A new on-line dual enzymatic method for simultaneous quantification of cholesterol and triglycerides in lipoproteins by HPLC

Shinichi Usui, Yukichi Hara, Seijin Hosaki, Mitsuyo Okazaki

Research output: Contribution to journalArticle

261 Citations (Scopus)

Abstract

We describe an on-line dual detection method using HPLC for lipoprotein analysis that allows simultaneous determination of cholesterol and triglyceride profiles from a single injection of sample. Two different gel permeation columns, TSKgel LipopropakXL and Superose 6HR, were applied to the dual detection system, evaluating analytical performance of the proposed method and the columns by analyzing serum samples from human and nonhuman subjects. Both TSK and Superose columns produced good within-day imprecision values less than 4.7% for cholesterol and 4.2% for triglyceride determination. Linear regression analysis showed the results from the Superose column (y) correlated well with those from the TSK column (x): y = 0.969x + 5.44 (r = 0.990) for total cholesterol (mg/dl), y = 1.08x - 11.14 (r = 0.985) for total triglycerides (mg/dl), and y = 1.093x - 0.06 (r = 0.978) for the ratios of triglycerides to cholesterol (mg/mg). Furthermore, the cholesterol and triglyceride profiles elucidated the differences in the resolution ability of the columns, which have not been apparent from a single lipid profile. We conclude that the dual detection concept with proper choice of column and enzymic reagents specific to the objectives of the particular study can facilitate studies of lipoprotein metabolism.

Original languageEnglish
Pages (from-to)805-814
Number of pages10
JournalJournal of Lipid Research
Volume43
Issue number5
Publication statusPublished - May 18 2002
Externally publishedYes

Keywords

  • Chylomicron
  • Gel permeation column
  • Hyperlipidemia
  • Lipoprotein profile

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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