Glucuronidation of Lipophilic Substrates: Preparation of 3-Benzo[a]pyrenyl-β-D-glucopyranosiduronic Acid in Multimilligram Quantities by Microsomal UDP-Glucuronyl Transferase

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Abstract
Convenient method for the enzyraic conversion of multimilli-gram quantities of 3-hydroxybenzo[a]pyrene to 3-benzo[a]pyrenyl-β-D-glucopyranosiduronic acid in 90% yield is described. Commer-cially available freeze-dried rabbit liver microsomes were incubated in the presence of UDPGA, 3-hydroxybenzo[a]pyrene, and Triton X-100 detergent (Figure 1). The course of the biosynthetic reaction was followed by fluorimetry. The glucuronide product was extracted from the acidified incubation supernate with ethyl acetate and the acid function of the glucuronide was utilized in an acid-base extraction procedure to purify the glucuronide from biological and unreacted starting material. The glucuronide pre-cipitated from ethyl acetate and was collected by centrifugation. High pressure liquid chromatography and spectroscopic techniques were used to verify the structure and purity of 3-benzo[a]-pyrenyl-β-D-glucopyranosiduronic acid.