Compositional analysis of raw meat:

Moisture, crude protein, fat and ash

percentages were estimated as described by
AOAC, (2002) with some modifications.

3.2.1. Moisture determination:

Moisture content of meat was determined

following the procedure described in
AOAC (2002). In order to determine the
moisture content the china dish was first oven
dried. Afterwards 5g minced sample was taken
in it and placed in hot air oven at 105±20C for
18 hours for drying. After 18 hours sample was
taken out and placed in desiccator for 5-10
minutes and weight. Then sample was weighed
and noted the weight. Then the sample
was again placed in oven till constant weight.
The following formula was used to
calculate the percentage of moisture.

Wt. of original
sample- Wt of dried sample ×
100

Moisture ( % ) ═

Wt. of original
sample

3.2.2. Crude protein

The nitrogen content in meat samples was determined by Kjeldahl’s method as described in
AOAC (2002). In this method nitrogen from protein and other nitrogenous constituents is
converted into to ammonium sulphate by the digestion of proteins with the concentrated H 2SO4.
The ammonium is distilled into standard boric acid after making the digest, alkaline. Percentage
of total nitrogen is calculated and the result is converted to “crude protein” by the multiplication
to a factor of 6.25.

Reagents

H2SO4 95-98%

Digestion tablets 1

Boric acid solution 4%

0.1N NaOH 40%

Hydrochloric acid solution 0.1N

Methyl red indicator

Procedure

i. Digestion

First of all, 3g of dried sample, 1 digestion tablet and 30mL of concentrated H 2SO4 were taken
into Kjeldal’s flask. The whole mixture was heated first at 40 oCfor 10 min and later at 60oC until
the mixture becomes greenish.

ii. Dilution
After temperature equilibration of digested material the contents were diluted to 250mL
with distilled water.
iii. Distillation
10mL of dilution made and 10mL of 40% NaOH was diluted against 4% boric acid
solution (containing methyl red as indicator) in distillation assembly. Ammonia escaped
was collected in boric acid solution with color changing from pink to golden yellow as
end point.

iv. Titration
 After distillation the mixture was titrated againt 0.1N sulphuric acid and volume of
sulphuric acid used was measured. Percentage nitrogen (%) was calculated asunder.
Crude protein was estimated by using the factor of 6.25 for conversion of nitrogen to
protein.

Nitrogen (%) = Titer of 0.1 N H2SO4 used × 0.0014 × 250 x 100

Weight of sample × Vol. of sample

Crude protein content = N % × 6.25

3.2.3. Crude fat determination

The Soxhlet apparatus was used for the determination of crude fat in each sample according to
AOAC (2002). 1g oven dried sample was used to extract the fat content with n-Hexane. Five
siphons back were done for complete extraction of fat from the meat samples. The n-Hexane was
evaporated and recovered with the help of rotary evaporator. The defatted sample was weighed
and crude fat was calculated by the given formula.

Crude Fat (%) = Wt. of sample - Wt. of defatted sample x 100

Wt. of sample

3.2.4. Ash determination

Ash content of meat sample was determined by using method as described in AOAC, (2002).
Meat sample 2±0.005g was taken in a crucible and placed on flame for charring of sample. After
that the sample was placed in muffle furnace at temperature 550 to 650oC until the greenish
material. The sample was cooled in a desiccator and weighed. The ash in the sample was
calculated as under.

Ash (%) = weight of ash in sample (g) x 100

Weight of sample (g)