Quantitative detection method of triglycerides in serum lipoproteins and serum-free glycerol by high-performance liquid chromatography

Mitsuyo Okazaki, Norikazu Komoriya, Hitonobu Tomoike, Noriko Inoue, Shinichi Usui, Sekiko Itoh, Seijin Hosaki

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We have developed a simple and reliable method for quantitative detection of triglycerides (TG) in serum lipoproteins and serum-free glycerol (FG) by high-performance liquid chromatography (HPLC). After separation of serum constituents using a new gel-permeation column (TSK gel Lipopropak XL, Tosoh) and a new eluent (TSK eluent LP-2, Tosoh), TG and FG were detected by on-line reaction using a modified reagent which contained glycerol kinase, glycerol-3-phosphate oxidase and lipoprotein lipase. HPLC patterns showed five peaks corresponding to chylomicrons, very-low-density, low-density, high-density lipoproteins and FG. Absolute concentrations of TG in each lipoprotein fraction and serum FG were calculated from the corresponding peak areas using standard FG as a calibrator. Due to its very high sensitivity of peak detection, this method has become desirable for the analyses of lipoproteins of very low concentrations such as in cell culture systems. This technique will contribute to a better understanding of lipoprotein TG and serum FG distribution in human and nonhuman subjects.

Original languageEnglish
Pages (from-to)179-187
Number of pages9
JournalJournal of Chromatography B: Biomedical Applications
Volume709
Issue number2
DOIs
Publication statusPublished - May 29 1998

Keywords

  • Free glycerol
  • Lipoproteins
  • Triglycerides

ASJC Scopus subject areas

  • Chemistry(all)

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