sticking to the pan. The milk was heated till it attained a
semi-solid consistency of dough. Thereafter, the product
was removed from the fire and rolled up into solid mass
known as Khoa-pat. The proximate composition of the
freshly prepared khoa in terms of moisture, crude fat, crude
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J Package Technol Res (2017) 1:2531
Table 1 Proximate composition of fresh Khoa
Parameters Content (w/w, %)
Moisture 20.68
Crude fat 40.85
Crude protein 23.08
Nitrogen free extract 31.1
Total ash 4.97
protein, nitrogen free extract, total ash is given in Table 1.
Benzoic acid (AR grade, Vikas Pharmaceuticals Labora-
tories, Mumbai) at 0.02% on dry weight basis was dis-
solved in a little quantity of boiling distilled water and
added to a portion of khoa (&3 kg) when the product was
still hot (about 8590 C). However, the maximum per-
mitted level is 300 ppm i.e. 0.03% [2]. Remaining khoa
was used as control (without preservative) for the study.
Packaging Materials
Rigid coextruded Polyvinyl Chloride/Ethylene Vinyl
Alcohol/Polyethylene (PVC/EVOH/PE) sheet (Thickness:
400 l, Water Vapour Transmission Rate (WVTR): 1.59 g/
m2/day, Oxygen Transmission Rate (OTR): \0.05 ml/
m2/day at 100% O2 concentration) was used to make the
trays supplied by AVI Global Plast Pvt. Ltd., Mumbai,
India and the coextruded plastic film having the structure of
Polypropylene/Polyamide/Ethylene Vinyl Alcohol/Poly-
ethylene (PP/PA/EVOH/PE) (Thickness: 100 l, WVTR:
4.32 g/m2/day, OTR: \0.05 ml/m2/day at 100% O2 con-
centration) as was used from Veripack Solutions India Pvt.
Ltd., Mumbai, India. In the food packaging applications,
these packaging materials are widely used due to its high,
gas and water vapour barrier characteristics for preserving
food freshness for up to many days.
Packaging of Khoa Under MAP
Khoa was divided into two batches, one batch (&3 kg)
contained preservative and the other (remaining &3 kg)
without preservative. Approximately 50 g of Khoa sample
was placed in each tray (dimension: 8.5 9 6.0 9 4.1 cm).
Modified atmosphere packaging of Khoa was done using
Veripack Roll-Stock Thermoform Fill Seal Machine
(Model: FLEXI, Italy). Modified gases combination of
10% CO2 and 90% N2 was used for the present study.
Samples packed without MAP was also used and marked
as ordinary packaging. Samples were designated as ordi-
nary packaging without preservative (Ord WP), ordinary
packaging with preservative (Ord P), MAP packaging
without preservative (MAP WP) and MAP packaging
with preservative (MAP P).
J Package Technol Res (2017) 1:2531
Storage
The packaged Khoa samples were stored at 27 2 C and
relative humidity of 65 2%. Khoa was taken out at an
interval of 24 h and then withdrawn samples were evalu-
ated for sensory evaluation, chemical analysis (moisture,
pH, titratable acidity), and microbial contamination (stan-
dard plate count and yeast and mould count).
Analytical Methods
Sensory Evaluation
Sensory evaluation of fresh and stored samples of Khoa
was done by subjective test based on 9-point hedonic scale.
Ten panelists (6 males and 4 females), in the age group
between 25 and 57 years, were selected from among the
faculty and technical staffs through personal communica-
tion and willingness to undertake this test. Instructions
were given to the panelists to rinse mouth with
potable water after evaluation of every sample. A 9-point
hedonic score system was used with the following indi-
vidual scores: 1Dislike extremely, 2Dislike very
much, 3Dislike moderately, 4Dislike slightly, 5
Neither like nor dislike, 6Like slightly, 7Like mod-
erately, 8Like very much and 9Like extremely [27].
They were asked to carefully examine and score the fol-
lowing attributes of khoa samples as per their level of
liking for: colour, flavor, texture and overall acceptability.
Chemical Analysis
Moisture
The moisture content present in Khoa samples was ana-
lyzed using the AOAC [28] method. It was expressed as %
of moisture content. Measurements were taken in triplicate
for each sample.
pH
The pH of Khoa was measured by using the calibrated
digital pH meter (DBK Instruments, Mumbai). The elec-
trode of the pH meter was calibrated against standard
buffer solution of pH 7.0 and 4.0 (Thermo Fisher Scien-
tific Pte Ltd., Singapore) before use. About 10 g of Khoa
sample was taken in a 50 ml beaker and mixed thor-
oughly with 10 ml of luke warm (4045 C) distilled
water, mixed thoroughly and the pH of Khoa was mea-
sured by inserting the electrode into the slurry and the pH
recorded [29]. Analysis was done in triplicate for each
treatment/sample.
27
Titratable Acidity
The titratable acidity in terms of percent lactic acid was
determined following the method as described in standard
IS: 4883-1980 [30]. Measurements were taken in triplicate
for each sample.
Microbial Analysis
Microbiological analysis of Khoa samples during storage
was examined for standard plate count (SPC) and yeast and
mould count (YMC) as per the standard methods as
described in IS 5402:2012 [31] and IS 5403:1999 [32]
respectively. Analysis was done in three replications for
each treatment/sample.
Statistical Analysis
The changes obtained due to sensory evaluation, chemical
characteristics and microbial contamination and were
analyzed statistically by following analysis of variance (F-
test) at 1 or 5% levels of significance using single factor
experiment with completely randomized design with equal
replications [33]. Significant F-test assures that the
observed difference among the treatment means is real and
not due to chance. All these calculations including standard
deviation were done using Microsoft Excel 2007 (Mi-
crosoft Corp., USA).
Results and Discussion
Sensory Evaluation
The average score for all sensory attributes (shown in
Fig. 1) decreased significantly (P \ 0.01) with storage
period. On the 0th day all the treatments secured nearly
equal score for all sensory characteristics. Samples packed
in Ord (WP), Ord (P), MAP (WP) and MAP (P) were found
to be unacceptable after 4, 8, 8 and 12 days of storage
respectively; hence further evaluation was discontinued.
Maximum sensory score for all the sensory properties was
obtained by samples packed under MAP (P) for the entire
storage period of 12 days. Flavor deterioration was greater
in Ord (WP) as compared to Ord (P) followed by MAP
(WP) and MAP (P). A slight rancid flavor was observed for
the Ord (WP) during the storage period which might be due
to the hydrolysis of fat by the lipase enzyme secreted by
bacteria. Similar type of observation was also reported by
Gaikwad and Hembede [34] for Ujani basundi stored at
different temperature. Presence of any off-odour in other
three treatments was not reported by any of the judges
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28
(a)
(b)
Fig. 1 Changes in sensory scores. a Flavour, b colour and appearance, c texture and d overall acceptability of khoa (with and without
preservatives) packed in different packaging conditions
during the storage. However, the decrease in flavor scores
may be attributed due to slight loss of freshness of the
product. The colour and appearance score decreased
(P \ 0.01) marginally (Fig. 1) from 8.20 to 8.02 which
might be due to evaporation of moisture from the product.
The colour score was highest for MAP (P) followed by
MAP (WP), Ord (P) and Ord (WP) for the whole storage
period. Similar observation was also reported during stor-
age of doda burfi [35]. The mean value for body and tex-
ture differed significantly (P \ 0.01) among all samples.
The decrease (8.268.04) in body and texture score was
observed in all samples but the trend was much progressive
in ordinary packaging (Ord P [ Ord WP) as compared to
the modified atmosphere packaging (MAP P [ MAP WP).
Due to moisture loss the product became slightly thicker;
however texture scores were not much influenced with
increase in storage period. During storage intervals overall
acceptability showed a significant (P \ 0.01) declining
trend from an initial score of 8.66.73. Mean overall
acceptance score indicated that changes were faster for Ord
(WP) followed by Ord (P), MAP (WP) and MAP (P) sam-
ples. Probably, this was caused by textural and flavor
123
J Package Technol Res (2017) 1:2531
(c)
(d)
changes taken place in the samples. The scores for texture
and colour remained in liked very much range throughout
the storage period for all the samples, whereas for flavor
and overall acceptability the scores remained between liked
slightly to like very much range.
Chemical Analysis
Moisture Content
Effect of storage on moisture content, pH, and titrat-
able acidity of the Khoa samples (with and without
preservative) is shown in Fig. 2. During storage, the loss of
moisture content from all the samples was observed where
the values dropped from 20.68 to 18.35% for Ord (WP),
17.67% for Qrd (P), 18.78% for MAP (WP) and 18.55%
for MAP (P) samples. However, the loss of moisture was
rapid in Ord (WP) samples compared to all other samples.
Statistically the decrease in moisture content was found to
be highly significant (P \ 0.01) throughout the storage
period. Similar trend of decreasing moisture content was
also observed in different products such as pinni, carrot
J Package Technol Res (2017) 1:2531 29

(a) place in vacuum packaging, but at a slower pace, as

25 observed in paneer tikka.
ORD WP ORD P MAP WP MAP P
24 Fresh Khoa samples showed a gradual but statistically
23 significant decreasing trend (P \ 0.01) in pH for all stored
22 samples. The final pH decreased from 6.47 to 6.37, 6.22,
Moisture, %

21 6.31 and 6.26 for Ord (WP), Ord (P), MAP (WP) and MAP
20 (P) samples, respectively. Suresh and Jha [42] and Chawla,
19 Singh and Patil [35] reported similar decrease in pH
18 observations incase of kalakand and doda burfi respec-
17 tively. The decrease in pH might be due to lactic acid
16 production by spoilage organisms [43].
15 The initial titratable acidity of fresh Khoa sample was
0 2 4 6 8 10 12 0.58 which gradually increased to (P \ 0.01) 0.71 for Ord
Storage Time, Days (WP) in 4 days, 0.8 for Ord (P) in 8 days, 0.74 for MAP
(b) (WP) and MAP (P) in 8 and 12 days respectively. In the
6.60 present study it was observed that the rate of increase in
Ord WP ORD P MAP WP MAP P
6.55 acidity was slower in Khoa samples with benzoic acid as
6.50 preservative, irrespective of packaging conditions.
Mahalingaiah et al. [44] reported similar kind of behavior
6.45
for kunda. The changes in pH cannot be directly correlated
pH

6.40
with change in acidity; still in the present study it was
6.35 observed that with the increase in acidity, the pH went on
6.30 decreasing. This type of behavior is also reported by
6.25 Londhe, Pal and Raju [6] while studying shelf-life of
brown peda. Several researchers [19, 34, 37, 45] reported
6.20
0 2 4 6 8 10 12
that increase in acidity is an indication of spoilage in dairy
Storage Time, Days products.

(c) 0.90 Microbiological Analysis

ORD WP ORD P MAP WP MAP P

Microbiological analysis of the stored Khoa sample was

0.80 examined which showed significant (P \ 0.01) increase in
Titratable Acidity

SPC and YMC count during the entire storage period.

0.70
0.60
0.50
0 2 4 6
Storage Time, Days
Fig. 2 Effect of storage on a Moisture content, b pH, and
c titratable acidity of the Khoa samples
milk cake, burfi kalakand and lal peda by Saxena et al.
[36]; Bajwa et al. [37]; Palit and Pal [38]; Rao and Goyal
[39] and Jha et al. [40] respectively. In case of Khoa
samples with preservatives it could be possible that addi-
tion of chemical additive in the Khoa had acted as
humectant and thereby made the free water unavailable for
easy escape from the product [35]. However, Verma, Goyal
and Bhatt [41] reported that loss of moisture also takes
Initially i.e. on 0 day, no microbial growth was observed
both in terms of TPC and YMC. However, the count
increased during the entire storage period irrespective of
the treatment/packaging conditions applied. According to
FSSAI [2], the maximum SPC limit for Khoa and Khoa
based products is 5 9 104 cfu/g. Hence, from Table 2 it is
observed that the values are within the desired range up to
8 10 12
4 days for Ord (WP), 8 days for Ord (P) and MAP (WP),
and 12 days for MAP (P) samples, further which it was
unsafe for consumption. It is revealed from the results that
addition of preservative retard the rate of microbial pro-
liferation in both the packaging conditions. Several other
workers had also reported about the increase in SPC for
burfi, peda and various other products [35, 4650].
Yeast and mould growth is one of the major factors for
spoilage of milk based products which limits their shelf
life. In the present study, no YMC was detected in the
initial or 0 day samples. However, with increase in
storage time significant (P \ 0.01) increase in YMC was
observed for all the samples. According to FSSAI [2], the

123
30 J Package Technol Res (2017) 1:2531

4.98 9 104 0.06

maximum YMC limit for Khoa and Khoa based products is

8.5 9 101 0.06

1 9 102 cfu/g. Hence, it is evident from Table 2 that up to
4 days for Ord (WP), 8 days for Ord (P) and MAP (WP),
and 12 days for MAP (P) samples, Khoa was considered
safe for consumption; further, higher values of YMC were
12

encountered which was considered as the spoilage of the
product. Similar type of findings was also reported by

4.78 9 104 0.08

Chawla, Singh and Patil [35], Palit and Pal [38], Rajarajan

6 9 101 0.07
et al. [51] and Kumbhar et al. [52]. The results revealed
that addition of preservative retard the rate of microbial
proliferation in both the packaging conditions.
10

Conclusion
4.79 9 10 0.03

4.17 9 104 0.05

3.64 9 104 0.06

9.5 9 101 0.04
10 9 101 0.05

4 9 101 0.03

Based on the test results and interpretation, it is concluded

4

that addition of chemical preservative (benzoic acid) in the

ordinary packaging condition increased the shelf life of the
product from 4 to 8 days, whereas for modified atmosphere


8

packaging (with mixture of gases) and addition of benzoic

3.93 9 10 0.04

3.76 9 104 0.06

2.45 9 104 0.06

acid showed an increased shelf life from 8 to 12 days.

7 9 101 0.06

4 9 101 0.05

2 9 101 0.05

Overall, from the results it is evident that shelf life of the

4

product was increased in MAP technology as compared to

ordinary packaging. Hence, it is recommended that MAP
Table 2 Changes in microbial quality of Khoa (with and without preservative) samples during storage

technology with chemical preservative is suitable for the

6

extension of shelf life of Khoa as compared to ambient

condition.
4.97 9 104 0.06

2.67 9 10 0.06

2.23 9 104 0.07

1.78 9 104 0.03

1.5 9 101 0.06
10 9 101 0.03

5 9 101 0.07

3 9 101 0.06
4

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