NAME: HAMMAD HUSSAIN SHAH ENROLLMENT NO: 5-2/2021/098

ASSIGNMENT: BIOCHEMISTRY
Q1. Explain the principle of estimation of glucose in urine by benedict’s test?

A1. As benedict’s test used to give confirmation of reducing sugar, as glucose is a reducing sugar with
aldehydic group present in it, When Benedict's reagent solution and reducing sugars are heated together,
the solution changes its color to orange-red/ brick red, this color change proves the presence of glucose in
urine by benedict test.

Q2. What is titration, titre factor, titrant and titrand?

A2. Titration is a common laboratory method of quantitative chemical analysis to determine the
concentration of an identified analyte, A titer is defined as the quotient of the nominal concentration of a
volumetric solution and the actual concentration. The calculated factor is then used as the correction
factor to the titrant and a titrand, a substance whose concentration is to be determined by titration.

Q3. What is the principle of detecting ketone bodies in given sample of urine?

A3. Acetoacetic acid or acetone reacts with nitroprusside in an alkaline solution to form a purple-colored
complex, Rothera’s test is sensitive to 1-5 mg/dl of acetoacetate and 10-25 mg/dl of acetone.

Q4. Explain Rothera’s nitroprusside test and Gerhardt reaction?

A4. Nitroprusside in alkaline medium reacts with a ketone group to form a purple ring. It is given by
acetone and acetoacetate, but not by Beta hydroxy butyric acid. Ketone bodies react with sodium
nitroprusside to produce purple coloring at the junction of the two layers, this test is also known as
Rothera’s nitroprusside test, And the second is Gerhardt reaction in which a reddish or purplish color
is given by aceto acetate.. If acetoacetic acid is present in the urine.This test is only given by acetoacetate
and not by beta hydroxy butyric acid directly.
Q5. Explain the process of the qualitative analysis of the given sample of human saliva?

A5. Rinse your mouth with clean water and then take a mouth full of water hold and hold the water in
your mouth for 2 minutes and then take it in a beaker, and then test the sample of human saliva in
following test:

1 MOLISCH TEST: which gives the conformation of carbohydrate in human saliva..

2 BIURET TEST: which gives the conformation of protein or peptide bonds in the sample.

3 MUCIN TEST: which gives the conformation of synovial fluid precipitation in acetic acid.

4. SALIVARY AMYLASE TEST: which gives the conformation of the amount of amylase in your blood
or urine.

5. THIOCYNATE TEST: which gives the conformation of thiocynate in the sample.