Rapid HPLC determination of total homocysteine and other thiols in serum and plasma: Sex differences and correlation with cobalamin and folate concentrations in healthy subjects

Donald W. Jacobsen, Vytenis J. Gatautis, Ralph Green, Killian Robinson, Susan R. Savon, Michelle Secic, Ji Ji, Joanne M. Otto, Lloyd M. Taylor

Research output: Contribution to journalArticlepeer-review

398 Scopus citations

Abstract

High-performance liquid chromatography with fluorescence detection has been utilized for the rapid determination of total homocysteine, cysteine, and cysteinylglycine in human serum and plasma. Our earlier procedure (Anal Biochem 1989; 178:208), which used monobromobimane to specifically derivatize thiols, has been extensively modified to allow for rapid processing of samples. As a result, >80 samples a day can be assayed for total homocysteine, cysteine, and cysteinylglycine. The method is sensitive (lower limit of detection ≤4 pmol in the assay) and precise (intra- and interassay CV for homocysteine, 3.31% and 4.85%, respectively). Mean total homocysteine concentrations in plasma and serum were significantly different, both from healthy male donors (9.26 and 12.30 μmol/L, respectively; P <0.001) and healthy female donors (7.85 and 10.34 μmol/L, respectively; P <0.001). The differences in total homocysteine between sexes were also significant (P = 0.002 for both plasma and serum). Similar differences were found for cysteine and cysteinylglycine. We found a significant inverse correlation between serum cobalamin and total homocysteine in men (P = 0.0102) and women (P = 0.0174). Serum folate also inversely correlated with total homocysteine in both sexes.

Original languageEnglish (US)
Pages (from-to)873-881
Number of pages9
JournalClinical Chemistry
Volume40
Issue number6
StatePublished - 1994
Externally publishedYes

ASJC Scopus subject areas

  • Clinical Biochemistry

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