CREATININE

Jaffe’s Kinetic Method 6x50 ml

Quantitative determination of Creatinine in serum or urine by means of 5. Run patients neat serum, 1:50 diluted urine and control samples by
photometric method. following the same procedure.

Only for in vitro use in clinical laboratory (IVD). Calculation

∆A U
Clinical Significance Serum/Plasma Creatinine (mg/dl) = x 2 (Standard Conc.)
Creatinine is the catabolic product of creatinine phosphate, which is used by the ∆A S
skeletal muscle. The daily production depends on muscular mass and it is ∆A U
excreted out of the body entirely by the kidneys. Urine Creatinine (mg/dl) = x 2 x 50 (Dilution Factor for Urine)
Elevated Creatinine levels are found in renal dysfunction reduced renal blood flow ∆A S
(shock, dehydration, congestive heart failure) diabetes acromegaly. Decreased
Creatinine levels are found in muscular dystrophy.
Quality Control
Principle Commercially available normal and pathological control sera are recommended to
monitor the performance of the procedure.
Creatinine reacts with the alkaline picrate (Jaffe reaction) producing a reddish
brown chromogen. The speed of this reaction under controlled conditions is a If control values are found outside the defined range, check the instrument,
measure of the creatinine concentration in the sample, as it behaves as a first reagents and calibrator for problems
order kinetic reaction for creatinine. Besides, it has been proved that non- Serum controls are recommended for internal quality control. Each
creatinine chromogens, which interfere with most of the conventional techniques, laboratory should establish its own Quality Control scheme and corrective
react within 30 seconds from the beginning of the reaction. Thus, the color actions if controls do not meet the acceptable tolerances.
increase there after is only due to creatinine.

Reagents Reference Values

Each Creatinine kit contains Male Serum 0.9 - 1.4 mg/dl
RI - Picric Acid Urine 1000 - 2000 mg/24 hrs.
R II - Alkali Female Serum 0.7 - 1.3 mg/dl
Creatinine STD - Creatinine Standard ( 2 mg/dl) Urine 600 - 1500 mg/24 hrs.
It is suggested that each laboratory establish its own reference range.
Reagent Preparation
Reagents are ready to use.
Mix one Volume of Reagent (R I) with one volume of Reagent (R II) (According to Reagent Performance
the requirement).
Measuring Range
Reaction is linear up to 20 mg/dl of Creatinine under the described assay
Stability conditions.
The provided reagents are stable till the date of expiry when stored as stated on For higher values, dilute sample 1:2 or 1:4 with saline solution and repeat
the labels. testing. Correct calculations multiplying by the dilution factor used.
Working reagent is stable for 30 days at 25o ± 5o C. Do not use the reagent if
turbid. Test Parameters
Visible turbidity in reagent or standard indicates reagent deterioration. Discard in
Mode Two Point Kinetic
such case.
Do not use reagents over the expiration date. Wavelength (nm) 505
Sample Volume (µl) 100
Specimens Working Reagent Volume (µl) 1000
Serum or urine. Lag Time (Sec.) 20
a) Collection: obtain serum in the usual way. 24-hour urine should be Measuring Time(Sec.) 60
collected in a thoroughly cleaned container, which should be refrigerated Standard Conc. (mg/dl) 2
(2°-8° C) during collection. Measure diuresis, take an aliquot and perform Reaction Temperature (° C) 37
a 1:50 dilution. to calculate the amount of creatinine eliminated during 24 Reaction Direction Increasing
hours. Normal Low (mg/dl) 0.9
b) Additives: none required. Normal High (mg/dl) 1.4
c) Known interfering substances: sera with hemoglobin concentration>30 Blank with Reagent
mg/dl or Bilirubin >10 mg/dl yield falsely increased values. No interference Units mg/dl
is observed from mild or moderate hemolysis, hyperlipemia, Linearity Limit 20 mg/dl
disproteinemia or non-creatinine chromogens (such as glucose, ascorbic
acid, ketoacids, glutathione or ergothioneine). For detailed interferences
refer to Young, et. al. 3 .
Literatures
1. Jaffe M. Physiol Chem 10:391, 1886
d) Stability and storage instructions: creatinine in serum is stable up to 24 2. RJ Henry et al, Clin. Chem.- Principles and Technics, 1974, pp 549-550.
hours and in urine upto 4 days, when refrigerated. 3. Young,D.S., et al, Clin.Chem. 21:1D (1975).

Procedure
In three colorimeter test tubes labeled B (Blank), S (Standard) and U
(Unknown), place: Manufactured by
Blank (B) Standard Unknown PRISM DIAGNOSTICS PVT. LTD.
(S) Sample (U) 7&8, Shiv Shakti Laghu Udyog Sankul,
Mumbai – Nashik Road,
Working Reagent 1000 µl 1000 µl 1000 µl Asangaon, Dist. Thane
Dis. Water 100 µl -- --
Standard -- 100 µl -- Marketed by
Unknown Sample -- -- 100 µl SYNERGY BIO, A Division of
Euro Diagnostic Systems Pvt. Ltd.
1. Set photometer cuvette temperature to 30o or 37oC. Zero photometer ‘Millennium House’ M. K. Srinivasan Nagar Main Road
with reagent blank at 505 nm. No. 144, Old Mahabalipuram Road, Perungudi, Chennai-600 096
2. After exactly 20 secs read and record absorbance A1 . Tamil Nadu, India, Phone: 044-2496 1064, 044-2496 0541
3. At exactly 60 secs after A1 , read and record absorbance A2 . Fax: 044-2496 2825 , email: eurods@vsnl.net, www.eurods.in
4. Calculate change in absorbance ∆A by subtracting A2 - A1 .