CREATININE REAGENT KIT
(ALKALINE PICRATE METHOD)
INTENDED USE:
The reagent kit is intended for the “in vitro” quantitaive
determination of Creatinine in Serum and Urine.
PROCEDURE :
Pipette into a clean dry test tube labeled as Standard (S)and Test
(T):
SUMMARY:
Creatinine is the catabolic product of creatinine phosphate, which Addition Sequence S T
is used by the skeletal muscle. The daily production depends on Working Reagent 1.0 ml 1.0 ml
muscular mass and it is excreted out of the body entirely by the Standard 50 µl -
kidneys. Elevated levels are found in renal dysfunction, reduced Sample - 50 µl
renal blood flow (shock, dehydration, congestive heart failure)
diabetes acromegaly. Decreased levels are found in muscular Mix well and read the initial absorbance (A 1) for the standard and
dystrophy. Test after exactly 30 seconds. Read another absorbance (A2) of
standard and Test exactly 120 seconds later. Calculate the
PRINCIPLE: change in absorbance DA for both the standard and Test.
Picric acid in an alkaline medium reacts with creatinine to form an
orange coloured complex with the alkaline picrate. Intensity of the Determine For Standard D AS = A2S - A1S
colour formed is directly proportional to the amount of creatinine
present in the sample. For Test D AT = A2T - A 1T
Creatinine + Alkaline Picrate Orange Coloured Complex
CALCULATION :
D AT
Creatinine Conc. mg/dl = X 2
CONTENTS: D AS
Reagent 1 : Creatinine Buffer Reagent D AT
Reagent 2 : Creatinine Picrate Reagent Urine Creatinine Conc. gm/L = X 2
Reagent 3 :Creatinine Standard 2 mg/dl D AS
Urine Creatinine gm/24hrs= Urine Creatinine in gm/L x Volume of urine
MATERIALS REQUIRED BUT NOT PROVIDED:- in 24hrs.
- Clean & Dry Glassware.
- Laboratory Glass Pipettes or Micropipettes & Tips. NORMAL VALUE :
- Bio-Chemistry Analyzer.
Serum : Male 0.9 - 1.4 mg /dl , Female - 0.8 -1.2 mg/dl
STORAGE & STABILITY Urine : Male 0.4 -1.8 gm/24h,Female - 0.35 -1.6 gm/24h
All reagents are stable at 2 - 8°C. Till expiry mentioned on the label. Each Laboratory should establish it's own normal range
representing its patient population.
SAMPLES:
Unhaemolysed Serum. Urine diluted (1:100) in saline. LINEARITY :
This procedure is linear upto 20 mg/dl.If value exceeds this limit
PREPARATION OF REAGENT & STABILITY : dilute the sample with normal saline (NaCl 0.9%) and repeat the
1. The reagent kit is stable at 2 - 8°C till the expiry date mentioned assay Multiply result by dilution factor.
on the bottles.
QUALITY CONTROL :
2. Once used the standard reagent should be stored at 2° -8° C.
For accuracy, it is advised to run known serum controls with
3. Step 1 : Working Reagent is prepared by combining equal each assay.
volumes of Reagent 1 and Reagent 2.
Step 2 : Mix by gentle swirling. LIMITATION & PRECAUTIONS :
Step 3 : Allow the reagent mixture to stand at R.T. For 5 1. Do not Freeze the Reagents.
minutes for equilibration.
2. During assay specified temperature has to be maintained.
The working reagent is ready for the use.
3. The time interval should be adhered as the kit reagent are
GENERAL SYSTEM PARAMETERS: Standardized accordingly
Reaction type : Fixed time (Increasing) 4. Do not pipette the reagent by mouth.
Wave Length : 505 nm (490 - 520 nm) 5. Use clean glassware free from dust or debris
Temperature : 37°C BIBLIOGRAPHY :
Delay time : 30 Sec Henry, R.J., Clinical chemistry, Principles and Techniques,
Interval : 120 Sec. 2nd Edition, Harper and Row, P. 525, 1974.
Reagent volume : 1.0 ml
CODE NO. PACK SIZE Reagent 1 Reagent 2 Reagent 3
Sample volume : 50 µl
S26 2 x 50 ml 1 x 50 ml 1 x 50 ml 1 x 3.0 ml
Standard concentration : 2.0 mg/dl
S26A 2 x 100 ml 2 x 50 ml 2 x 50 ml 1 x 3.0 ml
Zero setting : Deionised water
Light path : 1 cm IVD
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