Clinical Chemistry
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Clinical Chemistry 27: 1171-1175, 1981;
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Clinical Chemistry, Vol 27, 1171-1175, Copyright © 1981 by American Association for Clinical Chemistry

Response-surface-optimized, zinc-enhanced assay for serum alkaline phosphatase

JC Thompson, CT Hodges, GL Dobler and JA Williamson Jr

An optimized assay for alkaline phosphatase (EC 3.1.3.1) is reported. A discrete analyzer, the DuPont Automatic Clinical Analyzer (aca), was used for this study. The assay is based on results of response-surface experimental co-optimization techniques, and response is enhanced over the present aca assay. A key feature is the incorporation of zinc ions, both to fully optimize the assay and to reduce the sensitivity of measured activity to zinc-binding impurities in the buffer, 2-amino-2- methyl-1-propanol. In addition, a simple technique is described for measuring relative concentrations of zinc-binding impurities in this buffer. These features should be considered in the design of any assay for alkaline phosphatase that is based on p-nitrophenyl phosphate as substrate and 2-amino-2-methyl-1-propanol as buffer.




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Copyright © 1981 by the American Association for Clinical Chemistry.