Glucuronidation of aliphatic alcohols in human liver microsomes in vitro.

The glucuronidation of several short-chained aliphatic alcohols in vitro, with the use of human liver microsomes (HLM) as catalyst, was performed, and the kinetics were studied. The concentrations of the glucuronides were determined by gas chromatography-mass spectrometry after derivatization to the trimethylsilyl compounds. Alcohols from ethanol to pentanols were found to couple with activated glucuronic acid in HLM to a widely varying amount. For analytic reasons the glucuronide of methanol, which is formed after methanol consumption in human beings in vivo, could not be determined. The length of the alkyl chain played the decisive role in the maximum turnover rate of the glucuronidation. The affinity of the alcohols to UDP-glucuronosyltransferase (UDPGT), which catalyzes the glucuronidation reaction, was shown to depend strongly on their structure. Alcohols with a very short alkyl chain and secondary alcohols were glucuronidated much more slowly in comparison with findings for the longer chain primary alcohols and showed less affinity to UDPGT. The alcohols mutually inhibited glucuronidation. However, ethanol inhibited the glucuronidation of isopentanol or n-pentanol only in high concentrations, whereas the two pentanols inhibited each other to a high degree. The glucuronidation of aliphatic alcohols is probably catalyzed by only one of several very similar enzymes of the UDPGT. This finding was indicated by the fact that the Michaelis-Menten constants of the alcohols--with the use of different lots of the HLM from different liver donors--had nearly the same values.