Idli

ABSTRACT
Idli, a traditional breakfast food of India based on rice-black gram combination, is a

fermented, leavened, soft, spongy textured product. Idli batter has very short shelf-life
because of its high moisture content and live fermentation. There is a demand for ready-to-
cook idli batter in packaged form with moderate shelf-life. Thermal preservation is not
feasible as the batter co-agulates or idli is formed on heat application. Study was
undertaken to evaluate some of the non thermal methods of preservation (sonication,
irradiation and biopreservatives) for extension of shelf-life of fresh idli batter.
Fresh idli batter after fermentation and packing was sonicated at different power
levels (60, 100 and 182 µm) for different exposure times (5, 8, 12, and 15 min) or irradiated
(0.5, 0.75, 1.0 and 1.25 kGy) or preserved by adding Nisin (7.5 ppm) and Potassium
Sorbate (2000 ppm). The quality responses in terms of pH, acidity, standard plate count and
the overall acceptability were evaluated as a function of the preservation treatments. All the
three preservation techniques enhanced the shelf-life of idli batter substantially.
The shelf-life increased with the increase in sonication amplitude for most the
sonication times. The shelf-life also increased with the increase in irradiation
frequencies and with the addition of preservatives.
Preservation Ambient Storage Refrigerated storage

(30±2˚C) (7±1˚C)
No treatment 1 day 8 days
Sonication at 100 µm for 6 days 20 days

8min
Irradiation at 1.25 kGy 7 days 64 days
Addition of Nisin 10 days 25 days

(7.5ppm) + Potassium
Sorbate (2000ppm)
Keywords: Idli, fermented cereal-legume food, preservation, idli batter, sonication,

irradiation, preservatives, shelf-life.
i
CONTENTS
Sr. no Title Pg.no
ABSTRACT i
ACKNOWLEDGEMENT ii
LIST OF TABLES v
LIST OF FIGURES vi
ABBREVIATIONS AND SYMBOLS ix
I INTRODUCTION 1
II REVIEW OF LITERATURE
2.1 Formulation of idli batter 4
2.2 Fermentation of cereal-legume mix for idli 9
2.3 Preservation techniques for idli 11
III MATERIALS AND METHODS
3.1 Raw materials 17
3.2 Preparation of idli batter 18
3.3 Preservation of idli batter 19
3.4 Experimental design 22
3.5 Quality evaluation of idli batter 22
IV RESULTS AND DISCUSSION
4.1 Effect of sonication on preservation of idli batter 25
4.2 Effect of irradiation on preservation of idli batter 45
4.3 Effect of addition preservative on preservation of idli batter 58
V SUMMARY AND CONCLUSIONS 66
VI REFERENCES 69
VII APPENDIX
ACKNOWLEDGMENT
I believe in absolute oneness of God and also humanity. I bow my

head and express gratitude with all reverence and devotion to my “Almighty”
for showering his blessings.
I take this opportunity to express my deep sense of gratitude to

Dr.D.C. Joshi, Dean, College of Food Processing Technology and Bio-Energy
and Chairman of my Advisory Committee for his valuable suggestion,
constructive criticism coupled with excellent counsels and pains taking efforts
throughout the investigation, who provided me an opportunity for this
manuscript.
I feel a matter of great pride to record my reverence to the members

of my Advisory Committee Dr.A.G.Bhadania, Professor and Head,
Department of Dairy Engineering, Dr.S.S Kapdi, Professor and Head,
Department of Bio-Energy, Dr. Namrata Sutar, Assistant Professor, and
Er.Ramesh Modi Assistant Professor, for their timely support and
encouragement.
I am also thankful to all my teachers of Dr.R.V. Prasad, Dr.B.H. Joshi

and Mr. J.K. Momin and other staff members, who are responsible for the
drop of knowledge I may possess in the ocean of science. I wish to
acknowledge Mrs. Nimita Runajkar extending help during laboratory.
I acknowledge my lovely mother (Mrs.Shashikala), father

(Parthasarathi) and my brother (Preetam), for extending their sacrifice, faith,
unconditional love and everything that made me come this far.
Friendship being one of greatest blessing on the earth and it is a great

pleasure for me to acknowledge the help and support I received from my
friends. No language and printed words can express my feeling to my very
best friends Kavya, Swarana, Sunilkumar, Sridar, Nayan, Sriharsha, Pradeep,
and Kavita for being with me in moments of stress and tension, their selfless
ii
encouragement, patience, kindness and confidence, heartened me at most
crucial hours.
I am elated to avail this wonderful opportunity to record my

gratitude to all my seniors, Mamta, Ankita, Payal, Dipali, Viral, Akshay,
Tarang, my dearest and nearest friends Hetal, Payal, and all my
classmates and my well wisher juniors Gayatri, Srishti for their guidance
and cooperation.
I am thankful to Mannubhai, Poonamkaka and Vasavakaka for their

supportive help during the study.
I wish to extent special thanks to all of my beloved seniors, batch

mates and juniors for their encouragement and moral support during
research work.
Anand (Prarthana S.P.)

July,2013
iii
COLLEGE OF FOOD PROCESSING TECHNOLOGY & BIO-ENERGY
ANAND AGRICULTURAL UNIVERSITY,
ANAND – 388110
Dr. D.C.Joshi Tel. No. (02692) 261302

Principal Email: dcjoshi@aau.in
CERTIFICATE
This is to certify that the thesis entitled “DEVELOPMENT OF THE
PRESERVATION TECHNIQUE FOR IDLI BATTER” submitted by
Ms. PRARTHANA S P in partial fulfillment of the requirement for the award
of the degree of Master of Technology in Food Processing Technology to the
Anand Agricultural University is a record of bonafide research work carried
out by her under my guidance and supervision.
Place : Anand (D. C. Joshi)

Date : 16/07/2013 Major Advisor
CHAPTER I
INTRODUCTION
Food is a basic component of eco-system and human beings to select foods from the available
bio-resources which are edible. Traditional foods are popularly consumed and form an integral
part of our diet since early history. These are prepared in the household or in cottage industry
using relatively simple techniques and equipments (Aidoo et al., 2006).
India is traditionally rich in fermented foods. In the Indian sub-continent, fermented foods
using local food crops and other biological resources are very common. But the nature of the
products and the base material varies from region to region (Sekar and Mariappan, 2007).
Fermented foods such as idli and dahi were described as early as 700 BC. At present, there are
hundreds of fermented foods with different base materials and preparation methodology. Each
fermented food is associated with a unique group of micro-biota, which increases the level of
proteins, vitamins, essential amino acids and fatty acids in the food product. However, fermented
foods are still produced traditionally by spontaneous fermentation and only limited knowledge
has been obtained regarding the micro-biota of these products (Jeyaram et al., 2009).
There are various problems associated with the indigenous fermentation. They are uncontrolled
and often unhygienic, labor intensive, seen as primitive by some people, are normally not
integrated into the economic mainstream, have limited export potential, and in some cases, the
impact on nutritive value and safety is questionable (Singhal, 2005).
Cereal-based fermented foods are considered as staple diets in their respective regions. Most of
the foods such as idli, dosa, dhokla, koozhu, nan, parotta, ambali, pazhaiya soru are consumed on
the daily basis by the local population. Mostly they are made at household level and have short
shelf-life (Satish kumar et al., 2012).
Idli, a traditional breakfast food of India based on cereal-legume combination, is a white,

fermented (leavened), soft, spongy textured product. It’s widely consumed in entire South India
(Sridevi et al., 2010). Idli is a fermented, thick suspension made by blend of rice (Oryza sativa)
Intrpduction
and dehulled black gram (Phaseolus mungo). Traditionally, rice and black gram are soaked
separately. After draining water, rice and black gram are ground independently, with occasional
addition of water during the process. The rice is coarsely ground and the black gram is finely
ground. Then the rice and the black gram batters are mixed together with addition of a little salt
and allowing it to ferment overnight at room temperature. Finally, the fermented batter is placed
in special idli pans and steamed for 5–8 min (Blandino et al., 2003). The idli so prepared is
served hot.
Idli batter has very short shelf-life because of its high moisture content and live fermentation.
There is whey separation due to collapse in its volume after a certain period of fermentation
which further worsens with storage. Dried idli mixes are the alternatives put in the market.
However, because of the inferior texture of the product and its lower organoleptic quality, the
instant powders are not popular. Hence, there is a demand for ready-to-cook idli batter in
packaged form with moderate shelf-life. In general, the thermal techniques of preservation will
be suitable for enhancing the shelf-life. However, in case of idli, thermal preservation is not
feasible as the batter co-agulates or idli is formed on heat application. Hence only limited
alternative technologies can be applied in this case. No systematic studies are available on
preservation of idli batter. Hence, the present study was undertaken to evaluate some of the non
thermal methods of preservation for extension of shelf-life of fresh idli batter. The specific
objectives of the study were as follows;
1. To evaluate the effect of some non thermal methods of preservation on the shelf-life extension of
idli batter.
2. Evaluating the changes in microbial and organoleptic quality of preserved idli batter during
storage.
3. Determining the shelf-life of the preserved idli batter.
Ultra-sonication is considered to be an emerging and promising technology for industrial food

processing. It’s a non-thermal processing alternative for many liquid food products.
Its mechanical power is being used for a gentle yet effective microbial and particle size
reduction. Ultrasound, or sound of frequency ≥20 kHz, is generally associated with damage to
cells and is widely used in laboratory protocols for breaking cell walls to release intracellular
2
Intrpduction
products. Enzymes and other fragile macromolecules are known to be susceptible to damage by
ultrasound (Sulaiman et al., 2011). When food is exposed to ultrasonication, most of the yeast
cells are destroyed. Yeast cells that survive sonication generally lose their ability to grow. This
reduces the rate of fermentation substantially.
Food irradiation is another potential non-thermal food preservation process that involves
exposing the food to ionizing radiation. Radiation source normally used for food irradiation is:
Gamma-rays from the nuclides Co-60 or Cs-137. Although the mechanism of bacterial
inactivation by irradiation is not completely understood, it is believed that irradiation inactivates
microorganisms mainly by causing lesions in DNA. Irradiation is considered the only known
technique capable of ensuring the hygienic quality of raw food in a fresh or frozen state
(Loaharanu, 1995). According to established good manufacturing practices, irradiated food
products can be considered safe and nutritionally adequate (Olson, 1998). Irradiation enhances
the shelf-life of foods and ensures their innocuousness because most microorganisms in
vegetative form are extremely sensitive to irradiation at low doses (Radomyski et al., 1994).
Addition of bio-preservatives is another possible way of extending shelf-life of idli. The

preservation factors elaborated by LAB have been identified as peptides, generally known as
’Bacteriocins’. Nisin is commercially exploited bacteriocin used for a wide range of food
preparation for preservation (Jeevaratnam et al., 2005).
Considering above points of view, attempts were made to enhance the shelf-life of fresh idli
batter by sonication, irradiation and by adding bacteriocin.
3
CHAPTER II
REVIEW OF LITERATURE
2.1 Formulation of Idli Batter
Idli is a fermented food of India which is prepared by steaming a fermented black gram
(Phaseolus mungo L.) and rice (Oryza sativa L.) batter. It makes an important contribution to the
diet as a source of protein, calories and vitamins, especially B-complex vitamins, compared to
the raw unfermented ingredients. It can be produced locally and used as a dietary supplement in
developing countries to treat people suffering from protein calorie malnutrition and kwashiorkor.
Other legumes such as soybeans and Great Northern beans could be substituted for black gram in
preparation of an idli (Reddy et al., 1982).
Ghosh and Chattopadhyay (2011) prepared idli batter by soaking polished parboiled rice and
decorticated black gram for 4 h at 30±1°C in water. The blend ratios of rice to black gram in 2:1,
3:1 and 4:1 (w/w) were allowed for fermentation for different periods (7, 8, 9, 10 h) with the
addition of 2% (w/w) of salt. The density, pH, and percentage total acidity of the batter during
fermentation for different blend ratios ranged between 0.93 and 0.59 g cm-3, 4.21 and 5.9 and
0.44 and 0.91%, respectively. During fermentation, the maximum production of riboflavin and
thiamine were found to be 0.76 mg/100 g and 0.73 mg/100 g in 3:1 blend ratio of idli batter; and
the folic acid content was found to be the maximum of 0.75 mg/100 g of idli batter after 10 h of
fermentation.
Balasubramanian and Viswanathan (2007) while studying the physicochemical properties of

idli batter during fermentation, prepared idli batter by soaking polished parboiled rice and
decorticated black gram for 4 h at 30±1°C in water. Batters with rice to black gram ratios of 2:1,
3:1 and 4:1 (v/v) were fermented for 0, 6, 12, 18 and 24 h, adding 2% salt. Bulk density, pH and
percentage total acidity were in the ranges 0.94-0.59 g/cm3, 4.1-5.9 and 0.443-0.910%,
respectively. The consistency coefficient at any fermentation time increased as the rice to black
gram ratio was increased.
To optimize the ratios of rice to black gram dhal and fermentation time for idli, instrumental
Texture Profile Analysis (TPA) was conducted to optimize the textural properties of idli by
Manohar and Shetty (2012). A desirable score of 0.7439 was obtained for sensory attributes
4
Review of Literature
of idli made with the ratio of 3: 1.475 for IR20 idli rice and ADT3 variety black gram (with husk
removed after soaking) fermented for 10.2 h. It was concluded that at an optimum ratio of rice
and black gram dhal (3:1.575 w/w) with 14 h fermentation, Idli with desirable texture properties
can be prepared. The Table 2.1 shows the comparative data on the effect of ratios.
Table. 2.1: Effect of ratios of rice to black gram on pH, acidity and density of idli batter
Parameter 2:1 3:1 4:1 Source
pH 4.21 5.27 5.9
Acidity (%) 0.44 0.62 0.91

Ghosh and Chattopadhyay (2011)
Density (g cm-3) 0.93 0.71 0.59
pH 4.1 5.33 5.9
Balasubramanian and Viswanathan (2007)

Acidity (%) 0.443 0.73 0.91
Density (g cm-3) 0.94 0.74 0.59
Nazni and Shalini. (2010) prepared millet idli from rice and black gram dhal by incorporating
pearl millet. The developed products were analyzed for physical and chemical parameters and
organoleptic evaluation. The scores were compared with standard. The developed millet idli
were highly accepted and notable change in physical parameters of both millet incorporated
batter and idli was observed when compared to the standard. Except carbohydrate, remarkable
increase was observed in the nutrients such as protein, fat, fiber, calcium and iron in the mixed
and pearl millet idli compared to the standard.
Kaw and Linda (1987) prepared idli batters from rice of varied amylose contents namely: IR8
(high amylose), IR64 (intermediate amylose), IR24 (low amylose) and IR29 (non amylose or
5
waxy). The pH decreased and acidity (expressed as lactic acid) and non protein nitrogen
increased as fermentation progressed. However, the changes in these parameters were faster in
the batter prepared from varieties with higher and intermediate amylose. Volumes of batter from
low and non-amylose rice varieties were higher than those with intermediate and high amylose
content.
Rekha and Vijayalakshmi (2011) worked to reduce the natural fermentation period of idli by
adding under-utilized soy residue okara. Black gram was partially substituted with okara. They
reported that the pH, total acidity, amount of CO2 and viable count of yeast, mold and lactic acid
increased with time at the end of 10 h fermentation. Okara fortified idli was soft and spongy
compared to control idli.
Akolkar and Parekh (1983) prepared idli with ground rice flour, soya dal flour and salt. The
idli was prepared by steaming for 5 min in steamer both with unfermented and fermented batter
at 30˚ C for 16 h with addition of fenugreek leaves and lime powder. Their results showed that
fermented soy idli with lime and fenugreek leaves, improved growth and liver composition of
rat. Also, it was highly acceptable by pre-school children.
The effect of varietal differences and polishing of rice on quality parameters of “idli,” were
studied by Chandini et al. (2005). Two varieties of raw rice, “Jaya” and “Minilong,” and one
variety of parboiled rice “Ponni” with two degrees of polishing (high and low) were used.
Variations were observed in water and fat absorption capacities of two samples. Emulsification
capacity ranged from 102 to 110 mL/100 g. Foam capacities at different pH were similar, but
foam stability differed as a function of time. Rice with a lesser degree of polishing fermented
better with higher batter volume and microbial count, lesser shear value and gave softer idli.
However, sensory analysis revealed that idli prepared with low-polish rice scored significantly
lower for appearance and color quality compared with products prepared with high-polish rice.
They concluded that the quality characteristics of idli were influenced by the variety of rice and
the degree of polishing, but the two types of black gram used, whole and split, had no effect.
Geetha and Geetha (2012) studied new rice variety (TRY3) for idli making. They concluded
that rice variety, exhibiting high amylose and intermediate alkali spreading value should be
preferred for idli making. High viscosity and set back values indicated that in variety TRY3,
6
swelling granules is high and facilitates higher breakdown to a greater extent and more quantity
of batter. The organoleptic evaluation and consumer preference test confirmed that the idlis made
out of new variety rice TRY3 was superior in taste, palatability and stomach filling compared to
the market variety CR1009.
Ingredient composition and method of processing of instant rava idli mix (RIM) were
optimized and storage stability in polypropylene and paper-aluminium foil-polyethylene
laminate pouches at room temperature (RT) was studied by Madhura et al. (1998). During
storage of RIM, the concentration of free fatty acids and acidity value increased; hardness of idli
also increased, with a downward trend of acceptability. Results showed that instant RIM has a
shelf-life of 6 months at 30˚ C when packed in paper-aluminum foil-polyethylene pouches and 4
months at 37 °C when packed in polypropylene pouches.
A study was carried out by Shanthi et al. (2000) on the processing and evaluation of instant idli
mixes. They prepared idli mix using ground rice to Bengal gram ratio (4:1), 2% salt, 0.05% citric
acid and 0.2% sodium bicarbonate. The instant idli mix samples were packed in polypropylene
and metallised polyster and kept for 180 days. They reported that moisture, pH and thamine
content of idli mixes decreased during storage. Also organoleptic characteristics of mixes did not
change during storage.
Perez and Mishra (1995) studied flow behavior of regular and peanut-fortified idli batters.
They found that the regular and peanut-fortified idli batters showed shear-thinning behavior
before and after fermentation. Fermentation increased batter viscosity and degree of shear
thinning of regular and peanut-fortified batters. Fortification with defatted peanut flour (DPF) up
to 20%, significantly increased viscosity and the batters became more shear thinning at the 25–
30% DPF fortification level.
Germinated powders of high-quality millets were mixed and incorporated with other basic
traditional ingredients for preparing idli and dosa by Krishnamoorthy et al. (2013). The millet-
based idli contained high proportions of protein (15–18%), fat (5.0–6.2%) and carbohydrate (72–
74%) compared to the rice-based idli. The ash content was in the range of 1–2% and crude fibre
(3.0–4.9%). The millet-based dosa contained high proportions of protein (15–18%), fat (8.5–
9.8%) and carbohydrate (69–72%) compared to the rice-based dosa. Also the processing steps
like decortications, germination and fermentation significantly reduced the phytic acids (69%)
7
and tannin (78%) content in millet-based foods. The sensory evaluation results showed that the
overall acceptability of millet based product is ‘Like moderately’ with score of 7.7 ± 0.5.
Singh et al. (2007) studied the effect of substituting rice with extrusion cooked (75, 100 and
125°C) rice flour at 10, 20, 30 and 40% level while preparing idli. Both the levels of substitution
and temperature of extrusion had significant effects on the specific gravity, acidity and pH of the
idli batter, and textural and sensory quality of the idlis. The idlis prepared from idli mix
containing rice substituted by 20–30% of rice flour extruded at 125˚C were most acceptable.
Effect of addition of xanthan to instant idli batter was studied by Thakur et al. (1995). Xanthan
at the rate of 0.01, 0.05, 0.1, 0.2 or 0.3% was added to the batter mix in each of the following
ways: prior to addition of water; by dispersion in 50% of the water required to prepare the batter;
or by dispersion in the total amount of water required to prepare the batter. Effects of xanthan on
texture of idli depended upon concentration and addition method. The desired texture of idli was
obtained when approximate 0.1% pre-swollen xanthan was added to the batter mix.
Mahajan and Chattopadhyay (2000) studied development of chemically leavened cereal-

legume based instant mix (dhokla). They found that the proportion of rice: bengal gram: black
gram (0.6:1:0.3) was the best in terms of minimum bulk density and hardness in traditional
method while, the instant mix prepared from 45:45:10 proportion with added 1.8% sodium
bicarbonate, 1.28% citric acid, 4% sugar, holding for 15 min and 20 min steaming showed
increased acceptability for taste as well as sponginess.
Kaur et al., (2000) studied the effect of extruded flour and fermentation time on some quality
parameters of idli. Fermentation time showed the most pronounced effect on specific gravity, pH
and aqueous dispersion viscosity of batter and taste scores of idli. Substituting rice flour with
extruded rice flour at different levels has significant effect on expansion and appearance scores
of idli. The textural scores and compression force was significantly affected by extrusion
temperature. The formulation containing rice substituted by 30% extruded rice flour extruded at
175˚C and fermented for 24 h produced idli with highest expansion and overall acceptability.
8
2.2 Fermentation of Cereal-legume Mix for Idli
Fermentation is the process which involves the action of micro-organisms or enzymes which
cause desirable changes and significant modification to the food (Geoffrey, 1994). Fermented
foods are of great significance as they provide and preserve vast quantities of nutritious products
in a wide diversity of flavors, aromas and textures which enrich the human diet (Steinkraus,
1994). Fermented foods are important components of diet in part of world, especially Southeast
Asia, Near East and parts of Africa. They form an appreciable part of daily diet of people as a
main source of protein, calories and certain vitamins. The fermentation helps not only in
improving the organoleptic quality of legumes but also enhances nutritional quality (Reddy et
al., 1982; Vanveen and Steinkraus, 1970).
Mukherjee et al. (1965) found that predominant microorganism responsible for souring as well
as for gas production in idli batter, was Leuconostoc mesenteroides and leavening action was by
lactic acid bacterium. Caplice and Fitzgerald (1999) have reported that a strain of yeast
Torulopsis holmii besides lactic acid bacteria were responsible for fermentation of idli batter.
Venkatasubbaiah et al. (1985) reported the involvement of Trichosporan pullulans, Torulopsis
holmii and Torulopsis candida for fermentation in idli batter besides lactic acid bacteria.
Soni et al. (1986) found that Leuconostoc mesenteroides, Streptococcus faecalis, Lactobacillus
fermentum and Bacillus amloliquefaciens were the predominant bacteria responsible for souring
along with leavening of dosa batter and helped in the saccharification of starch.
Yadav and Khetarpaul (1995) studied the effect of fermentation period and temperature on anti
nutrients and in vitro digestibility of starch and protein of Wadi- a fermented legume product.
They concluded that higher the temperature and longer the period of fermentation, greater the
extent of phytic acid reduction. Fermentation at 35˚ C for 18 h reduced the phytate content to
half. Protein digestibility of Wadi batter fermented at 35˚ C was higher than at 25˚ and 30˚ C as
well as protein digestibility improved with fermentation.
Yajurvedi (1980) reported that fermentation temperature in the range of 25-30˚ C was the most
suitable, temperature above 40˚ C gave rise to some undesirable smell in idli batter. Also, he
reported that fermentation does not improve nutritive value of protein but increase vitamins. The
digestibility of idli was increased, along with decreased cooking time. Fermentation has not been
9
found to significantly affect the protein content of idli and khaman (Reddy et al,. 1981;
Rajalakshmi and Vanaja, 1967).
Soni et al. (1985) studied dosa with respect to some biochemical changes associated with
fermentation. They observed overall increase in microbial load, batter volume, total nitrogen,
soluble proteins, reducing sugars and decrease in pH, after 30 h fermentation. Amylases after
showing an increased activity in early stages, declined gradually with the progress in
fermentation.
Bruntha Devi et al. (2012) studied enhancement of nutritional value of finger millet-based
food (Indian dosa) by co-fermentation with horse gram flour. Natural fermentation of finger
millet–horse gram flour blend in different proportions (2:1, 3:1, 4:1 and 5:1) was performed for
24 h. Biochemical analysis showed reasonable drop in pH (6.6–4.2) and starch content (25.52%)
with considerable augment in titratable acidity (0.168–1.046%), soluble proteins (1.1-fold) and
free amino acids (2.6-fold) at 16 h. Total essential amino acids increased 1.1-fold at 16-h
fermentation with protein containing 48.68% of essential amino acids over total amino acids.
Lysine increased from 5.87 to 6.73 g of amino acid/100 g of total amino acids. Dosa, prepared
from 16-h fermented batter, showed better sensory attributes for 4:1 ratio.
Effect of fermentation on amino acid composition of cereal and pulse based foods was studied
by Riat and Sadana (2009). They showed that the fermented batter of idli and dosa contained
higher amount of methionine, cystine and lysine. After processing, maximum retention of lysine,
methionine and cystine was observed in steamed idli while fried sample recorded least lysine and
methionine.
Sridevi et al. (2010) conducted study on selection of starter cultures for idli batter fermentation
and their effect on quality of idli. They prepared idli batter using different cultures and packed in
polyethylene terephthalate – low density polyethylene (PET-LDPE) laminated pouches, high
density polyethylene (HDPE) tins and aluminum foil laminated LDPE pouches and stored at 30˚
C. Enterococcus faecium (Ef) + Candida versatilis (Cv) culture performed better and their result
showed rise in batter volume, level of CO2 production, titrable acidity (lactic acid) and pH.
Aluminum foil laminated LDPE pouches were found to be better in extending the keeping
quality of the batter.
10
A study was carried out by Agrawal et al. (2000) on flavor profile of idli batter prepared of
Candida versatilis CFR 505 and Pediococcus pentosaceus CFR 2123, packed in polyester
polylaminate pouches at 30˚ C. Desired flavor compounds were present up to 8 days of storage at
30 ± 2 ˚C, whereas undesirable flavors appear after 6 days of storage at 30 ± 2 ˚C.
A study was done by Kelkar et al. (1996) on the effect of processing on in-vitro carbohydrate
digestibility (IVCD) of cereals and legumes such as rice, bajra, jowar, wheat semolina, green
gram, red gram, black gram and some of the selected products such as Papad, Shev, Idli and
Dhokla. Fermentation increased IVCD of rice by 13-25%, 15-28% of legumes and 10-39% of
fermented products such as Idli and Dhokla. This increase may be due to the degradation of
starch into smaller fragments and formation of reducing sugars.
Joshi et al. (1989) studied microbial and biochemical changes during dhokla batter
fermentation with special reference to flavor compounds. Their result showed that change in
volume of the batter commence at the 10th h of fermentation. The total count of micro-organism
increased as fermentation time was increased. Also, the chemical changes such as pH of the
batter and steamed product decreased, total titrable acidity of batter and product increased. The
sensory qualities of product and characteristic pleasant flavor exhibited by the product of batter
fermented for 12 h while medium sour taste with 14 h fermentation and highly spongy texture
was obtained with 16 h fermentation.
The effect of amylase enzyme on idli batter fermentation was studied by Iyer and
Ananthanarayan (2007). It was observed that addition of α-amylase @ 5-25 U per 100 g of batter
increased the rate of fermentation and shortened the fermentation period. The batters to which 5
and 15 U amylase was added and fermented for 8 h at 30ºC yielded idli having good sensory
acceptability. Higher levels of amylase addition (25 U) adversely affected batter viscosity and
batter volume. In terms of bulk density and texture, these idli were comparable to the product
made from naturally fermented batter. Thus, the fermentation time was brought down to 8h
(vs.14 h for conventional).
2.3 Preservation Techniques for Idli
The stabilization of the idli batter at room temperature and refrigerated storage by using
various hydrocolloids and surface-active agents was studied by Nisha et al. (2005).
11
Hydrocolloids gave good stabilization, surface-active agents failed to stabilize the batter
although they reduced whey separation. Among the various hydrocolloids, 0.1% guar gum gave
best batter stabilization, and idli made from it after 10 days at room temperature and 30 days of
refrigerated storage of batter was found to be of acceptable quality.
Using preservatives and temperature are the two important hurdle technologies for preserving
idli batter. Addition of chemical preservatives potassium meta bisulphate, sodium benzoate and
potassium sorbate in the limits permitted, Nisin, a microbial polypeptide and the combination of
these two at temperatures of 28–30°C (room temperature), 4–8°C (refrigerated) and -18°C
(frozen) storage were studied by Nisha et al. (2007). A combination of 7.5 ppm nisin and 2000
ppm potassium-sorbate could preserve idli batter for 10 days at room temperature, 30 days at
refrigerated temperature and 45 days under frozen storage.
Idli was dried by Kanchana et al.(2008) using microwave drying (MD), vacuum assisted MD
and hot-air drying. Moisture, bulk density, water activity (aw) and instrumental texture quality
were studied. Moisture content of dehydrated idli ranged between 6.7 and 9.4% and the water
activity (aw) was between 0.166 and 0.441. Low power density and low temperature dehydrated
idli was acceptable. Rehydration of idli was better for product from hot-air oven drying followed
by vacuum assisted MD and domestic MD.
A study carried out by Yusuf and Varadaraj (1998) on inherent viable bacterial populations of
mesophilic aerobes and lactic in idli batter. The addition of Plantaricin LP84, a bacteriocin
produced by Lactobacillus plantarum NCIM 2084 to idli batter at 1% level was able to retard the
growth of the inoculated pathogens during fermentation.
Piyasena et al. (2003) studied inactivation of microbes using ultrasound. They found that the
pressure change cause cavitation and gas bubbles. B. subtilis spores using treatment at 20 KHz
and 150 µm at 500 KPa for 12 min was lethal. L. monocytogenes using treatment at 20 KHz and
117 µm at 200 KPa reduced D-value to 1.5min. Yersitia entercolitica at 30 KHz and 150 µm
from 0-600 KPa resulted in D-value decrease from 1.52 to 0.2 min.
Subramanian et al. (2007) studied ultrasonic honey processing and found that most of the yeast
cells were destroyed. Yeast cells that survive sonication generally lose their ability to grow. This
reduces the rate of honey fermentation substantially. It aids liquefaction at substantially lower
12
process temperatures of approx 35°C and can reduce liquefaction time to less than 30 seconds.
Sonication at a frequency of 20 KHz liquefied the crystals in the honey, completely. Due to the
minimal heat exposure, ultrasonic liquefaction results in a greater retention of aroma and flavor.
High amplitude ultrasound has a very good homogenization effect on milk with yogurt starter
compared with conventional homogenization (Hongyu et al, 2000). Sonication after inoculation
reduced the total fermentation time by 0.5 h. Increasing the ultrasound amplitude level before
inoculation significantly improved the water holding capacity and viscosity and reduced
syneresis. Sonication after inoculation did not decrease the syneresis effect but increase the water
holding capacity and viscosity.
Effect of pulsed electric field on inactivation of microorganism was studied by Kristina et al.
(2001). The S. cervevisiae was the most sensitive organism, followed by E.coli, when they were
exposed to 30 kv/cm, 20 pulses with 4 µsec duration. The most resistant organisms were Listeria
innocua, Leuconostocs mesentroides, with only a 3 log reduction by increasing the parameters to
35 kv/cm, 40 pulse and 4 µsec pulse duration; marked viability reduction.
The combined effects of heat treatment and gamma-irradiation on the inactivation of major
lactic acid bacteria associated with Kimchi fermentation was studied by Byun et al. (1989). The
radiosensitivities (D10 values) of LAB in case of irradiation treatment were 0.61 kGy in
Lactobacillus brevis, 0.60 kGy in Lactobacillus plantarum, 0.50 kGy in Leuconostoc
mesenteroides, 0.4 kGy in Pediococcus cerevisiae and 0.39 kGy in Streptococcus faecalis. The
combined treatment could be applied for the radiation preservation of Kimchi, minimizing the
side-effects like physical changes induced by the high dose irradiation or heat treatment.
Gamma irradiation up to 10 kGy in the early stage of Kimchi fermentation had a dose-
dependent effect on the inactivation of fermentative microbes, lowering the lactate
dehydrogenase (LDH) activity and delaying acidification. Although gamma irradiation on the
mid-fermentation stage of Kimchi inactivated the fermentative microbes effectively, LDH
activity remained high and acidification continued. Kimchi irradiated at 10 kGy had lower
acceptability scores than those of control, 2.5 and 5 kGy irradiated product. Therefore, gamma
irradiation up to 5 kGy in the early fermentation stage was recommended for aging control and
the improvement in the shelf-life of Kimchi (Hyun et al., 2004).
13
Effects of gamma irradiation on the physiological activity of Korean soybean fermented food
Chungkookjang (the whole cooked soybean product) and Doenjang (soybean paste) was studied
by Myung et al. (2002). The physiological activity was evaluated by angiotensin converting
enzyme inhibition, xanthine oxidase inhibition, tyrosinase inhibition and radical scavenging
ability and results indicated that the treatment at ≤ 10 kGy did not show any significant change
on physiological activities by irradiation.
An irradiation effect on biogenic amines (BAs) and microbiological populations of Korean

fermented soybean paste was studied by Kim et al. (2003). Soybean paste was prepared and
irradiated with doses of 5, 10, and 15 kGy, and then fermented at 25°C for 12
weeks. Bacillus spp. and LAB decreased by irradiation but increased during fermentation.
Biogenic amines (BA) content has no significant difference between control and irradiated
samples immediately after gamma irradiation. However, 4 kinds of BA’s, putrescine, tryptamine,
spermidine, and histamine, showed significant reduction by irradiation during fermentation.
Fermented sausage was prepared from batter which had been processed with gamma radiation
to reduce the microflora. Doses up to 5 kGy reduced the total aerobic bacteria in commercial
batter up to 2.2 log cycles. Irradiation reduced the coliform and staphylococci counts to levels
that are considered safe for public health. These reductions allowed the use of a lower inoculum
and a longer fermentation. The reductions shifted the percent inoculums relative to the total
population from 1% in the controls to 70% in the 5 kGy samples, and the shift in population
resulted in a more uniform fermentation and final product (Dickson and Maxey, 1985).
Salted shrimp was irradiated at 2 different stages: 1) irradiated immediately after processing, 2)
irradiated at optimum fermentation period, and fermented at 15°C for 10 weeks. During
fermentation, volatile basic nitrogen (VBN) content increased as the salt concentration and
irradiation dose was decreased. The combination of low salt concentration (15% or 20%) and
gamma irradiation (5 or 10 kGy) was effective in processing low-salted and fermented shrimp
(Lee et al., 2002).
Nisin is a natural antimicrobial peptide produced by strains of Lactococcus

lactis subsp. lactis that effectively inhibits Gram-positive and Gram-negative bacteria and also
the outgrowth of spores of Bacilli and Clostridia. It has been used as a biopreservative and a
potential agent in pharmaceutical, veterinary and health care products (Arauz et al., 2009).
14
Bacteriocins have improved synergistically the inhibitory effects of thermal and non-thermal
treatments, such as high-intensity pulsed electric fields (HIPEF) and high pressure (HP). The
combination of bacteriocins with heat may facilitate the application of mild thermal treatments,
which may diminish the typical cooked flavor of milk and reduce the cost of the heating
operation. In particular, the enhancement of the lethal effect of HIPEF and HP, when combined
with bacteriocin addition, may provide a suitable alternative for traditional thermal treatments,
since it causes minimal alteration to sensory properties of the product (Lopez, and Belloso.
2008).
Biochemical studies revealed that the bacteriocin were heat-stable even at autoclaving
temperature (121 °C for 15 min) and were active over a wide pH range (2–10). The bacteriocins
were inactivated by α-chymotrypsin and proteinase K but not other proteases. The antimicrobial
spectrum and some characteristics of this bacteriocin were identical to nisin. The ability of the
bacteriocin produced by Lc. lactis WNC 20 may be useful in improving the food safety of the
fermented product (Noonpakdee et al., 2003).
Preservative ability of LAB in foods is attributed to the production of anti-microbial

metabolites including organic acids and bacteriocins. Bacteriocins generally exert their anti-
microbial action by interfering with the cell wall or the membrane of target organisms, either by
inhibiting cell wall biosynthesis or causing pore formation, subsequently resulting in death. The
incorporation of bacteriocins as a biopreservative ingredient into food systems were studied
extensively and been shown to be effective in the control of pathogenic and spoilage
microorganisms. However, a more practical and economic option of incorporating bacteriocins
into foods can be the direct addition of bacteriocin-producing cultures into food (Sullivan et al.,
2002).
Lacticin 3147 is a novel heat-stable bacteriocin, produced by Lactococcus lactis DPC 3147,
that exhibits a broad-range inhibition spectrum similar to nisin. The effect of lacticin 3147 and
nisin on the shelf-life of fresh pork sausage and their ability to control pathogens (Clostridium
perfringens DSM 756, Salmonella Kentucky AT1) and nonpathogenic Listeria innocua DPC
1770 were investigated. The following preservative regimens were evaluated, both in broth and
sausage systems: (i) 450 ppm of sodium metabisulphite; (ii) 500 IU g-1 or ml-1 of nisin, (iii) 2500
15
arbitary units (AU) g-1 or ml-1 of lacticin 3147; (iv) 2% sodium lactate and 500 IU of nisin; (v)
2% sodium citrate and 500 IU g-1 or ml-1 of nisin; (vi) 2% sodium lactate and 2500 AU g-1 or ml-
of lacticin 3147, (vii) 2% sodium citrate and 2500 AU g-1 or ml-1 of lacticin 3147, (viii) 2%
sodium lactate, and (ix) 2% sodium citrate. There was no significant difference in the activity of
nisin and lacticin 3147 against any of the target strains used, both bacteriocins performing
significantly better than sodium metabisulfite against gram-positive strains in broth systems. In
addition, lacticin 3147 combined with either sodium citrate or sodium lactate maintained
significantly lower (P < 0.05) total aerobic plate counts and may function as an alternative to
sodium metabisulfite in the preservation of fresh pork sausage (Scannell et al., 2000).
16
CHAPTER III
MATERIALS AND METHODS
3.1 Raw Materials

Bulk samples of 25 kg milled rice (Oryza sativa) var. IR 20, and 10 kg dehulled black gram
(Phaseolus mungo) splits were procured from the local market (Fig. 3.1) and stored under
ambient conditions.
(a) (b)
Fig. 3.1: Bulk sample of (a) milled rice and (b) split black gram used to prepare Idli
The physico – chemical analysis of the raw material was carried out using standard procedures
and the values are depicted in Table 3.1. Prior to each experiment, rice and pulse splits were
washed with clean water.
Table 3.1(a) Physico – chemical property of rice.
Parameter Kernel elongation Volume expansion Alkali value Amylose(%) Protein
ratio (%)
1.6 4.2 4.5 24.3 8.3
IR 20
Table 3.1(b) Physico – chemical property of split black gram dhal.

Parameter Moisture Protein Fat Ash Total carbohydrate
(%) (%) (%) (%) (%)
10.9 24.0 1.6 0.9 62.8
Black gram dhal
17
Materials and Methods
3.2 Preparation of Idli Batter

Idli batter was prepared using the mixture of milled rice and dehulled black gram split (dhal) in
3:1 ratio. The ratio was selected based on the published literature (Ghosh and Chattopadhyay,
2011; Manohar and Sheety, 2012). The ingredients (rice and dhal) were processed for making
idli batter using good manufacturing practices (Agrawal et al., 2000). The process flow chart is
depicted as Fig 3.2.
Milled rice Black gram dhal
Wash and soak (6-8 hrs) Wash and soak (6-8 hrs)
Grind finely Grind coarsely
Combine slurries into a thick batter and mix well
Addition of salt @ 2%
Incubate at (30±2ºC) for 8–10 hrs
Fig 3.2 Process flow chart for prepration of idli batter (Agrawal et al, 2000).
18
Soaking was done in potable water for 6-8 h (Varadaraj et al., 1999). After draining the water,
rice and black gram were ground, during grinding water was added @ 1.5 to 2.0 times the
initial weight of the rice and black gram. The rice was coarsely ground and the black gram was
finely ground. The slurries were combined, salt @ 2% was added and stirred manually to form
a thick batter. The batter was allowed for fermentation for 8-10 h at room temperature (30±2˚
C), (Blandino et al., 2003). The Fig 3.3 shows the fresh idli batter prepared.
Fig. 3.3 Freshly prepared idli batter
3.3 Preservation of Idli Batter

Three different non-thermal techniques such as sonication, irradiation and addition of bio-
preservatives were attempted to enhance the shelf-life of idli batter. The preservation treatments
were given to the freshly prepared and fermented idli batter before storage.
3.3.1 Sonication of idli batter
Ultrasound generators (700 W and 230 W powers at 20 kHz) with an 8 and 6-mm diameter probe
(Model Q700, Q Sonica Sonicators Company, USA, and Model VCX 130, Sonics Vibra Cell
Company, USA) were used to sonicate idli batter (Fig 3.4). The amplitude levels selected were
60, 100 and 182. Exposure times were 5, 8, 12 and 15 min. Two sonicators were used because in
Q Sonica equipment, amplitude level more than 100 µm is not possible. The batter was evaluated
for its quality at the end of storage under either ambient temperature (30±2˚ C) or refrigerated
19
(7±2˚ C) storage. The samples were kept upto 8 days under ambient and 20 days for refrigerated
storage. The freshly prepared and fermented sample of (50 – 250 ml) of idli batter as described in
section 3.2 was taken in a glass beaker and kept on sonicator platform filled of water, and the
amplitude level and exposure time was set manually in digital panel fitted to the equipment.
After sample got sonicated the batter was filled in LDPE bag (300 gauge) and kept for storage
studies. The quality of fresh/the stored batter was evaluated by the methods described later in the
chapter.
(a) (b)
Fig 3.4: Sonication equipment (a) Q Sonica, Q700, (b) Sonic vibra cell, VCX 130.
3.3.2 Irradiation of idli batter
The idli batter was irradiated in Cobalt- 60 gamma radiation chamber (Model GC 5000,
ANGRU, Hyderabad) at 0.5, 0.75, 1.0 and 1.25 kGy at the rate of 9 kGy h-1 with an activity of
518 TBq (Fig 3.5). The programmed dose of radiation was controlled by removing the sample
from the radiation source at the appropriate time. The freshly prepared and fermented sample of
idli batter as described in section 3.2 was taken in LDPE bag (300 gauge) and put in irradiation
chamber. The chamber was closed and frequency was set manually and time was automatically
set by the equipment as per the strength of the irradiation probe. Irradiated sample was kept for
storage studies under both the ambient and refrigerated conditions. The irradiated batter was
20
studied at the end of 2, 4, 6 and 8 days of storage at ambient temperature (30±2˚ C). For
refrigerated (7±2˚ C) storage, samples were kept for 5, 10, 15 and 20 days. The quality of the
batter was evaluated using prescribed procedure described later on.
Fig. 3.5 Irradiation chamber GC 5000 used for treating idli batter
3.3.3 Chemical preservation of idli batter
The selective bio-preservatives were incorporated in the batter after fermentation. Weighed
amount of Nisin (7.5 ppm) and Potassium Sorbate (2000 ppm) were mixed (Fig 3.6) in a small
amount of water and solubilized. The solution was mixed well in the idli batter and homogenized
with a hand mixer to distribute in batter uniformly. After mixing the solution in the batter, it was
filled in LDPE bags (300 gauge) and kept for storage under ambient as well as refrigerated
conditions. The batter was evaluated at the end of 2, 4, 6 and 8 days of storage at ambient
temperature (30±2˚ C). For refrigerated (7±2˚ C) storage, samples were kept for 5, 10, 15 and 20
days.
21
Fig. 3.6 Nisin + Potassium Sorbate used as preservatives
3.4 Experimental Design
Variables Observations
1. Sonication Quality of treated and stored

Idli batter in terms of its,
 Amplitude: 60, 100 and 182 µm
 pH
 Exposure time: 5, 8, 12 and 15 min
 Acidity
2. Irradiation
 Standard plate count
 Frequency: 0.5, 0.75, 1.0 and 1.25 kGy
 Overall acceptability
3. Chemical preservation
 Nisin(7.5ppm)+Potassium sorbate(2000ppm)
4. Storage of idli batter
 Storage temperature and period
 30±2˚C for 2,4,6 and 8 days

 7±1˚C for 5,10,15 and 20 days
22
All experiments were conducted with three replications and the data were subjected to statistical
analysis using Factorial Complete Randomized Design and using analysis of variance
(ANOVA). Differences were identified as significant or non-significant based on mean squares
and F-test for significance at 5 % level of each treatment.
3.5 Quality Evaluation of Idli Batter
All the experiments were conducted and quality responses such as pH of batter, total titrable
acidity, total microbial count of batter and overall acceptability scores were determined.
Following techniques were adopted to analyze the above quality parameters.
3.5.1 pH of batter
The pH of the idli batter was checked individually using a microprocessor based digital pH meter
(pH Tester 30, Elico LI 610, Singapore) as per the procedure described by Ranganna (1986).
3.5.2 Acidity
The acid content in the term of total titrable acidity (TTA) of the batter was measured by titration
of 25 g of batter in 50 ml of distilled water against freshly prepared 0.1 N sodium hydroxide
solution using phenolphthalein as indicator. The TTA of fermented batter was calculated using
following formula and expressed as % of lactic acid (Iyer and Ananthanarayan, 2008).
TTA (%) = Titre×Normality of alkali× Volume made up× equivalent wt of acid×100

Vol. of sample taken for estimation × vol. of sample taken× 1000
3.5.3 Standard plate count
The standard plate count of the fermented batter was analyzed in terms of the total microbial
count. The test was performed on standard plate count agar using pour plate method. The plates
were incubated at 37˚ C for 24 h in incubator. The colonies which appeared after incubation were
counted as colony-forming units cfu/g in colony counter (Model: Galaxy 230, Make: Taurus
scientific, India).
23
3.5.4 Overall acceptability
Idli was prepared from the fresh, treated and stored batter as described above and was
evaluated for its organopletic quality. Five panelists were given a proforma for sensory
evaluation and asked to give the score for each sample based on the overall acceptability. A 9-
point hedonic test was employed for the attribute evaluated where 9 denoted “likely extremely”
and 1 indicated “disliked extremely” (Appendix - I).
3.5.5 Determination of shelf-life
The shelf-life of the sample was evaluated based on the quality parameters evaluated as above.
The cut-off period for the storage considered to be safe was determined considering the overall
acceptability score of the sample. The samples having overall acceptability score more than 6.0
were considered as acceptable and increase in the shelf-life was accordingly decided from the
combination of parameters.
The exact value of the shelf-life in days was calculated from the mathematical equations
obtained from each curve drawn between the overall acceptability and the variable.
24
CHAPTER IV
RESULTS AND DISCUSSION
This chapter deals with the results obtained from various experiments carried out
during the investigation. The data collected were statistically analyzed and the results are
discussed and interpreted for certain conclusions. The results obtained during the
investigation on different aspects are covered under following sub sections;
4.1 Effect of sonication on preservation of idli batter,

4.2 Effect of irradiation on preservation of idli batter, and
4.3 Effect of addition of preservatives on preservation of idli batter.
4.1 Effect of Sonication on Preservation of Idli Batter
The fresh and fermented samples of idli batter were sonicated at amplitude levels
of 60, 100 and 182 µm for 5, 8, 12 and 15 min exposure time. After the sample was
treated, it was packed in LDPE bag (300 gauge) for storage studies at both the ambient
and refrigerated conditions. The sonicated and stored idli batter was evaluated for
different quality attributes as a function of the sonication and storage conditions, Tables
4.1 through 4.4 show the data of different quality characteristics such as pH, acidity, SPC
and overall acceptability of stored idli batter after sonication. The ANOVA for each
response is also appended with the tables.
The samples at 182 µm sonication and for time more than 5 min got charred
during the sonication (Fig 4.1). Therefore, the data for 8, 12 and 15 min at 182 µm
treatment could not be collected. Hence practically only two levels of sonication (60 &
100 µm) were compared. Similarly when the treatment was inadequate, the samples
during storage got spoiled. The spoilage was in the form of either mold growth or gas
formation (Fig 4.2).
The quality evaluation of the charred or the spoiled samples could not be done
and hence those data in the Tables and Figs are missing.
25
Results and Discussion
Fig 4.1: Charred sample at 182 µm amplitude, exposing for more than 5 min
(a) (b)
Fig 4.2: (a)Spoilage due to mold growth and (b) Gas formation
26
Table 4.1: Effect of sonication and storage on pH of idli batter
Sonication 60 µm 100 µm 182 µm

amp litude & t ime No 5 min 8 min 12 min 15 min 5 min 8 min 12 min 15 min 5 min 8 min
Storage
Period (days)
sonication
Ambient 0 4.71 4.62 4.64 4.72 4.76 4.68 4.74 4.81 4.89 4.72
sonication time more than

(30±2˚c) 2 ** 4.44 4.48 4.51 4.57 4.60 4.67 4.72 4.76 4.70
Sample charred for

4 ** ** ** ** ** 4.58 4.61 4.68 4.73 4.67
6 ** ** ** ** ** ** 3.35 3.56 3.78 3.98
5 min
8 ** ** ** ** ** ** ** ** ** **
Refrigerated 5 4.56 4.32 4.41 4.54 4.67 4.36 4.46 4.62 4.74 4.54
(7±1˚c) 10 4.01 3.93 4.14 4.35 4.23 4.01 4.22 4.46 4.58 4.26
15 3.87 3.78 3.96 4.02 3.99 3.95 4.06 4.21 4.36 4.19
20 ** 3.02 3.34 3.46 3.61 3.38 3.57 3.68 3.88 4.05
Source D.F M.S F Value S.Em Test
Treatment 58 3.91 9.25 0.35 *
ANOVA Error 113 4.516
C.V % 13.09
* Significant at 5 % level of significance
** Samples spoiled
27
Table 4.2: Effect of sonication and storage on acidity (% TAA) of idli batter
Sonication amplitude & No 60 µm 100 µm 182 µm

time sonication 5 min 8 min 12 min 15 min 5 min 8 min 12 min 15 min 5 min 8 min
Storage
Period (days)
Ambient 0 2.741 2.704 2.679 2.598 2.562 2.667 2.578 2.458 2.421 2.531

(30±2˚c) 2 ** 2.977 2.851 2.695 2.606 2.812 2.713 2.649 2.593 2.712
Sample charred for

4 ** ** ** ** ** 3.196 2.894 2.787 2.712 2.963
6 ** ** ** ** ** ** 3.257 3.025 2.918 3.271
5 min
8 ** ** ** ** ** ** ** ** ** **
Refrigerated 5 2.918 2.985 2.961 2.752 2.642 2.761 2.672 2.544 2.451 2.512
(7±1˚c) 10 3.203 3.497 3.168 2.991 2.712 3.384 3.075 2.964 2.647 2.891
15 3.853 3.832 3.461 2.925 2.863 3.749 3.338 3.295 2.743 3.113
20 ** 4.389 3.974 3.815 3.541 4.112 3.767 3.611 3.266 3.287
Treatment 113 0.86 38.64 0.356 *
C.V % 17.01

** Samples spoiled
28
Table 4.3: Effect of sonication and storage on SPC (107 cfu/g) of idli batter

time sonication 5 min 8 min 12 min 15 min 5 min 8 min 12 min 15 min 5 min 8 min
Storage
Period (days)
Ambient 0 17.8 7.9 5.6 3.9 2.6 4.7 3.3 3.5 1.8 1.2

(30±2˚c) 2 ** 10.2 8.3 6.7 5.1 5.3 4.6 4.1 3.7 3.9
Sample charred for

4 ** ** ** ** ** 16.7 10.5 8.4 5.7 9.6
6 ** ** ** ** ** ** 14.8 10.1 9.8 13.8
5 min
8 ** ** ** ** ** ** ** ** ** **
Refrigerated 5 36.8 16.7 14.8 12.7 7.9 14.9 12.7 9.4 6.7 6.7
(7±1˚c) 10 75.7 20.1 18.5 15.9 13.5 18.2 15.9 11.8 9.3 13.3
15 140.3 32.5 27.9 22.8 20.1 28.6 24.3 19.7 14.6 20.1
20 ** 40.9 39.4 34.2 29.7 37.1 35.3 25.0 20.4 36.9
Treatment 112 0.21 32.40 0.358 *
C.V % 23.27
** Samples spoiled
29
Table 4.4: Overall acceptability score of sonicated and stored idli batter

time sonication 5 min
Storage
8 min 12 min 15 min 5 min 8 min 12 min 15 min 5 min 8 min
Period (days)
Ambient 0 8.9 8.0 8.1 8.2 8.4 8.5 8.6 8.6 8.8 8.6

(30±2˚c) 2 ** 7.8 8.0 8.1 8.2 7.9 8.3 8.4 8.6 8.4
Sample charred for

4 ** ** ** ** ** 6.4 6.5 6.6 6.8 7.2
6 ** ** ** ** ** ** 5.8 5.5 5.3 4.2
5 min
8 ** ** ** ** ** ** ** ** ** **
Refrigerated 5 8.1 6.8 7.1 7.5 8.2 8.5 8.6 8.7 8.8 8.9
(7±1˚c) 10 6.4 5.7 6.0 6.3 6.9 6.3 6.4 6.9 7.3 7.4
15 4.9 5.2 5.5 5.8 6.1 5.8 6.1 6.4 6.7 6.5
20 ** 3.7 4.8 5.4 5.7 3.9 5.0 5.6 5.9 5.4
Treatment 53 2.15 1.13 0.20 *
ANOVA Error 1
C.V % 22.98

** Samples spoiled
30
The variations in the quality parameters of the sonicated idli batter during storage
are also presented in the graphical way through Figs. 4.3 to 4.14. From the stastical
analysis, it was observed that all the three parameters (amplitude level, sonication time
and storage period) significantly (at 5% level) affected the quality and the shelf- life of the
idli batter.
For understanding the response of each quality attribute to the preservation

treatment given, the influence of sonication conditions on pH, acidity and SPC is
described in details here under.
4.1.1 Effect of sonication and storage on pH of idli batter
(a) Effect of sonication amplitude

As the amplitude of sonication was decreased from 100 to 60 µm, the pH of the
stored idli batter decreased for many samples for all the times of sonication and for both
the ambient and refrigerated storages (Fig 4.3 a & b). The pH values for the samples
treated at 182 µm were slightly higher than the other treatments in many of the samples.
The pH which was about 4.71 before sonication, immediately dropped to about 4.62
when sonicated for 5 min at 60 µm amplitude (Table 4.1).Though, the trend in varation is
not definite, in majority of the cases, the pH value increased on increasing severity of
sonication. This may be because of the possible liberation of acid from the microbes
which might have been destructed during sonication.
4.8
4.7
4.6 Amplitude level
pH
4.5
60 µm
4.4
4.3
100 µm
4.2
5 8 12 15
182 µm
Sonication time, min
Fig. 4.3(a): Effect of sonication amplitude and time on pH of idli batter after 2 days
of ambient storage.
31
5
Amplitude level
4.8
4.6
pH 60 µm
4.4
100 µm
4.2 182 µm
4
5 8 12 15
Fig.4.3(b): Effect of sonication amplitude and time on pH of idli batter after 5th days
of refrige rated storage
(b) Effect of sonication time
As the time of sonication was increased, the pH of the treated and stored idli batter
increased in most of the cases (Table 4.1). The trend was similar for all most all the
amplitude levels and all the storage periods. As mentioned earlier the pH value of the
sample decreased immediately after sonication.
4.8
Sonication
time
4.6
5 min
pH
4.4 8 min
12 min
4.2
60 100 182 15 min
Sonication amplitude, µm
Fig. 4.4(a): Effect of sonication time and amplitude level on pH of idli batter afte r 2
days of ambient storage
32
As the batter was treated at 60 µm, there was increasing in trend of pH for all sonication
times (Fig 4.4 a). Again, during the refrigrated storage, the pH of the sample increased
with the increase in time of sonication (Fig 4.4 b). This might be possibly due to the fact
that as the sonication treatment was prolonged, the microbial destruction might have been
more resulting in relatively less reduction in pH during storage.
4.8 Sonication time
4.6 5 min
pH
4.4 8 min
12 min
4.2
15 min
4
60 100 182
Fig. 4.4(b): Effect of sonication time and amplitude level on pH of idli batter after
5th days of refrigerated storage
(c) Effect of storage conditions
The pH value of the sonicated sample decreased during storage both under
ambient and refrigerated conditions (Fig 4.5 a & b). This was true for almost all the
combinations of sonication. Again, the pH of the non-sonicated ample also reduced
during storage (Table 4.1).
The value of the pH decreased to the level of about 3.02 form the original of 4.71.
the reduction in pH during the storage which is obvious due to the fact that the acidity of
the fermented product during the storage will increase. However, from the data, it is clear
that the reduction in pH is quite fast under ambient storage as compared to the
refrigerated samples treated with similar sonication. This implies that the fermentation
continued, though at slower rate during storage.
33
4.8
4.5 Storage period
4.2
pH 3.9 2 days
3.6
3.3 4 days
3
5 8 12 15 6 days
Fig. 4.5(a):Effect of storage (ambient) period on pH of idli batter sonicated at 100

µm for different times
4.8
4.5
Storage period
4.2
5 days
pH
3.9
3.6 10days
3.3
15 days
3
5 8 12 15 20 days
Fig. 4.5(b):Effect of storage (refrige rated) period on pH of idli batter sonicated at

100 µm for different times.
4.1.2 Effect of sonication and storage on acidity of idli batter

As the amplitude of sonication was increased from 60 to 100 µm, the acidity of
the stored idli batter decreased for all samples for almost all times of sonication and for
both the ambient and refrigerated conditions (Fig 4.6 a & b). The samples treated at 182
µm, behaved slightly differently. Again for longer sonication treatment (15 min), the
trend reversed in some of the samples (Table 4.2). The acidity which was about 2.74
before sonication, immediately dropped to about 2.70 when the sample was sonicated
34
3 Amplitude
level
2.7
Acidity (%)
2.4 60 µm
100 µm
2.1
182 µm
1.8
5 8 12 15
Fig. 4.6(a): Effect of sonication amplitude and time on acidity of idli batter after 2
days of ambient storage
3.2
3 Amplitude
level
2.8
Acidity (%)
2.6 60 µm
2.4 100 µm
2.2 182 µm
2
5 8 12 15
Fig. 4.6(b): Effect of sonication amplitude and time on acidity of idli batter after
for 5 min at 60 µm. There was clear indication that at 60 µm sonication, the treatment
was not effective to arrest the increase in acidity of the sample during storage.

The acidity of the treated and stored idli batter decreased in most of the cases
(Table 4.2) as the time of sonication was increased. The trend was similar for all most all
the amplitude levels and all the storage periods.
35
3.2
Sonication
2.9 time
Acidity (%) 5 min

2.6
8 min
2.3 12 min
15 min
2
60 100 182
Fig. 4.7(a): Effect of sonication time and amplitude level on acidity of idli batter
after 2nd days of ambient storage
3.2
Sonication
2.9 time
Acidity (%)
5 min
2.6
8 min
2.3 12 min
15 min
2
60 100 182
Fig. 4.7(b): Effect of sonication time and amplitude level on acidity of idli batter
after 5th days of refrigerate d storage
This might have been because of the possible destruction of more microbes at
longer time of sonication, which in term during the storage generated less a mount of
acidity. The acidity levels of the samples stored under refrigerated conditions were
lower at the corresponding period of storage as compared to for those samples stored for
similar period of storage under ambient conditions. The fermentation of batter might
have been slower under refrigerated storage.
36
(c) Effect of storage conditions
The acidity value of the sonicated sample increased during storage under both
ambient and refrigerated conditions (Fig 4.8 a & b). This was true for almost all the
combinations of sonication. Again, the acidity of the non-sonicated sample also
increased during storage (Table 4.2). The value of acidity increased to the level of about
4.3 from the original of 2.74. The trend is obviously due to the possible continuation of
fermentation during the storage. The rapid increase in the acidity of the treated idli batter
during storage indicates inadequacy of the preservation treatment given. The situation is
true particularly for the low amplitude levels and for shorter time of sonication and
under ambient storage.
3.4
Storage
3.2
period
Acidity (%)
3
2 days
2.8
4 days
2.6
6 days
2.4
5 8 12 15
Fig. 4.8(a): Effect of storage(ambient) period on acidity of idli batte r sonicated at

100 µm for different times
4.4
Storage
4
period
Acidity (%)
3.6 5 days
3.2
10 days
2.8
15 days
2.4
2 20 days
5 8 12 15
Fig. 4.8(b): Effect of storage(refrigerated) pe riod on acidity of idli batter sonicated

at 100 µm for different times
37
The rate of increase in acidity during refrigerated storage was slower as compared
to that under ambient storage. This might be due to the slower fermentation at lower
temperature of the batter.
4.1.3 Effect of sonication and storage on SPC of idli batter.

As the amplitude of sonication was increased, the total microbial count of the
stored idli batter decreased for all most all times of sonication and for both the ambient
and refrigerated storages (Fig 4.9 a & b). The microbial count in the non-sonicated idli
batter was quite higher than the treated samples.
10
Amplitude
level
SPC, 107 cfu/g
60 µm
100 µm
182 µm
1
5 8 12 15
Fig. 4.9(a): Effect of sonication amplitude and time on SPC of idli batter after 2 days
of ambient storage
100
Amplitude
level
SPC, cfu/g
10 60 µm
100 µm
182 µm
1
5 8 12 15
Fig. 4.9(b): Effect of sonication amplitude and time on SPC of idli batter after 5th
days of refrigerated storage
38
This indicates that amount of the microbial activity has been retarded due to the
sonication. This may be because of the destruction of cell wall of microbes due to
cavitation occur inside the cell wall of microbes during sonication. At the higher level of
amplitude, more microbes might have been destructed.
The microbial count of the treated and stored idli batter decreased as the time of
sonication was increased in all the cases (Table 4.3). The trend was similar for all most
all the amplitude levels and all the storage conditions (Fig 4.10 a & b). The data indicate
that the microbial count in the treated samples decreased when the amplitude of the
sonication was increased and also as the sonication time was prolonged. The possible
reasons have been mentioned earlier. However, the effect of sonication is clearly seen
when the data are compared with the non-sonicated samples.
10
Sonication
time
SPC, 10 7 cfu/g
5 min
8 min
12 min
15 min
1
60 100 182
Fig. 4.10(a): Effect of sonication time and amplitude level on SPC of idli batter after
2 days of ambient storage
From the Fig 4.10 (a & b), it is evident that the total microbial count in the
refrigerated stored samples were quite low than that in samples stored under ambient
conditions for similar period. The sonicated samples could be very well stored for
adequate time, particularly under refrigerated conditions (Table 4.3).
39
100
Sonication
time
SPC, 107 cfu/g

5 min
10
8 min
12 min
15 min
1
60 100 182
Fig. 4.10(b): Effect of sonication time and amplitude level on SPC of idli batter after
(c) Effect of storage
The SPC of the sonicated sample increased during storage both under ambient and
refrigerated conditions (Fig 4.11 a & b). This was true for almost all the combinations of
sonication. Again, the SPC of the non-sonicated samples were higher than that of the
treated samples for the corresponding storage period (Table 4.3).
100
Storage
period
SPC, 10 7 cfu/g
10 2 days
4 days
6 days
1
5 8 12 15
Fig. 4.11(a): Effect of storage (ambient) period on SPC of idli batter sonicated at
100µm for different times
The trend is obviously due to the possible continuation of fermentation during the
storage. The rapid increase in the SPC of the treated idli batter during storage indicates
40
inadequacy of the preservation treatment given as in some of the samples. The situation
is true particularly for the low amplitude levels and for shorter time of sonication and
under ambient storage.
The rate of increase in SPC during refrigerated storage was slower as compared to
that under ambient storage. This might be due to the slower fermentation at lower
temperature of the batter during storage.
100
SPC, 107 cfu/g
5 days
10
10 days
15 days
20 days
1
5 8 12 15
Fig. 4.11(b): Effect of storage (refrige rated) pe riod on SPC of idli batte r sonicated at
100 µm for different times
4.1.4 Effect of sonication and storage on overall acceptability of idli batter
As the amplitude of sonication was decreased from 100 to 60 µm, the overall
acceptability of the stored idli batter decreased for many samples for all the times of
sonication and for both the ambient and refrigerated storages (Figs 4.12 a & b). The
overall acceptability scores for the samples treated at 182 µm were slightly higher than
the other treatments in many of the samples. The overall acceptabilityfor fresh batter was
about 8.9 score before sonication, immediately dropped to about 8.0 when sonicated for 5
min at 60 µm amplitude.Though, the trend in varation is not definite, in majority of the
cases, the overall acceptability score deccreased on sonication.
41
Overall acceptability
Sonication
8.5 amplitude
60 µm
8
100 µm
7.5 182 µm
7
5 8 12 15
Sonicaion time, min
Fig 4.12 (a): Effect of sonication amplitude and time on overall acceptability of idli
batter afte r 2nd day of ambient storage
9
Sonication
8.5
amplitude
8 60 µm
7.5 100 µm
7 182 µm
6.5
5 8 12 15
Sonicaion time, min
Fig 4.12 (b): Effect of sonication amplitude and time on overall acceptability of idli
batter afte r 5th day of refrige rated storage
As the time of sonication was increased, the overall acceptability of the treated and stored
idli batter increased in most of the cases (Table 4.4). The trend was similar for all most
all the amplitude levels and all the storage periods. As mentioned earlier the overall
acceptability score of the sample decreased immediately after sonication.
As the batter was treated at 60 µm, there was increasing in trend of overall acceptability
score for all sonication times (Fig 4.13 a). Again, during the refrigrated storage, the
overall acceptability score of the sample increased with the increase in time of sonication
(Fig 4.13 b).
42
9
Sonication
8.5 time
5 min
8 8 min
12 min
7.5
15 min
7
60 100 182
Sonicaion amplitude, µm
Fig 4.13(a): Effect of sonication time and amplitude level on overall acceptability of
idli batter after 2 days of ambient storage
9
Sonication
8.5 time
5 min
8
8 min
7.5
12 min
7
15 min
6.5
60 100 182
Sonicaion amplitude, µm
Fig 4.13(b): Effect of sonication time and amplitude level on overall acceptability of
idli batter after 5th days of refrigerated storage
(c) Effect on storage conditions
The overall acceptibility score of the sonicated sample decreased during storage bot h
under ambient and refrigeration conditions (Figs 4.14 a & b). This was true for almost all
the combinations of soniaction. Again, the scores of the non-sonicated sample also
decreased during storage (Table 4.4). The trend is obviously due to the decrease in pH
and increase in acidity of the batter during storage. The rate of decrease in score during
refrigerated storage was slower as compared to that under ambient storage.
43
8.5 Sonication
overall acceptability
8 time
7.5 5 min
7
8 min
6.5
6 12 min
5.5
15 min
2 4 6 8
Storage period, days
Fig. 4.14(a): Effect of storage (ambient) pe riod on overall acceptability of idli batter
sonicated at 100 µm for different times
8.5
Sonication
time
overall acceptability
7.5
5 min
6.5
8 min
5.5
12 min
4.5
15 min
3.5
5 10 15 20
Fig. 4.14(b): Effect of storage (refrigerated) period on ove rall acceptability of idli
batter sonicated at 100 µm for different times
4.1.5 Shelf-life of sonicated idli batter
The shelf- life of the treated idli batter with different conditions of sonication and
subsequently stored under both the ambient and refrigerated conditions was calculated
based on the procedure described in the earlier chapter. The value of the storage period
for which the overall acceptability of the stored sample was more than 6.0 was computed
44
in days using the mathematical relationships. The data obtained are presented in Table
4.5.
Table. 4.5 Shelf-life (days) of sonicated idli batter under ambient and refrigerated
storage
Sonication Storage conditions

time
(min) Ambient Refrigerated
5 8 12 15 5 8 12 15
Sonication
amplitude
(µm)
No 1 day 8 days
sonication
60 2 2 2 2 9 11 14 16-17
100 4-5 5-6 5 5 12 15-16 16-17 20
182 5 * * * 15 * * *
The shelf- life of the idli batter increased on sonication. The increase was higher
under refrigerated conditions. The shelf- life also increased with the increase in sonication
amplitude for most the sonication times.
The longest shelf- life obtained was 6 days (100 µm & 8 min) and 20 days (100µm
& 15 min) under ambient and refrigerated storage, respectively as compared to only 1day
and 8 days for samples without sonication. It is substantial increase in shelf- life.
If the fresh- fermented idli batter is sonicated immediately at 100 µm for about
8min, then the shelf- life could be increased almost six times at ambient storage itself.
This will help in increasing the production and consumption of idli batter.
45
4.2 Effect of Irradiation on Preservation of Idli Batter
The fresh and fermented samples of idli batter were packed in LDPE bags (300
gauge) and were irradiated at different frequencies (0.5, 0.75, 1.0 and 1.25 kGy). After
treatment, the samples were kept for storage studies both under ambient and refrigerated
conditions. Tables 4.6 through 4.9 show the data of different quality characteristics such
as pH, acidity, SPC and overall acceptability of idli batter after irradiation and during
storage. The ANOVA for each response is also appended with the tables. The variations
in the quality parameters of the irradiated idli batter during storage are also presented in
graphical way through Figs 4.15 to 4.22. The influence of irradiation conditions on each
quality parameter (pH, acidity, SPC and overall acceptability) is described in details here
under.
Table. 4.6 Effect of irradiation and storage on pH of idli batter

Irradiation frequency No 0.50 0.75 1.00 1.25
(kGy)
irradiation
Storage
Period (days)
Ambient 0 4.62 4.58 4.61 4.76 4.84

(30±2˚c) 2 ** 4.42 4.44 4.56 4.69
4 ** 4.31 4.42 4.52 4.54
6 ** 3.72 3.94 4.08 4.25
8 ** 3.32 3.65 3.82 3.97
Refrigertaed 5 4.58 4.44 4.46 4.55 4.59
(7±1˚c) 10 4.06 4.36 4.38 4.42 4.48
15 3.92 4.31 4.36 4.40 4.45
20 ** 4.23 4.34 4.38 4.42
Source D.F M.S F value S.Em Test
ANOVA Treatment 9 0.027 2.25 0.030 *
Error 0 0.086
CV % 6.95
*** Non-significant at 5 % level of significance, ** Samples spoiled
46
Table. 4.7 Effect of irradiation and storage on acidity (% TAA) of idli batter
Irradiation frequency Non- 0.50 0.75 1.00 1.25
(kGy)
Storage
irradiation
Period (days)
Ambient 0 2.68 2.72 2.71 2.59 2.68
(30±2˚c) 2 ** 2.74 2.72 2.63 2.59
4 ** 2.88 2.86 2.82 2.77
6 ** 3.07 2.93 2.90 2.84
8 ** 3.44 3.35 3.14 3.01
Refrigertaed 5 2.64 2.48 2.44 2.38 2.27
(7±1˚c) 10 2.93 2.69 2.65 2.61 2.53
15 3.76 2.92 2.84 2.81 2.75
20 ** 3.22 3.03 3.01 2.88
ANOVA Treatment 2 0.060 1.03 0.038 ***
Error 0 0.14
CV % 12.89
*** Non-significant at 5 % level of significance ** Samples spoiled
Table. 4.8 Effect of irradiation and storage on SPC (107 cfu/g) of idli batter
Irradiation frequency Non- 0.50 0.75 1.00 1.25

(kGy) irradiated
Storage
Period (days)
Ambient 0 20.7 BDL BDL BDL BDL
(30±2˚c) 2 ** 1.1 1.5 1.3 2
4 ** 3.8 1.8 2.3 2.7
6 ** 9.6 7.3 8.5 8.2
8 ** 16.3 12 14.3 10.6
Refrigertaed 5 40.5 BDL 0.8 BDL BDL
(7±1˚c) 10 83.7 1.8 2.6 2.3 2.2
15 131.8 9.6 7.8 9.6 6.4
20 ** 14.7 13.5 10.8 5.8
ANOVA Treatment 2 17.7 12.86 0.348 ***
Error 0 96.6
CV % 18.5
*** Non-significant at 5 % level of significance ** Samples Spoiled BDL = Below
Detectable Level
47
Table. 4.9 Overall acceptability of score irradiated and stored idli batter
Irrad iation frequency No 0.50 0.75 1.00 1.25

(kGy )
irradiation
Storage
Period (days)
Ambient 0 8.0 7.8 8.0 8.4 8.6

(30±2˚c) 2 ** 7.5 7.8 8.0 8.4
4 ** 6.9 7.4 7.8 8.2
6 ** 5.2 5.5 5.9 6.4
8 ** 4.7 4.9 5.2 5.3
Refrigertaed 5 7.6 7.8 8.0 8.2 8.3
(7±1˚c) 10 5.5 7.4 7.6 7.9 8.0
15 4.3 7.2 7.5 7.8 7.9
20 ** 6.7 7.0 7.4 7.7
ANOVA Treatment 31 1.23 11.59 0.20 *
Error 1
CV % 15.61
** Samples spoiled *Significant at 5% level of significance
4.2.1 Effect of irradiation and storage on pH of idli batter
(a) Effect of irradiation frequency
As the frequency of irradiation was increased from 0.5 through 0.75, 1.0 to 1.25
kGy, the pH of the stored idli batter slightly increased for many samples stored both
under ambient and refrigerated conditions (Fig 4.15 a & b). However, the increase in pH
of the stored idli was very little on increasing the irradiation frequency. In fact, the
variation was non-significant at 5% level.
The increase in pH value might be because of the possible breaking of water

molecule due to bombarding of radiated particle which might have produced free ions
(OH- & H+).
48
5
Storage
4.6 period
4.2
pH 0 days
3.8
2 days
3.4
4 days
3
6 days
0.5 0.75 1 1.25
8 days
Irradiation frequency, kGy
Fig. 4.15(a): Effect of irradiation fre quency on pH of stored (ambient) idli batter
5 Storage
4.8 period
0 days
4.6
pH
5 days
4.4
10 days
4.2
15 days
4
0.5 0.75 1 1.25 20 days
Fig. 4.15(b): Effect of irradiation fre quency on pH of stored (refrigerated) idli batter
(b) Effect of storage conditions
The pH value of the irradiated sample decreased during storage both under
ambient and refrigerated conditions (Figs 4.16 a & b). This was true for all the
frequencies of the irradiation. The pH of the non- irradiated samples also reduced during
the storage (Table 4.6). The value of the pH decreased to the level of about 3.32 from
4.62 during the storage for the samples under ambient condition and 4.23 from the
original of 4.58 under refrigerated conditions.
The rate of decrease in pH with the increase in storage period was steady for
almost all the irradiation treatments. However, the decline in pH value was quite faster
under ambient conditions of storage as compared to the refrigerated samples. This is
49
5
Irradiation
4.6 frequency
4.2
pH 0.5 Kgy
3.8 0.75 Kgy
3.4 1.0 Kgy

1.25 Kgy
3
0 2 4 6 8
Fig 4.16(a): Effect of storage (ambient) period on pH of idli batter irradiated at

different frequencies
5
Irradiation
4.8 frequency
4.6
0.5 Kgy
pH
4.4 0.75 Kgy
4.2 1.0 Kgy

1.25 Kgy
4
0 5 10 15 20
Fig 4.16(b): Effect of storage (refrige rated) period on pH of idli batter irradiated at
obviously due to the effect of low temperature on further fermentation of the idli batter
during storage.
4.2.2 Effect of irradiation and storage on acidity of idli batter
As the frequency of irradiation was increased from 0.5, through 0.75, 1.0 to
1.25kGy, the acidity of the stored idli batter slightly decreased for many samples for both
the ambient and refrigerated storages (Fig 4.17 a & b). However, there was no definite
50
trend in the variation of acidity with irradiation frequency. In fact the variation was non-
significant at 5% level. There was decrease in acidity with the increase in frequency for
the longer storage period under ambient conditions (Fig 4.17 a).
4
3.8
Storage
3.6 period
3.4
Acidity (%)
0 days
3.2
3 2 days
2.8 4 days
2.6
6 days
2.4
2.2 8 days
0.5 0.75 1 1.25
Fig. 4.17(a): Effect of irradiation fre quency on acidity of stored (ambient) idli batte r
3.4
Storage
3.2
period
3
Acidity (%)
0 days
2.8
5 days
2.6
10 days
2.4
2.2 15 days
2 20 days
0.5 0.75 1 1.25
Fig. 4.17(b): Effect of irradiation frequency on acidity of stored (refrigerated) idli

batter
The acidity values for the refrigerated samples were relatively lower as compared
to those stored under ambient conditions. This is obvious as the lower temperature
conditions do not favor increase in acidity.
51
The acidity value of the irradiated sample increased during storage both under
ambient and refrigerated conditions (Fig 4.18 a & b). This was true for all the frequencies
of irradiation. The acidity of the non- irradiated sample also increased during the storage
(Table 4.7). The value of the acidity increased to the level of about 3.74 from 2.68 during
the storage for the samples stored under ambient conditions and 3.22 from the original of
2.64 under refrigerated conditions.
3.9 Irradiation
3.7 frequency
3.5
Acidity (%)
3.3 0.5 kGy

3.1
0.75 kGy
2.9
2.7 1.0 kGy
2.5 1.25 kGy
2.3
0 2 4 6 8
Fig 4.18(a): Effect of storage (ambient) period on acidity of idli batter irradiated at
3.4
3.2 Irradiation
frequency
3
Acidity (%)
2.8 0.5 kGy

2.6 0.75 kGy
2.4 1.0 kGy
2.2 1.25 kGy
2
0 5 10 15 20
Fig 4.18(b): Effect of storage (refrigerated) period on acidity of idli batter irradiated
at diffe rent frequencies
52
The rate of increase in acidity with the increase in storage period is steady for
almost all the irradiation treatments. However, the increase in acidity value was quite
faster under ambient conditions as compared to the refrigerated samples. This is
obviously due to the effect of low temperature on further fermentation of the idli batter.
The phenomena of reducing acidity for the initial period of refrigerated storage
was observed for all the irradiated samples.
4.2.3 Effect of irradiation and storage on SPC of idli batter
The non- irradiated sample showed plate count initially at 20.7×10-7 cfu g-1 (after
fermentation) but there was sudden decline in the total microbial count immediately after
treating the idli batter to irradiation (Table 4.8). The total microbial count was almost
below 1×107 cfu/g for the first day of ambient storage and almost upto 5 days under
refrigerated samples. Hence, the particular storage data are not drawn in Fig 4.19 (a & b).
Decrease in microbial population after irradiation might be due to the post-

irradiation effect where surviving cells that had been damaged by gamma-radiation
gradually might have died, not adapting to the surrounding environment.
Storage
10 period
SPC, 10 7 cfu/g
2 days
4 days
6 days
8 days
1
0.5 0.75 1 1.25
Irradiation frequency, KGy
Fig. 4.19(a): Effect of irradiation fre quency on SPC of stored (ambient) idli batte r.
53
Storage
SPC, 107 cfu/g

10 period
10 days
15 days
20 days
1
0.5 0.75 1 1.25
Irradiation frequency, KGy
Fig. 4.19(b): Effect of irradiation frequency on SPC of stored (refrigerated) idli

batter
However, the effect of irradiation frequency on the total microbial load of the
sample was non-significant at 5% level (Table 4.7). The trend was similar for samples
stored under both the storage conditions, refrigerated storage with relatively lower values.
The total microbial count of the irradiated samples increased during storage both
under ambient and refrigerated conditions (Fig 4.20 a & b). This was true for all the
frequencies of irradiation. The microbial count of the non- irradiated sample also
increased during storage (Table 4.7).
Irradiation
10 frequency
SPC, 10 7 cfu/g
0.5 Kgy
0.75 Kgy
1.0 Kgy
1 1.25 Kgy
0 2 4 6 8
Fig 4.20(a): Effect of storage period on SPC of stored (ambient) idli batter
54
Irradiation
10
SPC, 10 7 cfu/g
frequency
0.5 Kgy
0.75 Kgy
1.0 Kgy
1 1.25 Kgy
0 5 10 15 20
Fig 4.20(b): Effect of storage period on SPC of stored (refrigerated) idli batter
The SPC reduced drastically immediately on irradiation (Table 4.7). However, the
microbial count rose further rapidly during storage under ambient conditions than under
refrigerated. The microbial growth under ambient storage was obviously faster as
compared to that under refrigerated storage (Fig 4.20 a & b).
4.2.4 Overall acceptability of irradiated idli batte r
The variation in overall acceptability score with respect to irradiation treatment

and storage are depicted in Table 4.9.
9
8.5
Overall accepetability
8 Storage
7.5 period
7 2 days
6.5 4 days
6
5.5 6 days
5 8 days
4.5
0.5 0.75 1 1.25
Fig 4.21(a): Effect of irradiation frequency on overall acceptability of stored

(ambient) idli batter
55
8.5 Storage
Overall accepetability period
8
5 days
7.5 10 days
7 15 days
6.5 20 days
6
0.5 0.75 1 1.25
Fig 4.21(b): Effect of irradiation frequency on overall acceptability of stored

(refrige rated) idli batter
As the frequency of irradiation was increased from 0.50 through 0.75, 1.00 and
1.25 kGy, the overall acceptability score of the stored idli batter increased for all most all
the samples, for both the ambient and refrigerated storages (Fig 4.21 a & b). The overall
acceptability scores for the samples treated at 0.50 kGy were lesser than the other
frequencies.
The overall acceptability score of the irradiated sample decreased during storage
both under ambient and refrigerated conditions (Figs 4.22 a & b). This was true for all the
frequencies of the irradiation. The overall acceptability score of the non-irradiated
samples also reduced during the storage (Table 4.9). The value of the overall
acceptability score decreased of about 4.7 from 8.0 during the storage for the samples
under ambient condition and 4.3 from the original of 8.0 under refrigerated conditions.
The rate of decrease in overall acceptability score with the increase in storage
period was steady for almost all the irradiation treatments. However, the decline in
overall acceptability score was quite faster under ambient conditions of storage as
56
8.5
8 Irradiation
7.5 frequency
7 (kGy)
6.5
0.5
6
5.5 0.75
5
4.5 1
4
2 4 6 8 1.25
Fig 4.22(a): Effect of storage period on overall acceptability of stored (ambient) idli
batter
9 Irradiation
8.5 frequency
8 (kGy)
7.5
7 0.5
6.5
6 0.75
5.5
5 1
5 10 15 20
1.25
Fig 4.22(b): Effect of storage period on overall acceptability of stored (refrige rated)
idli batter
compared to the refrigerated samples. This is obviously due to the effect of low
temperature on further fermentation of the idli batter during storage.
4.2.5 Shelf-life of irradiated idli batter
The shelf- life of the treated idli batter with different frequencies of irradiation and
subsequently stored under both the ambient and refrigerated conditions was calculated
based on the procedure described in the earlier chapter. The value of the storage period
57
for which the overall acceptability of the stored sample was more than 6.0 was computed
in days using the mathematical relationships. The data obtained are presented in Table
4.10.
Table. 4.10 Shelf-life (days) of irradiated idli batter under ambient and refrigerated
storage
Storage pe riod(days) Storage conditions

Irradiation Ambient Refrigerated
frequency(kGy)
No irradiation 1 8
0.50 5 30-31
0.75 5-6 37
1.00 6 49
1.25 6-7 64
The shelf- life of the idli batter increased on irradiation. The increase was higher
under refrigerated conditions. The shelf- life also increased with the increase in irradiation
frequencies.
The longest shelf- life obtained was 7 days and 64 days under ambient and
refrigerated storage, respectively as compared to only 1 day and 8 days for samples
without irradiation. The enhancement in shelf- life is quite big and very well usefull for
improving the production and consumption of idli.
4.3 Effect of Addition of Preservatives on Preservation of Idli Batter
The preservatives were added to fresh idli batter after fermentation at the rate of
Nisin-7.5ppm and Potassium Sorbate-2000ppm. The idli batter was packed in LDPE bags
(300 gauge) and was kept for storage studies at ambient and re frigerated conditions.
Table 4.11, shows the data of different quality characteristics such as pH, acidity, SPC
and overall acceptability of idli batter during storage. The ANOVA for each response is
also appended with Tables 4.12 through 4.15. The variation in the quality parameters of
preserved idli batter during storage are also presented in graphical way through Figs 4.23
to 4.26.
58
Table 4.11 Effect of addition of preservative on quality of idli batte r during storage
Storage pH Acidity (%) SPC (107 cfu/g) Overall acceptability

period
Ambient
With Without With Without With Without With Without
preservative preservative preservative preservative preservative preservative preservative preservative
0 4.5 4.5 2.26 2.26 1.7 3.4 8.7 8.7
2 4.43 * 2.48 * 2.4 * 8.4 *
4 4.36 * 2.71 * 2.7 * 7.9 *
6 4.27 * 2.99 * 2.9 * 7.2 *
8 4.19 * 3.17 * 3.1 * 6.4 *
Refrigerated
5 4.41 4.32 2.29 2.45 5.56 40.9 8.5 7.8
10 4.34 4.09 2.58 2.96 14.4 86.3 7.6 6.0
15 4.26 3.34 2.79 3.57 21.7 149.7 7.2 5.6
20 4.13 * 3.11 * 31.4 * 6.6 *
*Samples spoiled
59
4.3.1 Effect of preservatives and storage on pH of idli batter
The samples spoiled within 24 h, if no preservative was added. However, during

the refrigerated storage, the samples could be stored for some days without preservation.
As the preservative was added, the pH could be maintained to relatively higher level
during the storage. This indicates that the preservatives were effective in retarding the
batter fermentation to some extent. The influence was more visible under refrigerated
samples (Fig 4.23 a & b).
4.6
4.5
4.4
4.3
4.2
pH
4.1 Nisin+ Potassium

4 sorbate
3.9
3.8 Without preservative
3.7
3.6
0 2 4 6 8
Fig. 4.23(a): Effect of storage (ambient) period on pH of idli batter preserved by

addition of preservatives
4.6
4.4
4.2
4
pH
3.8 Nisin+ Potassium

3.6 sorbate
3.4 Without preservation
3.2
3
5 10 15 20
Fig. 4.23(b): Effect of storage (refrigerated) pe riod on pH of idli batter preserved by

60
Table 4.12. ANOVA for pH of preserved idli batter

ANOVA Treatment 2 0.10 1.71 0.050 ***
Error 7 0.060
CV % 5.72
***Non-significant at 5% level of significance
Though, the variation in pH as the function of addition of preservative and storage

period was non-significant at 5% level of significance (Table 4.12).
4.3.2 Effect of preservative and storage on acidity of idli batter

As the preservative was added, the acidity could be maintained to relatively lower level
during the storage. This indicates that the preservatives were effective in retarding the
batter fermentation to some extent. The influence was more visible under refrigerated
3.5
3
2.5
Acidity (%)
2
1.5 Nisin+ Potassium sorbate
1 Without preservative
0.5
0
0 2 4 6 8
Fig. 4.24(a): Effect of storage (ambient) period on acidity of idli batter preserved by
61
3.4
3.2
3
Acidity (%) 2.8 Nisin+ Potassium
2.6 sorbate
2.4 Without preservation
2.2
2
5 10 15 20
Fig. 4.24(b): Effect of storage (refrigerated) pe riod on acidity of idli batte r preserved
by addition of preservatives
Table 4.13. ANOVA for acidity of preserved idli batter

ANOVA Treatment 2 0.28 61.33 0.036 *
Error 7 0.26
CV % 2.54
*Significant at 5% level of significance
Though, the variation in acidity as the function of addition of preservative and

storage period was significant at 5% level of significance (Table 4.13).
4.3.3 Effect of preservative and storage on SPC of idli batter
As the preservative was added, the SPC could be maintained to relatively lower
level during the storage. This indicates that the preservatives were effective in retarding
the batter fermentation to some extent. The influence was more visible under refrigerated
Table 4.14. ANOVA for SPC of preserved idli batter

ANOVA Treatment 2 0.46 21.44 0.032 *
Error 7 0.022
CV % 15.37
62
Though, the variation in SPC as the function of addition of preservative and

storage period was non-significant at 5% level of significance (Table 4.14).
SPC,10 7 cfu/g 10
Nisin+ Potassium
sorbate
Without preservative
1
0 2 4 6 8
Fig. 4.25(a): Effect of storage (ambient) period on SPC of idli batter preserved by
100
SPC, 107 cfu/g
Nisin+ Potassium
10 sorbate
Without
preservation
1
5 10 15 20
Fig. 4.25(b): Effect of storage (refrigerated) pe riod on SPC of idli batte r preserved
by addition of preservatives
4.3.4 Overall acceptability of preserved idli batter

As the preservative was added, the overall acceptability score could be maintained to
relatively higher level during the storage. This indicates that the preservatives were
63
effective in retarding the batter fermentation to some extent. The influence was more
visible under refrigerated samples (Fig 4.26 a & b).
8.5
7.5
With preservation
7
Without preservation
6.5
6
0 2 4 6 8
Fig 4.26(a): Effect of overall acceptability of preserved idli batter stored (ambient)
9
8.5
8
7.5
7
With preservation
6.5
Without preservation
6
5.5
5
0 5 10 15 20 25
Fig 4.26(b): Effect of ove rall acceptability of preserved idli batter stored
(refrige rated)
Table 4.15. ANOVA for ove rall acceptability of preserved idli batter

ANOVA Treatment 7 0.60 12.8 0.04 *
Error 0
CV % 3.78
64
Though, the variation in overall acceptability as the function of addition of

preservative and storage period was non-significant at 5% level of significance (Table
4.15).
4.3.5 Shelf-life of preserved idli batter
The shelf- life of the treated idli batter with the addition of different preservatives
and subsequently stored under both the ambient and refrigerated conditions was
calculated based on the procedure described in the earlier chapter. The value of the
storage period for which the overall acceptability of the stored sample was more than 6.0
was computed in days using the mathematical relationships.
The increase was higher under refrigerated conditions. The longest shelf- life
obtained was 10 days and 25 days under ambient and refrigerated storage, respectively as
compared to only 1 day and 8 days for samples without preservative.
4.4 Comparative of Preservation Techniques
As described above, all the three preservation techniques (sonication, irradiation

and addition of preservatives) were effective in enhancing the she lf- life of the idli batter.
The suitability of the particular technique depends upon the final requirement of the
shelf- life besides the cost factor. However, the comparative shelf- life for all the three
methods at their best operating parameters is depicted below in Table 4.16.
Table 4.16 Comparative shelf-life (days) of idli batter preserved through different
methods.
Preservation Ambient Storage Refrigerated storage

(30±2˚C) (7±1˚C)
No treatment 1 day 8 days
Sonication at 100 µm for 6 days 20 days
8min
Irradiation at 1.25 kGy 7 days 64 days
Addition of Nisin (7.5ppm) 10 days 25 days
+ Potassium Sorbate
(2000ppm)
65
66
CHAPTER V
SUMMARY AND CONCLUSIONS
Cereal-based fermented foods are considered as staple diets in their respective regions.
Most of the foods such as idli, dosa, dhokla, koozhu, nan, parotta, ambali, pazhaiya soru are
consumed on the daily basis by the local population. Idli, a traditional breakfast food of India
based on cereal-legume combination, is a white, leavened, soft, spongy textured product.
Idli is a fermented, thick suspension made by blend of rice and dehulled black gram.
Idli batter has very short shelf-life because of its high moisture content and live fermentation.
There is a demand for ready-to-cook idli batter in packaged form with moderate shelf-life.
Dried idli mixes are the alternatives put in the market. However, because of the inferior
texture of the product and its lower organoleptic quality, the instant powders are not popular.
Thermal preservation is not feasible as the batter co-agulates or idli is formed on heat
application. Study was undertaken to evaluate some of the non thermal methods of
preservation (sonication, irradiation and biopreservatives) for extension of shelf-life of fresh
idli batter. The specific objectives of the study were as follows;
1. To evaluate the effect of some non thermal methods of preservation on the shelf-life
extension of idli batter.
2. Evaluating the changes in microbial and organoleptic quality of preserved idli batter
during storage.
3. Determining the shelf-life of the preserved idli batter.
The fermented idli batter was sonicated at different amplitude levels (60, 100 and 182
µm) for different exposure times (5, 8, 12 and 15 min). Similarly the, idli batter was also
irradiated at different frequencies (0.5, 0.75, 1.0 and 1.25 kGy). The addition of
preservative ( Nisin @ 7.5 ppm combined with Potassium Sorbate @2000 ppm was also
attemted for extending shelf-life of idli batter. The freshly prepared batter after
fermentation was packed in LDPE bags, treated either with sonication or irradiation or
preservatives and stored both under ambient as well as refrigerated conditions.
66
Summary and Conclusions
The batter was evaluated at the end of 2, 4, 6 and 8 days of storage at ambient temperature
(30±2˚ C). For refrigerated (7±2˚ C) storage, samples were kept for 5, 10, 15 and 20 days for
all the treatments given above. The quality responses in tern of the pH, acidity, SPC and the
overall acceptability of the idli were recorded for all the variables and their combinations.
Based on the results obtained from various experiments following conclusions were
drawn;
1. As the sonication amplitude and sonication time was increased, the pH of the stored
idli batter increased under both ambient and refrigerated conditions. The reduction in pH
was quite fast under ambient storage as compared to the refrigerated samples treated with
similar sonication.
2. The acidity of the idli batter stored under both ambient and refrigerated storage is
reduced on the increasing in the sonication amplitude and time of sonication.The acidity
of the sonicated idli batter increased during storage.
3. As the sonication amplitude and sonication time was increased, the SPC of the stored
idli batter reduced under both ambient and refrigerated conditions. The SPC of the
sonicated batter increased during the storage.
4. The overall acceptability of the idli prepared from sonicated and stored batter,
improved when the batter was sonicated at higher amplitude for longer period. The
sensory quality deterioted during the storage of sonicated batter.
5. The shelf-life of idli batter increased on sonication.
6. The irradiation of the fresh and fermented idli batter enhanced its shelf-life both under
ambient and refrigerated conditions.
7. The effect of irradiation frequency on pH, acidity, SPC and overall acceptability was
non-significant at 5% level of significance. However, the quality of the irradiated batter
deterioted during storage.
67
Summary and Conclusions
8. The addition of preservatives could enhance the shelf-life of idli batter. The reduction
in pH during storage of the idli batter with added preservative was non-significant.
However, the increase in acidity and SPC was significant at 5% level.
9. The sonication at 100 µm for 8 min could enhanced the shelf-life of idli batter to 6
days and 20 days, irradiation at 1.25 kGy to 7 and 64 days and addition of Nisin @
7.5ppm +Potassium Sorbate @ 2000ppm to 10 and 25 days as compared to only 1 day
and 8 days under ambient and refrigerated storages, respectively.
68
Appendix
APPENDIX- I
Sensory evaluation score card for Idli
DATE:
Evaluator’s name:
Product name:
Sr. No. Product Appearance Color Taste Overall acceptability

code
1
2
3
4
5
6
7
Based on 9 point hedonic scale

9- Like extremely
8- Like very much
7- Like moderately
6- Like slightly
5- Neither like nor dislike
4- Dislike slightly
3- Dislike moderately
2- Dislike very much
1- Dislike extremely
Remarks/ Comments:
Signature
LIST OF TABLES
Table Page
No Description No.
2.1 Effect of ratios of rice to black gram on pH, acidity and 5
density of idli batter
3.1 (a) Physico-chemical property of rice 17
(b) Physico-chemical property of split black gram dhal
4.1 Effect of sonication and storage on pH of idli batter 27
4.2 Effect of sonication and storage on acidity (% TAA) of idli 28
batter
4.3 Effect of sonication and storage on SPC (107 cfu/g) of idli 29
batter
4.4 Overall acceptability score of sonicated and stored idli 30
batter
4.5 Shelf-life (days) of sonicated idli batter under ambient and 45
refrigerated storage
4.6 Effect of irradiation and storage on pH of idli batter 46
4.7 Effect of irradiation and storage on acidity (% TAA)of idli 47
batter
4.8 Effect of irradiation and storage on SPC (107 cfu/g) of idli 47
batter
4.9 Overall acceptability score of irradiated and stored idli 48
batter
4.10 Shelf-life (days) of irradiated idli batter under ambient and 58
refrigerated storage
4.11 Effect of addition of preservative on quality of idli batter 59
during storage
4.12 ANOVA for pH of preserved idli batter 61
4.13 ANOVA for acidity of preserved idli batter 62
4.14 ANOVA for SPC of preserved idli batter 62
4.15 ANOVA for overall acceptability of preserved idli batter 64
4.16 Comparative shelf-life (days) of idli batter preserved 65
through different methods
v
LIST OF FIGURES
Figure Page
No Description No.
3.1 Bulk sample of (a) milled rice and (b) split black gram used to 17
prepare Idli
3.2 Process flow chart for preparation of idli batter 18
3.3 Freshly prepared idli batter 19
3.4 Sonication equipment. (a) Q Sonica, Q700, (b) Sonic vibra cell, 20
VCX 130
3.5 Irradiation chamber, GC 5000 21
3.6 Nisin + Potassium Sorbate used as preservatives 22
4.1 Charred sample at 182 µm amplitude, exposing for more than 5 26
minutes
4.2 Spoilage due to (a) mold growth and (b) gas formation 26
4.3 (a) Effect of sonication amplitude and time on pH of idli batter 31
after 2 days of ambient storage
(b) Effect of sonication amplitude and time on pH of idli batter 32
after 5th days of refrigerated storage
4.4 (a) Effect of sonication time and amplitude level on pH of idli 32
batter after 2 days of ambient storage
(b) Effect of sonication time and amplitude level on pH of idli 33
batter after 5th days of refrigerated storage
4.5 (a) Effect of storage (ambient) period on pH of idli batter 34
(b) Effect of storage (refrigerated) period on pH of idli batter 34
4.6 (a) Effect of sonication amplitude and time on acidity of idli 35
(b) Effect of sonication amplitude and time on acidity of idli 35
4.7 (a) Effect of sonication time and amplitude level on acidity of 36
idli batter after 2 days of ambient storage
(b) Effect of sonication time and amplitude level on acidity of 36
idli batter after 5th days of refrigerated storage
4.8 (a) Effect of storage (ambient) period on acidity of idli batter 37
(b) Effect of storage (refrigerated) period on acidity of idli batter 37
4.9 (a) Effect of sonication amplitude and time on SPC of idli batter 38
after 2 days of ambient storage
(b) Effect of sonication amplitude and time on SPC of idli batter 38
after 5th days of refrigerated storage
vi
4.10 (a) Effect of sonication time and amplitude level on SPC of idli 39
(b) Effect of sonication time and amplitude level on SPC of idli 40
4.11 (a) Effect of storage (ambient) period on SPC of idli batter 40
(b) Effect of storage (refrigerated) period on SPC of idli batter 41
4.12 (a) Effect of sonication amplitude and time on overall 42
acceptability of idli batter after 2nd day of ambient storage
(b) Effect of sonication amplitude and time on overall 42
acceptability of idli batter after 5th day of ambient storage
4.13 (a) Effect of sonication time and amplitude level on overall 43
acceptability of idli batter after 2 day of ambient storage
(b) Effect of sonication time and amplitude level on overall 43
acceptability of idli batter after 5th day of refrigerated storage
4.14 (a) Effect of storage (ambient) period on overall acceptability of 44
idli batter soniacted at 100 µm for different times
(b) Effect of storage (refrigerated) period on overall 44
acceptability of idli batter soniacted at 100 µm for different times
4.15 (a) Effect of irradiation frequency on pH of stored (ambient) idli 49
batter
(b) Effect of irradiation frequency on pH of stored (refrigerated) 49
idli batter
4.16 (a) Effect of storage (ambient) period on pH of idli batter 50
irradiated at different frequencies
(b) Effect of storage (refrigerated) period on pH of idli batter 50
4.17 (a) Effect of irradiation frequency on acidity of stored (ambient) 51
idli batter
(b) Effect of irradiation frequency on acidity of stored 51
(refrigerated) idli batter
4.18 (a) Effect of storage (ambient) period on acidity of idli batter 52
(b) Effect of storage (refrigerated) period on acidity of idli batter 52
4.19 (a) Effect of irradiation frequency on SPC of stored (ambient) 53
idli batter
(b) Effect of irradiation frequency on SPC of stored 54

4.20 (a) Effect of storage (ambient) period on SPC of idli batter 54
(b) Effect of storage (refrigerated) period on SPC of idli batter 55
vii
4.21 (a) Effect of irradiation frequency on overall acceptability of 55
stored (ambient) idli batter
(b) Effect of irradiation frequency on overall acceptability of 56
stored (refrigerated) idli batter
4.22 (a) Effect of storage period on overall acceptability of stored 57
(ambient) idli batter
(b) Effect of storage period on overall acceptability of stored 57
4.23 (a) Effect of storage (ambient) period on pH of idli preserved 60
(b) Effect of storage (refrigerated) period on pH of idli preserved 60
4.24 (a) Effect of storage (ambient) period on acidity of idli preserved 61
(b) Effect of storage (refrigerated) period on acidity of idli 62
preserved
4.25 (a) Effect of storage (ambient) period on SPC of idli preserved 63
(b) Effect of storage (refrigerated) period on SPC of idli 63
preserved
4.26 (a) Effect of overall acceptability of preserved idli batter stored 64
(ambient)
(b) Effect of overall acceptability of preserved idli batter stored 64
(refrigerated)
viii
ABBREVIATIONS AND SYMBOLS
ANOVA Analysis of variance
CV Coefficient of variation
cfu Colony forming counts
˚C Degree celcius
df Degree of freedom
et al et ally
g Gram
h Hour (s)
kg kilogram
kGy kilogray
LDPE Low density polyethlyene
ms mean square
µm micrometer
min minute (s)
mL milliliter (s)
ppm parts per million
SPC Standard plate count
TTA Total titrable acidity
ix

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ABSTRACT

Idli, a traditional breakfast food of India based on rice-black gram combination, is a

Preservation Ambient Storage Refrigerated storage

No treatment 1 day 8 days

Sonication at 100 µm for 6 days 20 days

Irradiation at 1.25 kGy 7 days 64 days

Addition of Nisin 10 days 25 days

Keywords: Idli, fermented cereal-legume food, preservation, idli batter, sonication,

I believe in absolute oneness of God and also humanity. I bow my

I take this opportunity to express my deep sense of gratitude to

I feel a matter of great pride to record my reverence to the members

I am also thankful to all my teachers of Dr.R.V. Prasad, Dr.B.H. Joshi

I acknowledge my lovely mother (Mrs.Shashikala), father

Friendship being one of greatest blessing on the earth and it is a great

I am elated to avail this wonderful opportunity to record my

I am thankful to Mannubhai, Poonamkaka and Vasavakaka for their

I wish to extent special thanks to all of my beloved seniors, batch

Anand (Prarthana S.P.)

Dr. D.C.Joshi Tel. No. (02692) 261302

This is to certify that the thesis entitled “DEVELOPMENT OF THE

PRESERVATION TECHNIQUE FOR IDLI BATTER” submitted by

Ms. PRARTHANA S P in partial fulfillment of the requirement for the award

of the degree of Master of Technology in Food Processing Technology to the

Anand Agricultural University is a record of bonafide research work carried

out by her under my guidance and supervision.

Place : Anand (D. C. Joshi)

Idli, a traditional breakfast food of India based on cereal-legume combination, is a white,

Ultra-sonication is considered to be an emerging and promising technology for industrial food

Addition of bio-preservatives is another possible way of extending shelf-life of idli. The

2.1 Formulation of Idli Batter

Balasubramanian and Viswanathan (2007) while studying the physicochemical properties of

Parameter 2:1 3:1 4:1 Source

pH 4.21 5.27 5.9

Acidity (%) 0.44 0.62 0.91

pH 4.1 5.33 5.9

Balasubramanian and Viswanathan (2007)

Density (g cm-3) 0.94 0.74 0.59

Mahajan and Chattopadhyay (2000) studied development of chemically leavened cereal-

2.2 Fermentation of Cereal-legume Mix for Idli

2.3 Preservation Techniques for Idli

An irradiation effect on biogenic amines (BAs) and microbiological populations of Korean

Nisin is a natural antimicrobial peptide produced by strains of Lactococcus

Preservative ability of LAB in foods is attributed to the production of anti-microbial

MATERIALS AND METHODS

3.1 Raw Materials

Table 3.1(b) Physico – chemical property of split black gram dhal.

3.2 Preparation of Idli Batter

Milled rice Black gram dhal

Grind finely Grind coarsely

Combine slurries into a thick batter and mix well

Incubate at (30±2ºC) for 8–10 hrs

Fig. 3.3 Freshly prepared idli batter

3.3 Preservation of Idli Batter

3.3.1 Sonication of idli batter

3.3.2 Irradiation of idli batter

3.3.3 Chemical preservation of idli batter

Fig. 3.6 Nisin + Potassium Sorbate used as preservatives

3.4 Experimental Design