Journal of Food Processing and Preservation ISSN 1745-4549

DEVELOPMENT OF SHELF STABLE READY-TO-EAT VEGETABLE

PULAV USING RADIATION TECHNOLOGY
SUSHAMA MARATHE1, RAJALAKSHMI DESHPANDE, JYOTI TRIPATHY and SAHAYOG N. JAMDAR
Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085, India

1
Corresponding author. ABSTRACT
TEL: 191 022 25593962;
FAX: 191 022 25505151; The objective of this study was to develop a shelf stable ready-to-eat vegetable pulav,
EMAIL: samarathe5@gmail.com using radiation processing. The shelf stability of the product was assessed by micro-
bial and sensory analysis during storage. A minimum dose of 7.5 kGy was found to
Received for Publication November 11, 2015
be effective in controlling microbial growth in the product without compromising
Accepted for Publication May 6, 2016
the sensorial quality up to 1 year of storage at the ambient temperature. Radiation
doi:10.1111/jfpp.13104 processing up to 25 kGy dose did not alter proximate composition of the product.
Marginal increase in antioxidant activity was observed in the samples irradiated at a
dose of 25 kGy with concomitant increase in the free phenolic content. GCMS stud-
ies showed increase in the content of few aroma notes such as 2-octanol, eugenol
and eugenyl acetate for the samples irradiated at 10 kGy; however, they were not
perceived by the taste panelist during sensory analysis.

PRACTICAL APPLICATIONS
Vegetable pulav is an Indian ethnic delicacy consumed all over the country. How-
ever, the product has very limited shelf life and thus needs to be prepared fresh,
which is very time consuming. Present study defines process parameters for devel-
opment of microbiologically safe ready-to-eat vegetable pulav, using radiation proc-
essing. The findings of this study show radiation processing; a nonthermal
treatment could be successfully applied for improving the microbial quality, with-
out affecting the sensory attributes of the product and to improve the shelf stability
up to 1 year.

INTRODUCTION
Consumption of prepared food in developed and developing
countries is increasingly frequent. Thus, “Ready-to-eat” or
“convenience” foods are in great demand in those countries.
These products provide readily available meals to the con-
sumer. Moreover, such foods that have appropriately low
microbial counts and requisite nutritional profile could be
used to address the dietary needs of target groups such as
army, astronauts, travellers as well as immuno-compromised
patients (ICPs). The later includes patients undergoing cancer
therapy, organ transplant procedure as well as HIV-infected
people. Since food is the major source of pathogens and poses
high risk of gastrointestinal infections upon consumption of
unprocessed or partial processed food, ICPs are prohibited to
range of food and restricted to those with low microbial
count. However, a variety of foods like salads, sprouts as well
as other ethnic cuisines with exotic flavors and high nutri-
tional value can be made available to them by improving the
microbiological quality of these products.
Though the retort processing has been widely used as a food
processing technique to produce microbiologically safe prod-
ucts, the application of extreme heat for a long period reduces
nutritional value and alters the sensory quality of the food
product. However, these limitations could be overcome by
using a cold process such as radiation technology for the elimi-
nation of pathogenic and spoilage causing organisms. Ionizing
radiations have great potential to inactivate food borne patho-
gens as well as spoilage microorganisms in a variety of food
products without raising temperature and can serve as a final
critical control point to ensure the microbiological safety
of ready-to-eat foods. Thus, the development of radiation
pasteurized ready-to-eat foods for general population, army,

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DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV
travellers, astronauts and immuno-compromised patients is
an attainable goal for the radiation processing industry.
Recently, many studies have shown successful use of radiation
technology for ensuring microbial safety as well as for exten-
sion of shelf life of ready-to-eat foods (Lee et al. 2005, 2012;
Teets and Were 2008; Sharma et al. 2009; Song et al. 2009; Park
et al. 2012; Yoon et al. 2012; Yun et al. 2012; Zhu et al. 2012).
However, except for the few products like “Kimchi” or
“Noodles,” the process was mostly used for meat or chicken
products. Very few storage studies have been reported on
microbiological and organoleptic properties of irradiated
ready vegan meals.
Vegetable pulav is a type of rice preparation, which is
nutritionally balanced and wholesome, popular Indian cui-
sine. It is consumed throughout India with minor varia-
tions. However, this preparation has short shelf life of less
than 12 h at ambient conditions due to its high moisture
content as well as available nutrients. Kumar et al. (2011)
have shown that a combination of thermal processing (F0
value of 2.0) and gamma irradiation (4.0 kGy) could achieve
commercial sterility of vegetable pulav. However, the effi-
cacy of gamma irradiation alone, for preserving this product
has not been evaluated. Therefore, the objective of the pres-
ent study was to assess the effectiveness of radiation process-
ing as means to achieve commercial sterility and thus
improve the shelf stability of vegetable pulav without com-
promising nutritional and sensorial properties of the prod-
uct. The shelf stability was assessed in terms of its
microbiological and organoleptic quality during the storage
period. The study also includes evaluation of nutritional
properties, antioxidant activity as well as release of aroma
compounds in radiation processed vegetable pulav.
MATERIALS AND METHODS
Preparation and Irradiation of Vegetable
Pulav
The ingredients used for preparation of vegetable pulav were
rice (Basmati variety: 600 g), cut vegetables (onion, potato,
carrot, French beans, green peas: each 225 g), green gram
(soaked, 300 g), sunflower oil (55 g), dry spices (cumin
seeds, clove, pepper, cardamom, bay leaves, cinnamon: each
3.0 g) and salt (18 g). The preparation of pulav was carried
out in nonstick utensil of 5 kg capacity. Initially, the oil
is heated in pan to frying temperature to which spices were
added and stir fried for 1 min. To this onion slices were
added and saute for 5 min. After that, cut vegetables
were added and stirred fried for 5 min, to same mixture
washed and pre-soaked rice (rice soaked in water at 25C for
30 min) was added and stir fried for 5 min. To this,
1,200 mL of boiling water (100C) and salt were added. Rice
2 Journal of Food Processing and Preservation 00 (2016) 00–00 V
S. MARATHE ET AL.
was cooked on low flame for 15 min. It was allowed to settle
for 30 min and afterward spread on flat plate for cooling at
ambient temperature for 30 min. The total weight of the
final product was about 3.25 kg.
The vegetable pulav was packed (200 g per pack) in multi-
layer (12 mm polyester/12 mm aluminum foil/75 mm cast-
polypropylene) laminated pouches (dimension 15 cm 3
20 cm). The sealed packets were then subjected to gamma
radiation under frozen condition (278C, using dry ice) in
cobalt-60 gamma irradiator (Gamma Chamber 5000, Board
of Radioisotope and Technology, Mumbai, India) at the
dose rate of 2.5 kGy/h. The samples were exposed to gamma
radiation for 2, 3, 4, 6 and 10 h for achieving the dose of 5,
7.5, 5, 10, 15 and 25 kGy, respectively. Nonirradiated vegeta-
ble pulav was used as control for all the subsequent experi-
mental procedures. The irradiated samples were stored at
ambient temperature (28C) with relative humidity (RH) of
72%, while control samples were stored at 4, 8 and 28C in
order to assess the storage stability of the untreated product
at different storage temperatures.
Absorbed-dose mapping of the Gamma Chamber 5000
was performed with simulated product at room tempera-
ture in order to establish relations between reference dose
point with maximum and minimum dose points. A beaker
of volume 2 L was used to define the irradiation volume. A
reference dose point was identified at the outer surface of
the irradiation volume and was isolated from temperature
gradients in the actual product. Fricke dosimeters were
placed at the reference dose point and inside the irradiation
volume at 15 different positions. Rice was used as the
medium of absorbed dose and exposed for adequate time to
measure the dose within the measurable range of Fricke sys-
tem. The maximum dose position was observed at the front
surface of the irradiation volume and the minimum dose
position was at the body center. According to ISO/ASTM
51204:2004(E) during routine processing of the chilled or
frozen product, dosimeters were placed at the reference
position and minimum and maximum absorbed doses to
the product at sub-ambient temperature were determined
using the following established relations Dmin 5 0.7079 3
Dref and Dmax 5 0.8517 3 Dref.
For the chemical analysis such as proximate, antioxidant
activity, phenolic contents and GCMS studies, vegetable
pulav samples were packed in multilayer pouches, irradiated
as described earlier and ground to make smooth paste, so as
to have uniform distribution of the ingredients.
Storage Study
Microbiological Assessment. The irradiated vegetable
pulav samples stored at ambient conditions were analyzed
periodically at 0 h, 1 week, 2 weeks, 4 weeks, 1 month,
3 months, 6 months, 9 months and 12 months for microbial
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S. MARATHE ET AL.
count. The shelf life of nonirradiated vegetable pulav at 4, 8
and 28C was assessed after 24 h, 1, 2 and 4 weeks of storage
period according to Pizzo et al. (1982). For microbiological
analysis, each sample (25 g) was suspended in 225 mL of
physiological saline (0.85% NaCl) in sterile stomacher bag
and then homogenized using stomacher (Stomacher lab
blender, model 400, Seward, London, UK) for 1min at
150 rpm. Bacteriological Analytical Manual (BAM) was
referred for the evaluation of aerobic plate count, aerobic
spore count, anaerobic spore count and fungal (yeast and
mold) count by using Plate Count Agar (PCA), Soyabean
Casein Digest Agar (SDA), Reinforced Clostridium Agar
(RCA) and Rose Bengal chloramphenicol Agar (RBA),
respectively. Pour plate technique was used for all the analy-
sis except for yeast and mold count. The PCA and RBA
plates were incubated at ambient temperature for 48 h,
while SDA and RCA plates were incubated at 37C for 48 and
120 h, respectively, before taking the count.
Sensory Evaluation. Organoleptic evaluation of irradi-
ated and control, vegetable pulav was conducted using 9-point
hedonic scale according to Stone et al. (2012) and by following
ASTM guidelines (ASTM 2002). The scale was divided
between two extremes: “extremely good” to “extremely poor”
from 9 to 1 and with “fair” as the midpoint. Characteristics
such as appearance, color, texture, aroma, taste and overall
acceptability were judged. The samples were brought to 50C
before serving to the panelist (Microwave heating for 1 min at
900 W). Frozen control (untreated vegetable pulav samples
stored at 220C) brought to 50C served as control. The panel-
ists were selected on the basis of their liking for the product
and a product-specific preliminary training for the different
attributes was given to all the sensory panelist.
Proximate Analysis. Proximate composition of control
as well as irradiated samples was determined by AOAC
method (AOAC 1990).
Chemical Analysis. The samples were freeze dried using
lyophilizer (scanvac, cool safe, 554 pro, Lynge, Denmark)
before analysis of antioxidant activity and phenolic content.
Antioxidant Activity
Extract Preparation. The freeze dried samples (2 g) were
extracted with 25 mL of 80% aqueous methanol (v/v) at
25C with continuous shaking for 2 h (using an orbital
shaker, 150 rpm). Extracts were stored at 220C (MDF
U537D Sanyo, Japan) until analysis.
Estimation of Free Phenolics. The concentration of
free phenolics in the extracts was measured using Folin–
Ciocalteu assay (Hung and Yen 2002) with some modifica-
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DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV
tion. Appropriately diluted 0.2 mL extract was added to
0.05 mL Folin–Ciocalteu reagent (1:1 diluted with water)
and mixed thoroughly, and after 3 min, 1.0 mL of 2.0%
Na2CO3 solution was added. The absorbance of the mixture
was measured after 30 min at 750 nm using spectrophotom-
eter (Varian DMS 100). Standard curve was constructed
using gallic acid. Free phenolic content were expressed as
mg of gallic acid equivalent (GAE)/g legume.
Estimation of Radical (DPPH•) Scavenging Activity.
DPPH• radical scavenging activity was measured as per the
method by Brand-Williams et al. (1995). DPPH• stock solu-
tion was prepared by dissolving 270 mg/L of methanol. An
aqueous methanolic (80%) extract, diluted appropriately
was added to the methanolic solution of DPPH
radical and
the volume was made to 1 mL, the final concentration of
DPPH• was maintained at 75 mM. The mixture was shaken
vigorously and incubated in dark at 25C, for 30 min. The
absorbance of the resulting solution was then measured at
515 nm against methanol. The % scavenging was deter-
mined using the following formula:


% Scavenging 5 Acontrol 2Asample =Acontrol 3 100:
The antioxidant activity was expressed as units/g and one
unit was defined as the amount of antioxidant necessary to
scavenge 50% of initial concentration of DPPH• (equivalent
to 37.5 nmol) (Marathe et al. 2011).
Analysis of Volatiles
Each sample (10 g) was added with NaCl (10 g) and distilled
water (10 mL). The resultant mixture was homogenized
(B400, Buchi, Switzerland), while maintaining chilled con-
ditions using an ice bath. The homogenate obtained was fil-
tered through double layered muslin cloth. The extract
obtained was centrifuged at 12,000 rpm at 20C for 15 min.
To the sample, 3-hexen-1-ol (1.28 lg) was added as an inter-
nal standard. Samples were equilibrated at 30C for 30 min
with continuous stirring and the headspace was subse-
quently extracted by preconditioned PDMS/DVB/CAR
fibers (Supleco) at 30C for 20 min. After the extraction, the
fibers were desorbed (270C) in the injection port of the GC/
MS equipment. Quantification was performed by compar-
ing the peak areas of compounds with that of internal stand-
ard and results obtained were expressed as lg/kg.
Statistical Analysis
The data are reported as mean 6 standard deviation.
Analysis of variance was performed by using Origin Pro 8.0
(Origin Lab Corp.) and P < 0.05 was considered to be statis-
tically significant.
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DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV S. MARATHE ET AL.

TABLE 1A. MICROBIAL COUNT (log CFU/g) OF NONIRRADIATED VEGETABLE PULAV DURING STORAGE
Cell counts during storage (log CFU/g)

Control (4C) Control (8C) Control (28C)

0h 24 h 1 week 2 weeks 1 month 0h 24 h 1 week 0h 24 h

TBC 1.78 1.78 2.34 3.65 >6.00 1.78 1.78 >6.00 1.778 >6.00
YMC ND ND ND ND NA ND ND ND ND ND
TASC ND ND 1.30 1.89 NA ND ND ND ND ND
TANSC ND ND ND ND NA ND ND 1.6021 ND 1.000

NA, not analyzed as samples discontinued due to spoilage. ND, not detected (detection limits <1 log CFU/g), TBC, total bacterial count; YMC, yeast
and mold count; TASC, total aerobic spore count; TANSC, total anaerobic spore count.

The GC/MS data obtained for aroma volatiles (total ion
current [TIC] v/s retention time) was exported in ASCII for-
mat to Microsoft Excel using inbuilt software of GC/MS.
The corresponding ASCII data were then subjected to prin-
cipal component analysis (PCA) using XLSTAT 2012 soft-
ware (Addinsoft Inc.). Loading and score plots were drawn
using principal components 1 and 2. To know the nature of
the constituents responsible for the differences among con-
trol and radiation processed samples, factor loading data
obtained from PCA analysis were used.
RESULTS AND DISCUSSION
Storage Study
Microbiological Assessment. In spite of observing
good manufacturing practices (GMPs), fresh vegetable pulav
samples exhibited total bacterial counts of 1.78 log CFU/g
(Tables 1A and 1B). This could be due to aerial contamina-
tion of the samples during handling, cooling or packing.
When these samples were stored for 24 h, a microbial count
of >6 log CFU/g was recorded. The lower initial count
is mainly due to the involvement of cooking step (100C for
15 min), which eliminated major microbial populations.
Thus, the fresh vegetable pulav itself can be considered as
“low bacterial food” without any additional treatment (Pizzo
et al. 1982). Making “fresh food” available all the time
especially during travel as well as to aforementioned specific
target groups is not easy.
The microbial count of control and irradiated samples
during storage is depicted in Tables 1A and 1B. Aerobic
spore count, anaerobic spore count and fungal (yeast and
mold) count were below detectable limit at 0 h, in control
samples. Although cooking process significantly reduced the
microbial load, product sterility is not guaranteed in the
final product. Moreover, as no aseptic conditions were
observed during preparation of the product, aerial contami-
nation of the product could be possible during handling,
cooling and packaging. Thus, we expect the main source of
contamination is either from air or from the ingredients
used for the preparations. Since the product is cooked, pres-
ence of heat tolerant spores like Bacillus spp. could be
expected in the product. Anaerobic spores of Clostridium
spp. are common food contaminants in packed foods and
causes food poisoning (Addis and Sisay 2015), hence anaer-
obic spore count was carried out in order to make sure that
no such pathogenic organisms were present in the product
during storage.
Total bacterial counts in control samples stored for
1 week at 4 and 8C were 2.34 and >6 log CFU/g, respec-
tively, while anaerobic spore count was 1.60 log CFU/g in
sample stored at 8C. Control samples stored at 4C, however,
showed steady increase in total bacterial count and reached
up to 3.65 log CFU/g at the end of 15 days. The products did
not show any off odor or change in texture. According to the

TABLE 1B. MICROBIAL COUNT (log CFU/g) OF IRRADIATED VEGETABLE PULAV DURING STORAGE
Cell counts during storage (log CFU/g) (28C)

Irradiated (5 kGy) Irradiated (7.5, 10. 15 and 25 kGy)

0h 24 h 1 week 0h 24 h 1 week 1 month 3 months 6 months 9 months 12 months

TBC ND ND >6.00 ND ND ND ND ND ND ND ND
YMC ND ND ND ND ND ND ND ND ND ND ND
TASC ND ND 1.301 ND ND ND ND ND ND ND ND
TANSC ND ND 1.602 ND ND ND ND ND ND ND ND

NA, not analyzed as samples discontinued due to spoilage. ND, not detected (detection limits <1 log CFU/g), TBC, total bacterial count; YMC, yeast
and mold count; TASC, total aerobic spore count; TANSC, total anaerobic spore count.

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S. MARATHE ET AL.
FIG. 1. SENSORY EVALUATION OF RADIATION PROCESSED
VEGETABLE PULAV
Subjective sensory evaluation was carried out for different attributes by
trained taste panel (n 5 40). A 9-point hedonic scale was used where
each point was described as: 1 5 extremely poor, 2 5 very poor;
3 5 poor; 4 5 below fair above poor; 5 5 fair; 6 5 below good above
fair; 7 5 good; 8 5 very good; 9 5 extremely good.
international guidelines for microbiological standards, food
in which all the components of the food have been cooked
during manufacturing or preparation procedure and hence
low in microbial count as such is considered as level 1. The
acceptable limit for total aerobic plate count of these prod-
ucts is <4 log CFU/g, while for total aerobic spore count is
<2 log CFU/g. The pathogenic organisms like Salmonella,
Listeria monocytogenes and Campylobacter spp. should not
be detected in 25 g of the samples (https://fsrio.nal.usda.
gov/sanitation-and-quality-standards/microbiological-
standards-and-guidelines). The microbial count in control
samples at 4C were within acceptable limit till 15 days, while
at 8C, the count was much higher after a week. The spoilt
samples were also identified by sensorial properties like off
odor and slimy texture of the sample. Extensive spoilage as
observed for samples stored at 8C for 1 week or ambient
condition for 24 h also led to bulging of sample packets.
Based on established microbial guidelines (https://fsrio.nal.
usda.gov/sanitation-and-quality-standards/microbiological-
standards-and-guidelines) and results of sensory evaluation
conducted on samples, the shelf life of untreated vegetable
pulav stored at 28, 8 and 4C was less than 24 h, 1 week and
4 weeks, respectively.
Results from Table 1B reveal that gamma irradiation up
to 5.0 kGy was not sufficient for the elimination of initial
microbial load. As such, these samples were spoilt in 1 week
Journal of Food Processing and Preservation 00 (2016) 00–00 V
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DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV
time when stored at 28C. However, vegetable pulav irradi-
ated at 7.5 kGy did not show microbial growth during 1
year storage at 28C. None of these sample packs showed
bloating, which affirmed lack of microbial activity. The
results thus suggest that gamma irradiation treatment at
doses 7.5 kGy could take care of residual microbial load of
fresh vegetable pulav, which survived even after the heat
treatment given to the product during preparation of the
product as well as the post preparation microbial contami-
nation. Although effect of gamma radiation processing of
uncooked rice of different varieties on microflora as well as
keeping quality has been evaluated (Hiroshi and Hitoshi
1968; Wang et al. 1983; Sabularse et al. 1991), similar infor-
mation about radiation processed cooked rice is scarce.
Sensory Evaluation. Since samples irradiated at 5 kGy
were spoiled in 1 week time, sensory analysis of stored sam-
ples was carried out only for those irradiated at 7.5 kGy.
The sensory characteristics of the vegetable pulav samples are
shown in Fig. 1. The attributes studied were appearance,
color, odor, texture, taste, after taste and overall acceptability.
All irradiated samples were acceptable after 12 months stor-
age. The control sample, at 0 h, showed the highest mean
scores among all the samples for all the attributes studied,
while samples processed at highest dose of irradiation (25
kGy) showed significant decrease (P < 0.05). The mean scores
for samples treated at 15 kGy, although numerically lower,
were not significantly different (P  0.05) from control sam-
ples. However, all the samples were in the acceptable range.
The storage of samples resulted in slight decreased score for
all the attributes, however, they were in acceptable range. Kil-
cast (1991) has reported that the production of off-flavor was
a limiting factor for acceptability of a range of hot and cold
irradiated (1–4 kGy) ready meals. Nevertheless, he suggested
that these changes could be kept to a minimum when items
of strong flavor were included in the meals. The presence of
spices in vegetable pulav might have played the important
role in maintaining good acceptability score. The acceptabil-
ity of irradiated vegetable pulav is similar to those reported
for some of the Korean food namely Nutrition bar, Ramen
(ready-to-cook noodle) and two Korean traditional foods
(Kimchi, fermented vegetable; Sujeonggwa, cinnamon bever-
age) (Song et al. 2009).
Chemical Analysis
Proximate Analysis. Nutrient composition of the vege-
table pulav samples irradiated at different doses is presented
in Table 2. Proximate composition of nonirradiated sample
was 2.01, 27.9, 1.14, 67.58 and 1.37% for protein, carbohy-
drate, lipid, moisture and ash, respectively. No statistically
significant difference (P  0.05) was observed in proximate
compositions of control and irradiated samples, even for
5
DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV S. MARATHE ET AL.

TABLE 2. PROXIMATE COMPOSITION OF READY-TO-EAT VEGETABLE PULAV SAMPLES

Radiation processed vegetable pulav at different doses (kGy)

0 5 7.5 10 15 25
Moisture 67.6 6 2.1 66.5 6 3.2 67.8 6 1.9 65.8 6 3.4 67.8 6 2.5 66.9 6 3.2
Protein 2.01 6 0.13 2.02 6 0.17 2.02 6 0.21 2.07 6 0.15 2.02 6 0.24 2.09 6 0.23
Ash 1.39 6 0.13 1.50 6 0.15 1.50 6 0.21 1.54 6 0.22 1.57 6 0.29 1.39 6 0.17
Fat 1.14 6 0.09 1.11 6 0.12 1.10 6 0.15 1.14 6 0.20 1.32 6 0.21 1.16 6 0.14
CHO* 27.1 28.9 27.6 29.5 27.3 28.5

*CHO refers to carbohydrate content, calculated by difference method. The values in the row were not significantly different (P > 0.05).
The results are mean of six determinations 6 S.D. and expressed as relative percentage per g of dry weight.

those processed at 25.0 kGy. Our results are in agreement
with those reported by Farag (1998), who have shown that
high dose irradiation (up to 60 kGy) of full fat soybean
under ambient conditions did not affect its protein and
other macronutrient composition. No loss in nutritional
value after irradiation was also confirmed by performing
animal (broiler chicks) feeding studies using irradiated soy-
bean as part of chick feed, which exhibited no deleterious
effect on the growth of chicks.
Antioxidant Activity. There is a growing scientific inter-
est on the influence of radiation processing on biological
activities and compounds responsible for such activities.
The chemical changes taking place during irradiation are
either result of the direct effect of radiation on the food
components or by indirect action, through reactive inter-
FIG. 2. ANTIOXIDANT ACTIVITY AND PHENOLIC CONTENT OF
CONTROL AND IRRADIATED VEGETABLE PULAV
Antioxidant activity was measured by DPPH free radical scavenging
assay and expressed as unit/g dry pulav, where one unit was defined as
the amount of antioxidant necessary to scavenge 50% of initial
concentration of DPPH (equivalent to 37.5 nmol). The phenolic content
was expressed as mg of gallic acid equivalent/g dry pulav. The values
presented are mean of three independent experiments 6 S.D. Different
letters above the individual bar exhibits significant difference in the
values in given data set (P < 0.05).
mediates formed by radiolysis of water (Diehl 1995). Effect
of gamma radiation processing on the radical scavenging
activity of vegetable pulav is depicted in Fig. 2. Irradiated
samples showed slightly increased radical scavenging activ-
ity. Moreover, the content of free phenolics was also
observed to increase in similar manner. Thus, the increase in
antioxidant activity could be attributed to increase in free
phenolics, which are known for their antioxidant potentials.
It is reported in earlier studies that the extractability of phe-
nolics is increased due to radiation treatment which results
in increased antioxidant activity (Mohammad et al. 2009).
Analysis of Aroma Compounds
Aroma quality plays a critical role in deciding the consumer
acceptance of a food product (Variyar et al. 2003). To study
the effect of radiation processing on the aroma constituents
of control and radiation treated vegetable pulav was there-
fore of interest. For the analysis of aroma compounds sam-
ples irradiated at 5 and 25 kGy were not used, as samples
irradiated at 5 kGy were not shelf stable, while those irradi-
ated at 25 kGy exhibited lower acceptability relative to those
irradiated at 7.5, 10 and 15 kGy doses.
A total of 29 aroma compounds were detected in control
and irradiated vegetable pulav samples (Table 3 and Fig. 3).
The detected compounds were classified under four main
chemical classes: alcohols (5), aldehydes (10), terpenes (10)
and esters (4). Analysis of GC spectral data by principal
component analysis (PCA) revealed the changes in the vola-
tile composition as a result of radiation processing. The con-
trol sample and samples irradiated at 7.5 kGy were located
on the left side of F1, indicating no remarkable changes in
aroma composition at 7.5 kGy as compared to control. On
the other hand, samples irradiated at 10 and 15 kGy were
segregated from these samples and grouped together (on the
positive side of F1). It indicated that higher doses (10 and 15
kGy) resulted in significant changes in aroma constituents
as compared to control samples; however, they were not sig-
nificantly different from each other. Careful interpretation
of GC/MS data revealed that there were quantitative changes

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S. MARATHE ET AL. DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV

TABLE 3. QUANTITATIVE DISTRIBUTION (mG/kG OF VEGETABLE PULAV) OF VOLATILE OIL COMPONENTS IDENTIFIED IN VEGETABLE PULAV
IRRADIATED AT DIFFERENT DOSES
S. No. KI Compound Control 7.5 kGy 10 kGy 15 kGy
ad b c
1 735 3-methyl-1-butanol (C1) 1.62 6 0.03 1.07 6 0.14 0.51 6 0.05 1.73d 6 0.68
2 800 3-Hexenal (C2) 1.77a 6 0.09 1.63a 6 0.19 2.96b 6 0.11 3.07b 6 0.18
3 805 Butanoic acid, ethyl ester (C3) 6.58a 6 0.42 4.33b 6 0.54 2.69c 6 0.01 3.31d 6 0.27
4 815 Hexanal (C4) 6.59a 6 0.51 4.73b 6 0.55 3.52 c6 0.2 5.54d 6 0.31
5 833 Acetic acid, butyl ester (C5) 1.83a 6 0.19 2.28b 6 0.29 1.56a 6 0.24 2.26b 6 0.27
6 868 1-Hexanol (C6) 7.48a 6 0.27 6.74ab 6 0.88 6.25b6 0.15 10.48c 6 0.4
7 902 Heptanal (C7) 2.16a 6 0.09 1.77b 6 0.26 1.36c 6 0.01 2.48d 6 0.07
8 931 a-Pinene (C8) 3.98a 6 0.74 2.13b 6 0.12 0.76cd 6 0.56 0.46d 6 0.05
9 961 Benzaldehyde (C9) 3.88a 6 0.14 2.99b 6 0.06 4.38c 6 0.13 8.81d 6 1.65
10 985 1-Octen-3-ol(C10) 2.8a 6 0.24 2.87 a6 0.26 2.56b 6 0.11 5.09c 6 0.94
11 987 2-Octanol(C11) 28.71a 6 3.44 32.67ba 6 1.39 33.28cb 6 0.37 45.67d 6 9.3
12 995 Hexanoic acid, methyl ester (C12) 4.39a 6 0.16 2.85b 6 0.5 1.32c 6 0.1 2.2d 6 0.11
13 1032 l-Limonene (C13) 6.69a 6 0.17 5.87b 6 0.56 4.83cb6 1.24 8.59d 6 0.98
14 1039 Trans-b-ocimene (C14) 1.13a 6 0.04 1.99b 6 0.15 1.18a 6 0.02 6.72c 6 0.33
15 1086 Terpinolene (C15) 1.36a6 0.13 1.98b 6 0.15 0.93c 6 0.36 2.89d 6 0.26
16 1093 Nonanal (C16) 5.02a 6 0.22 5.43ab 6 0.3 5.46b 6 0.16 9.99c 6 1.61
17 1098 Linalool (C17) 0.24a 6 0.03 4.09b 6 0.64 3.95b 6 0.73 12.74c 6 0.41
18 1125 2-Caren-10-al (C18) 1.22a 6 0.17 2.92b 6 1.16 5.54c 6 1.22 4.54c 6 1.06
19 1135 Camphor (C19) 3.00a 6 0.14 1.31bc 6 0.33 1.50c 6 0.16 2.79da 6 0.41
20 1145 Cis-sabinol (C20) 0.39a 6 0.02 1.39bc 6 0.79 1.45c 6 0 1.98d 6 0.35
21 1152 3-Phenylpropionaldehyde (C21) 1.20a 6 0.26 1.19a 6 0.37 1.22a 6 0.98 1.29a 6 0.76
22 1165 p-Cymen-7-ol (C22) 3.30ab 6 0.81 2.90b 6 0.34 2.14cb 6 0.73 3.95dab 6 1.09
23 1175 Terpineol-4 (C23) 4.93a 6 0.47 21.36b 6 8.5 18.02b 6 3.62 18.93b 6 2.55
24 1209 Decanal (C24) 2.22ab 6 0.43 2.43a 6 0.37 1.78b 6 0.0 2.88ca 6 0.74
25 1225 Cumin aldehyde (C25) 4.44a 6 0.64 6.09b 6 0.3 9.47c 6 1.34 10.07c 6 1.45
26 1261 E-2-decenal (C26) 1.23ab 6 0.19 1.23ab 6 0.14 1.00b 6 0.13 2.54c 6 0.44
27 1275 Cinnamic aldehyde (C27) 35.53a 6 5.92 12.56a 6 0.55 17.92c 6 0.38 22.9dc 6 6.84
28 1355 Eugenol (C28) 563.65a 6 58.92 820.46b 6 120.25 1095.62c 6 41.01 1274.61d 6 192.35
29 1520 Eugenyl acetate (C29) 28.28ab 6 4.62 27.65b 6 0.76 100.6cd 6 22.59 124.3d 6 23.9

Values in the row, superscript with different letters differ significantly (P < 0.05).

FIG. 3. PRINCIPAL COMPONENT ANALYSIS OF AROMA COMPOUNDS OF CONTROL AND IRRADIATED (7.5, 10 and 15 kGy) VEGETABLE PULAV
SAMPLES
(A) Score plot depicting distribution of vegetable pulav samples irradiated at various doses with first two principal components. (B) Score plot
depicting distribution of various aroma compounds with F1 and F2 (refer Table 3 for compound information).

Journal of Food Processing and Preservation 00 (2016) 00–00 V

C 2016 Wiley Periodicals, Inc. 7
DEVELOPMENT OF READY-TO-EAT VEGETABLE PULAV
in the aroma compounds identified which could be respon-
sible for the segregation of samples in the PC plots.
A significant increase in the content of hexanol (green
aroma), benzaldehyde (nutty aroma), 1-octen-3-ol (green/
oily), 2-octanol (spicy green), eugenol (spicy/clove like) and
eugenyl acetate (fresh clove) was observed. An increase of
40% in hexanol, 127% in benzaldehyde, 81% in 1-octen-3-
ol and 59% in the content of 2-octanol was noted at a dose
of 15 kGy (Control 28.71 lg/g; 15 kGy–45.67 lg/g), while
no significant changes were observed at lower doses (7.5
kGy–32.67 lg/g; 10 kGy–33.28 lg/g). A radiation dose
dependent increase of 45, 94 and 126% was noted in the
content of eugenol at 7.5 kGy (820.46 lg/g), 10 kGy (1.95.62
lg/g) and 15 kGy (1274.61 lg/g), respectively; while eugenyl
acetate exhibited 3–4-fold increase at 10 kGy (100.6 lg/g)
and 15 kGy (124.3 lg/g). Also terpenes–linalool (floral),
terpineol-4 (spicy), cis-sabinol and 2-caren-10-al exhibited
a significant dose dependent increase. The content of linal-
ool was estimated to be 0.24 lg/kg in nonirradiated control
samples, which increased to 4.09, 3.95 and 12.74 lg/kg in
samples irradiated at 7.5, 10 and 15 kGy, respectively. While
the content of terpineol-4 was 4.93, 21.36, 18.02 and 18.93
lg/kg in control and samples irradiated at 7.5, 10 and 15
kGy, respectively. Three to fivefold increase in the contents
of cis-sabinol and 2-caren-10-al were noted as a result of
radiation treatment. The aforementioned compounds
which exhibited increase in their content in irradiated vege-
table pulav samples are known to be present in spices like
cardamom (small variety), cumin, clove, pepper, cinnamon
and bay leaves (Parthasarathy et al. 2008). Moreover, these
spices are the main ingredients responsible for aroma of the
vegetable pulav. Thus, enhanced release of these compounds
due to gamma irradiation of vegetable pulav might result in
the improved sensory quality of the produce per se or by
masking off flavors generated due to gamma irradiation of
food products (Kilcast 1991). The existence of glycosidic
precursors of alcohols including terpene alcohols is widely
reported in food products (Variyar et al. 2003). Radiation
induced breakdown of these glycosidic precursors and cor-
responding increase in the free aglycones resulting in
enhanced aroma quality has been reported earlier in several
food products such as coffee, nutmeg and fenugreek (Var-
iyar et al. 2003; Ananthakumar et al. 2006; Chatterjee et al.
2009). In case of nutmeg, four different glycosidic precur-
sors of p-cymene-7-ol, eugenol, methoxyeugenol and a-
terpineol have been shown and dose dependent breakdown
of all glycosides was reported (Ananthakumar et al. 2006).
In another study, radiation dose dependent breakdown of
phenol glycoside in fenugreek seeds was documented (Chat-
terjee et al. 2009). Further, a similar phenomenon was
also observed for glycosidic precursors of isoeugenol and
4-vinylguaiacol in coffee beans (Variyar et al. 2003).
8 Journal of Food Processing and Preservation 00 (2016) 00–00 V
S. MARATHE ET AL.
Thus, results obtained in the present study are in accordance
with the published data.
Also, radiation induced decrease in the contents of certain
compounds such as alpha-pinene, methyl ester of hexanoic
acid and cinnamic aldehyde was observed. The content of
alpha pinene (herbal) was 3.98 lg/kg in nonirradiated con-
trol samples, which decreased to 2.13, 0.76 and 0.46 lg/kg in
samples irradiated at 7.5, 10 and 15 kGy, respectively. The
content of hexanoic acid methyl ester was 4.39, 2.85, 1.32
and 2.2 lg/kg in control and samples irradiated for 7.5, 10
and 15 kGy, respectively. While cinnamic aldehyde (cinna-
mon like aroma) content varied from 35.53 to 22.9 lg/kg in
control and samples irradiated at dose of 15 kGy. Radiation
induced decrease in volatiles has been reported earlier in
orange juice (Fan and Gates 2001) and ginger rhizomes (Wu
and Yang 1994). Despite the changes in the content of some
of the aroma compounds as a result of radiation processing
as noted instrumentally and statistically, no remarkable
change in aroma quality was perceived by the sensory panel
in the irradiated vegetable pulav samples. The radiation
processed vegetable pulav samples up to a radiation dose of
15 kGy were therefore acceptable to the sensory panel.
CONCLUSION
A ready-to-eat vegetable pulav could be developed using
gamma irradiation dose between 7.5 and 15 kGy under fro-
zen conditions (278C) for achieving the commercial steril-
ity of the product. The product remained stable at ambient
conditions and accepted sensorially even after 12 months.
The nutritional properties of the vegetable pulav remained
unaltered even after processing at 25 kGy. The sensory scores
of the product processed at 7.5, 10 and 15 kGy were compa-
rable to that of unprocessed vegetable pulav, while a dose of
25 kGy slightly reduced the acceptability of the product.
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