A FLUOROMETRIC COUPLED ENZYMATIC METHOD FOR THE DETERMINATION OF AD Part I.
A new fluorometric, coupled, enzymatic method for the determination of adenosine triphosphate is described. The procedure uses the hexokinase reaction coupled with three other enzymes in a system of phosphorylation and double reduction to form NADPH. The NADPH is measured in a cheap oakley sunglasses for sale reaction catalyzed by diaphorase in which the nonfluorescent material, resazurin, is converted to the highly fluorescent compound, resorufin. This oakley optical kinetic method can determine adenosine triphosphate at the picomole level. Intra and inter assay CV's were both less than 3% and recoveries were quantitative. No significant interference was observed from EDTA or okley sale heparin. Correlation with an established method was significant (r = 0.99). Adenosine triphosphate was measured in platelets. Mean ATP levels in the platelets of "normal" subjects were found to be 2.17 (+OR ) 0.76 nmol/10('8) platelets (14.22 (+OR ) 6.32 nmol/mg protein). Part II. A new procedure was developed for the determination of non enzymatically glycated albumin (GA). Albumin was separated from serum or plasma using Sepharose blue dextran affinity chromatography. The "fructosamine assay" was oakley sale sunglasses improved and used to determine GA. The stable ketoamine linkage in GA reduced 3 (4,5 dimethylthiazol 2 yl) 2,4 diphenyltetrazolium bromide (MTT) to a colored formazan. Optimum conditions for the assay were established using the simplex optimization technique. The reagent blank value was minimal and MTT had a 3 fold greater molar absorptivity than nitroblue tetrazolium used in the original "fructosamine assay". The assay was adapted for use in a centrifugal analyzer (Flexigem('tm)) and GA was used as the standard. The within and between run CV's were 4.6% and 8.5%, respectively. Recovery of GA was quantitative.
The reference range for this method was 8.94 11.19% GA and normal and diabetic populations can be clearly discriminated (p.
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