Enantioselective immunoaffinity extraction for simultaneous determination of optically active bufuralol and its metabolites in human plasma by HPLC

Shigeo Ikegawa, Kenji Matsuura, Takehiro Sato, Ni Made Ria Isriyanthi, Toshifumi Niwa, Shinichi Miyairi, Hideo Takashina, Youichi Kawashima, Junichi Goto

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

A combined method of immunoaffinity extraction with high-performance liquid chromatography has been developed for the enantioselective determination of bufuralol and its metabolites in human plasma. The antibodies having high affinity toward the asymmetric center at the C-1 position of bufuralol and its 1'-oxidized metabolites and low affinity to their antipodes were elicited by immunization of rabbits with immunogens, (1R)- and (1S)-1'-oxobufuralol O-carboxymethyloxime-bovine serum albumin conjugates, respectively. 0.5 ml of the immunoaffinity adsorbent (7.6 mg · ml-1 for anti-(1S)-antibody and 28.8 mg · ml-1 for anti-(1R)-antibody) prepared by immobilization of an antibody was capable of retaining up to 1 μg of (R)- and (S)-bufuralol and up to 500 ng of other metabolites. The adsorbates were recovered quantitatively by elution with methanol-10 mM ammonium acetate buffer (pH 5) (95:5, v/v) without any interfering peaks on the high-performance liquid chromatogram. The proposed method was evaluated to be useful for the simultaneous determination of optically active bufuralol and its metabolite in plasma with acceptable recovery and precision.

Original languageEnglish
Pages (from-to)1-9
Number of pages9
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume17
Issue number1
DOIs
Publication statusPublished - 1998 May

Keywords

  • Bufuralol
  • Chiral separation
  • High-performance liquid chromatography
  • Immobilized antibody
  • Immunoaffinity extraction

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

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