An enzymatic method for the determination of butyrobetaine via conversion to carnitine after isolation by high performance liquid chromatography

Clin Chim Acta. 1988 Aug 15;176(1):17-27. doi: 10.1016/0009-8981(88)90170-2.

Abstract

An improved procedure for the determination of butyrobetaine [4-(N,N,N-trimethylammonio)butanoate] in plasma and tissue is described. Butyrobetaine was isolated by ion-exchange chromatography and high performance liquid chromatography. The isolation procedure was internally standardized with [3H]butyrobetaine. The recovery of butyrobetaine was greater than 90%. Following isolation butyrobetaine was enzymatically converted to carnitine using butyrobetaine hydroxylase and the resulting carnitine was assayed using carnitine acetyltransferase and [14C]acetylcoenzyme A. The conversion of butyrobetaine to carnitine and of carnitine to [14C]acetylcarnitine was greater than 98% as determined by high performance liquid chromatography. Using this method was analysed human sera (healthy controls) and tissues (autopsy) and found the following values: serum, 4.67 nmol/ml; kidney 17.6 nmol/g; liver, 26.5 nmol/g. The serum butyrobetaine values of twins suffering from carnitine deficiency were normal (3.78 and 3.87 nmol/ml), while the carnitine supplementation therapy caused an increase. Animal samples were analyzed and the values were 3-4 times higher than previously reported by others.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Betaine / analogs & derivatives*
  • Betaine / analysis
  • Carnitine / metabolism*
  • Child, Preschool
  • Chromatography, High Pressure Liquid
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Methods
  • Rats
  • Rats, Inbred Strains

Substances

  • Betaine
  • gamma-butyrobetaine
  • Carnitine