“DEVELOPMENT OF PROTEIN

ENRICHED PANCAKE MIX”

PROJECT WORK

SUBMITTED TO DEPARTMENT OF PG STUDIES IN FOOD SCIENCE

AND NUTRITION,

BESANT WOMEN’S COLLEGE, MANGALORE

IN THE PARTIAL FULFILMENT OF REQUIREMENT FOR THE AWARD OF THE

DEGREE OF MASTER OF SCIENCE IN FOOD SCIENCE AND NUTRITION

Submitted By,

FATHIMATH AFEEFA

Reg.no:193041963

Department of PG Studies in Food Science and Nutrition,

Besant Women’s College, M.G. Road, Kodialbail,

Mangalore -575003

October 2021

1
 CERTIFICATE

This is to certify that the project work entitled “DEVELOPMENT OF

PROTEIN ENRICHED PANCAKE MIX” is an authentic record of
independent research work done by Miss. FATHIMATH
AFEEA(Reg.no:193041963) under my supervision during the period of JULY
2021 to OCTOBER 2021, submitted to Mangalore university for the partial
fulfillment for the award of the degree of Master Of Science In Food Science
and the present work has not been previously formed the basis for the award of
degree, diploma, fellowship, associateship or other titles.

PLACE: VITLA, BANTWAL

DATE: 30 October 2021

RESEARCH GUIDE

Mrs. ROOPA RAO K

Assistant professor

Department Of PG Studies in Food Science and Nutrition

Besant Women’s College

M.G. Road, Kodialbail

Mangalore 575003

2
 BESANT WOMEN’S COLLEGE,

M.G. ROAD, KODIALBAIL, MANGALORE-575003

(Accredited by the NAAC with ‘A’ Grade)

CERTIFICATE

This is to certify that the project work entitled “DEVELOPMENT OF PROTEIN

ENRICHED PANCAKE MIX ”, submitted to the Department of PG Studies in Food
Science and Nutrition, by Miss. FATHIMATH AFEEFA (Reg.no:193041963) towards
the partial fulfillment of the degree of Master Of Science In Food Science And Nutrition,
is a faithful record of original work carried out by her in the academic year 2020-2021.

Place: VITLA, BANTWAL

DATE: 30 October 2021

HEAD OF THE DEPARTMENT

EXAMINERS:

1.

2.

3
 DECLARATION

I, FATHIMATH AFEEFA , hereby declare that project work entitled “DEVELOPMENT

OF PROTEIN ENRICHED PANCAKE MIX ” is a bonafide record of research work done
by me under the supervision of Mrs. ROOPA RAO K., Assistant professor , Department
of PG Studies In Food Science and Nutrition, Besant Women’s College, Mangalore.

The information depicted in the current report is the result of my own work, except where the
reference is made. The information provided in the report is authentic as per my knowledge.

The results embodied in the project work have not been submitted to any other university or
institution for the award of any degree, diploma, associateship, fellowship or similar titles.

Place: VITLA, BANTWAL

DATE: 30 October 2021

FATHIMATH AFEEFA

4
 ACKNOWLEDGEMENT

I would like to express my gratitude to almighty God for without his grace and blessings this
study would not have been possible.
I express my sincere thanks to Dr. SATHISH KUMAR SHETTY.P, Principal, Besant
Women’s College, Mangalore.
I pay my deep sense of gratitude to Ms. Roopa Rao K of Food Science And Nutrition
Department, Besant women’s college, Mangalore whose valuable guidance and kind
supervision given to me throughout the project work which shaped the present work as its
show. I am immensely obliged to my friends for their elevating inspiration, encouraging
guidance and kind supervision in the completion of my project.
I feel to acknowledge my indebtedness and deep sense of gratitude to my lecturers Ms.
SHWETA SINGH, Ms. ASIYAMATH SHAHDA FIRHATH.M.A, and Mrs INDIRA
Department of Food Science And Nutrition, Besant Women’s College, Mangalore to
encourage me to the highest peak and to provide me the opportunity to prepare the project.
Last, but not the least, my parents are also an important inspiration for me. So with due
regards, I express my gratitude to them

Place: VITLA, BANTWAL

DATE:30 October 2021

5
 TABLE OF CONTENTS

CHAPTER CONTENT PAGE

NUMBER NUMBER
Certificate 1
Certificate 2
Certificate 3
Declaration 4
Acknowledgement 6-8
Table of Contents 9
List of tables 10
Abstract 11-13
Chapter 1 Introduction 14-20
Chapter 2 Review of literature 14
2.1.Protein intake pattern 15

2.2.Nutrition composition 16
2.3. Germination 16
2.4. Health benefits 17
2.5. Prevalence of malnutrition 18
2.6. Product development 19
2.7 . Wheat flour 20
2.8 Jaggery 20
2.9 Packaging 20
Chapter 3 Materials and Methodology 21-26
3.1.Materials 22
3.1.1.Raw Ingredients 22
3.1.2. Equipment and glassware 22
3.1.3. Reagents and chemicals 22
3.2METHODOLOGY 23

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 3.2.1. Adulteration Test 23
3.2.2 Standardisation of Processing methods. 23
3.2.3 Variations of products 24
3.2.4. Determination of Protein content by Lowry’s 24
method
3.2.5. Preparation of Pancake mix 25
3.2.6. Sensory Evaluation by preference test 25
3.3.PROXIMATE ANALYSIS 26
3.3.1 Estimation of moisture content by Hot air oven 26
method
3.3.2 Estimation of the fibre content 26
3.4. SHELF LIFE STUDY OF THE PRODUCT 27
Chapter 4 RESULTS AND DISCUSSION 27-31
4.1. Adulteration test of raw ingredients. 28
4.2. Standardisation- processing methods 29
4.3. Protein estimation of Pancake mix. 29
4.4. Sensory evaluation by Preference test. 30
4.5. Valuation of final product 31
4.6. Proximate analysis 31
4.7. Shelf life study of the Pancake mix 32
Chapter 5 SUMMARY AND CONCLUSION 33-35
Summary 34
Conclusion 35
References 36-38
Appendix 39-44

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 LIST OF TABLES

SERIAL PARTICULARS PAGE

NUMBER NUMBER
1. Raw ingredients are used in the pancake 22
mixture
2. Adulteration test of raw ingredients 23
3. Different variations of the ingredients 24
4. Result -Adulteration test of raw ingredients 28

5. Processing method 29

6. protein content of different variations of 30

pancake mixer

7. Proximate analysis of protein rich pancake 31

mixer

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 ABSTRACT

Protein energy malnutrition (PEM) is a fairly common world wide in both children and
adults. With an ageing population, dietary approaches to promote health and independence
later in life are needed. In part, this can be achieved by maintaining muscle mass and strength
as people age. dietary recommendations for protein intake may be insufficient to achieve this
goal and that individuals might benefit by increasing their intake and frequency of
consumption of high-quality protein.
Pancake is a breakfast dish which is liked by all age group. Protein rich pancake product
provide adequate amount of protein and also contain fibre which provide bulk to the diet.
Developed pancake is a healthy choice that can be incorporated into the diet to prevent
protein deficiency.

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 CHAPTER 1:
INTRODUCTION

1.0.Introduction:
A healthy eating habits, despite of age involves consumption of high quality protein
meals throughout the day. Adequate consumption of dietary protein is important for
maintaining health status, growth and development functions throughout the life.
They also support physical activity, weight loss and prevention risk of disease. Food
protein quality is traditionally dependent on its amino acids content, protein contains
20 different amino acids are linked by peptide bonds. Amino acids helps in providing
nitrogen, hydrocarbon skeletons and sulphur and impossible to replace by any other
nutrients (such as lipids and carbohydrates) because nitrogen and sulphur are not
synthesised in the body. Amino acids are the precursors which is, most important for
the synthesis of proteins, peptides and low molecular weigh substances( nitric oxide,
RNA and DNA) with extensive physiological importance.
The protein under nutrition leads to anaemia, stunting growth, edema, physical
weakness, vascular dysfunction and impaired immunity.
The pancake mixer play a major role in market and they have great
demand because of variety of product and their convenience in preparation. Pancake
is weight based products are consumed globally by all the age groups.
Here the main aim of the project is to prepare a pancake mixer which is
nutritious. The ingredients are horse gram powder, wheat flour and jaggery powder.

Horse-gram powder:
Horse gram( Macrotyloma uniflorum) Is rich source of protein. It also contains
calcium and iron. Simple processing such as roasting soaking of or scrum were used
to reduce anti nutritional factor. in the processing method, the horse gram was roasted
for 10 minutes cool down and then powdered. Where the anti nutritional factors such
as tan in san fights reduced on processing, nutrition such as protein calcium iron and
phosphorus did not change when compared to that contained in raw horse gram.
Horse gram occupies an important place in human nutrition. Besides nutritional
importance, it has been linked to reduce the risk of various diseases due to the
presence of non negative bio active substances. The a recent review attributed with

10
recent scientific knowledge towards the possibility of exploring the horse gram, as a
source of food and nutraceutical compound.

Wheat flour:
Wheat floor is the most important food which contributes more calories, protein and
carbohydrates to the diet. It is also considered as a good source of minerals, B-group
vitamins and dietary fibre. It is low in fat and cholesterol. Wheat is highly
controversial because it contains a protein called gluten, which can trigger a harmful
immune response in predisposed individuals. People who are intolerant to gluten need
to eliminate weight from their diet . Wheat flour improve digestion and help prevent
colon cancer.

Jaggery powder:
Jaggery powder Containing less than 3 mm particle size is used to give a v good
flavour to pancake. The jaggery has a longer shelf life for upto two years. Jag contains
more nutrients than sugar because Sugar contains only empty calories whereas
jaggery contains iron, magnesium, potassium and manganese also contains small
amount of B vitamins and minerals, including calcium, zinc, phosphorus and copper.
Health benefits include improve digestive health, anaemia prevention liver
detoxification and improved immune function.

11
 CHAPTER 2
Review of literature

2.1. Protein intake pattern

Sumathi Swaminathan et al., (2012) stated that 60% of Indian diet of protein is derived
from cereals. From the last 25 years the National Nutrition Monitoring Board (NNMB) took
several surveys of protein intake and diet in urban and rural areas and also in slum area. The
survey shows that in Indian population the protein intake that is mainly from cereals is about
1gm/kg/day form slum, tribal and sedentary rural area. According to protein digestibility
corrected amino acid score (PDCAAS) in particular rural area the intake of protein is less .
The one third of the rural population appear to have inadequate quality of their protein intake
which is at risk of not meeting their daily requirements.

Sylvie Rousset et al., (2003) studied that sufficient level of protein required to maximise the
gradual body protein loss is noticed throughout aging. Age, sex and life conditions that may
modify protein intake and distribution. But the accurate amounts, also their daily distribution
that affects the protein utilisation and N retention which are not known in both young and
elderly individuals. Protein intake and protein distribution over daily meals is different for
both young and elderly individuals. For men the average protein intake was lower in the older
age group whereas opposite trend was seen in women. The protein intake distribution was
different between two age groups that is for lunch 56.5% of daily protein was eaten by the
elderly and only 47% by the younger subjects. So the protein intake pattern differs between
sexes and age groups.

Sumedha Minocha et al., (2017) analysed that the protein requirement associated to its
equality or digestibility to meet human essential amino acid requirements. In India they
consume cereal based food which has low quality protein leads to risk of quality protein
deficiency. From the study the percentage of the population at risk of quality protein
deficiency that vary between 4% and 26% among different age groups and between the urban
or rural sector . To reduce these risk requires greater intake of high quality protein such as

12
pulses, and that food subsidy policies move beyond cereals and become more quality
conscious .

2.2Nutritional Composition
SS Kadam et al., (1985) studied that the horse gram are grown mostly under dry land
agriculture . Like other legumes horse grams are deficient in methionine and tryptophan.
Horse gram is rich source of iron and molybdenum whereas it has higher trypsin inhibitor and
hem-agglutinin activities and polyphenols. The germination, dehusking, roasting and cooking
are shown to produce beneficial effect on nutritional quality.

M Marimuthu et al., (2013) evaluated the flour functionality and nutrient composition of
horse gram seeds were examined of their flour characteristics. The main aim is quantifying
physiochemical and functional properties which may serve as a guide to utilise its benefits for
human and animal nutrition . In this study they may provide a guide line for the use of horse
gram seed flour are good functional food for nutrition, food formulation and utilization.

Yadahally N Sreerama et al., (2008) investigated that the use of horse gram and its flour is
limited in legume composite flours and products due to the presence of anti nutritional
components, poor functional and expansion properties of horse gram make easier its use as
ingredient in food processing. New popped or expanded horse gram come from xylanase-
mediated depolymerisation of cell wall polysaccharides of horse gram. This popped horse
gram is resulted in increased length (5.3-6.8mm) and higher yield of expanded grain(63-98%)
. The xylanase treated and expanded horse gram flour had higher water (204.3g/100g) and oil
absorption capacities (98.4g/100g) than unprocessed flour which had 135.8g/100g and
74.6g/100g respectively at ambient conditions. This study shows that flour functionality and
nutritional value of horse gram can be improved by processing it into new product that can
be used as an ingredient in food processing
Iron was 8.71 -9.16 mg %(DMB)

Aishwarya V. Patil et al., (2019) stated that horse gram is a potential grain legume which
has excellent nutritional quality. Seven released varieties and five breeding lines of horse
gram were acquired from All India Coordinated Research project dry land project Vijaypur.
The horse gram were evaluated for crude protein, in vitro protein digestibility, antioxidant

13
activity and polyphenol content . There was no significant difference among the varieties was
observed.

2.3Germination
RS Pal et al., (2016) investigated the change in antioxidant activity, chemical composition
and mineral content of horse gram after dehulling and germination. When compared to the
raw and germinated horse gram the dehulled samples contain higher protein content. After
dehulling (29.31%) and germination (98.37%) the total soluble sugars (TSS) content increase
whereas the total lipid increases(10.98%) after dehulling and decreases after germination .
The phytic acid, tannin, and oxalic acid are decreased after dehulling and germination. Here
they studied that the bulk density(11.85%) and oil absorption capacity (18.92%) of flours
significantly increased after germination. After germination combined with dehulling
increase the nutritive value of horse gram and reducing anti nutrients.

Atinder Ghumman et al., (2016) studied the effect of germination on protein profiling,
functional and pasting properties of lentil and horse gram lines. The random coils and alpha
helix proportion of beta sheets was lowest whereas proportion of the beta sheets and beta
turns are highest . Germination of horse gram does not affect trypsin inhibitor. Amylose
content decreased during germination. Uncooked starch shows the increase in proportion of
small granules and decrease in proportion of large granules.

Vanshika Handa et al., (2017) optimised the soaking and germination conditions of horse
gram to decrease the anti nutritional factors as well as maintaining the nutritional properties
of horse gram. Under different illumination condition that is light and dark the horse gram is
soaked for 6, 12 and 18hr followed by germination for 0,24 and 48hr . Then this soaked and
germinated are dried at 55 degree Celsius in laboratory. The physiochemical and functional
characteristics are affected by both soaking and germination where the germination which is
done in dark and light conditions effect the ascorbic content, total protein, total phenols ,
antioxidant activity and tanin content . In this study it was found that the germinated sample
in the presence light decreases anti nutritional factors and there was increase in ascorbic acid
and total protein content.

14
2.4. Health benefit
Saroj Kumar Prasad et al., (2015) studied that horse gram is grown in wide range of
adverse climatic conditions which is an under-utilised crop. It plays an important role in
human nutrition and has rich source of protein , minerals and vitamins. Bioactive substances
such as phenolic acid , phytic acid, fibre, proteinase inhibitors have notable metabolic or
physiological effects . Horse gram is recognised by traditional medicine as they are the
potential therapeutic agent to treat kidney stones , urinary diseases piles common cold , fever
etc.

A Bhartiya et al., (2015) discussed the horse gram (Mycrotyloma uniflorum) belongs to
family Fabaceae are among under-utilised legumes which is lesser known neglected legume
that are cultivated in Asian and African countries. Horse gram has best therapeutic properties
and are used to cure kidney stones, asthma, bronchitis, piles , heart disease , urinary discharge
etc. other than this it also have anti diabetic , anti ulcer activity and also help in dietary
management of obese people due to the presence of beneficial bioactive compounds.

2.5.Prevelance of malnutrition
Dechelen Tshering Bhutia(2014) discussed that in India Protein Energy Malnutrition is a
major health problem in India. This affect is mainly seen in children which leads to
permanent impairment in later life. Underweight, stunted growth are the symptoms of PEM.
The widespread of stunting among under five is 48% and wasting is 19.8% and the
underweight is 42.5% is highest in the world. This PEM is also seen in children who is
suffering from poverty where they cannot meet their dietary nutritional requirements .

Aditya Vedantam et al., (2009) estimated the frequency of malnutrition among elderly in
rural people of south India. Nutritional status was evaluated using the Mini Nutritional
Assessment (MNA) questionnaire which had an eighteen- item nutritional screening
instrument used in elderly. Here as examined by the MNA, 14% of the 227 subjects were
malnourished and 49% are at risk. The majority are living with their children with low
income .In south India more than 60% subjects the low MNA scores shows the lower intake
of protein and require more attention.

15
Bobby Joseph et al., (2002) identified the prevelance of malnutrition in the rural areas of
Karnataka among 256 children who attended anganawadis. The children from 1-5years are
from villages located at the outskirts of Bangalore city. The children with wasting , stunting
and wasting and stunting was 31.2%, 9.4% and 29.2% respectively. To detect acute
malnutrition they used reference weight and calculated using Weech’s formula. The study
showed that mid-upper arm circumference (MUAC) should be used carefully because they
are not susceptible to detect all the malnutrition cases, although the MUAC for height can be
used because they are more susceptible. For the calculation of stunting Weech’s formula can
be used, however the sensitivity is not very high.

2.6.Product development
Thirukkumar.S et al., (2014) evaluated that horse gram is the inexpensive sources of
protein calcium and iron. Simple processing of horse gram used to reduce the anti nutritional
factors that is soaking and roasting. One portion of horse gram was washed and soaked in
required amount of water for 12 hours with periodical change of water every four hours. Then
horse gram is dried and then powdered. In another method horse gram was roasted on low
flame for 10 minutes , cooled and powdered whereas the tannins and phytates are anti
nutritional factors which are reduced during processing. The nutrients protein, calcium, iron
and phosphorus will not change during processing. In Wheat based chappathi to increase the
nutrient content of the product this processed horse gram is added. 10% soaked and dried or
15% roasted horse gram flour is added to chappathi prepared from wheat flour was highly
acceptable.

Shashi Jain et al., (2012) studied that the seed of horse gram are of to varieties that is ‘AK-
21’ and ‘AK-42’ were examined for their physiochemical and nutritional properties using
five product such as kasar, laddoo, mathri, biscuits and Khakra. The hydration capacity for
both varieties was 0.03g /seed at 6 hr similarly 0.04g/seed at 12,18 and 24 ht. When
compared to AK-42 the seed count , seed weight and density is higher in AK-21. After 18hr
after soaking in plain water the cooking time of Ak-42 was 50 min whereas cooking time for
AK-21 was 60 min. The protein content in AK-21 was 15.10g% and the protein content
inAK-42 was 15.32g%(DMB). The fat content of Ak-42 is less when compared to AK-21.
The fibre content ranged between 4.57-5.15g% . The amount of energy ranged from 376.12
to 377.21 kcal/100g and the iron was 8.71 -9.16 mg %(DMB)

16
Homi Joshi et al., ( 2020) described that horse gram is a cheap source of protein and nutrient
rich pulse. Horse gram is most suitable for commercial foods and its flour is used in
preparation of food products. Here they aimed at developing functional biscuits by adding
horse gram flour to wheat flour and to check the physical property and sensory
characteristics of sensory gram flour biscuits. Sensory evaluation shows that product
developed by adding 25% horse gram flour to wheat flour had good taste , flavour , texture
and overall acceptability. This study proves that horse gram flour can be successfully added
to get improved nutritional value.

Masaharu Seguchi(1990) assessed that pancakes baked with wheat flour that is heat treated
are decreased in their gumminess and increase in springiness. The temperature dependent
Brabendar Amylograph test of this heat treated wheat flour at 120 degree Celsius for
0,1.0,2.0,3.0 and 5.0hr shows that when the time of heat treatment is increased the first
viscosity of flour slurries starts at low temperature. The change in property by heat treatment
are consistent with properties of chlorinated wheat flour.

2.7.Wheat flour
Saeed Akhtar et al., (2011) studied that government and world organisations focused on
deficiencies of iron , zinc and vitamin A . Wheat flour is suitable for micronutrient
fortification. This fortification of wheat flour has been accepted as an appropriate strategy to
reduce micronutrient deficiencies.

Miki Ozawa et al., (2006) examined that the dry wheat flours divided into water soluble ,
gluten , prime starch and tailings component by an acetic acid fractionation technique. At
each temperature with a gradual increase in the time duration the recovery of the prime starch
component decreased whereas tailing component increased the other two components did not
change . These dry heated wheat flours gave pancake springiness. The interaction between
the tailing component and prime starch component in wheat flour produced by dry heating
was highly associated to improvements in the pancake springiness.

Monica whent et al., (2012) studied that from five wheat cultivars the whole wheat flour
was estimated for phenolic, carotenoid and tocopherol composition and also anti

17
inflammatory and anti proliferative activities against HT-29 cells. The total ferulic acid is
primarily present in the insoluble bound form that is ranged from 452 to 731 micro gram per
gram . Here the carotenoid is Lutein and was ranged from 1.5 to 4 micro gram / gram and
alpha tocopherol levels ranged from 12 to 61 micro gram per gram. The research display that
whole wheat flour of the five cultivars varied notably and there is possibility that wheat
varieties are selected based on possible health benefits.

2.8.Jaggery
J Singh et al., (2013) described that the 65% people of India lives in rural villages. The rural
population suffers due to malnutrition as they cannot meet their nutritional requirements.
Healthy food should be consumed so that the consumption of the food may prevent diseases .
Jaggery, is a product from sugarcane which is rich in important minerals. The magnesium
present in jaggery strengthens our nervous system also take care of our blood vessels. The
potassium and the low amount of sodium maintain the acid balance in the body cells. Jaggery
is also used as medicine for migraine in Ayurvedic field.

Parth Hirpara et al., ( 2020) explained that jaggery is a medicinal sweet where it contains
nutrients like protein vitamin and minerals like iron and copper . Texture of jaggery is hard
and has golden yellow colour. Jaggery is also an energy food which can be used for blood
purification, regular functioning of live and also keep blood healthy. It improves digestion ,
boosts energy, relieves constipation, it has anti toxic and anti carcinogenic properties,
treatment of bronchial or lung infections and pre menstrual syndrome
2.9.Packaging
Bernhard Saam (1999) studied that a package of stand up pouches in which a rectilinear
container holds the pouch inside with closed top portion , they are arranged between adjacent
base portion of the pouches occupying minimum space . The container can be opened by
separating the portions the line of weakening is broken.

18
 Chapter 3
Materials and Methodology

The main aim of the study is to develop a nutritious protein rich pancakes. They were
developed in different variations. The Horse-gram were analysed for their protein,
fibre and moisture content. Sensory evaluation was done and finally they were
checked for their shelf life.

19
 3.0. Materials and Methodology
3.1. Materials:
3.1.1. Raw ingredients:
The raw ingredients were bought from the local market in mangalore.
Table 1: Raw ingredients are used in the pancake mixture
Ingredients Amount(in grams)
Horse-gram 45
Wheat flour 35
Jaggery 20

3.1.2. Equipment and glassware:

Equipment: Weighing balance (Essae-Teraoka Pvt. Ltd ), Hot air oven (B.D.
instrumentation, Ambala Cantt), muffle furnace ( Rotek instruments, Kerala), 2375 Double
beam spectrophotometer, water bath (Labotech instruments, B.D. instrumentation, Ambala
Cantt).

Glassware: Beakers, Test tubes, Conical flasks, Glass rod, Burette, Standard flask,
measuring cylinder, Watch glass, Petri dish, Pipettes, Micropipettes, Conical flask.

3.1.3. Reagents and chemicals used: All reagents and chemicals used in the experiments
were of AR grade procured from Fisher Scientific, Mumbai; Medilise, Kerala; Emplura,
Mumbai.
 Bovine serum albumi
 alkaline copper sulphate
 0.1N sodium hydroxide solution
 1% sodium potassium tartarate
 2% sodium carbonate
 Folin Ciocalteau Reagent

3.2. METHODOLOGY:
3.2.1. Adulteration test:
Table 2: Adulteration test of raw ingredients

20
Ingredient Test Adulterant
Horse-gram Take 2-3 grams of horse-gram sample and place them in Extraneous matter
petridish then they are examined usually ( dust, stone,
insect and hair)
Jaggery To 1 gram of melted sample few drops of concentrated Chalk powder
hydrochloric acid were added. Mixed well.

3.2.2. Standardisation of processing method.

a) Roasting:
Horse-gram was roasted in a pan for 10 minutes in low flame, cooled and subjected to
milling.
b) Jaggery:
20gram of solid jaggery were ground to fine powder using mechanical grinder.
3.2.3. Variations of the product:

Table 3: Different variations of the ingredients

Variation 1 Variation 2
Horse-gram- 35gram Horse-gram- 45gram
Wheat flour- 45gram Wheat flour- 35gram
Jaggery- 20 Jaggery- 20

3.2.4. Determination of protein by Lowry’s method:

Powdered sample of one gram was mixed with 4ml of potassium phosphate buffer(pH is
7.4) and the sample was centrifuged at 500rpm for 15 minutes. 1ml of supernatant was used
for protein estimation. The standard sample was prepared using bovine serum albumin of
200μg/ml concentration with aliquots of 0.2-1.0ml which were made upto 1.0ml with
distilled water. 5 ml of alkaline copper sulphate solution was added and the test tubes were
allowed to stand for 10minutes, later 0.5ml of Folin-Ciocalteau Reagent(FCR) was added and
all the test tubes were incubated in dark for 30minutes. The absorbance was read at 660nm
using spectrophotometer.

21
3.2.4. Preparation of Pancake Mixer

Step1:Powdering of Horse-gram:
Roast the horse-gram and powder it.

Step2: Horse-gram was added to wheat

flour.

Step 3 : Addition of jaggery powder.

Step 4: Mixing

The amount of raw ingredients used in the product was depicted in the table 1.

3.2.5. Sensory evaluation using preference test:

The sample containing high amount of protein by Lowry’s method was subjected to the
preference test with random sampling using the sample size of 50.

3.3. PROXIMATE ANALYSIS:

Estimation of moisture and fibre of the product. The methods described by AOAC 2000.

3.3.1. Estimation of moisture content by hot air oven method:

In a sterile glass petriplate 5g of pancake mixer was taken whose weight was noted earlier.
The sample were dried in hot air oven at 125°C for 4 hours . Then the samples were cooled
in the desiccator for 30 minutes and the weight was noted that is (m1). The sample were
again placed in the oven for another 1 hour at 125°C. The samples were cooled for 30
minutes in the desiccator and the weight was taken that is (m2). The average of the both
weight were taken and was substituted in the formula to obtain the % of moisture.

22
Moisture (%) was calculated by the formula:

(weight of petridish+sample prior drying)(g) - ( weight of petridish+ sample

prior drying)(g) X100

Weight of the food sample(g)

3.3.2. Estimation of fibre:

In a 500 ml beaker, 2g of food sample (pancake mixer) was taken to that add 200ml of
0.255N of sulphuric acid solution and boil for 30 minutes. Then it was filtered using a muslin
cloth and washed with 200ml of boiling water, till all the acid was washed off. The content
were filtered in a filter paper washed with water and were dried for 2 minutes in hot air oven
for 250°C and the dried sample was scraped off to the weighed crucible and was dried again
for 100°C for 4 hours, it was cooled and weighed. The content were placed in the preheated
muffle furnace at 600°C for 30 minutes, cooled and weighed .
The crude fibre content was calculated by using formula;

Weight of the crucible before ashing - Weight of the crucible after ashing

Weight of the food sample(g)

3.4. Shelf-life study of the product:

Sensory evaluation of the pancake mixer was done by using 9 point hedonic scale rating on
the overall acceptability for 2month period with 4sessions of analysis on the 15th day , 30th
day, 45th day and 60th day.

23
 CHAPTER 4

RESULT AND DISCUSSION :

Lowry’s method was used to estimate the protein content of two different variations of
Protein rich pancake mixer.
Pancake mixer with high protein content was selected as final product. The final
product then checked for the presence of adulterant, moisture content, fibre content and shelf
life.

24
4.0. Results and discussion
4.1. Adulteration test:

Table 4: Adulteration test of raw ingredients

Ingredient Adulterant Observation Inference

Tested
Horse- Extraneous matter No extraneous matters Horse-gram was not adulterated.
gram (Dust, stone, were observed.
insect, hair)
Jaggery Chalk powder No effervescence was Jaggery was not adulterated.
observed.

All the ingredients used in the preparation of pancake mixer was free from adulterant.

4.2. Standardisation – Processing method:

Table 5: Processing method:

Processing Observation Inference

method
Roasting of  Horse-gram that were
horse-gram subjected to dry roasting had
acceptable colour, odour and
appearance.
 Horse-gram that were not Roasted horse-gram were
subjected to dry roasting had incorporated in the pancake mixer
unacceptable colour, odour preparation.
and appearance.

4.3. Protein estimation of pancake mixer by Lowry’s method.

25
Table 6: protein content of different variations of pancake mixer

Variation Compositions (total 100g Protein concentration

of pancake mixer) per gram of
sample(μg/ml)
Variation 1  Horse-gram=35g 580.23
 Wheat flour =45g
 Jaggery =20g
Variation 2  Horse-gram =45g 648.3
 Wheat flour=35g
 Jaggery =20g

Figure 1. protein content of different variations of pancake mixer

Protein concentrations of different

variations of pancake mixer.
Series1

648.3

580.23

variation 1 variation 2

From Figure 1, variation 2 has highest protein content when compared with variation 1.

26
4.4. Sensory evaluation by preference test:
Pancake made from two different pancake mixer were used for preference test. During
pancake preparation milk, baking soda , baking powder, cardamom powder were added to
improve the sensory quality of pancake . The sample size was 75, it was observed that out of
50 people 17of them liked variation 1 pancake mixer whereas 33of them preferred variation
2

Sensory evaluation by preference

test

34%
variation 1
variation 2
66%

Figure 2 Sensory evaluation by Preference test.

Figure 2 shows that the result Result of the preference test indicating that variation 2 was
most preferred for their sensory characteristics like appearance, flavour colour and odour . So
variation 2 was selected as final product.

4.5. Valuation of final product:

Sensory evaluation was done by preference test showed that variation 2 was liked by most of
the participants and Lowry’s method show that the protein content of variation 2 pancake
mixer was high and can be used by all the age groups.

27
4.6. Proximate analysis

The proximate analysis ( moisture, fibre and protein content) of protein rich pancake mixer
per 100 gms are given in Table 7 and Figure 5

Table 7: Proximate analysis of protein rich pancake mixer

Particulars Composition (100grams)

Moisture content 1.832g
Crude fibre 3.363g
Total protein 648.3mcg

 Total moisture content show the amount of water present in the protein rich was
 Crude fibre content is the non digestible component that provides bulk to the diet. The
crude fibre content of protein rich pancake mixer was found to be
 The total protein content of pancake mixer was found to be a protein rich
product.

28
4.7. Shelf life of protein rich pancake mixer:

Sensory characteristics of the pancake mixer were observed in 4 different sessions.

1st day – sensory characteristics of pancake mixer that is colour, odour, flavour and
appearance was examined.

15th day - The sensory characteristics of pancake mixer was same and acceptable.

30th day - There was no change in the sensory characteristics and it was acceptable.

45th day - There was no change in the sensory characteristics and it was acceptable.

60th day - there was a Noticeable change in the flavour, colour, odour and appearance was
observed

From this study , it is acknowledged that the shelf life of the protein rich pancake mixer was
60 days.

29
 CHAPTER 5

SUMMARY AND CONCLUSION

This section gives deep understanding of This study. The product developed was rich in
protein . The shelf life of the product upto 2 month.

30
5.1. SUMMARY

 The adulteration test asserted the raw ingredients used for the preparation of protein
rich pancake mixer Was free from contamination.
 Horse gram was subjected to roasting which is cost effective and improve the
nutritional properties and tastes.
 Two different variations was done to get the combination of the product.
 Various proportions were considered to finalise the perfect sample to get perfect
pancake.
 Approximate analysis and shelf life study was done.
 Most of the participants prefer the pancake mixer of horse gram 45g, wheat flour 35g
and jaggery 20g.
 The pancake mixer was then observed for their shelf life and it was remained til 2
months.

31
5.2. CONCLUSION:
Protein is very important to be included in the diet for all age groups. The pancake mixer is a
rich source of nutrients due to the presence of horse gram and wheat flour, which are locally
available and affordable. This pancake mixture is a good source of energy, protein and
carbohydrates. The shelf life of the product was shown to be for 2 months. consuming
adequate dietary protein is critical for maintaining optimal health, growth development which
is provided by this protein rich pancake mixer.

32
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36
APPENDIX

APPENDIX 1
Preparation of Reagents:
1) Estimation of protein by Lowry’s method:
 standard protein solution
20 mg of bovine serum albumin was weighed and transferred to a 100
ml standard flask. It was made up to mark with distilled water. The
concentration of the standard protein was 200mcg/ml.
 Alkaline copper sulphate solution
The alkaline copper sulphate was prepared by mixing 50ml of
solutionA and 1ml of freshly prepared solution B.
- 0.1N sodium hydroxide solution
4 grams of sodium hydroxide pellets were weighed in the analytical
balance and transferred to a 100ml standard flask using a funnel, it was
mixed with distilled water and shaken well till the pallets dissolved
completely and was made up to the mark .
-1%Sodium potassium tartrate solution
One gram of sodium potassium tartrate crystals were weighed and
were transferred to 100ml standard flask. It was made up to the mark
by distilled water. It is freshly prepared.

Solution A: 2% Sodium carbonate solution.

2 grams of sodium carbonate was wade and was transferred 100ml
standard flask. It was made up to the mark with 0.1N sodium
hydroxide solution.
Solution B: 0.5% copper sulphate solution.
0.5 grams of anhydrous copper sulphate crystals were weighed and
transferred into 100ml standard flask it was made up to the mark with
1% sodium potassium titrate solution.
 Folin ciocalteau Reagent (FCR):

37
 Okay 50ml of FCR was Prepared by taking 25ml of FCR in 50ml
measuring cylinder and making it up to 25ml by distilled water.
 Potassium phosphate buffer
3.03 grams of dipotassium hydrogen phosphate crystals and 1.035
grams of potassium di- hydrogen phosphate crystals were weighed
separately and were transferred to a 250ml standard flask it was made
up to the mark with distilled water.
The pH was adjusted by dipping the electrode of pH meter in the
buffer solution and adjusting the pH with 0.1 sodium hydroxide(If less
than 7.4) or with 0.1 hydrochloric acid(If more than 7.4)

2. Fibre estimation by AOAC method:

 0.255N sulphuric acid solution

7ml of concentrated sulphuric acid was measured in a volumetric cylinder and was
transferred into 1000ml standard flask using funnel, made up to mark with distilled water.

 0.313N sodium hydroxide solution

12.38 Gms of sodium hydroxide pellets were weighed using analytical balance and
transferred to a 1000ml standard flask and was added with distilled water until the pellets
dissolved. It was made up to mark with distilled water.

38
 APPENDIX-2
SENSORY EVALUATION OF PANCAKE USING 5-POINT HEDONIC
SCALE
FORM 1:-

NAME: _____________________
AGE: _______________________

Taste each of the following samples and tick how much you like it or dislike it.
Instructions: Rinse your mouth with water and have a piece of cracker before
tasting each sample.
V1 V2

Like
lot _____ _____

Like a little _____ ______

Neither like
nor dislike _____ _____

Dislike a little _____ _____

Dislikea
lot _____ _____

SIGNATURE: ________________________

DATE: _________________________

39
Form 2:-

You are receiving a set of samples of pancake. Kindly taste each of from the set and drink
water provided before tasting the next sample. Evaluate them by numbering on the basis of
attributes given below.
ATTRIBUTES pancake

V1 V2

APPEARANCE
COLOR
TEXTURE
AROMA
TASTE
OVERALL

ACCEPTABILITY
(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.)

SIGNATURE: ________________________
DATE: _________________________

40
 APPENDIX-3
SHELF LIFE STUDY OF PANCAKE MIX USING 9-POINT
HEDONIC SCALE

NAME :___________________
AGE :____________________
Taste each sample given and rate the characteristics given below on a scale of 1 to
9.
9-Like extremely 4-dislike slightly
8-Like very much 3- dislike moderately
7-Like moderately 2- dislike very much
6-Like slightly 1- dislike extremely
5-Neither like nor dislike
PANCAKE STORED :-

Sensory characteristics Day 1 Day 15 Day 30 Day 45 Day 60

Appearance

Colour

Texture

Aroma

Taste

Overall acceptability

SIGNATURE: ________________________
DATE:

41
DAY 1 _________________________
DAY 15 _________________________
DAY 30 _________________________

DAY 45 _________________________
DAY 60 _________________________

42
 APPENDIX 4
Optical density of solution used for calorimetric analysis:

 Estimation of protein by Lowry’s method

Aliquots of standard
S1 S2 S3 S4 S5 Blank
Volumes of standard
protein solution(ml) 0.2 0.4 0.6 0.8 1.0 -
Concentration of
standard protein 40 80 120 160 200 -
solution(mcg/ml)
Volume of distilled
water (ml) 0.8 0.6 0.4 0.2 - 1.0
Volume of copper
sulphate (ml) 5 5 5 5 5 5
Volume of Folin-
Ciocalteau 0.5 0.5 0.5 0.5 0.5 0.5
Reagent(ml)
Optical density of
standard solution at
660nm

43