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.
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 < or = 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 mumol/L, respectively; P < 0.001) and healthy female donors (7.85 and 10.34 mumol/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.
Jacobsen
D W
DW
Department of Clinical Pathology, Cleveland Clinic Foundation, OH 41195.
Gatautis
V J
VJ
Green
R
R
Robinson
K
K
Savon
S R
SR
Secic
M
M
Ji
J
J
Otto
J M
JM
Taylor
L M
LM
D000328:Adult; D000368:Aged; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D003545:Cysteine / Q000097:blood; D004151:Dipeptides / Q000097:blood; D005260:Female; D005492:Folic Acid / Q000097:blood*; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D008297:Male; D008832:Microchemistry; D008875:Middle Aged; D012016:Reference Values; D012680:Sensitivity and Specificity; D012727:Sex Characteristics; D013438:Sulfhydryl Compounds / Q000097:blood*; D014805:Vitamin B 12 / Q000097:blood*
D004151:Dipeptides; D013438:Sulfhydryl Compounds; D006710:Homocysteine; C028505:cysteinylglycine; D005492:Folic Acid; D003545:Cysteine; D014805:Vitamin B 12