All the components are stable until the stated expiration
date If stored tightly closed, refrigerated and protected
from light.
*Preparation and Stability of Working solution
Reagents
liquid and ready to use
CK NAC
Kinetic UV Method
Principle
Kinetic determination of CK according IFCC and DGKC
recommendation:
CK
Creatin phosphate + ADP
Creatine + ATP
HK
ATP + D - Glucose
D - Glucose-6-phosphate + ADP
G-6-PDH
D-Glucose - 6 - phosphate + NADP +
D-Gluconate - 6 - phosphate + NADPH + + H +
The Rate of NADPH formation, determined
photometrically at 340 nm, is direct proportional to CK
activity.
Note:
The activation of all CK fractions is due to the N - Acetil - L
- Cisteine (NAC) as recommended by IFCC and DGKC
*Reagents composition, Contents and Safety warnings
1. Buffer / enzyme reagent
Imidazole Buffer
pH 6.3
110 mmol/l
N - Acetil - L - Cisteine (NAC) 23 mmol/l
Magnesium Acetate
11 mmoll
EDTA
2.2 mmol/l
NADP
2.2 mmol/l
Exokinase
≥ 3300 U/l
AMP
5.5 mmol/l
Diadenosin pentaphosphate 0.025 mmol/l
D - Glucose
20 mmol/l
Sodium Azide
< 0,1 %
2. Coenzyme reagent
Creatinphosphate
0.19 mol/l
ADP
11.7 mmol/l
G - 6 - PDH
≥ 10000 U/l
EDTA
2.2 mmol/l
Sodium Azide
< 0,1%
According to the present laws the kit does not contain
substances classified as dangerous
Storage and Stability of Reagents
Store the kit at 2 - 8° C, avoid direct sunlight exposition
ONE REAGENT PROCEDURE
Mix 4 volumes of Buffer with 1 volume of Substrate
Working solution stability: 10 days at 2 - 8oC.
Performance Characteristics
A. LINEARITY LIMIT
The reaction is linear till to 1500 U/I
For higher value dilute sample 1:2 with normal saline,
repeat the test and multiply the value by 2
B. DETECTION LIMIT
Values less than 4 U/I give non - reproducible results
C. INTRA - ASSAY PRECISION (N = 20 replicates)
Mean (U/l)
SD
CV%
Control 1
166.25
3.389
2.04
Control 2
425.30
4.428
1.04
TWO REAGENTS PROCEDURE
See procedure
Bring reagents at Room Temperature before use
D. INTER - ASSAY PRECISION (20 replicates for 3 days)
Mean (U/l)
SD
CV%
Control 1
161.20
4.12
2.56
Control 2
420.07
7.19
1.71
Sample
Fresh serum, heparinized or EDTA Plasma.
Safety precautions
For in vitro diagnostic use only.
Do not pipette by mouth.
Exercise the normal precautions required for handling
laboratory reagents.
*Procedure
1. Wavelenght
340 nm
2. Cuvette
light path: 1 cm
3. Temperature
25/30/37oC.
4. Adjust the instrument to zero against distilled water
One Reagent Procedure
Sample
50 µl
Working Solution
1 ml
Two Reagents Procedure
Buffer
0.8 ml
Sample
50µl
Substrate
200 µl
Mix, wait 2 minutes, read the Absorbance and start the
stopwatch simultaneously.
Read again after 1, 2 and 3 minutes.
Calculate the average absorbance difference per minute (A/min)
*Calculation
E. ACCURACY
Comparation between this method (y) and another
commercial one (x) gave the following results:
N = 20
r = 0.9999177
y = 1.01x - 6.73
F. INTERFERENCES
1. Haemoglobin till to 50 mg/dl does not interfere
2. Bilirubin till to 30 mg/dl does not interfere
3. Triglycerides till to1000 mg/dl do not interfere
4. Ascorbic Acid till to 25 mg/dl does not interfere
Quality Control
Control sera are recommended to monitor the performance
of manual and automated assay procedures.
Bibliography
Mathieu M.et coll.: Recommendation pour la measure de
la concentrationcatalytique de la Cr.atinine Kinase dans le
s.rum humain. Ann. Biol. Clin. 40, 87 (1982)
For in vitro diagnostic use only.
The following symbols are used on labels
For in vitro diagnostic use
CK (U/I) = A/min x 4130
Use by (last day of the month)
Reference values
Temperature limitation
Male
Female
25°C
10 - 80 U/l
10 - 70 U/l
30°C
15 - 130 U/l
15 - 110 U/l
37°C
24 - 195 U/l
24 - 170 U/l
These values should only be used as a guideline.
Each laboratory should establish its Normal Reference
Range
Batch Code