Validated assay for the evaluation of multiple glucuronidation activities in human liver microsomes via liquid chromatography-tandem mass spectrometry

Abstract

A faster and more accurate LC-MS/MS method was established for the activity determination of multiple UGT isoforms in HLMs.

Cyp3a11‐mediated testosterone‐6β‐hydroxylation decreased, while UGT1a9‐mediated propofol O‐glucuronidation increased, in mice with diabetes mellitus

“…Simultaneous determination of 3‐EG, PG and AZTG by LC–MS/MS was performed as described previously . Briefly, LC separations were achieved using a Hypersil Gold C 18 (100 mm × 2.1 mm, 5 μm) with a guard column (10 mm × 2.1 mm, 5 μm) under a binary gradient, which included (A) 0.1% acetic acid, and 0.5 m m ammonium acetate in water and (B) acetonitrile (0 → 1.0 min: 20% B → 20% B; 1.0 min → 5.0 min: 20% B → 90% B; 5.1 min → 7.0 min: 20% B → 20% B) at a flow rate of 0.25 ml/min and at a temperature of 30 °C.…”

“…The reaction was initiated by adding UDPGA (7 m m ). After incubation at 37 °C for 20 min, the reaction was terminated and analysed as described previously .…”

Interaction between oblongifolin C and UDP-glucuronosyltransferase isoforms in human liver and intestine microsomes

“…The final volume of the organic solvents in the incubation mixture was not more than 1% (v/v). The LC-MS/MS system for the quantification of 4-MUG consisted of a Shimadzu Prominence UFLCXR system (Shimadzu Scientific Instruments, Columbia, MD) coupled to a Thermo Scientific TSQ Quantum Ultra triplequadrupole mass spectrometer (Shi et al, 2014).…”