CK NAC 30

Creatine kinase catalyzes the conversion of creatine phosphate and ADP to creatine and ATP. ATP phosphorylates glucose to glucose-6-phosphate in the pre sence of hexo kinase. GIucose-6-ph osp hate is ox idized to 6-phosphogIuconate, reducing NADP to NADPH in presence of G-6-PDH. The rate of increase in NADPH absorbance at 340 nm. is directly proportional to the activity of creatine kinase in serum/plasma.

Abbreviations

• ADP: Adenosine - 5 diphosphate
• ATP: Adenosine - 5 triphosphate
• G-6-PDH: Glucose-6-phosphate dehydrogenase CK Creatine kinase
• HK: Hexokinase

 PREPARATION OF WORKING SOLUTION

Prepare working solution by mixing Reagent R, and Reagent R2 in the ratio 4 : 1 as per requirement.

REAGENT STORAGE & STABILITY

The reagent kit should be stored at 2 8 C and is stable till the expiry date indicated on the label.

The working solution (4 R1 + 1 R2) is stable for 10 days at 2 - 8 Degree C.

COMPONENTS & CONCENTRATION OF WORKING SOLUTION 

Component	Concentration	Component	Concentration
Imidazole buffer pH 6.7	100 mmol/I	EDTA	2 mmol/I
Phosphocreatine	30 mmol/I	ADP	2 mmol/I
N-acetylcysteine (NAC	20 mmol/I	NADP	2 mmol/I
Magnesium acetate	10 mmol/I	AMP	5 mmol/I
Glucose	20 mmol/I
Di-(adenosine-5)-pentaphosphate	10 mmol/I
Glucose-6-phosphate dehydrogenase	1  1.5 KU/I
Hexokinase	1  2.5 KU/I

SPECIMEN COLLECTION & PRESERVATION

Blood should be collected in a clean dry container. Although serum is preferred, plasma with heparin or EDTA can be used. The assay should be carried out on the same day, as far as possible. Serum and plasma samples are stable for 1 week at 4 C and 1 month at -10 C. The samples should be brought to room temperature prior to use. Avoid use of haemolysed and grossly contaminated samples.

PROCEDURE

• Reaction type: UV - Kinetic
• Reaction direction: Increasing
• Wavelength: 340 nm.
• Flowcell  temperature: 37 Degree C
• Zero  setting with:  Distilled water
• Delay  time :  180 seconds
• No. of readings: 4
• Interval:  30 seconds
• Sample  volume: 0.04 ml (40 II)
• Working solution volume:  (4R  1R2)1.0 ml (1000 II)
• Factor: 4127
• Linearity:  2000 IU/I 

To ensure adequate quality control, it is recommended that each batch should include a normal and an abnormal commercial reference control serum. It should be realized that the use of quality control material checks both instrument and reagent functions together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glassware and accuracy of pipetting.