[High-performance liquid chromatographic assay for plasma total homocysteine].
OBJECTIVE
To establish a high-performance liquid chromatography (HPLC) assay with an internal standard for determining plasma total homocysteine.


METHODS
The Hcy-mixed disulfides or protein bound Hcy in the plasma were reduced with TCEP. Subsequently, the thiol-compounds in the samples were derivatized with a thiol-specific fluorogenic reagent, SBD-F. The derivate was then separated and detected by HPLC with fluorometric detection.


RESULTS
The detection limit was 0.625 mumol.L-1; the rate of recovery was 98.87%; the intra-assay coefficient of variation (CV) was less than 5.5%; and the inter-assay CV was less than 9.0%.


CONCLUSIONS
The method presented in this paper is simple, sensitive, precise, and stable.
Dai
Chong-wen
CW
Molecular Hematology Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Zhang
Guang-sen
GS
Wu
Xian-ping
XP
D000328:Adult; D000368:Aged; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005260:Female; D015590:Fluorophotometry / Q000379:methods; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012680:Sensitivity and Specificity
D006710:Homocysteine