Clinical Chemistry
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Clinical Chemistry 47: 966-967, 2001;
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(Clinical Chemistry. 2001;47:966-967.)
© 2001 American Association for Clinical Chemistry, Inc.

Technical Briefs

Evaluation of the Drew Scientific DS30 Homocysteine Assay in Comparison with the Centers for Disease Control and Prevention Reference HPLC Method

Ming Zhang1, Elaine W. Gunter1 and Christine M. Pfeiffera1

1 National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341

aauthor for correspondence; fax 770-488-4609, e-mail cpfeiffer@cdc.gov

Determination of total homocysteine (tHcy) in plasma is becoming an important diagnostic procedure in clinical chemistry because a slightly increased concentration of tHcy in plasma has been discussed as an important independent risk factor for atherosclerotic diseases (1). Many methods, mostly by HPLC, have been reported for measuring tHcy (2). However, these methods are relatively complex and require highly specialized equipment. Drew Scientific, Inc. (company named for identification purposes only; this evaluation does not constitute an endorsement by CDC) has developed the DS30 tHcy system for measuring tHcy in plasma. We evaluated this new system and compared it with our CDC reference HPLC method (3).

The DS30 tHcy system comprises a small HPLC system using a 5-cm reversed-phase column, a tHcy assay reagent set that contains the necessary reagents and calibrators [5 and 20 µmol/L homocystine (concentration was equivalent to the free thiol)] and a quality-control (QC) set containing two concentrations of Hcy. The sample preparation requires 200 µL of plasma. A batch of 30 samples can be processed within 90 min. After the addition of 10 µL of the internal standard (IS; 2-mercaptoethylamine) to 200 µL plasma, the mixture of disulfides, mixed disulfides, and protein-bound thiols is reduced using 20 µL of tris(2-carboxyethyl)phosphine (TCEP). Protein is precipitated from this solution with trichloroacetic acid, and 100 µL of the supernatant is then derivatized with a fluorescent thiol-specific dye [ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBDF) in EDTA/borate buffer] at 60 °C for 50 min. The thiol derivatives are separated in a subsequent step by HPLC and detected by . . . [Full Text of this Article]


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


Home page
 Clin. Chem.Home page
D. M. Hill, L. J. Johnson, P. J. Burns, A. M. Neale, D. M. Harmening, and A. C. Kenney
Effects of Temperature on Stability of Blood Homocysteine in Collection Tubes Containing 3-Deazaadenosine
Clin. Chem., November 1, 2002; 48(11): 2017 - 2022.
[Abstract] [Full Text] [PDF]


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