Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

The present investigation entitled “Studies on Utilization of Papaya Peel and Seed
Powder for Development of Functional Bakery Products’’ was carried out in the
Food Technology Laboratory of Department of Technology, Shivaji University,
Kolhapur, during year 2017-18.

3.1 Materials
3.1.1 Raw materials
Raw material preparation- waste from the papaya i.e papaya peel and papaya seed
powder was prepared on the basis of required quality parameters and necessary pre-
treatments such as washing, grading, sorting etc., was carried out. Refined wheat flour
and other ingredients was purchased on the basic of quality parameters from local market.
All raw materials were purchased on the basis of required quality parameter from
the local market of Kolhapur, Maharashtra.

1. Flour: Commercial wheat flour (maida) was procured from the local market.
2. Fat: Commercial shortening i.e hydrogenated vegetable oil for bakery purpose
was obtained from the local market.
3. Sugar: Whole cane sugar was purchased from the local market. It was ground and
sieved in lab to obtain uniform particle size and also to remove any physical
impurities if present.
4. Salt: Iodised salt was procured from the local market for its use in preparation of
bread.
5. Leavening Agent: Baking powder (Weikfield) was used as a leavening agent in
muffins and was procured from the local market.
6. Milk powder: Commercial SMP was procured from the local market.
7. Packaging Material: Aluminium Laminate (polyethylene/ aluminium foil/
polyethylene-150 gauge) was used for the packaging of cookies and Linear Low
Density Polyethylene (LLDPE-150 gauge) cling wrap was used for packaging of
muffins. Packaging materials used were procured from local market.

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

3.1.2 Chemicals
Chemicals required for the chemical analysis of raw materials and products prepared
were available in the Food Technology laboratory of Department of Technology, Shivaji
University, Kolhapur.
3.1.3 Utensils
Glass utensils such as burette, pipette, test tubes, petri plate set and other utensils such as
stainless steel pots, plates, knives, etc. were available in the laboratory.
3.1.4 Equipment’s and instruments
The equipment’s and instruments such as Soxhlet apparatus, Kjeldhal apparatus, oven,
muffle furnace, gas cylinder, etc. were used from the laboratory.

3.2 Methods
3.2.1 Preparation of papaya peel powder
Papaya

Washing in normal water

Peeling manually by using peeler

Peels cutting and washing

Sanitize by using of Zeolin and keep covered for 15 min

Keep in a tray and drying at 60°C for 24 hrs. in tray drier

Grinding and sieving ( 30mesh size)

Packing in polythene and Keep in sealed container

Figure 1. Flowchart of preparation of papaya peel powder (Bokaria and Ray, 2016)

Papaya Peel Powder was prepared by reference of Bokaria and Ray, 2016 with
slightly modification as shown in Figure 1. The papaya fruits were washed under running
water, disinfected and rinsed. The papaya peels were manually removed using stainless
steel knives and were weighed to determine the yield. Material preparation and the
physicochemical analyses were performed at the laboratory. After the peels were

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

weighed, they were cut into small pieces then dried in tray dryer at 60°C for 24 hours and
grinded. The crushed material was then sieved through a 30 mesh sieve to obtain powder.
The peel powder was again weighed to calculate the yield, then papaya peel powder was
vacuum packed and stored at 4°C for future analysis.

3.2.2 Preparation of papaya seed powder

Selection of papaya

Washing (remove dirt, dust)

Cutting

Remove seeds

Washing (removal of residue of pulp)

Drying at 60°C

Grinding and Sieving (30 mesh size)

Storage
Figure 2. Flow chart of production of papaya seed powder
(Shrivastava,2002)
Papaya seed Powder was prepared by reference of Shrivastava with slightly
modification as shown in Figure 1. First, the papaya (Carica papaya) seeds were removed
and washed in drinking water for removal of the residual pulp. Then, drying of the seeds was
performed in tray dryer at 60°C for about 24 hours. After drying, dried papaya seeds were
ground in the grinder to reduce the particle size. The ground material was allowed to pass
through sieve size 30 mesh, the larger particle on the sieve was again taken for grinding and
passed through the sieve to obtain fine powder. The papaya seed powder was vacuum packed
and stored at 4°C for future analysis.

3.2.3 Response Surface Methodology (RSM)

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

Response Surface Methodology (RSM) was used for optimization of papaya peel
and seed powder fortified functional cookies on the basis of following responses:
Sensory parameters: Color, flavor, texture, taste and overall acceptability.
Textural properties: Hardness of cookies were studied.

3.3 Physical and Functional Properties of raw material

3.3.1 Physico analysis of raw materials

1. Thickness: The thickness of the raw material was measured by a vernier caliper.
The average value of six determinations was taken and reported in millimeter
(AACC, 1967).
2. Diameter: The diameter of raw material was determined using the scale placing
them edge-to-edge. The average diameter of was reported in centimeter.
3. Weight: Weight of raw material and cookies were measured by weighing balance
(Shimadzu Ltd., Model ELB300). The average of three determinations was
reported in gram (g).
4. Color: The color of raw material was visually observed.

3.3.2 Functional Properties of raw material

1. Bulk density (BD):
Bulk density was determined as per the method by filling a measuring cylinder
with the flour with less than 12% moisture slightly above the liter mark. The cylinder was
tapped ten times till the products measured up to the liter mark. The weight of the flour
was taken and bulk density was calculated with the formula (Udachan and Sahoo, 2017).
Bulk density (g/ml)= Weight of flour (g)
Volume taken by flour (ml)

2. Water and oil absorption capacity (WAC and OAC)

The water absorption capacity (WAI) were measured according to the standard
reported method by (Udachan and Sahoo, 2017) with minor modifications. Each
sample (1 g) of ground sample was suspended in 20 ml of distilled water in a 50 ml
centrifuge tube and stirred with a glass rod, put in water bath for 30 min at 30°C
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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

temperature. Subsequently, the dispersions were centrifuged at 2000rpm for 10 min

using a centrifuge (RemiQ-8C, India). The supernatants were poured into dry test
tubes and stored overnight at 110 °C for the process of evaporation.
Water absorption capacity(g/g) = Weight of sediment
Weight of dry solids

3. Foaming capacity (FC) and foam stability (FS)

The method described by Narayana and Narasinga Rao (1982) was used for the
determination of foam capacity (FC) and foam stability (FS). Two grams of flour sample
was added to 50 ml distilled water at 30 ± 2oC in a 100 ml measuring cylinder. The
suspension was mixed and properly shaken to foam and the volume of the foam after 30 s
was recorded. The FC was expressed as a percentage increase in volume. The foam
volume was recorded 1 h after whipping to determine the FS as a percentage of the initial
foam volume.
FC = Volume after whipping - Volume before whipping X 100
Volume before whipping
4. Swelling power (SP)
The estimation of swelling power and water solubility index were carried out
according to the method of (Udachan and Sahoo,2012) with some modifications. A
suspension of 500 mg of starch 20 ml of distilled water was heated in a water bath at
50°C, 60°C and 70°C for 30 min. The suspension was then cooled rapidly at room
temperature and centrifuged at 5000 r. p.m. for 20 min. After this 10 ml Aliquot was
pipetted into a weighing dish and dried at 120°C for 2 hr to determine the soluble
content. The remaining supernatant was carefully removed by suction and weighed to
determine the water solubility index of starch granules. Swelling power (%) was
calculated with corrections for soluble.
Swelling power = Weight of paste x 100
Weight of sample

3.2.2.3 Chemical analysis

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

1. Moisture
Moisture content of raw material was analyzed by standard
(A.O.A.C.1990) method. Weighing the 5g ground sample and
subjected to oven drying at 110˚c for 4hours. It was again
weighed after cooling in desiccators until the constant weight. The
resultant loss in weight was calculated as moisture content in
percent. (A.O.A.C. 1990)

Initial weight of sample−Final weight of sample

% Moisture = ×100
Initial weight of sample
2. Ash
Ash content of raw material was determined by standard
(A.O.A.C.1990) method. 5g sample was weighed into crucible,
which was heated at low flame till all the material was completely
charred and cooled. Then it was kept in muffle furnace for about 4
hours at 600˚C. It was again cooled in desiccators and weighed
and repeated until two consecutive weights were constant. The per
cent ash was calculated by reading the difference between the
initial and final weight (A.O.A.C., 1990).
Weight of ash
% Ash = ×100
Weight of sample

3. Fat
The crude fat content was estimated by using Soxhlet
method. The oven dried sample was extracted by using petroleum
ether (boiling point 60ºC to 80ºC). The extraction was continued
till the completion of 6-15 cycles. Then ether was evaporated and
the flask was cooled by keeping in desiccators. After cooling, the

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flask was weighed and crude fat percentage was calculated by

using following formula (A.O.A.C., 1990).
Weight of ether soluble extract
% Crude fat = ×100

Weight of sample

4. Protein
Protein was determined by Micro-kjelahl method using
0.5g of ground sample by digesting the same with concentrated
sulfuric acid (H2SO4) containing catalyst mixture for 3-4 hours at
100˚C.Then it was distilled with 40 per cent NaOH solution and
liberated ammonia was trapped in 4 per cent boric acid and then it
was titrated with 0.1N HCL using mixed indicator (Methyl red:
Bromocresol green 1:5). The per cent nitrogen was calculated and
protein was estimated in the sample by multiplying with factor
6.25 (A.O.A.C., 1990).
14 × N.of HCl ×(Sample titer−Blank titer)× Vol.make up
% Nitrogen = ×100
Aliquote of digest taken × Wt.of sample taken× 100

% Protein=% Nitrogen ×6.25

5. Crude fiber

About 2g of moisture and fat free samples were weighted

into 500 ml beaker and 200 ml boiling 0.255N (1.25 W/V) H 2SO4
was added. The mixture was boiled for 30 minutes keeping the
volume constant by addition of water at frequent intervals. At the
end of this period, the mixture was filtered through a filter paper
and the residue washed with hot water till free from acid. The
material then transferred to the same beaker and 200 ml of boiling
0.313N NaOH solution added. After boiling for 30 minute, the
mix residue was washed with hot water till free from alkali

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

followed with same alcohol. It was then transferred to a crucible,

dried over night at 80ºC - 100ºC for 2-3 hr. then cooled and
weighted again. The difference in the weights represented the
weight of crude fiber (A.O.A.C. 2000).
[(W2−W1) − (W3−W1)]
% Crude fiber= ×100
Weight of sample

Where,
W= Weight of empty silica crucible
W2=Weight of crucible with residue
W3= Weight of crucible with residue after ashing
6. Carbohydrates
Carbohydrate content was calculated by difference method using following formula
(Ranganna, 1986).

% Carbohydrate = 100 – (% Moisture + % Ash + % Protein + % Crude fat)

7. Mineral determination

The minerals contents i.e. (sodium, potassium, magnesium, calcium, iron, copper,
manganese and phosphorus of wheat biscuits and functional cookies with papaya peel
and seed powder were determined according to the methods described in AOAC (1997).
The samples were wet acid digested using a nitric acid and perchloric acid mixture
(HNO3, HCLO4, 2:1 v/v). The amounts of iron, copper, and manganese in the digested
sample were determined using a GBC Atomic Absorption 906 A as described in AOAC
(1997). Sodium and potassium were determined by flame photometer 410. Calcium and
magnesium were determined using Double Beam Atomic Absorption. Phosphorus was
detemined according to the method described in A.O.A.C. (1997). (Hanan et al.,2012).

8. Antioxidant activity

Effect of the sample extracts on DPPH radical was measured by using a slightly
modified method previously described by Tang et al. (2002). Amount of 200 μl of the

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sample extract (0.62 – 4.96 mg/ml) or ascorbic acid (0.04 – 1.28 mg/ml) were added to 1
ml of 0.1 mM DPPH in 80% methanol. The mixture was shaken vigorously and left to
stand in dark room for 30 min at room temperature. Absorbance of the solution was
measured spectrophotometrically at 517 nm with deionized water as blank. The capability
of sample to scavenge the DPPH radical was calculated according to the equation as
follows:

Scavenging effect = Absorbance of coat 517nm - Absorbance of sample at 517nm x 100

(%) Absorbance of control at 517nm

3.3 Preparation of control cookies and functional cookies

The cookies preparation process is conventional and easy. Cookies were prepared
by method of Vaijapurkar K. et.al. 2015 with slightly modification as follows. Initially all
the raw materials such as wheat flour, papaya peel powder, papaya seed powder, baking
powder, milk powder and fat of good quality were procured from local market of
Kolhapur. All these ingredients were weighed as per the requirement and mixed
thoroughly using sieve so that all ingredients get uniformly mixed. After mixing of all
ingredients addition of water was done in order to form dough. Dough was knead in order
to turn it into soft mass and allowed to rest for 15 min. Then dough was rolled with the
help of floured rolling pin to a desirable thickness and cut with the help of cutter of
desirable shape and size. These pieces were placed in baking oven at temperature 120°C
for 30 min followed cooling and packing of cookies.

3.3.1 Control cookies

Control samples were prepared without any addition of papaya peel and seed
powder by using sugar, fat and baking powder

Raw material

Weighing all the ingredients

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

Creaming

Sieve all dry ingredients mix well

Preparation of dough (addition of water)

Kneading

Sheeting

Cutting

Baking
(120°c for 30min)

Cooling

Weighing and Packing

Figure 3. Flowchart of control cookies production (Vaijapurkar K. et al., 2015)

3.3.2 Preparation of functional cookies by addition of papaya peel and

seed powder

Weighing all the ingredients

Creaming

Sieve all dry ingredients mix well

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

Addition of papaya peel and seed powder

(Wheat Flour: Papaya Peel Power: Papaya Seed Powder,96.63:3.82:1.24 respectively)

Preparation of dough (addition of water)

Sheeting

Cutting

Baking
(120°C for 30min)

Cooling

Weighing and Packing

Figure 4: Flowchart of functional cookies production (Vaijapurkar K. et al., 2015)

3.4 Optimization of control cookies and functional cookies

With the help of response surface methodology (RSM), functional cookies was
optimized on the basis of sensory and textural properties.

The optimization of control cookies was carried out by varying proportion of different
components such as flour, sugar and fat.
3.4.1.1 Optimization of flour in control cookies samples
In case of control samples sugar and the amount of baking powder and fat added in
cookies are kept constant. Wheat flour content was varied to get an acceptable quality of
cookies. In preparation of control cookies the amount of wheat flour was varied as
90%, 95% and 100%.
Table 1. Optimization of flour in control samples
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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

Sample Wheat Fat (gm) Baking Water (ml) Sugar (gm)

code flour(gm) powder(gm)
Control 90 50 4 20 50
WF1
Control 95 50 4 20 50
WF2
Control 100 50 4 20 50
WF3

3.4.1.2 Optimization of sugar in control cookies samples

In case of control samples flour and the amount of baking powder and fat added in
cookies are kept constant. Sugar content was varied to get an acceptable quality of
cookies. In preparation of control cookies the amount of sugar was varied as 45%,
50% and 55%.

Table 2. Optimization of sugar in control cookies samples

Sample Wheat Fat (gm) Baking Water (ml) Sugar (gm)
code flour(gm) powder(gm)

Control S1 100 50 4 20 45

Control S2 100 50 4 20 50

Control S3 100 50 4 20 55

3.4.1.3 Optimization of fat in control cookies samples

In case of control samples flour and the amount of baking powder and sugar added in
cookies are kept constant. Fat content was varied to get an acceptable quality of cookies.

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The preparation of control cookies the amount of fat was varied as 45 %, 50%, and
55%.

Table 3. Optimization of fat in control samples

Sample Wheat Fat (gm) Baking Water (ml) Sugar (gm)
code flour(gm) powder(gm)

Control F1 100 45 4 20 50

Control F2 100 50 4 20 50

Control F3 100 55 4 20 50

The control cookies were prepared by using wheat flour, sugar, fat, baking
powder, SMP and water. In this case of wheat flour, sugar, fat were optimized i.e 100, 50,
and 50 gam respectively. Depending upon sensory evaluation cookies having good taste,
texture and overall acceptability were finalized.

3.4.2 Replacement of wheat flour by papaya peel and seed powder

For replacement of wheat flour by papaya peel and seed powder in which sugar, baking
powder, fat were kept constant. Response surface methodology used for optimization of
functional cookies by varying different runs by wheat Flour, papaya peel powder and
papaya seed powder. In this study, RSM was used to determine the optimum formulation
for a functional cookie with high protein and fiber level compared to a conventional
cookie.

Table 4. Optimization of raw material by using RSM

Runs Wheat flour Papaya peel powder Papaya seed

powder

1 97.5 2.5 1.5

2 97.5 2.5 1.5

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3 97.5 2.5 1.5

4 96.01 1.01 0.61

5 98.99 3.99 2.39

6 97.5 0 1.5

7 96.01 1.01 2.39

8 96.01 3.99 0.61

9 97.5 5 1.5

10 97.5 2.5 1.5

11 98.99 1.01 0.61

12 100 2.5 1.5

13 98.99 3.99 0.61

14 97.5 2.5 1.5

15 98.99 1.01 2.39

16 97.5 2.5 3

17 97.5 2.5 0

18 97.5 2.5 1.5

19 95 2.5 1.5

20 96.01 3.99 2.39

3.5 Physical and Chemical analysis of cookies

3.5.1 Physical properties
1. Thickness: The thickness of the cookies was measured by a vernier caliper.
The average value of six determinations was taken and reported in millimeter
(AACC, 1967).
2. Diameter: The diameter of cookies was determined using the scale placing
them edge-to-edge. The diameter was measured by laying cookies and

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measuring to the nearest mm (AACC 1967).The cookies were rotated at 900

and its diameter was remeasured as a check determination. The average
diameter of was reported in centimeter.
3. Weight: Weight of cookies measured by weighing balance (Shimadzu Ltd.,
Model ELB300). The average of three determinations was reported in gram
(g).
4. Color: The color of baked cookies were read as L*, a* and b* using Hunter
Lab. The Hunter lab colorimeter was calibrated by Hunter color standard prior
to the reading of cookies samples. Color analysis done at ICT Mumbai.
Surface color of cookies was determined by measuring tristimulus L
(lightness), a (redness), b (yellowness) values with a colorimeter calibrated
with a white standard plate.(Bhat & Ahsan,2015)

5. Spread ratio of cookies

The spread ratio of the baked cookies were determined from the ratio of width and
thickness of cookies (Suriya et al., 2017).

Cookie Spread Ratio was calculated by the following formula

Average diameter of 4 cookies

Spread Ratio= Average thickness of 4 cookies

3.5.2 Chemical analysis of cookies

The moisture, crude protein, crude fat, total ash and fiber content was analyzed as
described in chapter 3.2.2.3 above.

3.6 Textural analysis of cookies:

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Hardness of baked cookies was measured in terms of breaking strength using

Texture Analyzer (TA-HD plus, stable Micro systems). Breaking strength of cookies was
placed on the platform such that they are supported at two points and the blade was
attached to the crosshead of the instrument. The analyzer was set to return to start cycle,
at a speed of 1.5 mm/s and a distance of 15 mm. This experiment simulates the
assessment of hardness by consumer holding the cookie in hands and breaking by
bending. The peak force from the subsequent curve was measured as the breaking
strength of the cookies. Texture analysis instrument, capable of measuring virtually any
physical product characteristic such as hardness, factorability, adhesiveness, extensibility
of foods (Suriya et al. 2016).

3.7Sensory evaluation of cookies

Sensory evaluation was carried out by a panel of ten semi trained panel members.
Hedonic rating test was employed using 9-point hedonic scale. Sensory parameters such
as color, taste, texture and overall acceptability were evaluated (Ranganna, 1986). The
following were the numerical scores assigned:
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

3.8 Shelf-Life Study of cookies

3.8.1 Storage Temperatures and Sampling Times
The samples cookies were stored in LDPE at 25±1°C temperature. The samples
were analyzed at every 15 days of interval during storage up to 90 days.

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Appearance, color, flavor, taste, and overall acceptability of papaya peel and seed
powder fortified functional cookies and control cookies were judged by a panel of 10
semi-trained judges by using 9-point hedonic rating (1 = dislike extremely, 9 = like
extremely) scale.

3.8.2 Analysis of Chemical properties

3.8.2.1 Moisture
Cookies samples were ground with a mortar. The moisture content was determined
through weight loss after heating the sample in an oven at 105°C until constant weight
(AOAC, 2000). Determinations were made at least in duplicate.
3.8.3 Color and Flavor
Color was evaluated by visually appearance. Flavor was evaluated by using sensory
organs. (Siddiqui et al., 2012).
3.8.4 Total plate count
The total plate count method was used for microbial analysis (USFDA, 2001).

3.8.4.1 Preparation of food homogenate

1:10 dilution of the well mixed sample was prepared by aseptically transferring sample to
the desired volume of diluents. 1 g of sample was weighed into a sterile blender jar. And
9 ml of diluents was added and blended for 2 min at low speed (approximately 800 rpm).
3.8.4.2 Serial Dilution
Pipette 1 ml of food homogenate into a tube containing 9 ml of diluents. From the first
dilution transfer 1 ml to second dilution tube containing 9 ml of diluents. Repeat using a
third, fourth or more tubes until the desired dilution is obtained (USFDA, 2001).

3.8.4.3 Pour Plating and Incubation

Nutrient agar:
Table 5. Composition of Nutrient agar

Components g/lit
Peptone 5.0
Yeast Extract 1.5

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 Studies on Utilization of Papaya Peel and Seed Powder for Development of Functional Bakery

NaCl 5.0
Agar 15.0
Beef Extract 1.5
pH 7.4
12-15 ml nutrient agar was added (cooled to 45 ± 1°C) to each plate within 15 min of
original dilution. 1 ml of each dilution was pipette out into separate, duplicate,
appropriately marked petri dishes. Immediately sample dilutions were mixed with agar
medium thoroughly and uniformly by alternate rotation and back and forth motion of
plates on flat level surface. Agar was allowed to get solidify. Inverted solidified petri
dishes, were allowed to incubate promptly for 48 ± 2 h at 35°C (USFDA, 2001).
3.8.4.4 Total plate count and calculation
The results per dilution were counted. The observed colonies were multiplied by the
dilution factor and the results were recorded (USFDA, 2001).
Cfu /gram = No. of colonies × dilution factor

3.9 Packaging Material Analysis

Packaging material analysis was done at Indian Institute of Packaging, Mumbai
by following methods.
3.9.1 Tensile strength
1) Tensile Testing Machine - The tensile testing machine used shall maintain a rate of
traverse of one grip as constant as possible. The mean value of the speed of separation
shall be such that the initial strain rate on the test specimen is 10 mm/mm of the specimen
per minute (for 50 mm specimen gauge length the speed should be 500 mm/min). The
load scale shall be accurate to within one percent or 0‟1 n whichever less was the load
range shall be such that the breaking load of the test pieces fails between I5 percent and
85 percent of the full scale reading.
2) Preparation of Test Pieces - Gauge length of the specimens shall be 50 f 1 mm and the
width shall be 10 to 25 mm. Cut five test pieces from the sample in the lengthwise
direction and a further five in the crosswise direction. The total length of the specimen

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should be at least 50 mm longer than the gauge length. Measure the thickness using a
suitable micrometer.
3) Procedure - Condition the test pieces for not less than one hour at a temperature of 27
5 2°C and 65 f 5 percent relative humidity and clamp their necks in the machine at the
grips separated by 50 mm. Start the machine at the pre-adjusted speed of 500 mm/min
and note the load and elongation at break.
Calculation and Report
1) Tensile Strength at Break - The tensile strength at break shall be calculated in (kgcm2)
from the original area of cross section. The mean of five results shall be expressed for the
lengthwise and CCOSS- wise samples.
2) Elongation at Break - Elongation at break shall be expressed at percentage of the
original length between the reference lines. The mean of five results shall be expressed
for the lengthwise and crosswise sample.

3.9.2 GSM
GSM stands at gram per square meter. It is the weight of fabric in one gram per square
meter. Its unit is gm/m2

3.9.3 Thickness of Packaging material

Thickness of packaging material was calculated using micrometers in micron (um).

3.10 Statistical Analysis

Optimization Response surface methodology which involves design of experiments,
selection of levels of variables in experimental runs, fitting mathematical models and
finally selecting variable levels by optimizing the response (Khuri and Cornell, 1987)
was employed in the study. To optimize the various parameters for the product response
surface methodology (RSM) was used. The experimental data obtained from the design
were analyzed by the package Design-Expert® version 9.0.2 software, Stat-Ease. The full
quadratic equation of the response variables was derived by using RSM as following Eq.

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Y= β0 + β1A1 + β2B2 + β3C3 + β11A1 2 + β22B2 2 + β33C3 2 +

β12A1B2+ β13A1C3 + β23B2C3 … (1)
Where,Y= responses; β0 = constant; β1, β2, β3 = linear regression;
β11, β22, β33 = interaction regression; A1, B2, C3 = variables.

39