Practical # 1

Scheme for detection of Proteins

 General reactions of proteins

1. Lead acetate test
2. Sulpho salicyclic test
3. Esback’s test

Color Reaction of Protein

1. Biuret test
2. Xanthoprotiein test
3. Millon’s test
4. Formaline test
5. Sulphur test

Salicyclic acid test

EXPERIMENT OBSERVATION INTEREFERENCE

2ml (0.5) + 2 drops of sulpho- If ppt dissolve, clear solution BJP +ve
salicyclic acid
If ppt persists Other Proteins present (albumin
etc)

Color Reaction of Proteins

Biuret test:-

EXPERIMENT OBSERVATION INTEREFERENCE

2ml (5%) NaOH + 2 drops of 1% Solution containing protein Peptide linkage protein are
CuSO4. Divide the solution into 2 turns purple violet present
parts, T.T.A (test tube A) and
T.T.B(test tube B).
Add 1ml of distilled water(d/w)
in TTA
Add 1 ml of original solution in
TTB

Coagulation Test:-

EXPERIMENT OBSERVATION INTEREFERENCE

3ml (OS) and heat on low flame If coagulation appears Albumin and globulin present

Half Saturation Test

EXPERIMENT OBSERVATION INTEREFERENCE

Take 3ml of given solution in test If white ppt. appears Globulin is present
tube and add equal amount
(3ml) of saturated
ammonium(SO4). Now the
 solution becomes half saturated

Full Saturation Test

EXPERIMENT OBSERVATION INTEREFERENCE

Take 5ml of given solution in a If white ppt. appears Albumin is present
test tube and saturate it with
solid ammounium sulphate

Precipitation of Protein

Acetic Acid test for Casein

Reagent :- 1% Acetic Acid solution

EXPERIMENT OBSERVATION INTEREFERENCE

5ml (OS) + 1% Acetic acid drop White ppt +ve Casein +ve
by drop till solution changes to
white ppt.

Principle:-
Casein is precipitated out at its iso electric PH(4.6) when 1% Acetic acid is added dropwise.

CT(confirmation test) for Casein(Neuman’s test):-

Reagent:-
conc. H2SO4 : Con. HNO3 : Ammonium molybedate solution 5% : Methyl Red Indicator

Principle:- On heating e’ conc. H2SO4 e’ HNO3. Casein is digested and phosphorous is released, NH3 is added to provide
alkaline media.

Ammonium molybedate react e’ phosphorous in alkaline media form yellow ppt of phospho-ammonium molybedate

EXPERIMENT OBSERVATION INTEREFERENCE

Ppt. of acetic acid test + 1 drop Yellow ppt. of phospho- Casein Confirmed
of HNO3 + 12 drops of H2SO4, ammonium molybedate appear
then heat till white fumes
appear.
Add 1 drop of HNO3, heat till
clear solution appear, then cool
& dilute e’ 5ml D/w
Add 1 drop of methyl red of
strong NH3 till color becomes
light yellow + 2ml of ammonium
molybedate

Proteinuria:-
Pre Renal, Renal (Glomerulus) and Post Renal (bladder trouble, could be stones / trauma)