Use of N-methyl-D-glucamine as buffer in the determination of serum alkaline phosphatase activity.
We describe a method for determining serum alkaline phosphatase activity with use of N-methyl-D-glucamine buffer, Na+ is a definite activator, whereas NH4+ and Li+ inhibit enzyme activity. Optimum reaction conditions are: methylglucamine buffer, 0.35 mol/L, pH 10.2 +/- 0.1 (30 degrees C); NaCl, 70 mmol/L; MgCl2, 0.5 mmol/L; disodium 4-nitrophenyl phosphate, 15 mmol/L; reaction temperature, 30 degrees C; reaction time, 2 min. The assay conditions are optimum for all human serum isoenzymes.
D000293:Adolescent; D000328:Adult; D000368:Aged; D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000097:blood*; D000641:Ammonia / Q000494:pharmacology; D002021:Buffers; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000097:blood*; D008094:Lithium / Q000494:pharmacology; D008297:Male; D008536:Meglumine; D008875:Middle Aged; D012016:Reference Values; D013012:Sorbitol / Q000031:analogs & derivatives
D002021:Buffers; D007527:Isoenzymes; D013012:Sorbitol; D008536:Meglumine; D000641:Ammonia; D008094:Lithium; D000469:Alkaline Phosphatase