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Clinical Chemistry 42: 2028-2032, 1996;
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Clinical Chemistry, Vol 42, 2028-2032, Copyright © 1996 by American Association for Clinical Chemistry

Assay of plasma homocysteine: light sensitivity of the fluorescent 7- benzo-2-oxa-1, 3-diazole-4-sulfonic acid derivative, and use of appropriate calibrators

NP Dudman, XW Guo, R Crooks, L Xie and JS Silberberg
Center for Thrombosis and Vascular Research, University of New South Wales, Prince Henry Hospital, Little Bay, NSW, Australia. nickdud@ozemail.com.au

The recognition of homocysteine as a vascular risk factor has led to increased clinical interest in assaying plasma homocysteine concentrations. Our aim was to improve the reliability of a widely used assay based on HPLC of the fluorescent 7-benzo-2-oxa-1, 3-diazole-4- sulfonic acid (SBD) derivative. We found that SBD derivatives of homocysteine, cysteine, and N-acetylhomocysteine were highly unstable in light but essentially stable in the dark for several hours at either 0 degree C or 25 degrees C. As our primary calibrator, we chose homocystine added to human serum for more consistent results than homocysteine or homocystine in an aqueous buffer. N-acetylcysteine was effective as an internal recovery standard. We observed a previously unreported peak with a prolonged elution time in some plasma samples from subjects who had ingested methionine. Our findings suggest improvements in this and other assay procedures for plasma homocysteine.


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