Establishment of a two-dimensional chiral HPLC system for the simultaneous detection of lactate and 3-hydroxybutyrate enantiomers in human clinical samples

Shu Ling Liu, Tsubasa Oyama, Yurika Miyoshi, Shiow Yunn Sheu, Masashi Mita, Tomomi Ide, Wolfgang Lindner, Kenji Hamase, Jen Ai Lee

研究成果: 雜誌貢獻文章同行評審

10 引文 斯高帕斯(Scopus)

摘要

A two-dimensional chiral high-performance liquid chromatography system was established for simultaneous detection of lactate (LA) and 3-hydroxybutyrate (3HB) enantiomers in human clinical samples. d-LA is increased upon kidney damage but 3HB protected against kidney injury. Therefore, determining the concentrations of d,. l-LA and d,. l-3HB simultaneously would be useful for evaluating pathological conditions. LA and 3HB were pre-column-derivatized with the fluorescent reagent 4-(. N-chloroformylmethyl-. N-methylamino)-7-nitro-2,1,3-benzoxadiazole (NBD-COCl) at 60. °C for 15. min and separated in the first dimension with a capillary monolithic octadecylsilane column. The mobile phase consisted of 13% acetonitrile and 0.05% tirfluoroacetic acid in water. Chiralpak QD-AX and KSAACSP-001S enantioselective columns were used in the second dimension to separate LA and 3HB enantiomers, respectively. Mobile phases were mixed solutions of methanol and acetonitrile containing formic acid. The separation factors were 1.14 and 1.08, respectively. The detection limit of LA and 3HB enantiomers was 10. fmol/injection. This method was applied to human clinical samples; intra- and inter-day relative standard deviations of LA and 3HB enantiomers were, respectively, 1.04-3.25% and 1.61-5.12% in plasma, 9.19-11.2% and 4.60-5.89% in urine, and 7.12-8.90% and 2.86-6.97% in saliva. This novel analytical method is a powerful tool for investigating variations in LA and 3HB enantiomers under disease conditions.

原文英語
頁(從 - 到)80-85
頁數6
期刊Journal of Pharmaceutical and Biomedical Analysis
116
DOIs
出版狀態已發佈 - 12月 10 2015

ASJC Scopus subject areas

  • 分析化學
  • 藥學科學
  • 藥物發現
  • 光譜
  • 臨床生物化學

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