Experiment 1 Determination of titratable acidity of milk powder

Principle:-
Acidity of milk powder is estimated by titration with N/10 NaOH. The titratable acidity
test is employed to ascertain if milk powder is of such a high acidity as to reduce its
keeping quality and heat stability. The acidity of milk powder is not a true measure of
lactic acid present but in practice, gives a good indication of the quality of it. In practice
the titration method is employed and measures the buffering capacity of milk powder
and true acidity.
Apparatus:-
conical flask, glass rod, burette, pipette – 10 ml (volumetric), pipette – 10 ml
(graduated), pipette – 2 ml (graduated)
Reagents:-
Standard NaOH solution, phenolphthalein indicator
Procedure:-
Standard NaOH solution: – 0.1 N Prepare a concentrated stock solution of sodium
hydroxide by dissolving equal parts of sodium hydroxide (Sticks or pallets) in equal
parts of water in a flask. Tightly stopper the flask with a rubber bung and allow any
insoluble sodium carbonate to settle out for 3-4 days. Use the clear supernatant liquid
for preparing the standard 0.1 N solution. About 8 ml stock solution is required per litre
of distilled water.

Phenolphthalein indicator – Dissolved one gram of phenolphthalein in 100 ml of 95 %

ethyl alcohol. Add 0.1N sodium hydroxide solution until one drop gives a faint pink
colouration. Dilute with distil water to 200 ml.

Procedure:-
1. Weigh accurately about 1 g of the sample into a conical flask.
2. Add 10ml of boiling water and stir with the flat end of a glass rod until a perfectly
smooth liquid is obtained. Cool to room temperature.
3. Add 1ml of phenolphthalein indicator solution and titrate the contents against
standard sodium hydroxide solution added drop by drop from the burette until the
appearance of pink color.

Calculation: - Titratable acidity (% lactic acid) = 9 V1 N

V2
Where V1 = volume in ml of the standard sodium hydroxide required for
titration

N = normality of the standard sodium hydroxide solution and

V2 = volume in ml of milk taken for the test

Results: Titratable acidity of the given milk powder is................ %

 Experiment 2 Determination of moisture content in milk powder
Principle: - Moisture content is one of the most commonly measured properties of dried
foods. The moisture content is important due to the legal and labelling requirements and
to maintain the food quality, microbial stability and cost of the products. Knowledge of
the moisture content is also necessary to predict the behaviour of foods during
processing. According to FSSR, 2011 moisture content in the dried milk food should not
be more than 4% m/m for whole milk powder and 5% m/m for partially skimmed and
skimmed milk powder.

Apparatus:-
Flat-bottom dishes with lid: Dishes of nickel, aluminium or of other suitable metal not
affected by boiling water, 70 to 80 mm in diameter and not more than 25 mm deep,
provided with short glass stirring rod having a widened flat end. The dishes shall have
lids which fit well and can readily be removed, Hot air oven maintained at 102 ± 1°C/
Vacuum Oven: maintained at 50 – 55 0C, Desiccator: containing an efficient desiccant,
weighing balance

Procedure (by hot air oven):

1. Uncover a dish and place the dish and its lid in a hot air oven at 102 ± 2°C for 1 h.
Place the lid on the dish, transfer the covered dish from the hot air oven to the
desiccator. Allow it to cool to room temperature and weigh it.
2. Approximately 1.0 g of powder was weighed in a moisture dish, covered the dish
with lid and the weight was recorded.
3. Uncovered the dish and placed the dish and its lid in the oven at 102 ± 2°C for 2h.
4. After drying, replaced the lid and transferred the covered dish to the desiccator.
Allowed it to cool to room temperature and accurately weighed the dish. The
process was repeated until successive weighing do not differ by more than 0.5 mg.
Calculation:
𝑀₁ − 𝑀₂
Moisture content (%) = ( ) × 100
𝑀₁ − 𝑀
Where,
M = Mass of the empty dish (g)
M1 = Initial mass of the dish and lid with the material taken for analysis in g
M2 = Final mass of the dish and lid with the material after drying (g)
Experiment 3 Determination of total ash content in milk powder
Principle:-

Sample is kept in a muffle furnace maintained at 550+/-100C to remove organic matter

leaving behind the ash expressed as percentage.
Apparatus:-
Flat bottom dish-stainless steel, porcelain, silica or platinum. muffle furnace- maintained
at 550+/-10 0C and desiccator

Procedure:-
1. Weigh accurately about 3 g of the previously dried material in a dish.
2. Heat the dish gently on the flame till it became whitish in color.
3. Place the dish in a muffle furnace at 550+/-200c till gray ash results.
4. Cool the dish in a desiccator and weigh.
5. Repeat this process of heating for 30 minutes, cooling and weighing until the
difference between two successive weighing is less than one milligram.
6. Record the weight.

Calculation:-

Total ash (% by weight) = 100(W2-W1)

(W1-W)
Where W2 = weight in g of the dish with the ash,
W = weight in g of the empty dish,
W1 = weight in g of the dish with material taken for the test.

Results:-
Total ash of the given sample of milk powder is................ %
Experiment 4 Determination of fat content in milk powder by Mojonnier method
Principle:-
The milk powder is treated with hydrochloric acid solution and ethyl alcohol; the former
to dissolve the protein and the latter to help precipitate the proteins. Fat is extracted with
diethyl ether and petroleum ether. Mixed ethers are evaporated and the residue
weighed. This method is considered suitable for reference purposes. Strict adherence to
details is essential in order to obtain reliable results.

Apparatus:- Mojonnier fat extraction flask or any other suitable extraction tube (as per
IS specification), cork or stopper of synthetic rubber unaffected by usual fat solvents,
100 ml flat bottom flask with G/G joint or stainless steel or aluminium dishes of 5.5 cm
height and 9 cm diameter or glass bowl, analytical balance, drying oven, boiling water
bath

Reagents:-
Hydrochloric acid solution (density 1.125 g/ml at 20°C) – Dilute 675 ml of conc.
hydrochloric acid solution to 1000 ml; ethanol – 95 to 96 % v/v; diethyl ether - free from
peroxides, light petroleum ether boiling range 40 to 60°C.
Procedure:-
1. 2 gm of sample was transferred into Mojonnier flask (the test portion should be
delivered as completely as possible into the lower (small) bulb of the extraction
flask).
2. 10ml of hydrochloric acid solution was added to wash the test portion into the
small bulb of the extraction flask.
3. Mojonnier flask was gently heated in a boiling water bath (avoid charring) until
all the particles were dissolved completely. Allowed the flask to stand for 20-30
min in boiling water bath and then cooled them under running tap water.
4. 10 ml of ethanol was added and mixed gently but thoroughly by allowing the
contents of the flask to flow backward and forward between the two bulbs.
5. 25 ml of diethyl ether was added, closed the flask with pre wetted cork (dipped
in water at 60-700C for 10-15 min) and mixed the content properly.
 6. 25 ml of the petroleum ether was added and again repeated the previous step.
7. Allowed the content to stand for 30 – 45 min at room temperature (30°C) under
undisturbed condition till the upper layer is clear.
8. Carefully removed the cork (rinse it inside of the neck of the flask with mixed
solvent so that rinsing runs into the flask) and decanted the ether solution from
Mojonnier flask into the pre weighed conical flask.
9. Outside of the neck of extraction flask was rinsed with a little of the mixed
solvent and collected the rinsing in the conical flask.
10. Evaporated the solvent by keeping the conical flask over boiling water bath.
11. 5 ml of the ethanol was added to the contents of the Mojonnier flask and mixed
as described in step 4.
12. Second extraction was carried out by repeating the operations as described in
step 5 to 10.
13. Third extraction was carried out as described in step 5 to 10 but using 15ml of
diethyl ether and 15ml of petroleum ether.
14. After extraction, the solvent in the conical flask was evaporated in hot air oven
at 102 ± 2°C for 1 hour.
15. After incubation, the fat containing dish was removed from oven, cooled and
weighed.
16. Blank test was carried out simultaneously as described in the step 2 to 15, while
the test portion was replaced with 10 ml of water.

Calculation:-
Fat % (w/w) = weight of extracted fat /weight of powder X 100

Results: Fat % (w/w) in the given milk powder is................ %

 Experiment 5 Determination of fat content in powder by Gerber Method
Principle:-

When a definite quantity of sulphuric acid and amyl alcohol are added to a definite
amount of powder, the proteins will be dissolved and the fat globules will be set free,
which remains in liquid state due to heat produced by the acid. On centrifugation, fat
being light will be separated on top of the solution.

Apparatus and glassware:- Analytical balance, Butyrometer (0-10%, milk) with lock
stopper and key, Butyrometer (0 – 35 %,powder) with lock stopper and key, constant
temperature water bath, centrifuge machine, tilt measure – 10 ml and 1 ml, spatula, lock
stopper, lock stopper pin

Chemicals and Reagents:-

sulphuric acid (90-91%), 1.807-1.812 specific gravity at 270C, iso-amyl alcohol, 0.804 –
0.810, specific gravity at 200C, distilled water

Procedure:-
1. Take 10 ml of Gerber acid in milk / powder butyrometers.
2. Add carefully 0.5 to 1.0 ml water (70 0C) to Gerber acid.
3. Take 1.69 g powder for milk butyrometer / 2.5 g for powder butyrometer.
4. Add 1.0 ml of amyl alcohol.
5. Make up the volume with water (60-70 0C)
6. Shake carefully keeping the lock stopper away from the body.
7. Centrifuge at 1200 rpm for 5 minutes.
8. Read the fat column directly in case of powder butyrometer and in case of milk
butyrometer calculate fat as given below.

Calculations:-
% Fat = 6.67 X Reading of fat column in milk butyrometer

Results: Fat % (w/w) in the given milk powder is................ %

Experiment 6 Determination of solubility in milk powder

Principle:-

Milk powder is dissolved in water and centrifuged. Amount left in the supernatant is
dried and expressed as percentage.

Apparatus:-
Centrifuge tube, water bath, vortex vibrator, laboratory oven

Procedure:-
2. Take 1 g of powder in a glass centrifuge tube.
3. Add 10 mL of distilled water and stirred vigorously in vortex vibrator for 1 min.
4. It was then thermostated in a water bath at 37 °C for 30 min and centrifuged at
3000 rpm for 10 mins.
5. The obtained supernatant was collected and dried in a laboratory oven at 100°C
until constant mass was realised.

Calculations:-

Solubility was calculated according to the following equation:

Solubility= (ms/mp) x 100,
Where, ms is the mass (g) obtained by drying of the supernatant and m p is the mass (g)
of the powder taken for analysis

Results:-
Solubility of the given sample of milk powder is................ %
 7. Bulk Density
Bulk density is the measure of the mass of milk powder which occupies a fixed volume.
The bulk density of normal spray dried powder may vary between 0.5-0.8g/ml. It affects
the powder flowability and storage stability.
There are two types of bulk density
Loose Bulk Density
Procedure:-
1. A 100ml graduated cylinder is weighed and tare weight of the cylinder is taken.
2. A funnel with short stem is placed over the cylinder opening.
3. Dry powder is allowed to flow freely to the funnel up to 100ml mark.
4. The weight of the cylinder along with the powder is recorded.
5. The net weight of 100ml powder is calculated and the result is expressed as
g/ml.

8. Tapped Bulk Density

Procedure:-
1. After finding the loose bulk density, we have a cylinder with 100ml powder in it.
2. The powder is then tapped manually approximately 50 times on a marble solid surface
from a height of 10 cm to measure the final volume till a constant volume is obtained.
Calculation:
(W₂ − W₁)
𝐿𝑜𝑜𝑠𝑒 𝐵𝑢𝑙𝑘 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 (%) =
100
(W₂ − W₁)
𝑇𝑎𝑝𝑝𝑒𝑑 𝐵𝑢𝑙𝑘 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 (%) =
𝑉1
Where, W1= weight of graduated cylinder
W2 = weight of cylinder + powder
V1= volume after tapping
Bulk and tapped bulk density are associated with the powder's flowability and storage
stability. High bulk density is desirable because it involves reduced packaging, storage
and transport costs per kg of powder
9. Flowability
Flowability of powder refers to the ease with which the powder particles move with
respect to one another. This is an important but complex property. It is a measure of
free flow characteristics of powder. Proper flow of powder is important for manufacture
of powder especially for conveying and packaging, handling and measuring. In general,
powders with good flow properties are those with large agglomerates and few fines.
Using the Carr index and Hausner ratio (HR), the flowing characteristics of the
milk powder are determined. Carr's index measures the compressibility or free-flowing
property; while HR measures the powder's cohesiveness. The Carr's index (C) and
Hausner ratio (HR) were calculated using bulk and tapped density by the following
formulas:-
𝑇𝑎𝑝𝑝𝑒𝑑𝑏𝑢𝑙𝑘𝑑𝑒𝑛𝑠𝑖𝑡𝑦(𝜌T ) − 𝑏𝑢𝑙𝑘𝑑𝑒𝑛𝑠𝑖𝑡𝑦(𝜌B )
Carr ′ s Index (C) = ∗ 100
Tapped bulk density(𝜌T )

𝑇𝑎𝑝𝑝𝑒𝑑 𝑏𝑢𝑙𝑘 𝑑𝑒𝑛𝑠𝑖𝑡𝑦(𝜌T )

Hausner Ratio (HR) =
Bulk density(𝜌T )
Carr's index (% compressibility) and Hausner’s ratio are generally used as the index of
flowing properties of dry powders. Materials with a Hausner ratio of more than 1.34 are
regarded to be cohesive and consequently less free to flow. As per Carr, materials non-
free-flowing powders have Carr’s index of greater than 25 %.