Fifth Semester Practical Paper V- Biochemistry
1. Estimation of blood sugar by Folin-Wu method.
2. Estimation of creatinine in urine by Jaffe’s method.
3. Estimation of inorganic phosphate in food samples by Fiske-Subbarow method.
4. Estimation of total reducing sugar in honey by DNS method.
, Estimation of Blood Sugar by Folin Wu Method
Aim:
Estimation of blood sugar by Folin Wu method, being provided with a standard glucose solution
containing 200 μg/mL of glucose.
Principal:
In this method, the proteins in blood are precipitated by tungstic acid. The protein free filterate containing
glucose is treated with alkaline copper reagent. Cuprous oxide th thus
us formed is treated with
phosphomolybdic acid. Phosphomolybdic acid is reduced to phosphomolybdous acid which is blue in
colour.
Procedure:
Pipette out different aliquots (0.2 – 1.0ml) of standard glucose solutions (200 μg) into different test tub
tubes
and 1.0 ml of the blood filtrate into two different test tubes. Make up the unknown sugar solution given in
the standard flask up to the mark with distilled water and shake well. Pipette out 1 mL of this made-up
made
solution (as unknown) into each of two separate
separate tubes. Make up the volume to 2.0 ml in each case by
adding distilled water. Add 2.0 ml of alkaline copper reagent to each test tube. Mix the contents well &
boil in water bath for 10 minutes. Cool the test tubes & add 2.0 ml of phosphomolybdic acid to each of
test tubes. Mix the contents and allow the test tubes to stand for 2 minutes. Then add 4.0 ml of distilled
water to each mix well & read the absorbance at 490 nm against the blank.
Result: The amount of glucose in the given solution = ____________
________ μg
1. Estimation of blood sugar by Folin-Wu method.
2. Estimation of creatinine in urine by Jaffe’s method.
3. Estimation of inorganic phosphate in food samples by Fiske-Subbarow method.
4. Estimation of total reducing sugar in honey by DNS method.
, Estimation of Blood Sugar by Folin Wu Method
Aim:
Estimation of blood sugar by Folin Wu method, being provided with a standard glucose solution
containing 200 μg/mL of glucose.
Principal:
In this method, the proteins in blood are precipitated by tungstic acid. The protein free filterate containing
glucose is treated with alkaline copper reagent. Cuprous oxide th thus
us formed is treated with
phosphomolybdic acid. Phosphomolybdic acid is reduced to phosphomolybdous acid which is blue in
colour.
Procedure:
Pipette out different aliquots (0.2 – 1.0ml) of standard glucose solutions (200 μg) into different test tub
tubes
and 1.0 ml of the blood filtrate into two different test tubes. Make up the unknown sugar solution given in
the standard flask up to the mark with distilled water and shake well. Pipette out 1 mL of this made-up
made
solution (as unknown) into each of two separate
separate tubes. Make up the volume to 2.0 ml in each case by
adding distilled water. Add 2.0 ml of alkaline copper reagent to each test tube. Mix the contents well &
boil in water bath for 10 minutes. Cool the test tubes & add 2.0 ml of phosphomolybdic acid to each of
test tubes. Mix the contents and allow the test tubes to stand for 2 minutes. Then add 4.0 ml of distilled
water to each mix well & read the absorbance at 490 nm against the blank.
Result: The amount of glucose in the given solution = ____________
________ μg
