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Clinical Chemistry 48: 665-668, 2002;
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(Clinical Chemistry. 2002;48:665-668.)
© 2002 American Association for Clinical Chemistry, Inc.

Technical Briefs

Effects of Anticoagulant and Time of Plasma Separation on Measurement of Homocysteine

Natalia Louise Duarte1, Xing Li Wang2 and David Emil Leon Wilcken1a

1 Cardiovascular Genetics Laboratory, Department of Medicine, Prince of Wales Hospital, Randwick, NSW 2031 Australia;
2 Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX 78245-0549

aauthor for correspondence: fax 612-9382-4921, e-mail d.wilcken@unsw.edu.au)

Current interest in plasma total homocysteine (tHcy) measurements has increased with evidence that even mild hyperhomocysteinemia may be an independent risk factor for vascular disease (1) and that increases occur with vitamin deficiencies and reduced renal function. This has led to the investigation of problems associated with measurement, in particular the stability of tHcy after blood collection (2)(3). Transfer of homocysteine from red cells to plasma after venesection may occur and produce a 10% per hour increase in plasma tHcy concentrations (4), and thus influence assessments of the relative risk of disease (5). To reduce this increase, samples may be stored on ice and centrifuged within 1 h of collection. In epidemiologic surveys or even routine collection, this may not always be feasible.

We studied the stability of plasma tHcy measured by a fluorescence polarization immunoassay (FPIA) in samples from 9 cystathionine ß-synthase (CßS)-deficient homocystinuric patients and 13 healthy individuals. Blood was collected into both EDTA (1.8 g/L) and NaF (1.8 g/L EDTA and 3 g/L NaF) tubes. For six of the healthy individuals, we also used lithium heparin (14 kIU/L), sodium citrate (32 g/L trisodium citrate + 4.2 g/L citric acid; 3.2% solution), and ACD-B (13.2 g/L trisodium citrate + 5.25 g/L citric acid + 14.7 g/L glucose; 1.3% solution) tubes. The CßS-deficient patients were diagnosed and treated as described previously (6)(7), and the controls were healthy staff. Informed consent was obtained from all participants.

Within 15 min of collection, samples were divided into five aliquots. One aliquot from each sample was centrifuged (10 000g for 5 . . . [Full Text of this Article]


References




The following articles in journals at HighWire Press have cited this article:


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R. R. Calam, I. Mansoor, and J. Blaga
Homocysteine Stability in Heparinized Plasma Stored in a Gel Separator Tube
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S. Clark, L. D. Youngman, J. Sullivan, R. Peto, and R. Collins
Stabilization of Homocysteine in Unseparated Blood over Several Days: A Solution for Epidemiological Studies
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D. M. Hill, L. J. Johnson, P. J. Burns, A. M. Neale, D. M. Harmening, and A. C. Kenney
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A. P. Burke, V. Fonseca, F. Kolodgie, A. Zieske, L. Fink, and R. Virmani
Increased Serum Homocysteine and Sudden Death Resulting from Coronary Atherosclerosis With Fibrous Plaques
Arterioscler. Thromb. Vasc. Biol., November 1, 2002; 22(11): 1936 - 1941.
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