A general assay for UDPglucuronosyltransferase activity using polar amino-cyano stationary phase HPLC and UDP[U-14C]glucuronic acid

Anal Biochem. 1986 Nov 15;159(1):198-205. doi: 10.1016/0003-2697(86)90328-3.


The assay of UDPglucuronosyltransferase activity toward various substrates using UDP[U-14C]glucuronic acid is described. HPLC on a polar amino-cyano bonded phase column was used to separate radioactive glucuronides from unmetabolized UDP[U-14C]glucuronic acid and other labeled reaction products. Radioactivity was measured using flow-through scintillation counting. All the glucuronides analyzed, with one exception, chromatographed with the same retention time (9.0-9.6 min) under the conditions described. Glucuronide conjugates were identified by comparison with retention times of commercial glucuronide standards, using radioactive aglycones, or hydrolysis with beta-glucuronidase. The method provides a unified, sensitive (100-200 pmol of glucuronide product) and reproducible assay for a wide variety of UDPglucuronosyltransferase substrates, and could be extended to include many others.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, High Pressure Liquid / methods*
  • Glucuronates
  • Glucuronosyltransferase / analysis*
  • Male
  • Microsomes, Liver / enzymology
  • Rats
  • Rats, Inbred Strains
  • Substrate Specificity
  • Uridine Diphosphate Glucuronic Acid


  • Glucuronates
  • Uridine Diphosphate Glucuronic Acid
  • Glucuronosyltransferase