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Clinical Chemistry 34: 1607-1610, 1988;
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Clinical Chemistry, Vol 34, 1607-1610, Copyright © 1988 by American Association for Clinical Chemistry

Rapid determination of the hypoxanthine increase in ischemic exercise tests

PA Bolhuis, R Zwart, PR Bar, M de Visser and HJ van der Helm
Department of Neurology, Academic Medical Centre, Amsterdam, The Netherlands.

After ischemic exercise tests, performed to detect glycogenoses or myoadenylate deaminase (EC 3.5.4.6) deficiency, the increases in serum lactate and ammonia usually are measured. Determination of hypoxanthine instead of ammonia can also be used to show myoadenylate deaminase deficiency, but HPLC of hypoxanthine is time-consuming. As a substitute, we developed an indirect enzymatic equilibrium method for hypoxanthine based on coupling the chromogenic system 3,5-dichloro-2- hydroxy-benzenesulfonic acid/4-aminophenazone with formation of hydrogen peroxide by xanthine oxidase (EC 1.1.3.22). The pH optimum is at 7.8 and the absorbance maximum at 510 nm. The calibration curve is linear from 0 to 100 mumol/L and the detection limit is 0.9 mumol/L. Analytical variability (CV) was 1.5% to 3.6% within-run, 4.5% to 8.5% between-run. The assay can be performed with a standard spectrophotometer or a centrifugal analyzer. The coefficient of correlation was 0.68 between hypoxanthine and ammonia increases in plasma from controls who performed the exercise test.


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D. Duprez, M. De Buyzere, A. Van Wassenhove, and D. Clement
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Angiology, February 1, 1992; 43(2): 126 - 133.
[Abstract] [PDF]


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Copyright © 1988 by the American Association for Clinical Chemistry.