Chapter – 3 Materials and Methods

he present investigation entitled “Development and Quality Evaluation of Low

T Cost Weaning Formulations from Locally Available Food Materials” was carried
out at Institute of Home Science and Department of Food Science & Technology,
Faculty of Applied Sciences and Technology, University of Kashmir, Srinagar. The
materials and methods are described under the following headings.

3.1. Procurement of Raw Materials

The food commodities - Rice (Oryza sativa), Green gram (Vigna radiata), Banana
(Musa paradisiaca), Potato (Solanum tuberosum) and Apple (Malus domestica) were
used to formulate thirty six composite blends (diets). All the ingredients were purchased
from local markets of Srinagar city.

The diets were prepared by blending all the ingredients in different proportions. Rice,
potato and banana were used as major ingredients of the mixture. Protein source was
derived from green gram. Apple was supplemented to the mixture to provide vitamins
and minerals.

3.2. Preparation of Ingredients

3.2.1. Preparation of Rice Flour

Rice grains were cleaned to remove extraneous material and admixture of other food
grains. The grains were washed, soaked in water for 2 hours, drained, and dried in
ambient temperature followed by grinding in the electric grinder (Sujata, Model
Powermatic plus) to make fine flour and sieved by 80-100 mesh sieves and then
packaged in airtight plastic container till further use (Usha et al., 2010). The flow chart
for preparation of rice flour is given in Figure. 3.1.

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Chapter – 3 Materials and Methods

Rice

Clean and Sort

Soak (2 hours)

Air dried

Grinding

Sieving

Rice Flour

Fig. 3.1: Flow chart for preparation of rice flour

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Chapter – 3 Materials and Methods

3.2.2. Preparation of sprouted green gram flour

3.2.2.1 Cleaning

Green gram (Vigna radiata) was purchased from local market, cleaned to remove
extraneous material. Cleaning was done by winnowing and hand sorting.

3.2.2.2. Steeping/soaking

The cleaned green gram seeds were washed three times in excess distilled water. Then,
the cleaned and washed green gram seeds were soaked in a volume of water 3 times
the weight of seeds (3:1) for 12 hours in a container at ambient temperature (Almeida-
Dominguez et al., 1993).

3.2.2.3. Sprouting

The traditional sprouting procedures were adapted to laboratory conditions as follows:

the steeping water was drained off and the soaked green gram seeds were washed twice
using distilled water to prevent the growth of microorganisms during sprouting. The
soaked seeds were wrapped in damp muslin cloth to stimulate sprouting. The green
gram seeds were allowed to sprout for 42 hours. The content was left in ambient
conditions and watered 2-3 times a day to enhance the sprouting process. The sprouts
were dried overnight at room temperature by keeping under a fan.

3.2.2.4. Milling

The sprouted grains were ground in an electric grinder (Sujata, Model Powermatic plus)
to make fine powder and sieved by 80-100 mesh sieve. The milled sample was then
packed in airtight plastic container. The container was stored at room temperature until
further use. The procedure is as shown in Fig. 3.2.

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Chapter – 3 Materials and Methods

Green Gram

Clean and Sort

Soak (12 hour) Grinding

Draining of the Steeping Sieving

Water

Unsprouted Green Gram

Germinate (42 hour)
Flour

Air dried

Splitting & Dehusking

Grinding

Sieving

Sprouted Green Gram Flour

Fig. 3.2: Flow chart for preparation of green gram flour

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Chapter – 3 Materials and Methods

3.2.3. Preparation of potato flour

Potato (Solanum tuberosum) was washed, peeled and hand trimmed with stainless steel
knife. Peeled potatoes were cut into four quarters, then pressure cooked (Prestige TTK)
in water. Boiled pieces were mashed and dried overnight in an oven (Yorks hot air
oven) at 70ºC by spreading as a thin film. After complete drying, the mashed potatoes
were milled, passed through 80-100 mesh sieves to obtain fine flour of uniform size
(Gahlawat and Sehgal, 1998). The flour was packed in airtight plastic containers until
further use. The flour was packed in airtight plastic containers until further use (Fig.
3.3)

Potato

Wash/peel/slice

Pressure Cook

Mashing

Loading on Trays

Drying in hot air oven

Grinding

Sieving (80-100 mesh)

Potato Flour

Fig. 3.3: Flow chart for preparation of potato flour

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Chapter – 3 Materials and Methods

3.2.4. Preparation of Apple Pulp

Fresh, mature and healthy apple was harvested from local gardens. Red Delicious
variety of apples was used for the pulp. The fruit was thoroughly washed to remove
dirt, dust, pesticide residues and microflora on the surface of the fruit. It was washed,
peeled, cored and sliced. Apple was crushed with the help of home-scale mixer-cum-
juicer (Sujata Model Powermatic plus) for obtaining homogenized pulp (Fig. 3.4.) and
the pulp was subjected to different pretreatments that include:

Table 3.1: Treatment combinations were as follows:

Treatments Preservatives (g/kg) of apple pulp

T₁ Control (no preservative, no pasteurization)

T₂ 0.1% potassium metabisulphite + 0.1% citric acid

T₃ Pasteurization at 65°c for 30 min.

T₄ Pasteurization at 65°c for 30 min + preservative (0.1% potassium

metabisulphite + 0.1% citric acid)

The treated pulp samples were transferred to sterilized glass bottles and stored under
ambient conditions (25°C) for a period of 90 days and analysis were carried out after
every 15 days (Durrani et al., 2010).

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Chapter – 3 Materials and Methods

Apple

Washing

Peeling/Coring

Cutting

Pulping

Addition of 0.1% KMS + Bottling Addition of 0.1% KMS + Bottling

0.1% Citric acid 0.1% Citric acid

Corking Corking
Mixing thoroughly
Mixing thoroughly

Bottling
Bottling Wiping Thermal processing

Cooling
Corking
Corking Labelling
Wiping
Thermal processing
Wiping
Storage at Labelling
ambient Cooling
temperature
Labelling
Wiping Storage at
ambient
temperature
Labelling
Storage at
ambient
temperature
Storage at
ambient
temperature

Fig. 3.4: Flow chart for preparation of apple pulp

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Chapter – 3 Materials and Methods

3.2.5. Preparation of banana pulp

Undamaged bananas were purchased from local market of Srinagar. Bananas were
peeled manually and cut into pieces. The banana was pulped with the help of a
laboratory scale blender at lowest speed for 5 minutes (Sujata, Model Powermatic plus)
for obtaining homogenized pulp (Fig. 3.5.) and the pulp obtained was mixed with
chemical preservatives as per treatment combination presented in Table 3.2. The treated
pulp samples were transferred to sterilized glass bottles and stored under ambient
conditions (30-40°C) for a period of 90 days and analysis were carried out after every
15 days (Durrani et al., 2010).

Table 3.2: Treatment combinations were as follows:

Treatments Preservatives (g/kg) of banana pulp

T₁ Control (no preservative, no pasteurization)

T₂ 0.1% potassium metabisulphite + 0.1% citric acid

T₃ Pasteurization at 65°c for 30 minutes

T₄ Pasteurization at 65°c for 30 minutes + preservatives (0.1% potassium

metabisulphite + 0.1% citric acid)

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Chapter – 3 Materials and Methods

Banana

Washing

Peeling/Coring

Cutting

Pulping

Addition of 0.1% KMS + Bottling Addition of 0.1% KMS + Bottling

0.1% Citric acid 0.1% Citric acid

Corking Corking
Mixing thoroughly
Mixing thoroughly

Bottling
Bottling Wiping Thermal processing

Cooling
Corking
Corking Labelling
Wiping
Thermal processing
Wiping
Storage at Labelling
ambient Cooling
temperature
Labelling
Wiping Storage at
ambient
temperature
Labelling
Storage at
ambient
temperature
Storage at
ambient
temperature

Fig. 3.5: Flow chart for preparation of banana pulp

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Chapter – 3 Materials and Methods

3.3. Formulation of Weaning Foods

The formulation of weaning foods is generally based on this assumption that the child
consumes an average of 100g per day. Thirty-six formulations (Diet 1-Diet 36) of
weaning food were developed from the prepared flour/pulp from rice, potato, green
gram, apple, banana and sugar. Liquid base (water or milk) to ingredients was added to
obtain formulation of desirable consistency. The compositions of formulation are
shown in table 3.3, 3.4 & 3.5 and the flow charts are shown in figures 3.6, 3.7 & 3.8.

Table 3.3: Formulation of Rice based weaning foods

Weaning Water Milk Rice UGG SGG Apple Sugar

mixes ml/100g ml/100g
DIET 1 1060 - 70 10 0 10 10

DIET 2 960 - 60 20 0 10 10

DIET 3 880 - 50 30 0 10 10

DIET 4 1060 - 70 0 10 10 10

DIET 5 960 - 60 0 20 10 10

DIET 6 880 - 50 0 30 10 10

DIET 7 - 1060 70 10 0 10 10

DIET 8 - 960 60 20 0 10 10

DIET 9 - 880 50 30 0 10 10

DIET 10 - 1060 70 0 10 10 10

DIET 11 - 960 60 0 20 10 10

DIET 12 - 880 50 0 30 10 10

UGG: Unsprouted green gram flour, SGG: Sprouted green gram flour

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Chapter – 3 Materials and Methods

Rice Flour Green Gram Flour Apple Pulp

Milk or Water Blending Sugar

Mixing

Rice Based Weaning

Foods

Fig. 3.6: Flow diagram of weaning formulations prepared from rice, green gram,
apple, milk and sugar

Table 3.4: Formulation of Potato based weaning foods

Weaning Water Milk

Potato UGG SGG Apple Sugar
mixes ml/100g ml/100g
DIET 13 800 - 70 10 0 10 10
DIET 14 720 - 60 20 0 10 10
DIET 15 640 - 50 30 0 10 10
DIET 16 800 - 70 0 10 10 10
DIET 17 720 - 60 0 20 10 10
DIET 18 640 - 50 0 30 10 10
DIET 19 - 800 70 10 0 10 10
DIET 20 - 720 60 20 0 10 10
DIET 21 - 640 50 30 0 10 10
DIET 22 - 800 70 0 10 10 10
DIET 23 - 720 60 0 20 10 10
DIET 24 - 640 50 0 30 10 10
UGG: Unsprouted green gram flour, SGG: Sprouted green gram flour

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Chapter – 3 Materials and Methods

Potato Flour Green Gram Flour Apple Pulp

Milk or Water Blending Sugar

Mixing

Potato Based
Weaning Foods

Fig. 3.7: Flow diagram of weaning formulations prepared from potato, green
gram, apple, milk and sugar

Table 3.5: Formulation of Banana based weaning foods

Weaning Water Milk

Banana UGG SGG Apple Sugar
mixes ml/100g ml/100g
DIET 25 600 - 70 10 0 10 10
DIET 26 680 - 60 20 0 10 10
DIET 27 760 - 50 30 0 10 10
DIET 28 600 - 70 0 10 10 10
DIET 29 680 - 60 0 20 10 10
DIET 30 760 - 50 0 30 10 10
DIET 31 - 600 70 10 0 10 10
DIET 32 - 680 60 20 0 10 10
DIET 33 - 760 50 30 0 10 10
DIET 34 - 600 70 0 10 10 10
DIET 35 - 680 60 0 20 10 10
DIET 36 - 760 50 0 30 10 10

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Chapter – 3 Materials and Methods

Banana Pulp Green Gram Flour Apple Pulp

Milk or Water Blending Sugar

Mixing

Banana Based
weaning Foods

Fig. 3.8: Flow diagram of weaning formulations prepared from banana, green
gram, apple, milk and sugar

3.4. Proximate Analysis of Formulated Diets

Proximate analysis: Raw ingredients and formulated diets were analyzed for moisture,
fat, protein, ash, carbohydrate and energy. All analysis was done in triplicates.

3.4.1. Determination of Moisture Content

One of the standard reference methods for moisture determination is the oven drying
method (AOAC 1990, method 14.004). This involved drying to a constant weight at
100 oC and calculating moisture as the loss in weight of the dried samples. The
crucibles were thoroughly washed and dried in an oven at 100 oC for 30 min and
allowed to cool inside desiccators. After cooling, they were weighed using weighing
balance and their various weights recorded as (W1). Then, 2.0 g of the samples were put
into the crucibles and weighed to get W2. Thereafter, the sample plus crucible were
placed inside the oven and dried at 100 oC for 16-18 hours. After drying, the crucibles
were transferred to the desiccator to cool. The crucible and its dried sample were

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Chapter – 3 Materials and Methods

reweighed until the constant weight was reached to get W3. Then, the moisture content
of the samples was calculated from the equation:

( )
( )
( )

Where, W1 = Initial weight of empty crucible,

W2 = Weight of crucible + sample before drying and

W3 = Final weight of crucible + sample after

3.4.2. Determination of Ash Content

Total ash of the samples was determined by Furnace Incineration method (AOAC, 2000
method 14:006) based on the vaporization of water and volatiles with burning organic
substances in the presence of oxygen in the air to CO 2 at a temperature of 550 oC.
About 2g of sample was weighed into a porcelain crucible and incinerated at 550 oC for
6 hr in an ashing muffle furnace (Model 1184A Fisher Scientific, Houston, TX) until
ash was obtained. The ash was cooled in a dessicator and reweighed. The % ash content
in the samples was calculated from the equation:

( )

3.4.3. Determination of crude fat content

Fat content was determined by using the Soxhlet extraction method (AOAC, 1990). The
sample (1g) was taken into pre-weighed thimbles. The thimbles were plugged with a
thin layer of cotton and kept in a beaker for drying in an oven at 100 °C for 8h. The
thimbles were then transferred to the Soxhlet apparatus containing petroleum ether (60-
80 °C B.P.) along with the pre-weighed aluminum canisters. It was allowed to run for
about 2-3 hours at the rate of 2-3 drops/sec. The aluminum canisters were removed and
kept in hot air oven for evaporation of residual solvent and transferred to desiccators for
cooling and weighed. The process of drying and cooling was continued till constant
weight was achieved. The percent ether extract was then calculated and expressed as
percent Fat.

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Chapter – 3 Materials and Methods

Where, W1 = Net wt. (g) of sample

W2 = weight (g) of empty canister.

W3 = weight (g) of canisters with fat

3.4.4. Crude protein determination

The micro-Kjeldahl method for protein determination is employed for protein

determination. This is based on three principles:

Digestion: RNH2 + 2H2SO4  (NH4)2 + SO4 + CO2 + H2O

Distillation: (NH4)2 SO4 + 2NaOH  RNH3 + 2H2O + Na2SO4

Absorption: 3NH3 + H3BO3  (NH4)2BO3-

Titration: (NH4)3 BO3 + HCl  H3BO3 + 3NH4Cl

Digestion

The sample (0.5g) was weighed into the micro-Kjeldahl flask. Potassium sulphate
(5gm) and cupric sulphate (1gm) were added to the Kjeldahl flask. After placing
Kjeldahl tube on digestion bench and solution was heated till it became green. The
tubes in the rack were cooled in a fume hood till the solution became colorless.

Distillation

The digested sample was carefully transferred into the steam distillation apparatus
followed by addition of 20mls NaOH solution (40%) and boric acid (4%). Distillation
was carried out for 15 minutes and the liberated ammonia was trapped into boric acid in
a 100ml conical flask containing 2-3 drops of methyl red indicator.

Titration

The ammonia trapped was then titrated against 0.1N HCl solution. End point of titration
is indicated by appearance of wine colour, which indicates that, all the nitrogen trapped
as ammonium borate has been removed as ammonium chloride (AOAC, 1990).

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Chapter – 3 Materials and Methods

Calculation

The percentage nitrogen was calculated by using the formula:

( – )
( ) Eq. 3.4

The crude protein is determined by multiplying percentage nitrogen by a constant factor

of 6.25.

Protein (%) = N (%) x 6.25

3.4.5. Determination of total carbohydrate content

Total percentage carbohydrate was determined by the difference method as reported by

Omemu (2011). This method involves adding the total values of crude protein, crude
fat, and ash constituents of the sample and subtracting it from 100. The value obtained
is the percentage carbohydrate constituent of the sample. The formula for the
calculation of percentage carbohydrate is shown as:

Carbohydrate (%) = 100 - (fat% +protein% + ash% ) Eq. 3.5

3.4.6. Determination of gross energy content

The energy values of the weaning food formulations were determined by computation
and expressed in calories. It was calculated from fat, carbohydrate and protein contents
using the Atwater‟s conversion factors:

1Kcal/100g= [(4×carbohydrate) + (4×protein) + (9× fat)] Eq. 3.6

3.5. Physico-Chemical Analysis

3.5.1. Determination of pH

The pH of formulated diets was determined by using digital pH meter (Inolab WTW
Series 720). The pH meter was first calibrated using buffer of pH 4.0 and 7.0 at room
temperature. The sample was then taken in the 100ml beaker, stirred and electrode of
pH meter put in it and direct reading from pH meter was taken when the reading
stabilized.

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Chapter – 3 Materials and Methods

3.5.2. Determination of Titrable acidity

According to (AOAC, 2000), to determine the titrable acidity, 10 ml of sample was

taken and homogenized with distilled water in a blender. The whole mass was then
transferred to a 100 ml volumetric flask and the volume was made up to the mark with
distilled water. 10ml sample solution was taken in a conical flask. 2 or 3 drops of
phenolphthalein indicator were added and then flask was shaken vigorously. It was then
titrated immediately with 0.1 N NaOH solutions from a burette till a permanent pink
color appeared. The volume of NaOH solution required for titration was noted and
percent of titrable acidity was calculated.

( )
Eq.3.7

3.5.3. Determination of Total Soluble Solid

A hand-held refractometer (N-1E, °Brix 0-32%, ATAGO Co., Ltd, Tokyo, Japan) was
used to determine the total soluble solid by calibrating with distilled water followed by
dropping 1-2 drops of sample solution onto the clean surface of the refractometer
(Ahmed et al. 2011).

3.5.4. Determination of specific gravity

Specific gravity of the sample was determined according to AOCS method (1993). The
specific gravity bottle was placed in a water bath maintained at 25˚C and filled with
distilled water. It was removed, wiped dry (outside the bottle) and weighed. The bottle
was emptied, dried and again placed in water bath at 25˚C. The bottle was refilled with
the sample and made to stay in the water bath for 30 minutes. It was then removed,
cleaned and wiped (outside the bottle) completely dry and weighed. The specific
gravity at 25˚C was calculated using the equation
( – )
Eq. 3.8

3.5.5. Determination of sugars

Lane and Eynon (1923) method as detailed by Ranganna (1991) was followed.
Measured quantity of pulp (20ml) was taken in 250ml volumetric flask to which about
100ml distilled water was added and neutralized with 40% sodium hydroxide using
phenolphthalein as an indicator and clarified with 2ml neutral lead acetate (45%) for

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Chapter – 3 Materials and Methods

about 30 minutes. Excess of lead was removed with 5ml potassium oxalate (22%). Then
the volume was made to 250ml with distilled water and filtered through whatman No.4
filter paper. The filtrate was titrated against 10ml of standardized Fehling‟s solution
using Methylene blue as indicator to a brick red color, for determination of reducing
sugars.

Reducing sugars: The filtered sample was taken in 50ml burette and was titrated
against 10 ml of standardized Fehling‟s solution using Methylene blue as indicator to a
brick red color, for determination of reducing sugars and was calculated as

( ) Eq. 3.9

3.6. Determination of Invitro-Protein Digestibility (IVPD) by Pepsin-

Pancreatin Method

In-vitro protein digestibility was measured according to Saunders et al. (1973) with
some adjustments. 250 mg of the sample was precisely weighed and placed into a 50-ml
centrifuge tube. 15 ml of 0.1 N HCl containing1.5 mg of pepsin was added and the tube
was incubated at 37°C for 3 h. The suspension was then neutralized with 3.3 ml of 0.5
M NaOH and treated with 4 mg of pancreatin in 7.5 ml of 0.2 M phosphate buffer (pH
7.9). The mixture was then gently shaken and incubated at 37°C for 24 h. After
incubation, the sample was treated with 10 ml of 10% trichloroacetic acid and
centrifuged at 14,000 rpm in Beckman centrifuge for 5 min at 11°C. The supernatant
was discarded and the pellet was air dried over-night and used to analyze the indigested
protein fragment. The octuplicate was combined together, in order to obtain ~2 g of
sample. The nitrogen in the indigested fragments was estimated using the Kjeldahl
method as described before. The following calculation used for protein digestibility is
as given below.

Digestibility was calculated using the formula:

–
Eq. 3.10

The total protein content was obtained from the crude protein analysis.

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Chapter – 3 Materials and Methods

3.7. Determination of Color

The Color of the diets was measured using a Hunter‟s Lab colour analyzer (MiniScan
XE Plus, Model 45/0-S, Hunter Associates Laboratory Inc., Reston, VA, USA). The
equipment was calibrated using white and black standard ceramic tiles. In the Hunter‟s
lab colorimeter, the colour of a sample is denoted by the three dimensions, L*, a* and
b*. L*, refers to lightness of the colour of the diets and ranges from black = 0 to white =
100. A negative value of a* indicates a green colour where the positive value indicates
red-purple colour. A positive value of b* indicates a yellow colour and the negative
value a blue colour. ΔE* is a parameter that quantifies the overall color difference of a
given sample compared to a reference sample. It was calculated using the equation:

ΔE* = √ (ΔL*2 + Δa*2 + Δb*2) Eq. 3.11

Where, ΔL* = (L*₁− L*o); Δa* = (a*₁− a*o); and Δb* = (b₁*− bo*).

Subscript „o‟ depicts the color value for reference sample (untreated sample) and
subscript „1‟ depicts the color value for the sample being analyzed (Caivano, 2012). C*
coordinate is the chroma [Eq. (1) and H* coordinate the hue (Eq. (2)] (Wrolstad &
Smith, 2010).

Chroma C* = √ (a*) 2 + (b*) 2 Eq. 3.12

Hue angle H° = tan-1 { *} Eq. 3.13

3.8. Determination of Anti-nutrients

3.8.1. Oxalate analysis

Oxalate was determined by AOAC (1990) method. 1g of the sample was weighed into
100ml conical flask. 75ml of 3M H₂SO₄ was added and the solution was carefully
stirred intermittently with a magnetic stirrer for about 1h and then filtered using
whatman No. 1 filter paper. The sample filtrate (extract) (25 ml) was collected and
titrated against hot (80-90°C) 0.1N KMnO₄ solution to the point when a faint pink color
appeared that persisted for atleast 30 s. The concentration of oxalate in each sample was
obtained from the calculation: 1 ml 0.1 permanganate = 0.006303 g oxalate.

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Chapter – 3 Materials and Methods

3.8.2. Phytic acid analysis

The phytic acid (or phytate) content was measured by the method of Thompson and
Erdman, (1982). The ground sample, 2g was extracted with 40 ml of 0.5 mol/L HNO₃
for 2 hour and then filtered. The solution, 1 ml was used for the total phosphorus
determination. Ferric chloride was added to another 10 ml of solution and heated in
boiling water for 75 min. After centrifugation at 12,000 ×g for 15 min, the supernatant
was used to determine the soluble phosphorus. Total and soluble phosphorous levels
were determined colorimetrically using 0.05 mol/L ammonium thiocyanate and were
estimated using a phosphorus standard curve. The difference between total and soluble
phosphorus was insoluble phosphorus. The phytic acid content was calculated from the
insoluble phosphorus; assuming 1 ml of phytic acid contains 6 mol of insoluble
phosphorus.

3.9. Determination of Viscosity

Viscosity was measured using a Brookfield Viscometer (Model LV DV-E 230;

Middleboro, Massachusetts, USA). The cooked gruel was poured into the viscometer
beaker, cooled to 40°C and viscosity was measured in centipoises, cP, using spindle
number 62 at a shear rate of 30 revolution per minute (RPM). Within 3 minute, the
average of the maximum and minimum viscosity reading was recorded according to the
speed.

3.10. Sensory Evaluation

The prepared gruel samples were presented to 20 weaning mother panelists. The
panelists were given the fundamental information about the purpose and objective of
the test, so that the selection of panelists was based on basic requirements of a panelist,
such as availability for the entire period of evaluation, interest, willing to serve. The
health status of the panelists was also considered during panelist selection (not suffering
from colds and allergies that affect their sensitivity for the product). The gruels were
served to the panelists in white and transparent glass cups at about 40°C and asked to
rinse their mouth with fresh room-temperature water that was provided to them, before
next serving. The containers with the samples were coded in three digits and kept far
apart to avoid crowding and for independent judgment. The panel members were seated
in an open well illuminated laboratory (independent booth) and asked to rate the gruel

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Chapter – 3 Materials and Methods

samples based on appearance (color), flavor, odor, and texture (mouth-feel), using a
nine point hedonic scale (where 1 = dislike extremely and 9= like extremely). Overall
acceptability of the samples was also rated on same scale with 9 = extremely acceptable
and 1 = extremely unacceptable.

3.11. Estimation of Cost

The cost of formulated weaning foods were determined according to the price of the
materials used in preparing the foods, gas, electricity, jars, water and others. The cost of
the prepared weaning foods was compared with market price of the three imported
commercial baby foods at the same time.

3.12. Storage of Weaning Formulations

The samples were stored in the laboratory for 3 days in sealed plastic containers at
ambient and refrigerated temperature. The analysis of formulations for pH, titrable
acidity and various sensory parameters were conducted at 8 hr storage interval.

3.13. Statistical Analysis

Results were expressed as mean of triplicate analyses. A one-way & two-way analysis
of variance and Duncan's test were used to establish the significance of differences
among the mean values at the 0.05 significance level. The statistical analyses were
performed using SPSS software (Systat statistical program version 21 (SPSS Inc.,
USA).

72