Chemistry Laboratory 3

Kinetic analysis of creatine kinase procedure
Creatine Kinase reactions occur at zero order kinetics is when the enzyme is the only thing effecting rate, the velocity is constant and there is excess substrate.
The sources of error in the total Creatine Kinase procedure
interference due to chloride and sulfate contamination, and samples that exceed the maximum range need to be diluted. There was also human sources of error in that the procedure calls for measurements of absorbances at 60 second intervals which is difficult to get perfect.
Quality control
represented by level 1 and level 2 controls. They have a known value and a reference range. If the reported results are not within the reference range, the patient results cannot be released. They are used to evaluate the accuracy of the procedure, indicating proper technique and performance.
Isoenzyme immunochemical measurement
we use an antibody for the M monomer of the CK isoenzymes. Isoenzymes are the CK enzymes MM, MB, and BB. They are located in different areas of the body and can be measured as indicators of certain conditions. To measure CK activity we used the presence of an antibody to the CK-M monomer (to measure MM and MB). The antibody inhibits the activity of CK-MM and inhibits half the activity of CK-MB; the concentration multiplied by 2 represents the total activity of the CK-MB activity (which is indicative of a heart attack)
isoenzymes electrophoretically measurement
To measure isoenzymes electrophoretically you would use an electrophesis gel, which takes longer.
The possible sources of error in the CK-MB procedure
hemolyzed samples, which are not suitable due to the high levels of adenylate kinase, ATP, and G6P, and overestimation of CK-MB activity if CK-BB activity is high (more then 20% of total activity is represented by CK-MB)
Specimen requirements and storage for the creatine kinase procedure
A clear non hemolyzed sample is preferred, but plasma in a tube with heprin is acceptable. The serum should be stored at 2-6 degrees C for a max of 5 days.
Why do CK values differ in Male and Females?
CK values differ due to the average muscle composition difference in men and women. Man have more muscle so therefore their CK levels are higher
Three common methods used for separation and determination of CK isoenzymes?
Immunochemical method, electrophoretically, and via chromatography
What is Macro CK and why is it formed?
Macro CK is a macroenzyme formed from the normal CK enzyme and an immunoglobulin. Macro CK doesn’t show up differently on the CK assay and could cause an increased reading
Calculation of Total CK activity (U/L)
(Average change in absorbency/min)/Absorptivity x (total volume (mL)/sample volume (mL))
x

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