CREATININE REAGENT KIT
(Alkaline Picrate Method)
INTENDED USE
This reagent kit is used for the in vitro quantitative determination of creatinine in serum and urine.
SUMMARY
Creatinine is the end product of creatine phosphate metabolism in muscles. It is excreted in urine.
Serum creatinine level helps assess kidney (renal) function.
Elevated creatinine indicates renal impairment, muscle damage, or muscular dystrophy.
PRINCIPLE
Creatinine reacts with alkaline picrate to form an orange-colored complex.
The intensity of the color is directly proportional to the amount of creatinine present and is measured spectrophotometrically at 520 nm.
Reaction:
Creatinine + Alkaline Picrate → Orange-Coloured Complex
CONTENTS
Reagent 1: Creatinine Buffer Reagent
Reagent 2: Creatinine Standard Reagent
Reagent 3: Creatinine Picrate Reagent
MATERIALS REQUIRED
Clean dry glassware
Micropipettes & tips
Colorimeter or Spectrophotometer
Test tubes
SPECIMEN
Use serum or urine.
Serum should be diluted (1:100) in saline.
PREPARATION OF REAGENT & STABILITY
Reagents are supplied ready-to-use.
Store at 2–8°C.
Working reagent is stable for 7 days at room temperature (R.T.) or 1 month at 2–8°C.
PROCEDURE
| Pipette into Test Tubes | Blank | Standard | Test |
|---|---|---|---|
| Working Reagent | 1.0 ml | 1.0 ml | 1.0 ml |
| Standard | – | 0.1 ml | – |
| Sample | – | – | 0.1 ml |
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Mix well and read absorbance (A) of the Standard and Test after 30 seconds and again after 90 seconds at 520 nm.
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Distilled water is used as Blank.
CALCULATION
Where:
Aₛ = Absorbance of Standard
Aₜ = Absorbance of TestNORMAL VALUES
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Serum: Male – 0.6–1.4 mg/dL; Female – 0.6–1.2 mg/dL
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Urine (24 hours): 1.0–2.0 g/24 hr
(Note: Values may vary by method and laboratory.)
QUALITY CONTROL
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Use normal and abnormal serum controls to check assay performance.
LIMITATION & PRECAUTIONS
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Avoid hemolysed samples.
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Maintain precise timing and temperature.
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Do not use reagents after expiry date.
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For in vitro diagnostic use only.
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