Comparison of High Performance Liquid Chromatography and Fluorescence Polarization Immunoassay for Determination of Total Homocysteine in Human Plasma

Kang, Moon Soo; Choi, Jong Weon; Nahm, Chung Hyun; Lee, Jong Wook; Lee, Chul Hoon; Kim, Jin Ju; Pai, Soo Hwan

Abstract

BACKGROUND
It was purposed to estimate correlation between fluorescence polarization immunoassay (FPIA) and high performance liquid chromatography (HPLC), and precision of individual methods. It was also objected to describe distribution of plasma total homocysteine in Korean adults.
METHODS
The subjects were 100 adults admitted to Inha University Hospital during the month of October, 1998. The total plasma homocysteine concentration was measured by FPIA (IMx analyzer, Abbott Laboratories, IL, USA) and by HPLC (ACCLAIM Biogenic Amines Testing System, Bio-Rad Laboratories, CA, USA) using Bio-Rad Homocysteine.
RESULTS
Plasma homocysteine levels (mean+/-SD) from Korean healthy adults by FPIA and HPLC were 9.75+/-3.80micromol/L, 7.72+/-3.36micromol/L, respectively. Plasma homocysteine levels according to sex by FPIA were 11.79micromol/L for male, 7.71micromol/L for female, and those by HPLC were 9.47micro mol/L for male, 5.98micromol/L for female, respectively. Intra-assay coefficient variations (CVs) of low, medium, and high concentration by FPIA are 1.83%, 0.47%, and 1.66%, and those by HPLC are 5.53%, 5.37%, and 4.56%, respectively. Inter-assay CVs of low, medium, and high concentration by FPIA are 2.28%, 1.44%, and 1.29%, and by HPLC are 7.23%, 5.54%, and 4.95%, respectively.
CONCLUSION
Plasma homocysteine levels from male were significantly higher than female in Korean. Plasma homocysteine levels were increased according to increment of age. FPIA was more convenient, automatic, rapid, and reproducible than HPLC and also excellently correlated with HPLC. It is concluded that FPIA will potentially benefit for quantifying homocysteine in clinical laboratories.