CREATININE REAGENT KIT

(Enzymatic Method)
INTENDED USE:
The reagent kit Is intended for the “in vitro” quantitative
determination of Creatinine in Serum or Urine.
PROCEDURE FOR SERUM IN SEMI AUTO ANALYZER :
SUMMARY:
Pipette into a clean dry test tube labeled as Standard (S)and Test
Creatinine is formed in muscles from phosphocreatinine. It is an (T):
important form of energy by being store of high energy phosphate. Addition Sequence B S T
Creatinine determination have one advantage over urea determinat- Regent R1 450 µl 450 µl 450 µl
ion that it is not affected by high protein diet. Serum creatinine is more Standard 50 µl -
specific and sensitive indicator of renal function. Simultaneous esti- Sample - 50 µl
mation of serum urea and creatinine provides better information.
Mix & Incubate for 5 min at 37°C then add
serum urea nitrogen and creatinine ratio is >15 in prerenal failure
and <10 in renal failure. Decresed levels are found in muscle dystrophy. Regent R2 150 µl 150 µl 150 µl
PRINCIPLE: Mix well and incubate for 5 min at 37°C. Measure the absorbance
Creatininase of Test and Standard against the reagent blank.
Creatinine + H2O Creatine
Creatinase
Creatine + H2O Sarcosine + Urea CALCULATION :
Sarcosine Oxidase D Absorbance of sample
Sarcosine + O2 + H2O Glycine + HCHO + H2O2 Creatinine Conc (mg/dl) = X Standard Conc.
+1 +2 Peroxidase D Absorbance of standard
2H2O2 + 4-AA + TOOS Quinone pigment + 4H2O
LINEARITY :
*1 : 4-Aminoantipyrine,
*2 : N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine This procedure is linear up to 40 mg/dl.If value exceeds this limit
Creatinine concentration can be obtained by measuring quinone pigment photometrically. dilute the sample with normal saline (NaCl 0.9%) and repeat the
assay Multiply result by dilution factor.
CONTENTS:
Reagent 1 : Creatinine R1 PROCEDURE FOR SERUM IN FULLY AUTO ANALYZER :
Reagent 2 : Creatinine R2
Reagent 3 : Creatinine Standard (2 mg/dl) Sample : 8 µl
R1 : 270 µl R2 : 90 µl
MATERIALS REQUIRED BUT NOT PROVIDED:-
- Clean & Dry Glassware.
- Laboratory Glass Pipettes or Micropipettes & Tips. Measure Measure
- Bio-Chemistry Analyzer.
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37 C
STORAGE & STABILITY 10 (Min)
0 5
All reagents are stable at 2- 8°C till expiry mentioned on the label.
PROCEDURE FOR URINE :
SAMPLES:
Pipette into a clean dry test tube labeled as Standard (S)and Test
Serum or Urine (T):
Creatinine is stable in serum for 1 day at 2- 8°C. Addition Sequence B S T
Regent R1 450 µl 450 µl 450 µl
Urine for 24 hours collection is preferred. Dilute the specimen 1:9 Standard 10 µl -
with distilled / deionised water before the assay. Sample - 10 µl
Mix & Incubate for 5 min at 37°C then add
PREPARATION OF REAGENT & STABILITY : Regent R2 150 µl 150 µl 150 µl
Reagent 1, Reagent 2 and standard are ready to use

GENERAL SYSTEM PARAMETERS: Mix well and incubate for 5 min at 37°C. Measure the absorbance
Reaction type : End Point (Increasing) of Test and Standard against the reagent blank.
Wave Length : 546 nm CALCULATION :
Wave Length : 630 nm D Absorbance of sample
Temperature : 37°C Creatinine Conc (mg/dl) = X Standard Conc.
Incubation Time : 10 min. D Absorbance of standard
Reagent 1 : 450 µl
Reagent 2 : 150 µl LINEARITY :
Sample volume : 50 µl / 10 µl This procedure is linear up to 200 mg/dl.If value exceeds this limit
Standard concentration : 2.0 mg/dl dilute the sample with normal saline (NaCl 0.9%) and repeat the
Zero setting : Reagent assay Multiply result by dilution factor.
Light path : 1 cm
BIOLOGICAL REFERENCE RANGE :
Serum : Male : 0.6 - 1.1 mg /dl ,
: Female : 0.5 - 0.8 mg/dl
Urine : Male : 1070 - 2150 mg/dl (24 hrs accumulated urine)
: Female : 769 - 1200 mg/dl (24 hrs accumulated urine)

Each Laboratory should establish its own normal range representing

its patient population.

QUALITY CONTROL :
For accuracy, it is advised to run known serum controls with
each assay.

LIMITATION & PRECAUTIONS :

1. Do not Freeze the Reagents.
2. During assay specified temperature has to be maintained.
3. The time interval should be adhered as the kit reagent are
standardized accordingly
4. Do not pipette the reagent by mouth.
5. Use clean glassware free from dust or debris

BIBLIOGRAPHY :
Artiss, J.D., Mc Enroe, R.J., Zak, B.; Clin. Chem, 30 (1984)
1389
CODE NO. PACK SIZE
S31 80 ml
S31A 160 ml IVD www.jas-anz.com.au/register

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