DEVELOPMENT OF BAKERY AND CONFECTIONERY

PRODUCTS USING GHEE RESIDUE

By

P.VINODHINI
B.Tech (Dairy Technology)

THESIS SUBMITTED TO
ACHARYA N.G.RANGA AGRICULTURAL UNIVERSITY
IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE AWARD OF THE DEGREE OF
MASTER OF FOOD SCIENCE AND TECHNOLOGY

INTERFACULTY PG PROGRAMME

POST GRADUATE AND RESEARCH CENTRE

ACHARYA N.G.RANGA AGRICULTURAL UNIVERSITY
RAJENDRANAGAR, HYDERABAD-500 030.

OCTOBER, 2004
 CERTIFICATE

Miss. P. VINODHINI has satisfactorily prosecuted the course of

research and that the thesis entitled “DEVELOPMENT OF BAKERY AND

CONFECTIONERY PRODUCTS USING GHEE RESIDUE” submitted, is the

result of original research work and is of sufficiently high standard to warrant its

presentation to the examination. I also certify that the thesis or part thereof has not

been previously submitted by her for a degree of any University.

Date : (Ms. KANWALJIT KAUR)

Place : Hyderabad Major Advisor

 CERTIFICATE

This is to certify that the thesis entitled "DEVELOPMENT OF

BAKERY AND CONFECTIONERY PRODUCTS USING GHEE
RESIDUE’’ submitted in partial fulfillment of the requirements for the degree of
MASTER OF FOOD SCIENCE AND TECHNOLOGY for Acharya N. G.
Ranga Agricultural University, Hyderabad is a record of the bonafide research work
carried out by Miss. P. VINODHINI under my guidance and supervision. The
subject of the thesis has been approved by the student’s advisory committee.

No part of the thesis has been submitted for any other degree or diploma.
The published part has been fully acknowledged. All the assistance and help received
during the course of investigation have been duly acknowledged by the author of the
thesis.

(Ms. KANWALJIT KAUR)

Chairman of the Advisory committee

Thesis approved by the student Advisory Committee

Chairman : (Ms. KANWALJIT KAUR) _______________
Assistant Professor
Department of Food and Nutrition
College of Home Science
Rajendranagar, Hyderabad

Member : (Dr. S. SUMATHI) _______________

Associate Professor
Department of Food and Nutrition
Post Graduate Research Centre
College of Home Science
Rajendranagar, Hyderabad

Member : (Dr. K. KONDAL REDDY) _______________

Associate Professor
LPT Department
College of Veterinary Science
Rajendranagar, Hyderabad
 CONTENTS

Chapter No. Title Page No.

I INTRODUCTION

II REVIEW OF LITERATURE

III MATERIALS AND METHODS

IV RESULTS AND DISCUSSION

V SUMMARY AND CONCLUSION

LITERATURE CITED

APPENDIX
 DECLARATION

I, Miss. P.VINODHINI here by declare that the thesis entitled

“DEVELOPMENT OF BAKERY AND CONFECTIONERY PRODUCTS
USING GHEE RESIDUE” submitted to Acharya N.G.Ranga Agricultural
University for the Degree of MASTER OF FOOD SCIENCE AND
TECHNOLOGY is a result of original research work done by me. It is further
declared that the thesis or any part there of has not been published earlier in any
manner.

Date : (P. VINODHINI)

Place : Hyderabad
 LIST OF TABLES

Table Title Page No.

No.
1 Chemical composition of hand pressed ghee residue

2 Effect of method of preparation of on phospholipids

content of ghee residue

3 Composition of Phospholipids of different of ghee residue

4 Composition of ghee residue

5 Sensory evaluation- Mean scores of biscuits

6 Sensory evaluation-mean scores of chocolate toffees

7 Objective evaluation-compression and cutting strength of

biscuits

8 Objective evaluation-cutting strength of chocolate toffee

9 Nutrient composition of biscuits (Fresh)

10 Nutrient composition of chocolate toffee (Fresh)

11 Sensory evaluation-mean scores of biscuits (After one

month storage period)

12 Sensory evaluation-mean scores of chocolate toffee (After

one month storage period)

13 Objective evaluation-compression and cutting strengths of

biscuits (After one month storage period)

Table contd…
Table Title Page No.
No.
14 Objective evaluation-compression and cutting strengths of

chocolate toffee (After one month storage period)

15 Nutrient composition of biscuits (After one month storage

period)

16 Nutrient composition of chocolate toffees (After one month

storage period)
 LIST OF ILLUSTRATIONS

Figure No. Title Page No.

1. Schematic representation of the study

2. Mean sensory evaluation scores of biscuits

(Fresh)

3. Mean sensory evaluation scores of chocolate

toffee (Fresh)

4. Mean sensory evaluation scores of biscuits (After

one month storage period)

5. Mean sensory evaluation scores of chocolate

toffee (After one month storage period)

6. Moisture difference between fresh and stored

samples

7. FFA difference between fresh and stored samples

 LIST OF PLATES

Plate No. Title Page No.

1 Biscuit making – mixing flour into the creamed fat

and sugar

2 Biscuit making – baking of biscuits

3 A – Control biscuits

4 B – Biscuits with 10% replacement of fat with ghee

residue

5 C – Biscuits with 20% replacement of fat with ghee

residue

6 D – Biscuits with 30% replacement of fat with ghee

residue

7 A – Control chocolate toffee

8 B – Chocolate toffee with 10% replacement of fat

with ghee residue

9 C - Chocolate toffee with 20% replacement of fat

with ghee residue

10 D - Chocolate toffee with 30% replacement of fat

with ghee residue

11 Sensory evaluation of the products developed

12 Objective evaluation of the products developed

13 Nutrient analysis of the products developed

 LIST OF ABBREVIATIONS

% : Per cent
°C : Degree centigrade
μm : Micrometer
ANGRAU : Acharya N G Ranga Agricultural University
AOAC : Association of Official Agricultural Chemists
AOCS : American Oil Chemist Society
APDDCF : Andhra Pradesh Dairy Development Co-operative
Federation
BHA : Butylated hydroxy anisole
BHT : Butylated hydroxy toluene
CD : critical difference
FFA : Free fatty acids
GR : Ghee residue
Gr : Grams
HMF : 5-hydroxymethyl-2-furfural
Kg : Kilogram
Min : Minutes
N : Newton
NS : Non significant
PER : Protein efficiency ratio
rpm : Rotations per minute
SD : Significant difference
SMP : Skim milk powder
WVTR : Water Vapour Transmission Rate
 LIST OF APPENDICES

Appendix No. Title Page No.

I Method of preparation of Biscuits

II Method of preparation of Chocolate toffee

III Score card for sensory evaluation of biscuits

IV Score card for sensory evaluation of chocolate

toffee

V Estimation of moisture

VI Protein estimation

VII Estimation of Fat

VIII Estimation of Ash

IX Estimation of Acid Insoluble Ash

X Estimation of Free Fatty Acids

XI Crude fibre estimation

 ACKNOWLEDGEMENTS

I take this opportunity to express my heart full thanks and gratitude to

all those people who have helped immensely in the successful completion of my
research work and for making it a rich learning experience for me.

At the outset, I consider it a proud privilege to work under the able and
meticulous guidance of my Major Advisor Ms. Kanwaljit Kaur, Assistant
Professor, Foods an Nutrition Department, College of Home Science. I humbly
express my profound gratitude to her for being a source of inspiration,
constant encouragement, for her constructive criticism coupled with utmost
patience which has contributed immensely in making this modest endeavour a
reality.

I sincerely thank my Minor Advisors Dr. K. Kondal Reddy, Associate

Professor, Department of Livestock Products Technology, College of
Veterinary Science, Rajendranagar, Hyderabad and Dr. (Mrs.) S. Sumathi,
Associate Professor, Department of Foods and Nutrition, Post Graduate and
Research Centre, Rajendranagar, Hyderabad for their valuable suggestions,
able guidance and encouragement to complete my research work.

I am profoundly indebted to Dr. (Mrs.) V. Vimala, Programme

Director, Department of Foods and Nutrition, Post Graduate and Research
Centre, Rajendranagar, Hyderabad for providing me a milieu in the form of
necessary facilities conducive enough to my research work completion.

I deem it necessary to express my thanks to the staff of Post Graduate

and Research Centre, Rajendranagar and Department of Foods and Nutrition
and College of Home Science, Saifabad for their co-operation and helpful
suggestions during the entire period of my investigation.
 I would be failing in my duties if I don’t acknowledge the everlasting
love, unparalleled affection, constant encouragement and moral support given
to me by my ever loving parents Mr. P. Sanjeeva Reddy and Mrs. P. Shailaja
Reddy, brother Master P. Vishnu Vardhan Reddy

Diction is not enough to express my thanks to my would be

Mr. M. Shailender Reddy, for his moral support and unlimited love and
affection.
My grateful thanks to my friends, Uma, Deepti, Shilpa, Bharathi,
Surya, Bindu, Ranjitha, Padmaja, Srilakshmi, Srinivas, Narender, Vijaya
bhaskar, Krishna Mohan, Vijay, Rajanikanth, Vishnu for their moral support
and cooperation.
I also thank Mr. Prabhakar Rao, Quality Control Officer, APDDCF,
Vijaya Dairy, Hyderabad for providing me with ghee-residue for the
experiment.
I am thankful to Eeshwar Computers, Bhavani Colony, Rajendranagar
for the neat execution of the thesis.
My very special thanks to Baburao, Venkataiah, Yedukondalu, Ravi,
Zeenath for their help and coordination.
Finally, I thank all the people who directly or indirectly helped me to
complete my thesis successfully.

Date : ( P. VINODHINI )
 CHAPTER I

INTRODUCTION

Ghee residue is the by-product of ghee-manufacturing industry. During the

manufacture of ghee, the Solid-non-fat (SNF) present in the cream or butter

appears in the form of small particles known as ghee residue. It is obtained as a

moist, brownish sediment after molten ghee has been strained out. This is a source

of good quality protein and minerals. Nearly 27.5 per cent of total milk produced

in India is diverted for the production of ghee (Dairy India, 1997) and roughly

90,600 tonnes of ghee residue is annually produced in this country as a by-product

which is wasted.

In ghee production, the ghee-residue obtained at household level is

generally consumed by mixing it with cooked rice, by spreading over chapattis, by

mixing it with sugar or kneading into dough made for chapattis. However, in large

scale ghee manufacturing units, it is thrown away as an industrial waste because of

which there is a loss of appreciable quantity of nutrients in terms of fat, protein

and minerals.

Ghee is generally prepared by boiling cream or butter to a temperature of

110-115°C. The source i.e cream or butter used for manufacturing ghee can be of

different types :
 a) Desi Butter: Butter prepared after adding starter culture to cream or milk.

b) Creamery Butter: Butter made by direct churning of cream without adding

starter culture.

c) Sweet Cream : Fresh cream which is prepared or separated from milk with

the help of cream separator.

d) Sour Cream : Cream which has high acidity.

The yield of ghee residue depends upon the method of manufacturing ghee.

Santha and Narayana (1978) and Verma (1978) studied the yield of ghee residue

and found that ripening of cream prior to clarification of cream reduced the yield

of ghee- residue.

Chemical composition of ghee residue also varies depending upon the

source of preparation.

Table 1: Chemical composition of hand pressed ghee residue was given by

Verma (1997)

AVERAGE per FAT ASH

SOURCE OF MOISTURE PROTEIN LACTOSE
cent FAT OF per per
GHEE RESIDUE per cent per cent per cent
SOURCE cent cent
Desi Butter 77 13.4 33.4 32.8 15.2 5.2

Creamery 85 5.7 65 25.5 Traces 3.8

Butter
67 4.1 60.2 18 12.3 2.4
Sweet cream
67 8 38.8 41.6 7.3 4.3
Sour cream
 Ghee residue is a nutrient dense product contributing to high amount of

proteins, fat, carbohydrates and minerals. In addition to contributing to nutrients,

ghee residue is also a rich source of anti-oxidants. Santha and Narayana (1979)

stated that the anti-oxidant property of ghee residue is not due to one constituent

alone, but due to many constituents such as phospholipids, amino acids, free

sulphydryls, free sugars from lactose and their interaction products with protein

and phospholipids etc and they reported that out of all the phospholipid fractions,

cephalin has the greatest anti-oxidant activity.

Ghee residue can also be used as a flavoring agent as it is rich in chief

flavoring compounds including free fatty acids (FFA), carbonyls and lactones.

Galhotra and Wadhwa (1991) studied the levels of FFA, carbonyls and lactones

and found the levels as 11, 10 and 132 times higher than that of ghee respectively.

Keeping in view the nutritive value, anti-oxidant property and flavour

potential of ghee residue and also to reduce industrial waste and utilize the by-

product in a better way, there is a considerable need for the utilization of ghee

residue for product development as there is increasing demand for new products

dense in nutrients at affordable price. Therefore, the present study was aimed to

utilize ghee residue for development of bakery and confectionery products with

good nutritive value at less cost and help in the utilization of this industrial waste.

Therefore, the present study is taken up to develop biscuits and chocolate

toffees by partial replacement of fat with ghee residue at different levels of

replacement i.e. 10 per cent, 20 per cent and 30 per cent. Biscuits and chocolate
toffees are amongst the popular and the most commonly processed foods with

certain advantages like, they are highly nutritious products at affordable prices,

has good shelf life, available in different packs and sizes because of which it is

easy to use during travel or at home. Biscuits are amongst the lowest cost

processed foods and toffees on the other hand are on the popular list of all age

group people. So the present study is planned to develop products like biscuits and

chocolate toffees by partial replacement of fat with ghee residue.

Ghee residue is an industrial waste, which is left unused adding to the day

to day increasing garbage. Ghee residue being a valuable source of nutrient,

exhibiting excellent antioxidant property, and also contributing to the flavour

components there is a considerable need for the utilization of ghee residue for

product development to utilize the waste more fruitfully to develop a product

which is not only nutritive but within the affordable price.

The formula developed for the product can be commercialized by food

industries and add to the existing list of health foods, since this is the demand of

the present day. Earlier attempts were made to utilize ghee residue for

development of some bakery and confectionery products at laboratory level, but it

unfortunately never went into the commercial arena. Hence a study has been

undertaken for utilizing ghee residue for product development with the following

objectives:
 „ To develop biscuits and toffees with partial replacement of fat with ghee

residue.

„ To study the product quality, acceptability, proximate principles and free

fatty acid levels of the developed product and of ghee residue.

„ To study the keeping quality of the developed product.

This study opens new areas of research as ghee residue by virtue of its

chemical composition, physical characteristics and good shelf-life because of the

presence of anti-oxidants has great potential and is more amenable to exploit its

utilization in bakery and confectionery products, which would solve the problem

of disposal and reduce their cost of production and increase the nutritional quality

of the product.
 CHAPTER II

REVIEW OF LITERATURE

A comprehensive review of literature is essential for any good research

endeavour as it provides background information to aid the investigator in

designing and analyzing research work. Literature directly or indirectly related to

the development of food products using ghee residue has been examined and

presented below under following broad headings:

2.1 Nutritive value of ghee residue

2.2 Processing, yield, treatment and storage of ghee residue

2.3 Antioxidant property of ghee residue and its browning compounds.

2.4 Flavour potential of ghee residue

2.5 Utilization of ghee residue for product development

2.1 NUTRITIVE VALUE OF GHEE RESIDUE

Santha and Narayanan (1978) studied the composition of ghee residue

obtained from different sources and found that the composition from different

sources was, fat content ranging between 32-70 per cent, protein 12-13.9 per cent,

moisture 8-30 per cent, lactose 2-14 per cent and ash 1-8 per cent.
 Relwani (1978) studied the detailed composition of ghee residue and

found that the product is rich in good quality milk proteins and nitrogenous

compounds and so it can be used as human dietary supplement.

Santha and Narayanan (1978) studied the fatty acid composition of

phospholipids present in ghee residue and revealed that it has no fatty acids lower

than 12 carbon atoms and also found the values of total saturated fatty acids and

total unsaturated fatty acids to be 58.7 per cent and 41.3 per cent respectively.

Grewal (1979) studied the protein efficiency ratio (PER) of ghee residue

and found that it was very low. So, he concluded that the low value of PER is due

to the damage of some essential amino acids, especially lysine.

Kapoor and Pal (1979) studied the PER and casein values of ghee residue

and found it to be 2.55 and 2.62 respectively. He also concluded that GR can be a

good source of animal proteins for the vegetarian population of our country.

Malhotra (1980) studied and compared the nutritive value of ghee residue

and skim milk powder (SMP) and found that the nutritive value of ghee residue is

low as compared to SMP, but he also found that by supplementing ghee residue

with a combination of lysine (8 per cent), methionine (2.5 per cent) and trytophane

(1.4 per cent) increases its nutritive value even slightly higher than that of SMP.

A study conducted by Arumugham et al. (1989) to arrive at the nutritive

value of GR, revealed that GR is a good source of protein - 43.7 gross protein

value, ash 8.98 per cent, 0.88 per cent calcium, 0.5 per cent phosphorus, 14.4 per

cent nitrogen free extract with no crude fibre.

 Verma (1991) gave the chemical composition of ghee residue made by

different processes and found that ghee residue of creamery butter source had high

fat (65 per cent), whereas GR of sour cream had high protein (33 per cent), and

GR of desi butter had high lactose (15.5 per cent) and high ash (5.2 per cent).

2.2 PROCESSING, YIELD, TREATMENT AND STORAGE OF GHEE

RESIDUE

2.1.1 Yield

A study conducted by Santha and Narayanan (1978) on the yield of GR

with different sources revealed that there was a maximum yield in the case of

direct cream method -12 per cent and yield of both creamery butter and desi butter

almost gave the similar yield of around 3.7 per cent.

Verma (1997) also studied the yield of GR with different sources of buffalo

milk and found that the yields of GR with sweet cream, sour cream, washed sweet

cream, desi butter and creamery butter was 7.7 per cent, 5.1 per cent, 3.5 per cent,

1.6 per cent and 1.2 per cent respectively.

2.2.2 Treatment and Processing

Ghee residue has soft and smooth texture but gets progressively hardened

during storage. The change in the textural characteristics is much faster

particularly during the first 15 days and by the end of the month the grain becomes

very hard and gritty. In order to minimize the undesirable characteristics, it is

necessary to process GR so as to yield a soft and smooth texture essential for

edible preparations.

Prahlad (1954) studied and suggested a number of treatments of GR to

make it soft and smooth.

Treatment 1: Loosely tying the residue in the form of bundle and cooking

in boiling water 30 minutes.

Treatment 2: Cooking the residue in boiling solution of 1.0 per cent

sodium bicarbonate for 30 minutes.

Treatment 3: Washing the residue with 59 per cent alcohol and then

cooking in boiling water for 30 minutes.

Treatment 4: Washing the residue with 50 per cent alcohol and then boiling

in 1 per cent solution of Sodium bi- carbonate for 30 minutes.

Treatment 5: Autoclaving the residue from Treatment (3) at 15 lb / square

inch / 10 minutes after incorporating 2 per cent vinegar.

Treatment 6: Autoclaving the residue obtained from Treatment (4) after

incorporating 2 per cent vinegar.

Treatment No-4 was found to be desirable where residue was boiled with 1

per cent solution of sodium-bi-carbonate for 30 minutes after washing it with 50

per cent alcohol was found to be the best to remove excess fat from the GR and

improve the texture.

2.1.2 Recovery and Storage of GR

 Viswanathan et al. developed 2 methods for recovery of ghee from GR:

a) Centrifugal Process:

This consists of heating GR in water at 65° C so as to transfer the occluded

ghee in the residue water. Ghee is subsequently recovered by centrifuging the

water- fat phase. The method yields 25 per cent ghee and the efficiency of the

process is 46 per cent.

b) Pressure Technique:

This consists of subjecting the heated GR (65-70°C) to a limited pressure in

a hand screw or hydraulic press. The method yields 45 per cent of ghee and the

efficiency of the process is 67 per cent.

Subbulakshmi (1990) studied the shelf life of GR stored for a period of 90

days in plastic, glass and tin containers. It showed no significant changes in FFA

content, peroxide value and tintometer readings. Storability was better in glass and

tin containers as compared to plastic ones.

2.3 ANTI-OXIDANT PROPERTY OF GHEE RESIDUE AND
BROWNING COMPOUNDS

Kirigaya et al. (1968) reported that the anti-oxidative activity increased in

proportion to the colour intensity of the browning reaction product, whereas, the

reductones formed during the browning process contributed little to the activity.

Consequently, it was assumed that melanoidin pigment play an important role in

the anti-oxidative activity.

Kirigaya et al. (1971) observed that the browning compounds obtained by

condensation of amino acids and sugars have potent antioxidant properties, among

them the low molecular weight fractions have stronger anti- oxidative effect than

the high molecular weight fractions.

Pruthi et al. (1973) studied the anti-oxidant property of phospholipids

present in GR and described a heat processing method for the extraction and

fortification of ghee with ghee residue phospholipids. He found that the oxidative

stability of ghee could be increased by increasing its phospholipids content

through heat treatment of ghee residue with ghee or by the addition of solvent

extracted phospholipids.

A study conducted by Santha and Narayanan (1977) on the antioxidant

property of GR revealed that the anti-oxidant property of GR depends upon the

method of preparation of ghee. The anti-oxidant property of GR made by

creamery butter method was maximum followed by desi butter and sweet cream.
Effect of method of preparation on phospholipids content of ghee residue is given

in Table 2.

Table 2: Effect of method of preparation on Phospholipids content of ghee

residue

Phospholipids Phospholipids content

Type of ghee residue (per cent) (per cent)
(Mean ± S.D) (Mean ± S.D)
Unripened cream GR 0.92± 0.21∗ 1.57 ±0.40∗

Ripened cream GR 0.62± 0.26∗ 1.10± 0.47∗

Creamery Butter GR 6.27 ±2.60 17.39± 6.81

Desi Butter GR 1.78 ±0.56 4.95 ±1.85

∗ Statistically significant (P<0.05)

A study conducted by Santha and Narayanan (1978) revealed that the

antioxidant property of GR is due to the presence of phospholipids, amino acid

profile and other non-lipid constituents present in the ghee residue.

Santha and Narayanan (1978) studied the anti-oxidant property of GR and

concluded that the anti-oxidant property was mainly due to the phospholipids

followed by α-tocopherol and Vitamin A, among the lipid constituents. And

among the non-lipid constituents, the amino acids proline, lysine, cysteine

hydrochloride and tryptophan showed anti-oxidant property. He also gave the

composition of phospholipids of different types of ghee residue which is presented

in Table 3.
Table 3: Composition of Phospholipids of different sources of Ghee residue.

Source of Phosphatidyl Phosphatidyl Phosphatidyl Sphingomyelin Phosphatidyl LPC +

phospholipids Choline Ethanolamine Serine Inositol LPE ∗
(Wt per cent) (Wt per cent) (Wt per cent) (Wt per cent) (Wt per cent) (Wt per cent)
5.6
Unripened 28.7 23.3 4.5 29.1 4.8
Cream GR
4.2
Ripened 28.6 24.3 4.8 29.3 5.1
Cream GR
6.2
Creamery- 28.2 23.0 4.1 30.0 5.1
butter GR
6.8
Desi-butter 28.6 20.4 4.0 29.5 5.0
GR
1.6
Butter-serum 29.3 26.9 5.0 31.3 5.9

∗ - LPC: Lysophosphatidyl Choline ; LPE: Lysophosphatidyl ethanolamine.

Santha and Narayanan (1978) found that the anti-oxidant efficiency of GR

decreases with increase in the temperature of clarification and also found that,

among all the phospholipids fraction, cephalin has the greatest anti-oxidant

property.

Santha and Narayanan(1979) studied the anti-oxidant property and found

that the anti-oxidant property of GR is not due to one constituent alone, but due to

many constituents such as phospholipids, amino-acids, free sulphydryls, free

sugars from lactose and their interaction products with proteins and phospholipids.

El-Zeany and El-Khateeb (1982) studied the anti-oxidant activity of fat-

soluble and fat-insoluble brown pigments formed during browning of fatty foods

during storage. Results showed that the fat-soluble pigments revealed no anti-

oxidant properties whereas fat-insoluble pigments showed some anti-oxidant

activity. Their antioxidant activity was attributed to the presence of metal

inactivating groups and some other functional groups.

Pagote and Bhandhari (1988) incorporated 2 per cent ghee residue in ghee

and stored at 80°C and found that the development of peroxides in ghee samples

was slower than in control. So, they concluded that GR contained certain anti-

oxygenic substances.

Sripad (1988) estimated the amount of 5-hydroxymethyl-2-furfural (HMF)

which is a major browning compound present in ghee residue and having anti-

oxidant property and found that the HMF content of ghee residues obtained during

clarification of ghee at 110°C and 120°C were 113.93 and 460.64 μm per gram

respectively. He concluded that, as the temperature of clarification increased the

HMF values also increased.

Wadhwa et al. (1991) compared the anti-oxidant property of butylated

hydroxy anisole (BHA), butylated hydroxy toluene (BHT) and ghee residue and

found that their anti-oxidant property was parallel to each other. So they

concluded that GR can be used as a source of natural anti-oxidants for improving

the shelf-life of food products including dairy products, where use of synthetic

antioxidants is generally not preferred because of their toxic effects.

2.4 FLAVOUR POTENTIAL OF GHEE RESIDUE

Galhotra (1991) extracted ghee residue lipids by using methanol and found

the levels of FFA and carbonyls to be high in ghee residue than in ghee indicating
that the GR is a rich source of flavour compounds. He also observed that the gas

chromatograms of lactonic isolates of GR showed a complex profile of 47 peaks.

The homologous series of n-saturated delta and gamma lactones from C6-C16 and

C18 have been characterized in GR. Delta lactones compared to gamma lactones

are the major components and among lactones Delta C12, C14 and C18 were the

major lactones in GR.

2.5 UTILIZATION OF GHEE RESIDUE FOR PRODUCT

DEVELOPMENT

Incorporation of ghee residue in product development opens new areas of

research as GR is a good source of nutrients, anti-oxidants and contributes to a

good flavour. Incorporation in product not only reduces disposal problem and

increase by-product utilization productivity, but also reduces the cost of the

product. Literature related to the utilization of GR for product development has

been broadly classified into following sub-headings:

2.5.1 Ghee residue as a source of natural anti-oxidant

Sripad (1988) extracted browning compounds with ethyl alcohol from GR

and incorporated to ghee clarified at 110°C and 120°C at 0.4 to 1.0 per cent level

and stored at 50°C and 37°C to study the influence of these compounds on the

autoxidative stability and keeping quality of ghee. He found that the keeping

quality of ghee was enhanced by more than 2 months in case of cow ghee and
more than 1.5 months in case of buffalo ghee prepared at 110°C and concluded

that the antioxidant property of browning compounds in GR is effective in low

temperature clarified ghee.

Wadhwa et al. (1991) conducted a study on flavoured butter oil by

incorporating GR at the rate of 15-20 per cent followed by heating to 120°C / 3

min and found that the shelf life of flavoured butter oil was improved and it was

almost parallel to that as BHA and BHT. So they concluded that GR is a rich

source of natural anti-oxidants.

Ramamurthy et al. (1996) added ghee residue at 1.5 per cent level,

containing 6.012 per cent of phospholipids, to the ghee samples prepared from

cow and buffalo butter heated at 120°C. Cow ghee was more resistant than buffalo

ghee for oxidation. The added ghee residue enhanced the keeping quality in both

the cases. The increase in keeping quality was attributed to the phospholipids

present in ghee residue.

2.5.2 Ghee residue as a source of flavour concentrates

Wadhwa and Bindal (1995) utilized GR for flavouring bland products like

vanaspathi, butteroil etc. they obtained the best flavoured vegetable fat and butter-

oil by heating them with 10 per cent GR along with 20 per cent water to

120°C/flash followed by filtration through four folds muslin cloth and

centrifugation at 3000 rpm for 10 minutes. The products prepared had a score of 8,

thus eliminating the use of synthetic flavour.

2.5.2 Ghee residue in the development of confections

The physico-chemical properties of processed ghee residue are very

suitable for preparation of confections. It contains the major constituents in

suitable proportion and possesses fine texture that imparts requisite body to such

products. Further the treatment during processing of these confections involves

heating to such an extent that it completely arrests enzymatic activity and flavour

deterioration in the final product. The higher fat content in the residue quite often

obviates the need for addition of oils and fats in its preparation.

Wadhwa (1997) developed candies by replacing fat with GR. The candy

recipe he used for preparation of candy was GR-1kg, sugar 500-625gr, dry

coconut powder 125-250 gr. He prepared sugar syrup (50 per cent) and mixed GR

thoroughly into it with a ladle and heated simultaneously till it became sticky, then

added coconut powder and evenly spread on a plate and cooled to 5-10°C and cut

into cubes and wrapped in parchment paper. The product prepared gave good

acceptability score.

2.5.4 Ghee residue in the development of edible pastes

Prahlad (1954) prepared edible pastes for sandwiches by mixing processed

GR with salt at the rate of 2.5-3 per cent and marmite (a yeast product) at the rate
of 0.1-0.5 per cent heating on a low flame for about 5 minutes till it forms paste.

The developed product had good scores and had a shelf life of 2 months under

proper packaging.

2.5.5 Ghee residue in the development of burfi-type sweet

Verma and De (1978) prepared a burfi-type sweet called chocosidue burfi

from GR which was processed earlier in 0.5 per cent sodium carinate solution for

30 minutes. They prepared chocosidue burfi by using GR-425 grams, khoa-200

grams, chocolate-20 grams and sugar 170 grams and found that the sensory

evaluation scores were equivalent to that of the control burfi.

2.5.6 Ghee residue in the development of Bakery Products

Subbulakshmi (1990) developed cakes, biscuits and porridges by replacing

fat with GR and found that the cakes prepared by 100 per cent replacement of

butter with GR had acceptable scores and partial replacement of GR in biscuits,

porridges improved flavour and acceptability scores.

Borawake and Bhosale (1996) developed nankatai type cookies and sponge

cakes by partial replacement of fat with GR without affecting their physico-

chemical and sensory properties and found that hydrogenated vanaspati fat could

be satisfactorily replaced up to 30 per cent in nankhatai type cookies and up to 20

per cent in sponge cakes without adversely affecting the sensory qualities.
 CHAPTER III

MATERIALS AND METHODS

The present study was carried out to find the effect of partial

replacement of fat with ghee-residue. A detailed description of the procedure

followed for conducting the experiment is given in this chapter.

3.1 Procurement of raw materials

3.2 Standardization of the recipe and product development

3.3 Acceptability of the products by sensory and objective evaluation

3.4 Nutrient analysis

3.5 Statistical analysis

3.1 PROCUREMENT OF RAW MATERIAL

Ghee residue: Ghee residue was procured from Andhra Pradesh Dairy

Development Co-operative Federation (APDDCF) Vijaya dairy, Lalapet,

Hyderabad, where ghee is prepared by creamery butter method. Firstly, cream

is churned to convert it to Butter and then it is heated to 110-115°C to produce

ghee. The ghee residue which is left to settle down, the supernatant ghee is

recovered and the sediment i.e ghee residue is treated or processed in order to

recover as much ghee as possible from the residue by washing it with hot water

and then with chilled water. The washed water is kept aside so that the ghee

separates and forms a layer on the top which can be easily removed and the
ghee residue which settles down is thrown away as waste. The ghee residue,

was collected from the dairy and stored in an air tight container for the use of

experiment.

Proximate principles and FFA levels were estimated for ghee residue

before using for product development.

• Protein was determined according to the Microkjeldahl method (AOAC,

1990).

• Fat content was estimated with the Soxhlet apparatus (AOAC,1990).

• Moisture was estimated by oven drying method (AOAC, 1990).

• FFA level of the extracted fat was estimated by (AOCS, 1954) method.

The description of the procedures followed for the estimation of

proximate principles and FFA content are given in the Appendices V to X.

Other raw materials like sugar, vanaspati, whole wheat flour, skim milk

powder, cocoa powder were procured from the local market and care was taken

to purchase branded products to produce consistent product quality.

3.2 STANDARDIZATION OF THE RECIPE AND PRODUCT

DEVELOPMENT

Two commonly consumed bakery and confectionery products i.e

biscuits and chocolate toffee were selected for product development and

standardized.
 Standardization of the Biscuits: Many trails were carried out to

standardize the recipe for biscuits using different types of raw materials and

preparation methods. Biscuits were developed using both whole wheat flour as

well as refined wheat flour and evaluated for sensory attributes like texture,

colour, acceptability, taste etc. Biscuits developed using whole wheat flour

gave better sensory scores compared to refined wheat flour. Detailed

description of the recipe and the method of preparation of biscuit selected for

the study is given in Appendix- I.

Standardization of Chocolate Toffee: Different methods, recipes and

also raw material were used to develop best quality products. As for biscuits,

the toffee also was evaluated for sensory attributed like appearance, texture,

taste and acceptability and the method and recipe, and which gave the best

quality product was finalized for the study. Detailed description of the recipe

and the method of preparation of chocolate toffee selected for the study is

given in Appendix- II.

Both the products that were standardized, evaluated and assessed for

organoleptic qualities were again put for sensory evaluation before the selected

panel and finalized the recipe to be used for the study.

Product Development: Control and experimental samples of biscuits

and chocolate toffees were prepared in College of Home Science, Foods and

Nutrition Department, Hyderabad using the standardized recipe. After

preparation biscuits were wrapped in butter paper and then packed in

polyethylene pouches and sealed. Chocolate toffees were wrapped in

aluminium foil and then packed in polyethylene pouches and sealed. The

packed products were labelled with the relevant specifications of the type, the

level of replacement of fat with ghee residue and its date of manufacture. A

part of sample was utilized for the analysis, subjective and objective evaluation

and the other part was stored in a place, which is free from insects and rodants

for studying the storage stability.

3.3 ACCEPTABILITY OF THE PRODUCTS BY SENSORY AND

OBJECTIVE EVALUATION

3.3.1 Objective evaluation

Schimadzu E2 Tester, a universal testing machine available at College

of Home Science, Foods and Nutrition Department, Hyderabad was used to

evaluate the physical properties of the products prepared. The cutting strengths

and the compression strengths were determined for biscuits and cutting strength

was determined for chocolate toffee. The products were evaluated when fresh

and also after one month of storage period.

3.3.2 Subjective evaluation

Development of scorecard: The score card was prepared keeping in

view the quality characteristics of the products. Descriptive terms were given to

various quality attributes like taste, flavour, texture, colour and overall

acceptability and numerical scores were assigned. A five point Hedonic scale

was adopted to score each of the attribute. While framing score card highest
score (5) was assigned to most preferred characteristic and least score (1) to

undesired characteristic in the product.

Selection of panel member: To select the panel members a section of

the staff and students of College of Home Science and Post Graduate and

Research Centre, Foods and Nutrition Department were selected as panel

members. Among them screening was carried out and ten members who

showed high sensitivity and consistent performance were selected as panel

members. The score card developed was used for evaluation of the products.

Product Evaluation: The selected panel of judges were used in the

evaluation of the developed products. The evaluation was carried out at Post

Graduate and Research Centre, Rajendra nagar, ANGRAU, Hyderabad on two

different days for biscuits and for chocolate toffee. The panel members were

presented with:

1) Four samples of biscuits (Control, 10%, 20%, 30% replacement of fat

with GR).

2) Four samples of chocolate toffees (Control, 10%, 20%, 30%

replacement of fat with GR).

Samples were numbered as I, II, III and IV without giving detailed

identification of the products to get accurate results. All the products were

evaluated between 10 AM and 12 noon or between 2 PM and 4 PM when the

panelists were not hungry, to rule off the human errors. The results were taken

in duplicates and the average of the readings was taken as final score. The
products were evaluated when fresh for acceptability using score card and also

after one month of storage period to record the changes during storage.

3.4 NUTRIENT ANALYSIS

Nutrient analysis was conducted for the products developed. The

nutrients analysis was carried out at Post Graduate and Research Centre,

ANGRAU, Hyderabad,. when the samples were fresh and also after storage

period of one month.

• Protein was determined according to the Microkjeldahl method

(AOAC, 1990).

• Fat content was estimated with the Soxhlet apparatus (AOAC, 1990).

• Moisture was estimated by oven drying method (AOAC, 1990).

•Ash content was estimated by using (AOAC, 1990).

• FFA level of extracted fat was estimated by AOCS (1954) method.

•Crude fibre was determined by (AOAC,1984).

The detailed description of the procedures followed for the estimation of

proximate principles and FFA content of the extracted fat are given in the

Appendices V to XI

3.5 STATISTICAL ANALYSIS

Statistical analysis was carried out by the procedures laid down by

Snedecor and Cocharn (1989). The data was tabulated and subjected to
statistical analysis at the end of the study for mean, standard deviation and

significance.
Plate No 7: Nutrient Analysis of the product development
Plate No 5: Objevtive analysis of the products

Plate No 6: Sensory evaluation of the products

Plate No 1: Biscuit making-Mixing of flour into the creamed fat and
sugar.

Plate No 2: Biscuit making-Baking of Biscuits

Fig No.
Fig. No.
 CHAPTER IV

RESULTS

The results pertaining to the study on partial replacement of fat with

ghee residue in biscuits and chocolate toffee developed are sequentially

arranged and presented under the following headings:

4.1 Estimation of proximate principles and FFA level of the extracted fat of

ghee residue

4.2 Sensory evaluation of the products

4.3 Estimation of proximate principles and FFA level of the extracted fat of

the products

4.4 Storage stability of the products

4.1 PROXIMATE ANALYSIS AND ESTIMATION OF FFA LEVEL

OF THE EXTRACTED FAT OF GHEE RESIDUE

Table 4: Composition of ghee residue

Constituents of ghee residue Percentage

Moisture 45.7

Protein 12.8

Fat 15.5

Ash 0.96

Acid insoluble ash 0.39

FFA content of the extracted fat. 1.25

 The proximate principles and the FFA content of the extracted fat of

ghee residue are presented in Table 4.

4.2 SENSORY EVALUATION OF THE PRODUCTS

4.2.1 Subjective evaluation of the products

4.2.1.1 Biscuit:

The mean scores of sensory evaluation of biscuit samples when fresh are

presented in Table 5.

Table 5: Sensory evaluation – mean scores of biscuits

Variations Replacement Appearance Texture Taste Flavour Overall

level (%) acceptability
Control - 5.00±0.00 4.93± 0.28 4.81±0.28 4.88± 0.32 4.98± 0.18

10% 4.82± 0.46 4.91± 0.28 4.85± 0.35 4.89± 0.48 4.95± 0.25
Replaceme
nt level of 20% 4.60± 0.63 4.88± 0.42 4.82± 0.46 4.91± 0.68 4.92± 0.54
fat with
ghee 30% 4.46± 0.64 4.85 ±0.65 4.86± 0.61 4.95± 0.61 4.91± 0.61
residue

From the table it is clear that the control sample scored high scores for

appearance and texture, but the overall acceptability score of the 10 per cent

replacement was very close to the control i.e 4.95 and 4.98 respectively and the

difference also was not statistically significant.

4.1.1.2 Chocolate toffee:

The mean scores of the sensory evaluation of freshly prepared chocolate

toffee are presented in Table 6.

Table 6: Sensory evaluation – mean scores of chocolate toffee

Variations Replacement Appearance Texture Taste Flavour Overall

level (%) acceptability
Control - 5.00± 0.00 4.88± 0.32 4.72 ±0.28 4.89± 0.61 4.88± 0.18

Replacement 10 4.93± 0.25 4.81± 0.54 4.68± 4.51 4.91± 0.28 4.85± 0.29
level of fat
with ghee 20 4.89± 0.48 4.72± 0.35 4.61± 0.32 4.93± 0.54 4.83± 0.81
residue
30 4.83± 0.28 4.71 ±0.58 4.60 ±0.81 4.95± 0.68 4.82± 0.56

The mean sensory evaluation scores for chocolate toffee presented in

Table 6 showed that the overall acceptability of 10 per cent replacement in

chocolate toffee was very close to that of control i.e 4.85 and 4.88 respectively

and the difference was not statistically significant.

4.2.2 Objective evaluation of the products

The Schimadzu E2 tester, a universal testing machine was used to

evaluate the physical characteristics i.e. compression strength and cutting

strength, where jigs suited for the test were selected from a wide varieties of

jigs separately available and then mounting them to the machine.

4.2.2.1 Biscuit:

Table 7 presents the compression and cutting strength of fresh biscuits

which was determined.

Table 7: Objective evaluation- compression and cutting strength of

biscuits

Variation Replacement Compression Cutting strength

level (%) Test (Newton)
(Newton)

Control - 15.71 5.82

Replacement 10 15.75 5.83

level of fat with
ghee residue 20 15.76 5.88

30 15.89 6.11

The results of the objective evaluation showed that as the level of

replacement of ghee residue increased in biscuits, compression and cutting

strengths increased as compared to the control sample. Control sample had low

values for cutting and compression test (5.82 N and 15.71 N), whereas the

samples with 30 per cent replacement had high values (6.11 N and 15.89 N).

2.2.2 Chocolate toffee

Table 8 gives the cutting strengths of the control and experimental samples of

chocolate toffees.
Table 8: Objective evaluation – cutting strength of chocolate toffee

Variations Replacement level (%) Cutting test (Newton)

Control - 3.72

10 3.42
Replacement level of
fat with ghee residue 20 2.35

30 2.25

The results of the cutting test for chocolate toffee shown (Table 8)

showed that the cutting strength decreased as the replacement level increased.

Cutting strength value is highest for the control and lowest for 30 per cent

incorporation i.e. 3.72 Newton and 2.25 Newton respectively.

4.3 PROXIMATE ANALYSIS AND ESTIMATION OF FFA

CONTENT OF THE PRODUCTS

4.3.1 Biscuit

Nutrient composition and FFA content of the biscuits are presented in

Table 9.
Table 9: Nutrient composition of biscuits (Fresh)

Variation Replacement Moisture Protein Fat Ash Acid Crude FFA

level (%) % % % % insoluble fibre %
ash % %
Control 2.45 5.25 28.715 0.865 0.002 0.19 0.50
- (0.014) (0.014) (0.004) (0.007) (0.001) (0.014) (0.021)

2.68 7.00 25.45 1.015 0.006 0.20 0.60

10 (0.014) (1.414) (0.009) (0.001) (0.00) (0.014) (0.014)
Replacement
level of fat 2.83 8.745 24.765 1.027 0.006 0.175 0.62
with ghee 20 (0.014) (0.008) (0.009) (0.002) (0.00) (0.007) (0.007)
residue
2.95 9.62 22.37 1.1280 0.006 0.1950 0.65
30 (0.014) (0.014) (0.010) (0.001) (0.00) (0.007) (0.042)

Critical Difference (0.05) 0.0394 1.9631 1.1799 0.010 NS NS 0.020

NS- Non Significant

Table 9 shows increase in moisture, protein, ash, acid insoluble ash and

FFA as the level of incorporation increased from 10-30 per cent except for fat

which is decreased as the level of replacement increased.

The protein value is high in the 30 per cent replacement. Fat content of

the control sample was 28.72 per cent and that of experimental products ranged

from 22.3-25.5 per cent. There was a significant difference in the content of

protein and fat i.e C.D value was 1.9631 and 1.1799 respectively.

4.3.2 Chocolate Toffee

Nutrient composition of control and ghee residue replaced samples were

determined and presented in the Table 10.

Table 1O: Nutrient composition of chocolate toffee

Variations Replacement Moisture Protein Fat Ash Acid FFA

Level (%) % % % % insoluble %
ash %
15.95 2.64 12.2095 0.515 0.002 0.47
Control - (0.014) (0.014) (0.014) (0.007) (0.00) (0.028)

16.55 3.50 12.133 0.57 0.003 0.50

10 (0.017) (0.0141) (0.976) (0.028) (0.00) (0.012)
Replacement
level of fat 16.65 4.38 11.3345 0.55 0.003 0.52
with ghee 20 (0.015) (0.014) (0.111) (0.007) (0.00) (0.014)
residue
17.05 5.25 11.07 0.62 0.004 0.52
30 (0.16) (0.057) (0.015) (0.014) (0.00) (0.028)

Critical Difference(0.05) 0.0376 0.2132 0.0907 0.0460 NS 0.010

NS- Non Significant

From the Table10 the nutrient analysis showed that the moisture and ash

content of the control was 15.939 per cent and 0.51 per cent respectively, and

there was a slight increase in the moisture and ash content as the level of

replacement increased with a critical difference of 0.0376 and 0.0460

respectively. Protein content was highest in 30 per cent replacement and lowest

in the control i.e. 5.25 per cent and 2.64 per cent respectively. The fat content

was the high for the control and low for 30 per cent replacement sample with a

critical difference of 0.0907.

There was no marked difference in the levels of acid insoluble ash of

control as well as experimental. The FFA content of the control was 0.48 per
cent and that of experimental samples ranged from 0.50-0.52 per cent with

different level of replacement.

4.4 STORAGE STABILITY OF THE PRODUCT

Sensory evaluation i.e. objective and subjective evaluation and nutrient

analysis was conducted after a period of one month to test the storage stability.

4.4.1 Sensory evaluation

4.4.1.1 Subjective evaluation:

Mean sensory evaluation scores of the biscuits after one month storage

period are presented in Table 11.

Table 11: Sensory evaluation- mean scores of biscuits (After one month
storage period)

Variation Replacement Appearance Texture Taste Flavour Overall

Level % acceptability

Control - 5.00 ±0.00 4.88± 0.28 4.80 ±0.81 4.88 ±0.18 4.91± 0.71

Replace- 10 4.83± 0.51 4.78± 0.41 4.80± 0.61 4.79± 0.81 4.88± 0.25
ment level
of fat with
ghee 20 4.63 ±0.43 4.61± 0.61 4.81± 0.83 4.86± 0.68 4.87 ±0.35
residue

30 4.51± 0.68 4.60 ±0.78 4.83± 0.23 4.91± 0.72 4.81± 0.21

The results of the sensory evaluation of the biscuits after storage of one

month showed that there was a slight deteriotion in the texture, flavour, taste
and over all acceptability attributes, but the difference was not statistically

significant.

Mean sensory evaluation scores of he chocolate toffee after one month

of storage period are presented in Table 12.

Table 12: Sensory evaluation – mean scores of chocolate toffee (After one
month storage period)

Variation Replacement Appearance Texture Taste Flavour Over all

Level % Acceptability

Control - 5.00 ±0.00 4.81± 0.18 4.71 ±0.28 4.81±0.91 4.81± 0.61

Replacement 10 4.89± 0.18 4.71± 0.21 4.67± 0.31 4.83±0.27 4.79± 0.59
level of fat with
ghee residue
20 4.88± 0.71 4.68± 0.79 4.66±0.16 4.89±0.11 4.77± 0.88

30 4.81± 0.68 4.65± 0.56 4.56 ±0.20 4.90±1.83 4.71± 0.11

The overall acceptability scores were not much affected at any level of

replacement even after one month storage period and were desirable on 5- point

hedonic scale, and the difference was not statistically significant.

4.4.1.2 Objective evaluation after one month storage period:

Compression and cutting strengths of the biscuits and chocolate toffees

are given in the Table 13 and Table 14 respectively.

Table 13: Objective evaluation – mean scores of biscuits (After one month
of storage period)

Variations Replacement Compression test Cutting test

level % (Newton) (Newton)
- 15.64 5.79
Control
15.65 5.81
Replacement 10
level of fat with 15.68 5.85
ghee residue 20
15.77 6.10
30

The results of the compression and cutting test after one month of

storage period (Table 13) showed that there was no marked difference in the

compression and cutting strengths when compared to the fresh samples (Table

7) even after storage period of one month.

Table 14: Objective evaluation- mean scores of chocolate toffee (After

one month of storage period)

Variation Replacement level Cutting test

% (Newton)

Control - 3.713

10 3.421
Replacement level of fat
with ghee residue 20 2.30∗

30 2.23∗
∗ - Statistically significant
 The results of the cutting test for chocolate toffee (Table 14) showed

that there was no significant difference between the cutting strengths of the

fresh samples (Table 8) and samples stored for one month.

4.4.2 Nutrient analysis after one month storage period

Nutrient analysis was conducted for biscuits and chocolate toffee after

one month storage period and are presented in the tables 15 and 16.

Table 15: Nutrient composition of biscuits after one month storage

Variation Replacement Moisture Protein Fat Ash Acid Crude FFA

level (%) % % % % insoluble fibre %
ash % %
Control 3.88 5.25 28.715 0.865 0.002 0.19 0.81
- (0.608) (0.014) (0.004) (0.007) (0.001) (0.014) (0.014)

4012 7.00 25.45 1.015 0.006 0.20 0.52

10 (0.014) (1.414) (0.009) (0.001) (0.00) (0.014) (0.014)
Replacement
level of fat 4.38 8.745 24.765 1.027 0.006 0.175 0.48
with ghee 20 (0.14) (0.008) (0.009) (0.002) (0.00) (0.007) (0.028)
residue
4.44 9.62 22.37 1.1280 0.006 0.1950 0.53
30 (0.028) (0.014) (0.010) (0.001) (0.00) (0.007) (0.014)

Critical Difference (0.05) 6.9266∗ NS NS NS NS NS 3.6915∗

NS- Non Significant ; ∗- Significant

The results of the nutrient analysis of biscuits after one month storage

period showed a significant difference of 6.9266 for moisture and 3.6915 for

FFA levels of extracted fat in the product, whereas there was no significant

difference in protein, fat, ash, acid insoluble ash and crude fibre content

between the fresh and the stored samples of control and samples in which fat

was replaced by ghee residue.

Table 16: Nutrient analysis of chocolate toffee after one month storage

Variations Replacement Moisture Protein Fat Ash Acid FFA

Level (%) % % % % insoluble %
ash %
16.153 2.64 12.2095 0.515 0.002 0.68
Control - (0.191) (0.014) (0.014) (0.007) (0.00) (0.00)
16.883 3.50 12.133 0.57 0.003 0.52
10 (0.139) (0.0141) (0.976) (0.028) (0.00) (0.010)
Replacement
level of fat 16.956 4.38 11.3345 0.55 0.003 0.53
with ghee 20 (0.102) (0.014) (0.111) (0.007) (0.00) (0.001)
residue
17.205 5.25 11.07 0.62 0.004 0.53
30 (0.00) (0.057) (0.015) (0.014) (0.00) (0.00)

Critical Difference between 6.8754∗ NS NS NS NS 4.0976∗

fresh and stored samples

NS- Non significant; ∗- Significant

The results of the nutrient analysis conducted for chocolate toffee after

one month storage period showed a significant difference of 6.8754 for

moisture and 4.0976 for FFA. But there was no significant difference for

protein, fat, ash and acid insoluble ash of fresh and stored samples of all levels

of replacement.
 Experimental design of the study

Selection and procurement of raw materials

Standardization of recipes

Product Development

Control

10% replacement of fat with GR

Biscuits 20% replacement of fat with GR Chocolate toffees

30% replacement of fat with GR

Subjective and objective evaluation

Fresh Stored

Nutrient analysis

Fresh Stored

Fig.1: Schematic representation of the study

 5.10
control
5.00 10% replacement
20% replacement
4.90 30% replacement

4.80
5- point Hedonic scale

4.70

4.60

4.50

4.40

4.30

4.20

4.10
APPEARANCE TEXTURE TASTE FLAVOUR OVERALL
ACCEPTABILITY
Sensory attributes
Fig. 2: Mean Scores of Sensory Evaluation of Biscuits
(Fresh)  

 
 

5.10

control
5.00
10%
replacement
4.90
5- Point Hedonic scale

4.80

4.70

4.60

4.50

4.40
APPEARANCE TEXTURE TASTE FLAVOUR OVERALL
ACCEPTABILITY
Fig. 3: Mean Sensory evaluation
Sensory Attributes scores of Chocolate
Toffee (Fresh)  

 
 

5.10
control
5.00
10%
4.90 replacement

4.80
5-point Hedonic scale

4.70

4.60

4.50

4.40

4.30

4.20
APPEARANCE TEXTURE TASTE FLAVOUR
OVERALL ACCEPTABILITY
Sensory attributes
Fig. 4: Mean sensory evaluation of scores of Biscuits
(After one Month storage period)  

 
 

5.10

control
5.00
10%
4.90 replacement
5-point Hedonic scale

4.80

4.70

4.60

4.50

4.40

4.30
APPEARANCE TEXTURE TASTE FLAVOUR OVERALL
Sensory attributes ACCEPTABILITY
Fig. 5: Mean sensory evaluation scores of Chocolate
Toffee (After one Month storage period)  

 
 

0.9
Control
0.8 10% Replacement
20% Replacement
0.7 30% Replacement

0.6
Percentage

0.5

0.4

0.3

0.2

0.1

0
FFA (Fresh) FFA (One month) FFA (Fresh) FFA (One month)

Chocolate Toffees Variation Biscuits

Fig7: FFA difference between fresh and stored samples  

 
 

20

18 Control
10% Replacement
16
20% Replacement
30% Replacement
14

12
Percentage

10

0
Moisture (Fresh) Moisture (One month) Moisture (Fresh) Moisture (One month)

Chocolate Toffees Biscuits

Variation
Fig. 6: Moisture difference between fresh and stored
samples  
 CHAPTER V

DISCUSSION

The results obtained in the study are discussed in detail in this chapter

under the following headings:

5.1 Proximate principles and FFA content of ghee residue

5.2 Sensory evaluation of the products

5.3 Proximate principles and FFA content of the product

5.4 Storage stability of the products

5.1 PROXIMATE ANALYSIS AND FFA CONTENT OF GHEE

RESIDUE

According to the experiment the nutrients analysed in ghee residue were

moisture, protein, fat, ash, acid insoluble ash, and free fatty acids.

5.1.1 Moisture

Moisture is one of the most important parameter of any food product

which plays a vital role in the living process. It also plays an important role in

determining the textural qualities and shelf life of the product.

Moisture content of the processed ghee residue of APDDCF, Vijaya

Dairy, Hyderabad, was high i.e 45.7 per cent as it was processed by washing

with hot water twice followed by washing once with chilled water in order to

recover as much ghee as possible from the residue. According to Santha and
Narayana (1978), the moisture content of the untreated or unprocessed ghee

residue ranged between 8-30 per cent.

5.1.2 Protein

Proteins are the complex organic nitrogenous substances that are vital to

the living process and play an important role in many biochemical, biophysical

and physiological processes in the human body. The adequacy of proteins in

terms of quality and quantity in the diet is an important measure of

malnutrition.

The protein content of the ghee residue was found to be 12.8 per cent. A

study conducted by Kapoor and Pal (1979) showed that the quality and quantity

of protein in ghee residue was good with high PER values and that it can serve

as a good and cheap source of animal proteins to the vegetarian population

when supplemented in the diet.

5.1.3 Fat

Fat imparts palatability to the food and supplies more than twice the

energy furnished by either protein or carbohydrate per unit weight.

The fat content of the processed GR was found to be 15.5 per cent.

whereas, the fat content of the untreated ghee residue ranged from 32-70 per

cent according to Santha and Narayanan (1978a) and Verma (1997). So the

treatment given by the dairy industry in order to recover ghee was found
appreciable as the fat content of the treated or processed ghee residue was as

low as 15.5 per cent compared to the untreated.

5.1.4 Ash

Ash or minerals play a vital role in the proper functioning of the body

and form part of many hormones and enzyme systems.

The ash content of the ghee residue was also studied by Santha and

Narayanan (1978) and Verma (1997). They found that the value ranged

between 1-8 per cent depending upon the type of source by which it is

prepared. But in this experiment, where ghee residue was used, the ash level

was found to be less i.e 0.96 (Table 4) because of thorough processing of ghee

residue by the dairy industry where ghee residue was procured.

5.1.5 Acid insoluble ash

Ash that is insoluble in acid was estimated by heating ash with 5N

Hydrochloric acid. The value was found to be 0.39 per cent (Table 4).

5.1.6 Free fatty acids

Free fatty acid levels indicate the extent of rancidity of the fat and the

products.

The FFA levels of ghee residue was found to be very high i.e. 1.25 per

cent in the ghee residue procured. Galhotra and Wadhwa (1991a and 1991b)

found GR to be a rich source of free fatty acids which is eleven times higher
than that present in ghee. This high content of FFA is responsible for the

flavour of ghee.

5.2 SENSORY EVALUATION OF THE PRODUCTS

To assess the suitability of any new product for food use, carrying out

acceptability tests becomes the most important requirement (Burdon, 1989).

The acceptability of products where fat was replaced by GR at different levels

was carried out in comparison to the control product. Sensory evaluation was

carried out by subjective and objective analysis.

The results of the subjective analysis of biscuits and chocolate toffees

(Table 5 and 6) showed the scores for the sensory attributes like colour, texture,

appearance, taste, flavour and overall acceptability which are presented in the

figures 2 and 3 and are discussed below.

5.2.1 Subjective analysis

Colour

In the present study biscuits in which fat was replaced by GR developed

brownish specks on the surface after baking affecting colour and appearance.

The highest mean score for colour in biscuits was obtained by the fresh control

samples and the least was for the 30 per cent replacement of fat with GR. It has

been observed that there was a decrease in the colour score as the level of

replacement was increasd. This could be due to the inpalatable brown colour of

ghee residue. This is in accordance with the study of Borawake and Bhosale
(1996), where it was reported that increasing levels of replacement of fat with

GR decreased the colour and appearance scores in nankhatai type cookies and

sponge cakes. Similar observations were also made by Ramadevi and Khader

(1987) for the use of GR in cookies, where appearance was affected by the

replacement of fat with ghee residue.

Toffees prepared by replacing fat with GR also had small brownish

specks, but the brownish colour of the GR on the whole did not affect the

overall acceptability scores but there was a slight decrease in the scores for

colour. The highest score for colour was recorded by the fresh control samples

and the lowest was recorded by 30 per cent replacement sample with not much

critical difference as the colour of ghee residue was masked by the colour of

cocoa. This is in accordance with the study conducted by Subbulakshmi et al.

(1990) where it was reported that, though the distinct colour of the GR could

not be masked in most of the recipes except in chocolate cakes and coffee nut

cookies as they were acceptable in terms of appearance and colour due to the

colour of cocoa.

Flavour

Flavour is a combination of taste, smell and mouthfeel. In the mouth and

pharynx are many taste buds capable of detecting sweet, sour, salty and bitter.

As shown in the Table 5, highest scores for flavour were recorded by

samples of biscuits with 30 per cent replacement of fat with GR and the lowest

for the control. It is clear from the table 5 that as the level of replacement was
increased, the flavour scores also increased. This is probably because of the

high flavour potential of GR due to the presence of high FFA, carbonyls and

lactones (Galhotra and Wadhwa 1991a and 1991b). Similar findings were

observed by Subbulakshmi et al. (1990), who concluded that ghee flavour in

the residue improves the aroma of the products there by improving the over all

acceptability.

The results of sensory evaluation scores (Table 6) of chocolate toffees

for flavour were very similar to that of biscuits i.e as the level of replacement

increased, the flavour scores also increased. This is also due to the high flavour

profile of ghee residue due to presence of FFA, carbonyls and lactones

(Galhotra and Wadhwa 1991a and 1991b).

Texture

Texture is a physical state of food. It is a sensation that is associated

with palate, tongue and gums.

The texture attribute of biscuits can be judged on its crispiness,

shortness, brittleness and uniformity. It was observed that there was a slight

decrease in the scores of texture with increased level of replacement when

compared with the control but the difference was not statistically significant.

This may be due to the decrease in amount of fat and increase in the amount of

protein which made the biscuits little hard. Similar observations were made by

Subulakshmi et.al (1990) for use of ghee residue in biscuits, cookies, porridges
and chapathis where the texture of the products became little hard by replacing

or incorporating ghee residue.

The texture attribute of chocolate toffee can be judged on its

smoothness, softness and breaking and cutting strength. There was a slight

decrease in the scores of texture as the level of replacement increased.

Replacement of ghee residue made the toffee soft when compared with the

control but the difference was not statistically significant

Taste

The basic requirement of any food to be accepted is taste. Taste rules the

palate and decides the overall acceptability to a large extent.

In the biscuit samples, it was observed that the 30 per cent replacement

of fat with GR obtained the high score as the control sample obtained the least

score in taste attribute with no particular critical difference. It is due to the

flavour and aroma of ghee present in GR which gives a good mouthfeel which

was appreciated by the panel of judges. Similar findings were observed by

Wadhwa and Bhindal (1995), where they incorporated GR in vanaspathi, butter

oil etc and found that there was an improvement in the taste and flavour.

Similar results were also obtained by Verma and De (1978) where in burfi type

sweet called chocosidue burfi was prepared with khoa and ghee residue which

gave better taste when compared with the control.

 The results of the sensory evaluation showed no critical difference in

particular but the scores for taste attribute were slightly lower in the samples

replacing 30 per cent of fat with GR than in control. The scores for taste

showed a decreased value with increase in the level of replacement.

Overall acceptability

The overall acceptability of the product depends on the weight, size,

moisture content, alcohol insoluble solids, fibre content, density, colour,

acidity, sugar substances, peroxide value etc. (Anerine, 1973).

The overall acceptability scores of biscuits in control sample was high

and the least score obtained was by 30 per cent replacement sample. The

overall acceptability scores decreased as the level of replacement increased but

the difference was statistically not significant. Similar results were obtained by

Subbulakshmi et al. (1990), where the mean score obtained for the control was

2.16 which increased to 2.34 and 2.40 in 50 and 100 per cent ghee residue

substituted biscuits and cakes respectively on a three point scale.

Highest score for overall acceptability was obtained for the control

sample of chocolate toffee, but the scores decreased slightly as the level of

replacement of ghee residue increased which was statistically non- significant.

The acceptability with 10 per cent replacement sample were very close to that

of the control which indicates the best level of replacement.

5.2.2 Objective evaluation

 Objective evaluation was done by using SchimadzuE2 tester, a universal

testing machine for measuring textural characteristics like compression and

cutting strength. The results of objective evaluation for biscuits and chocolate

toffees given in tables 7 and 8.

The results of the compression and cutting tests for biscuit samples

indicates no marked difference between the control and experimental samples

but, the slight difference recorded suggests there were minute textural changes

after the replacement. Samples of 30 per cent replacement had the high values

for compression and cutting strengths (15.89 and 6.11) and control had the low

values i.e. 15.76 & 5.88 (Table 7) which suggests that the experimental

samples were slightly harder than that of control. This is due to the higher

protein content and low fat recorded in the experimental samples which

increased as the level of replacement increased. Similar results were observed

by Gandhi et al. (2001) who observed that increase in protein and decrease in

fat content of biscuits increased the compression and cutting hardness of

biscuits.

Cutting strength of the toffees was slightly decreased as the level of

replacement increased. Control sample recorded the high value (3.725) and the

least value was recorded by 30 per cent replacement sample (2.25) (Table 8).

This suggests that as the moisture content was increasing by replacing fat with

GR, the product became smooth and soft when compared to the control.

However, no studies on objective analysis of the products replacing fat with

ghee residue in toffees were conducted.

5.3 PROXIMATE ANALYSIS AND FFA CONTENT OF THE
PRODUCT

In both the products i.e. biscuits and chocolate toffee the estimation of

moisture, protein, fat, ash and acid insoluble ash and FFA. Crude fibre was

estimated in biscuits in biscuits only.

Moisture

Moisture content of the control was lower 2.45 per cent (Table 9) than

that of the samples in which fat was replaced by ghee residue. As the level of

replacement was increased the moisture level also increased with a critical

difference of 0.0394. This might be due to the high moisture content of ghee

residue which is 45.7 per cent (Table 4). Similar results were also obtained by

Borawake and Bhosale (1996) where fat was replaced by GR at the levels of

10, 20 and 30 per cent in Nankhatai type cookies and sponge cakes and found

that the moisture level increased in both sponge cakes and nankhatai type

cookies as the replacement level increased.

The results of chocolate toffee were very similar to that of biscuits i.e.

as the level of replacement was increased, the moisture content also increased.

The highest moisture level was recorded by 30 per cent replacement sample
and low for control i.e. 17.050 per cent and 15.95 per cent respectively with a

critical difference of 0.0376.

Fat

Fat content in both biscuits and chocolate toffees decreased as the level

of replacement increased. The fat content of the control was high i.e 28.715 per

cent for biscuits and 12.20 per cent for chocolate toffee (Table 9 and 10) and

low for 30 per cent replacement sample i.e 22.37 per cent for biscuits and 11.07

per cent for chocolate toffees (Table 9 and 10). This is due to lower fat content

of ghee residue 15.5 per cent (Table 4) and also high replacement of the fat.

Statistically there was a significant difference of 0.0907 for toffees and 1.1799

for biscuits.

Ash

High ash content was recorded in both biscuits and toffees which

replaced 30 per cent of fat with GR whereas, the low ash content was recorded

by the control i.e 0.620 per cent and 0.515 per cent respectively. As the level of

replacement increased the ash content also increased with a critical difference

of 0.0104 for biscuits and 0.046 for chocolate toffee. This is due to the fact that

the ash content of GR was 0.98 per cent (Table 4) and when it was replaced, it

increased the ash content of both the products as a whole.

From the table it is clear that the acid insoluble ash content was high for

the samples replacing 30 per cent of fat and low for the control i.e 0.004 per

cent and 0.002 per cent for toffee and 0.006 per cent and 0.002 per cent for
biscuits. But statistically there was no critical difference between the control

and the replacement samples.

Free Fatty Acids

Free fatty acids level in biscuits and chocolate toffees increased as the

level of replacement increased. The high value recorded for biscuits and toffees

was by 30 per cent replacement sample i.e 0.53 per cent and 0.52 per cent and

lowest was recorded by the control i.e. 0.50 for biscuits and 0.47 for toffees.

This is due to the higher levels of free fatty acids in GR which is eleven times

higher than that of ghee (Galhotra and Wadhwa, 1991a and 1991b).

Crude Fibre

From the study conducted it is seen that replacement of ghee residue has

no influence on the crude fibre as the levels of crude fibre were same as that of

control. This may be attributed to the absence of crude fibre in the ghee

residue.

5.4 STORAGE STABILITY OF THE PRODUCTS

Sensory evaluation and nutrient analysis was conducted after one month

storage period in order to check the storage stability of the products.

5.4.1 Sensory evaluation stored products

 Subjective evaluation: Figure 4 shows the subjective evaluation scores

of biscuits after storage period of one month. There was no difference observed

after storage in the appearance of the biscuits at all levels of replacement and

even in control when compare with the fresh samples. Texture attribute score

was less for all the samples of biscuits when compared with fresh as they lost

their crispness due to increase in moisture content which was mainly due to the

Water Vapour Transmission Rate (WVTR) of packaging material. Storage had

no effect on taste and flavour of the control samples, but the scores of taste and

flavour were slightly decreased in all levels of replacement as compared to that

of fresh samples as the flavour of ghee was lost due to improper packaging.

Fig. 5 shows the subjective evaluation scores of the chocolate toffees

after a storage period of one month. The results showed that there was no

difference in the appearance of the products before and after storage. But, the

texture attribute of all the stored chocolate toffees had low scores as compared

to that of the fresh as the moisture level increased which made the samples soft

and smooth as showed by the nutrient analysis. Control samples had no marked

difference in taste and flavour but the samples in which fat was replaced with

GR had slightly less scores after storage when compared to that of fresh

samples. This may be attributed to the high Water Vapour Transmission Rate

of packaging material which allowed air to pass through the packaging material

resulting in the loss of flavour of ghee slightly similar to that of biscuits. The

over all acceptability of all the samples of chocolate toffees decreased slightly

after storage but statistically there was no significant difference.

 Objective evaluation: Effect of storage on texture at all levels of

replacement in biscuits as presented in Table 13 indicate that the compression

and cutting hardness decreased when compared with fresh samples after

storage as the moisture level increased due to the WVTR of packaging material

resulting in the loss of crispness.

The results of the cutting hardness presented in Table 14 showed that

there was a slight decrease in the cutting hardness of the all the stored samples

of chocolate toffees as compared to that of fresh indicating the increase in

moisture after storage.

5.4.2 Proximate analysis and FFA content of the products after storage

Proximate analysis after storage period of one month showed that there

was no difference between the protein, fat, ash, acid insoluble ash and fibre

content of biscuits and chocolate toffees, but there was a significant difference

in the levels of moisture and FFA of both the products.

Moisture content of biscuits and chocolate toffees irrespective of

replacement level were found to increase after storage which is presented in the

Figure 6 and a significant difference of 6.9266 and 6.8754 was observed

between the moisture content of fresh and stored samples of biscuits and

chocolate toffees respectively. This may be due to the WVTR of the packaging

material which increased the moisture content of the biscuits and toffees. These

results were in agreement with Zabik et al. (1979), and Doncaster et al. (1986)
who reported moisture gain by biscuits in different types of packaging

materials used for packing and storage.

Changes in FFA content of both biscuits and chocolate toffees during

storage are represented in the figures 7. The FFA content of the control biscuits

and chocolate toffees after storage increased due to increase in moisture

content, which promoted fat oxidation during storage whereas there was no

significant difference in the FFA contents of the 10, 20 and 30 per cent

replacement samples even after storage of one month. This may be due to the

presence of anti-oxidants in ghee residue which inhibited oxidation inspite of

increase in moisture content (Santha and Narayana, 1978).

It is observed from the study undertaken that replacement of fat with

ghee residue had many beneficial effects like increasing protein and minerals

content, increasing the shelf life, decreasing the cost of production by nearly

10-14 per cent of the basic recipe at the household level depending upon other

ingredients, which will be much higher at the commercial scale of production

proving economical and also reduces the problem of disposal which makes the

study important and opens up a promising future for food industries.

 CHAPTER VI

SUMMARY AND CONCLUSIONS

Ghee residue is the brownish, moist sediment obtained as a by- product

during manufacturing of ghee. The Solid-non-fat present in the cream or butter

appears in the form of small particles and settles down forming ghee residue in

the process of ghee manufacturing. At household level it is utilized by mixing it

with rice or by spreading over chapattis but on large scale ghee manufacturing

dairy industries it is thrown away as an industrial waste because of which there

is a loss of appreciable quantity and quality of nutrients which vary as fat (15-

60%), protein (18-40%) in ghee residue depending upon the method used in

preparation of ghee. Ghee residue is also a rich source of anti-oxidants and

flavour compounds which makes it a useful by-product.

Recognizing the vital importance of ghee residue, the present study was

carried out to develop and evaluate the acceptability of the bakery and

confectionery products replacing fat with ghee residue at the level of 10, 20 and

30 per cent. Ghee residue utilized for the experiment was procured from

APDDCF, Vijaya Dairy, Hyderabad where, ghee is prepared by sweet cream

method. The ghee residue settled down during manufacture of ghee was

separated and washed twice with hot water and once with chilled water in order

to recover as much ghee as possible from the residue. The washed ghee residue

was procured and stored in a steel container for use of the experiment. Other

raw materials were procured from the local market and care was taken to
purchase good quality branded raw materials. Biscuits and Chocolate toffees

were selected as the products for experiment and, the recipe and method of

preparation was standardized before acceptability evaluation of the final

product. Biscuits and Chocolate toffees were developed using 10, 20 and 30 per

cent replacement of fat with ghee residue and compared with the control.

Sensory evaluation of the biscuits and chocolate toffees was conducted

using a score card developed using 5 point hedonic scale by a panel of ten

judges. The products were evaluated as fresh sample and also after on month of

storage period. Objective evaluation using Schimadzu E2 Tester was conducted

for measuring cutting and compression strengths for biscuits and cutting

strength for toffees when fresh and after one month storage period. Moisture,

protein, fat, ash acid insoluble ash, crude fibre and FFA content were also

determined in fresh and stored samples. The results of the study were

statistically analysed for mean, standard deviation and critical difference.

Sensory evaluation of the study revealed that there was a slight decrease

in the appearance, texture and overall acceptability of the biscuits and

chocolate toffees as the level of replacement increased. But there was an

improvement in flavour and taste attributes in both the products as the level of

replacement increased.

Objective evaluation conducted using Instron for measuring

compression and cutting strengths revealed that there was an increase in the

compression and cutting strengths of biscuits indicating that the biscuits

became hard as the replacement level increased due to increase in protein and
decrease in fat content. In the case of toffees the cutting strength decreased

indicating that the product became soft as the replacement level increased due

to higher moisture content.

Nutrient analysis conducted revealed that there was an increase in

moisture, protein, ash, acid insoluble ash and FFA content as the level of

replacement increased in both biscuits and toffees as ghee residue is a rich

source of moisture, protein and minerals. But, there was a slight decrease in the

fat content of both the products as ghee residue had a low fat content of 15.5

per cent and level of replacement of fat being high. Crude fibre content of the

biscuits was not affected by replacing fat with ghee residue as GR has no crude

fibre.

Storage studies after one month period revealed that there was a slight

decrease in the acceptability of both the products and at all levels of

replacement due to increase in moisture and slight loss of flavour of ghee

which affected the texture, taste and flavour attributes. This is due to the high

Water vapour transmission rate of packaging material, which allowed moisture

to pass through it. The FFA content was not affected even after storage in the

samples where fat was replaced by GR, whereas, the control sample had a

significant increase in FFA content indicating that the GR is a rich source of

natural anti-oxidants.

The present research conducted is a preliminary step towards

development of products replacing fat with ghee residue as it is economical

with a potential to develop into a good health food with better shelf life, as the
present day population becoming more health conscious exploring health foods

and its benefits. The potential of this by-product has to be exploited to its

maximum extent in the coming years as it is a rich source of nutrients, natural

anti-oxidants, flavours and its utilization in product development also reduces

the problem of disposal, for which further research has to be done, which might

find newer applications of this by- product even at commercial level.

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 APPENDIX - V

Estimation of Moisture

The moisture content of the samples was determined by using the

method of AOAC (1990).

Procedure

¾ 10 grams of the sample was taken into a petridish with lid and the exact

weight was noted down (W1).

¾ The sample was dried in an oven at 100 to 105°C till constant weight

was obtained.

¾ The sample was cooled in a dessicator and the final weight was taken

(W2).

Calculation

Moisture % = Initial weight (W1) - Final weight (W2) x 100

Initial weight (W1)

 APPENDIX - VI

Protein estimation

Protein content was estimated by Microjeldhal method by the procedure

of AOAC (1990).

Principle

The nitrogenous compounds of the sample are converted into

ammonium sulphate by boiling with concentrated Sulphuric acid. It was

subsequently decomposed by addition of excess of alkali and the liberated

ammonia was absorbed into boric acid solution containing bromocresol green

indicator by stream distillation. Ammonia formed a loose compound,

ammonium borate, with boric acid, which was titrated directly against

standardized hydrochloric acid.

Chemical reagents

1) 40% NaOH : 400 grams of NaOH was dissolved in 1000ml of distilled

water by steeping the beaker in running water in a sink.

2) 2% Boric acid : 10 grams of reagent grade boric acid was dissolved in

500 ml of hot distilled water.

3) Mixed indicator :

a) 0.2% bromocresol green in alcohol : 20 mg of bromocresol green was

dissolved in 10 ml alcohol.
 b) 0.2 % methyl red : 20 mg of methyl red powder was dissolved in 10

ml alcohol .

5 ml of (a) and 2 ml of (b) are mixed together and used as mixed

indicator.

4) 0.5 N HCl : 100 ml of 0.1 N HCl (accurately standardized against 0.1 N

NaOH) was dissolved in 500 ml in a volumetric flask.

5) Digestion mixture : 98 grams of potassium sulphate and 2 grams

sulphate are ground together.

Procedure

Digestion

500 mg of food sample was weighed into boiling tube. 5 grams of

digestion mixture and 10 ml of concentrated Sulphuric acid were carefully

added and the samples were digested in the digestion block for 1 hour at 375oC.

the tubes were removed and distilled water (approximately 50ml) was added

carefully from the sides while still hot.

Distillation

In a 100ml conical flask, 50ml of 4 per cent boric acid was added with

few drops of mixed indicator. The condenser outlet of the steam distillation

apparatus should dip below the surface of the boric acid solution. The digested

sample was completely transferred by means of repeated washings to the

chamber of the distillation apparatus. The chamber should be previously

cleared off any contaminated ammonia by repeated washings and steam

generation should be started. 50ml of NaOH solution was slowly added,

followed by one more rinsing of the digested sample. Sample should be

distilled for exactly 3 minutes after solution changed to a blue colour, the

receiving flask was lowered and steam generation was stopped. The condenser

outlet should be washed into the receiving flask. The contents of the flask were

titrated with 0.5 N HCl till the color changed to original pink. The blank was

also run simultaneously.

Calculations

1ml of 0.5 N HCl = 14.007 grams of nitrogen.

1 gram of nitrogen = 100/16 = 6.25 grams of protein.

Protein % = Titre value x 14.007 x 0.5 x 6.25 x 100

____________________
Wt of the sample (mg)
 APPENDIX - VII

Estimation of Fat

The fat content of the sample was estimated as crude ether extract of the

dry material.

Method

The dry sample (5-10 grams) was weighed accurately into a thimble

(made with Whatman no. 1 filter paper – AOAC technique) and was placed in

the soxhlet apparatus or soxtic labconco. It was extracted with petroleum ether

(60-80 C Boiling Point) for about 16 hours. The ether extract was filled into a

weighed beaker, the flask was rinsed four to five times with small quantities of

petroleum ether added to the beaker. Petroleum ether was removed by

evaporation and the flask with the residue was dried in an oven at 80 – 100°C

and later cooled in a dessicator and weighed.

Calculation

Weight of ether extract

Fat% (gram / 100 gram ) = ___________________ x 100
Weight of ether sample
 APPENDIX - VIII

Estimation of Ash

The ash content of the sample was estimated by AOAC (1990) method.

Procedure

¾ The temperature of the muffle furnace was set at 600oC and the crucible

was heated for 1 hour and transferred into a dessicator. After cooling to

room temperature the crucible was weighed (W1).

¾ 2 grams of sample (defatted) was weighed into the crucible of known

weight (W2).

¾ The sample was incinerated at 600 oC for 2 hours.

¾ The crucible was transferred into the dessicator and cooled to room

temperature and weighed (W3).

Calculation

Weight of the sample taken = W2- W1

Weight of the ash obtained = W3 – W

Weight of Ash
% Ash = _________________ x 100
Weight of the sample
 APPENDIX - IX

Estimation of Acid Insoluble Ash

Method

¾ After total ash determination, the ash contained in the crucible, add 25

ml of 5 N

¾ HCl and heat in a water bath / 10 minutes.

¾ Cool and filter with whatman filter paper No- 42 or its equivalent.

¾ Wash the filter paper with water until the washings are free from the

acid

¾ Keep it in an oven at 100oC for 3 hours.

¾ Ignite in a muffle furnace at 550 oC / one hour and record the weight.

Calculations

Weight of the empty crucible = W

Weight of crucible cum ash = W1

Weight of crucible cum acid insoluble ash = W2

Acid Insoluble ash % by weight = 100 (W2-W) / W1-W

 APPENDIX – X

Estimation of Free Fatty Acids

Method

¾ Take 10 ml of the sample in a conical flask

¾ Add 50-100 ml of ethyl alcohol to the conical flask and neutralize

¾ Heat the contents for 5 minutes

¾ Add phenolphthalein indicator to the flask and titrated against standard

alkali. (i.e. 0.1-0.5 NaOH)

Calculations

FFA in terms of Oleic acid % by wt = 28.2VN / W

FFA in terms of Lauric acid% bt wt = 20.0 VN / N

FFA in terms of Ricinoleic acid % by wt = 29.8 VN / W

FFA in terms of Palmitic acid %by wt = 25.6 VN / W

 APPENDIX - II

Method of preparation of Chocolate Toffee

Ingredients

Product Sugar SMP Fat GR Cocoa Water Liquid

(chocolate (grams) (grams) (grams) (grams) Powder (ml) Glucose
Toffee) (grams) (ml)

Control 200 50 27 - 15 150 5

10%
replacement 200 50 24.3 2.7 15 150 5
of fat with
GR
20%
replacement 200 50 21.6 5.4 15 150 5
of fat with
GR
30%
replacement 200 50 18.9 8.1 15 150 5
of fat with
GR

Method

™ Sugar and water were mixed and boiled until all the sugar gets

dissolved.

™ In the control sample only fat was added whereas in the

experimental sample fat and Ghee residue both were added.

™ When all the fat got dissolved, SMP, Liquid Glucose and Cocoa

powder were added and mixed thoroughly so that there was no lump

formation.
™ The contents were boiled till soft ball consistency.

™ The contents were removed from the flame and transferred into a

non-stick container and beaten thoroughly with the help of a wooden

ladle.

™ When it became thick, it was poured on to a greased tray and cut

into pieces.
 APPENDIX-I

Method of preparation of Biscuits

Ingredients

Product Flour Milk Sugar Fat GR Baking NH3 Vanilla

(Biscuit) (grams) (ml) (grams) (grams) (grams) Powder

Control 300 50-100 150 150 - 1/2 tsp 1/8 tsp Few
drops

Replacing 300 50-100 150 135 15 1/2 tsp 1/8 tsp Few
10% fat drops
with GR

Replacing 300 50-100 150 120 30 1/2 tsp 1/8 tsp Few
20% fat drops
with GR

Replacing 300 50-100 150 105 45 1/2 tsp 1/8 tsp Few
30% fat drops
with GR

Method

™ Flour, salt and baking powder were sieved.

™ Fat and sugar were creamed till light and fluffy for the control sample,

whereas for the experimental sample fat, sugar and ghee residue are

creamed.

™ Flour was added to creamed mixture and made into smooth dough by

adding milk.

™ This was roll out thick on a dusted platform and cut into shapes.

™ Cut biscuits were placed on a greased tray and baked at 180° C for 15-

20 minutes, cooled and packed.

 APPENDIX-IV

SCORE CARD FOR SENSORY EVALUATION OF CHOCOLATES

Name of the panel member: Date:

9 Please evaluate the following samples using the 5 point hedonic scale given below.

9 Tick the preferred characteristic as per your evaluation.

SENSORY CHARACTERISTICS Sample Sample Sample Sample

1 2 3 4

Colour and Appearance

Highly Appealing 5
Appealing 4
Moderately Appealing 3
Slightly Appealing 2
Not Appealing 1

Texture
Very Soft 5
Soft 4
Average 3
Hard 2
Very Hard 1

Taste
Very good 5
Good 4
Fair 3
Poor 2
Very poor 1

Flavour
Very Pleasant Flavour 5
Pleasant 4
Fair 3
Poor 2
Very poor 1

Overall Acceptability
Highly Acceptable 5
Acceptable 4
Moderately Acceptable 3
Fairly Acceptable 2
Not Acceptable 1

Suggestions and comments: Signature

 APPENDIX- III

SCORE CARD FOR SENSORY EVALUATION OF BISCUITS

Name of the panel member: Date:

9 Please evaluate the following samples using the 5 point Hedonic scale given below.

9 Tick the preferred characteristic as per your evaluation.

SENSORY CHARACTERISTICS Sample Sample Sample Sample

1 2 3 4

Colour and Appearance

Highly Appealing 5
Appealing 4
Moderately Appealing 3
Slightly Appealing 2
Not Appealing 1
Texture
Crisp and Crunchy 5
Hard and Brittle 4
Soft and Brittle 3
Hard 2
Not Acceptable 1
Taste
Very good 5
Good 4
Fair 3
Poor 2
Very poor 1
Flavour
Very Pleasant Flavour 5
Pleasant 4
Fair 3
Poor 2
Very poor 1
Overall Acceptability
Highly Acceptable 5
Acceptable 4
Moderately Acceptable 3
Fairly Acceptable 2
Not Acceptable 1

Suggestions and comments: Signature

 APPENDIX XI

Crude fibre estimation

The crude fibre present in biscuits was estimated by using AOAC, 1984.

Principle

¾ Take 5 grams of sample in a 250 ml beaker

¾ Add 200 ml of 1.2% Sulphuric acid in to the beaker.

¾ Boil for 30 minutes.

¾ Filter and wash until all the acid washes off and then scrap the residue.

¾ Take the residue in 250ml beaker again and add 200 ml of 1.2% alkali

and then boil.

¾ Filter and wash the residue till it is free from alkali

¾ Add acetone so as to absorb the moisture.

¾ Scrap and dry the residue and weigh.

¾ Ash it and record the weight.

Calculation

Crude fibre % by weight = 100(W1-W2) / W

W1= Weight in grams of crucible and contents before ashing

W2= Weight in grams of crucible and contents after ashing

W= Weight of material taken for the test