Assay of fatty acid omega-hydroxylation using high-performance liquid chromatography with fluorescence labeling reagent, 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one (BrMB)

Anal Biochem. 1991 Nov 15;199(1):132-6. doi: 10.1016/0003-2697(91)90280-7.

Abstract

A sensitive nonradioisotopic method is reported for measuring microsomal lauric acid omega-hydroxylation activity. The assay is based upon separation and detection of 12-hydroxylauric acid formed by means of high-performance liquid chromatography following fluorescence labeling of the carboxyl group with 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one (BrMB). The use of 10-hydroxycapric acid as an internal standard affords the accurate and reproducible assay. The differential effect of dehydroepiandrosterone, a peroxisome proliferator, on the omega-hydroxylation activity in the liver and kidney of rats is also reported.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzoxazines
  • Chromatography, High Pressure Liquid / methods
  • Dehydroepiandrosterone / pharmacology
  • Fluorescent Dyes
  • Hydroxy Acids / analysis*
  • Hydroxy Acids / metabolism
  • Hydroxylation
  • Kidney Cortex / drug effects
  • Kidney Cortex / enzymology*
  • Lauric Acids / analysis*
  • Lauric Acids / metabolism
  • Male
  • Microsomes / drug effects
  • Microsomes / enzymology*
  • Microsomes, Liver / drug effects
  • Microsomes, Liver / enzymology*
  • Oxazines*
  • Rats
  • Rats, Inbred Strains
  • Reference Values
  • Spectrometry, Fluorescence

Substances

  • Benzoxazines
  • Fluorescent Dyes
  • Hydroxy Acids
  • Lauric Acids
  • Oxazines
  • lauric acid
  • 3-bromomethy-7-methoxy-1,4-benzoxazin-2-one
  • Dehydroepiandrosterone