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OBJECT: To estimate the conc. Of creatinin in a

given serum sample

REAGENTS :

R1

-Sodium hydroxide

R2

-Picric acid

Standard: 2mg/dl (177 micromole/l)

Mix 4parts of R1 with 1part of R2, the mixture is known as monoreagent.

PRINCIPLE:

Creatinin is a waste product excreted by the kidneys mainly

by glomerular filtration. The concentration of creatinin in plasma of a healthy
individual is fairly constant, independent from water intake, exercise and
rate of urine formation. Therefore increased plasma creatinin values always
indicate decreased excretion I.e., impaired kidney function.

Creatinin forms a orange-red complex in an alkaline solution. The difference

in absorbance at fixed time intervals during conversion is proportional to the
conc. Of creatinin in the sample.

creatinin + picric acid -------→ creatinin picrate complex

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PROCEDURE:

Wavelength: 492 nm

Optical path: 1 cm

Temperature: 20-25 C

Measurement: Against reagent blank

BLANK SAMPLE STANDARD

SAMPLE ___ 100 micro ___

liter

STANDARD ___ ___ 100 micro liter

DISTILLED WATER 100 micro liter ___ ___

MONOREAGENT 2 ml 2 ml 2 ml

Mix and read absorbance A1 after one minute. Read absorbance A2 after two
minutes.

∆A = [ (A2 – A1) sample or standard ] - [(A2 – A1) blank ]

CALCULATIONS:
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Creatinin conc. = ∆A sample / ∆A standard X conc. Of standard

RESULT:

The conc. Of creatinin in the given sample is found out to be

-------------- mg/dl

INTERPRETATION: