CK-NAC

Presentation:
20 x 2,5 mL. Ref.: 30200
CREATINE KINASE
Store: 2 - 8 ºC. NAC. Kinetic UV.
Quantitative determination of creatin kinase (CK). CLINICAL SIGNIFICANCE
General laboratory equipment.
Creatine kinase is a cellular enzyme with wide tissue distribution in the
Only for in vitro use in clinical laboratory (IVD) body. Its physiological role is associated with adenosine triphosphate
TEST PROCEDURE
TEST SUMMARY (ATP) generation for contractile or transport systems.
1. Assay Conditions
Creatine kinase (CK) catalyses the reversible transfer of a phosphate group Elevated CK values are observed in diseases of skeletal muscle and
- Wavelength : ............................. 340 nm. 1,5,6
from phosphocreatine to ADP. This reaction is coupled to those catalysed by after myocardial infarction .
- Cuvette: ..................................... 1 cm light path. Clinical diagnosis should not be made on a single test result; it should
hexokinase (HK) and glucose-6-phosphate dehydrogenase (G6P-DH): - Constant temperature . . . . . . .. . .25ºC /30ºC / 37ºC.
CK integrate clinical and other laboratory data.
2. Adjust the instrument to zero with distilled water or air.
Phosphocreatine + ADP  → Creatine + ATP 3. Pipette into a Cuvette: REAGENT PERFORMANCE
HK
ATP + Glucose → ADP + Glucose-6-phosphate 25-30ºC 37ºC. - Measuring Range:
WR (mL) 1.0 1.0 From detection limit of 0.878 U/L. to linearity limit of 1300 U/L.,
G6P −DH
G6P + NADP   → 6-Phosphogluconate + NADPH + H
+ +
Sample (µL.) 40 20 under the described assay conditions.
The rate of NADPH formation, measured photometrically, is proportional to 4. Mix and incubate for 2 minutes. If results obtained were greater than linearity limit, dilute the
the catalytic concentration of CK present in the sample1,2. 5. Read the absorbance (A) of the sample, start the stopwatch and sample 1/10 with NaCl 9 g/L. and multiply result by 10.
read absorbance at 1 min. interval thereafter for 3 min. - Precision:
REAGENTS COMPOSITION 6. Calculate the difference of absorbance and the average absorbance Intra-assay n= 20 Inter-assay n= 20
Imidazol pH 7.0 100 mmol/L difference per minute (∆A/min.) Mean (U/L) 144 478 146 494
R1 Glucose 20 mmol/L SD 3.88 6.98 4.55 9.57
Magnesium acetate 10 mmol/L CALCULATIONS
Buffer CV (%) 2.69 1.49 3.11 1.94
EDTA 2 mmol/L 25 – 30ºC ∆A/min x 4127* = U/L of CK - Sensitivity: 1 U/L = 0.0001 ∆A/min
37ºC. ∆A/min x 8095* = U/L of CK - Accuracy: Results obtained LABKIT reagents did not show
ADP 2 mmol/L
systematic differences when compared with other commercial
AMP 5 mmol/L Units: One international unit (IU) is the amount of enzyme that transforms
reagents
di-Adenosine-5- pentaphosphate 10 mmol/L 1 µmol of substrate per minute, in standard conditions. The concentration The results obtained using 50 samples were the following:
R2 NADP+ 2 mmol/L is expressed in units per litre of sample (U/L). Correlation coefficient (r): 00997.
Substrate Hexoquinase (HK) 2500 U/L
Temperature conversion factors Regression equation: y= 1.031x – 0.5355.
Glucose-6-phosphate dehydrogenase (G6P-DH) 1500 U/L The results of the performance characteristics depend on the
To correct results to other temperatures multiply by:
N-acetyl cysteine 20 mmol/L analyzer used.
Creatine phosphate Assay Conversion factor to
30 mmol/L
temperature 25ºC 30ºC 37ºC
REAGENT PREPARATION AND STABILITY INTERFERING SUBSTANCES
25ºC 1.00 1.56 2.44
Working reagent (WR): Dissolve ( → ) 1 tablet of R.2 in 2,5 mL. of R.1. - No interferences were observed with bilirubin up to < 20 mg/dL
30ºC 0.64 1.00 1.56
and hemoglobin up to 10 g/L1,2
Cap and mix gently to dissolve contents. 37ºC 0.41 0.63 1.00
- A list of drugs and other interfering substances with CK
Stability: 10 days at 2-8ºC or 3 days at room temperature (15-25ºC). determination has been reported by Young et. al2,3.
QUALITY CONTROL
All the components of the kit are stable until the expiration date on the label
Control sera are recommended to monitor the performance of the
when stored at 2-8ºC, protected from light and contamination prevented NOTES
procedure, LABTROL H Normal Ref. 30950 and LABTROL H Pathological
during their use. 1. Use clean disposable pipette tips for its dispensation.
Ref. 30955. If control values are found outside the defined range, check
Do not use reagents over the expiration date. 2. Formulation to reach constant:
the instrument, reagents and calibrator for problems.
Do not use tablets if appears broken. Tv= Total volume in mL
Signs of Reagent deterioration: Serum controls are recommended for internal quality control. Each ∆A/min x 4127* or * Tv x 1000 ε NADPH = 6.22 at 340 nm
- Presence of particles and turbidity. laboratory should establish its own Quality Control scheme and 8095* = U/L CK ε x LP x Sv LP= Light path
- Blank absorbance (A) at 340 nm. > 1.60 corrective actions.
Sv= Sample volume in mL
All the reagents of the kit are stable up to the end of the indicated BIBLIOGRAPHY
REFERENCE VALUES1
month and year of expiry. Store tightly closed at 2-8ºC. Do not use 1. Abbot B et al. Creatinine kinase. Kaplan A et al. Clin Chem The C.V. Mosby
reagents over the expiration date. 30ºC 37ºC 25ºC
Men up to 80 U/L. 130 U/L. 195 U/L. Co. St Louis. Toronto. Princeton 1984: 1112-116.
2. Gerhardt W et al. Creatine kinase B-Subunit activity in serum after
SPECIMEN Women up to 70 U/L. 110 U/L. 170 U/L. immunohinhibition of M-Subunit activity. Clin Chem 1979;(25/7): 1274-1280.
Serum or plasma1: Stability 7 days at 2-8ºC, protected from light. (These values are for orientation purpose). 3. Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed AACC Press,
The creatin kinase activity decrease 10% after 1 day at 2-5ºC of after 1 hour 1995.
It is suggested that each laboratory establish its own reference range. 4. Young DS. Effects of disease on Clinical Lab. Tests, 4th ed AACC 2001.
at 15-25ºC.
5. Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.
MATERIAL REQUIRED BUT NOT PROVIDED 6. Tietz N W et al. Clinical Guide to Laboratory Tests, 3rd ed AACC 1995.
- Spectrophotometer or colorimeter measuring at 340 nm.
- Thermostatic bath at 25ºC, 30ºC o 37ºC (± 0.1ºC)
- Matched cuvettes 1.0 cm light path.

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