Testing Methods Of Casein protein powder | Solubility of Casein Protein Powder in Water

How to check casein Solubility in dm water and  ash content of casein and test fat content of casein.


a). Chemical Weighing Balance.

b). Stirrer with Regulator.

c). Glass Beaker.

d). Thermometer.

e). Glass Plate.

f). Separating Funnel.

g). Thermostatically controlled heating mantel.

How to make casein protein by curds at home?


a). Casein - 15.0

b). Borax - 3.0

c). Canguard BIT - 0.4

d). DM Water - 81.6

3. Procedure of Casein Solubility 

a). Weigh accurately required quantity of DM water in a glass beaker.

b). Add casein powder in it gradually under stirring and start heating.

c). Add borax at 50ºC and increase temperature up to 80ºC and maintain for 2hours.

d). After 2 hours take out the sample on glass plate and check the dispersion, if found OK, and then cool it.

e). Add Canguard below 40ºC and drain it and kept for ageing.

f). Check the insoluble by diluting with 4 times water & kept the solution in a separating funnel for overnight.

g). Next day check following______ If insoluble’s are there they will settle calculate the % insoluble’s must be in range.

h.) Compare it with control. 

Determination of ash and acid-insoluble ash | What determines acid insoluble ash

Recommended Testing procedures of Acid Insoluble Ash.

Determination of Casein Ash Content 

Place about 3 g of the ground material, accurately weighed, or the quantity specified in the monograph, in a suitable tared dish. (for example, of silica or platinum), previously ignited, cooled and weighed. Incinerate the material by gradually increasing the heat, not exceeding 450 °C, until free from carbon; cool, and weigh. 

If a carbon-free ash cannot be obtained in this way, exhaust the charred mass with hot water, collect the residue on an ash-less filter-paper, incinerate the residue and filter-paper, add the filtrate, evaporate to dryness, and ignite at a temperature not exceeding 450 °C. Calculate the content in mg of ash per g of air dried material.

ash Cu-risible

Determination of acid-insoluble ash

Boil the ash for 5 minutes with 25 mL of hydrochloric acid (70 g/l) TS; collect the insoluble matter in a sintered crucible, or on an ash-less filter-paper, wash with hot water, and ignite at about 500 °C to constant weight. Calculate the content in mg of acid insoluble. ash per g of air-dried material.



a) Analytical Balance-0.1g accuracy.

b) Muffle furnace capable of being controlled at 825+25 ºC.

c) Desiccator containing an effective desiccant.



a) Weigh accurately 3g casein in previously dried and-cooled silica dish.

b) Transfer the dish in muffle furnace maintained at825+25 ºC for 3 hrs.

c) Cool in a desiccator.

d) Repeat the process of drying cooling and weighing at 30 min. interval until the difference between two consecutive weighing is less than 1.0mg.

Ash content of casein (%) = (W1-W) x 100



W1- Weight in g of crucible containing Ash.

W2- Weight in g of crucible with the dried material taken for the test.

W- Weight in g of crucible.

Acid Casein Composition.

Ash %                                           3.8                                          3.6

Lactose, %                                   0.1                                          .0.1

Sodium, %                                    1.2-1.4                                     <0.1

Fat%                                              1.1                                            1.2

Determination of Fat Content in Casein 


a) Mojonnier tube

b) Water bath (01 to 120 ºC)

c) Measuring Cylinder (5 ml to 25ml)

d) Pipettes (graduated of capacity 10ml).

e) Separating funnel


a) Hydrochloric Acid-(AR) Dilute 675ml of concentrated Hydrochloric Acid (1.18 gm/ml, to 1000ml with water.)

b) Ethanol (99.9X)

c) Diethyl ether (AR)-Free from Peroxides

d) Light Petroleum- ether (Boiling range between 30 and 60 ºC)


a) Take 4gm casein in to Mojonnier tube.

b) Add 20ml of diluted HCL and mix. Well.

c) Heat it in water bath to 100 ºC to dissolve the casein content completely.

d) Cool it to below 20 ºC by running water.

e)Add 20ml of Ethyl Alcohol and add 40ml Diethyl ether, Shake it vigorously for 30 sec.

f) Add 40ml Petroleum ether again, and put the stopper on the Mojonnier tube, shake it for 10 min.

g) Leave it until the content is completely separated. Transfer the supernatant into pre weighted dry flask. 

h) Make extraction 3 times by repeating the above procedure using ether and light petroleum.( By help of separating funnel )

i) The composite extract is dried on water bath in flask.

j) Flask is to be dried at 100 ºC in a oven for one hour then transfer the flask into a desiccator for cooling and then weight it.

Fat % by weight = (A-B)x 100/W

Weight of sample taken


A = Weight of the flask with fat

B = Weight of the empty flask


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