CHEMISTRY REAGENT KITS
   ---> CREATININE E.P. (Jaffe's)- Colorimetric End-Point

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CLINICAL SIGNIFICANCE
Increased levels are associated with substantially impaired renal function.

PRINCIPLE
In an alkaline medium, creatinine reacts with picrate to produce and ORANGE YELLOW colour. The coloration is    proportional to the creatinine   concentration.

REAGENTS SUPPLIED

Cat. No. BC 227

Composition

1. Picric acid 2x80 mL
    Picric acid 30mMol/L
2. Sodium Hydroxide 50 mL
    Sodium Hydroxide 750mMol/L
3. Standard 15mL
    Creatinine 2mGs/dL

STABILITY
24 months at 18-25°C away from light. The working standard is stable for 3 days at 2-8°C away from light and 24 hours at25°C + 5°C. Standard to be stored at 2-8°C.

SAMPLE
Non-haemolysed serum or plasma (heparinofEDTA)
No prior patient preparation is needed. (All samples should be handled as potential infective agents as no laboratory methods make conclusive findings for its safety. Therefore, adequate protective laboratory measures should be taken while handling such materials).

URINE SAMPLE
If Urine to be tested use fresh sample.
Dilute urine 1 to 25 or 1 to 50 in distilled water.
Perform the test with the dilution as for the
serum.
Multiply the result by dilution factor (25 or 50)

WARNING
This reagent system isfor in vitro use only. This reagent system is contains preservatives and components that have not established for safety if contacted on broken skin or eye or taken orally. In case of such incidents wash off with plenty of water, or consulta physician.

MANUAL METHOD
STEP 1. PROTEIN PRECIPITATION
Pipette into a Centrifuge tube
Serum or Plasma.............................................1mL
PicricAcid Reagent No. 1....................................3mL
Mix well. Keep in a boiling water bath for one minute. Cool immediately and separate clear supernatant by centrifuge or filteration.

STEP2. COLOURING
   Pipette into 3 Test Tubes Blank mL Sample Standrd mL
   Distilled water 0.05 - -
   Creatinine Standard - 0.05 -
   Picric Acid ReagentNo.1 1.5 1.5 -
   Sodium Hydroxide Reagent No.2 0.05 0.05 0.05
   Supernatant from Step 1 - - 2.0

Mix well. Keep at room temperature for 15 mins. Read optical density (OD) at 500-540 nM or GREEN filteragainst blank. Final colour is stable upto 30 minutes.
NOTE: Reagent and sample volumes can be altered proportionately.
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RESULTS
CreatinineinmGs/dL
                                ODTest
                            ------------------   x 2
                              OD Standard

EXPECTED VALUES
Serum: 0.68to1.48mGs/dL
             (60to130µMol/L)
Urine:   1 to2Gms/24hrs
             (9to18mMol/24hrs)
As with all diagnostic methods, the final diagnosis should not be made on the result of a single test as well as laboratory diagnosis must be confirmed with clinical manifestations.

LIMITATIONS
Alkaline picrate is sensitive to various metalic elements. In vitro addition of metalic compounds or contaminations make false results. This assay is linear up to 15.0Gms/dL proteins. For values higher than 12 mGs/dL (1063 µMol/L), repeat the test diluting the serum with distilled water. Multiply the  results  by the dilution factor (i.e. 2foradilution of 1:1).

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QUALITY CONTROL
To ensure adequate quality control, each kit should be tested against a standard control sera. It should be realised that the use of quality control material checks both instrument and reagent function together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glasswares and accuracy of pippetting and timings.
It is appropriate to establish each laboratory's accuracy constant and interpret values. Similarly, laboratory findings should be established by clinical manifestations.

BIBLIGRAPHY
Brod, J. and Sirots (1948) J. Clin. Invest 27. 645 Bittles,A.H. Bells and Nelli, D.W.(1965) J. Clin. Path 18, 377 Bonsues R. W. and Taussky H.H. (1945) J. Bio chem. 159.581. Owen. J.A. lggo, B. Seandrett, F.J. and Stewart C.P. (1945) J. Biochem. 58.426. Torro, G.AndAckermann, PG. (1975) pract clin. Chem. Little Brown and Co. Boston