Single-shot multi-reaction monitoring of intact marker conjugates for quantitative profiling of human major microsomal glucuronidations and its utility to screen ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2016-11

AUTHORS

Yuqing Cheng, Jing Zhou, Meiling Wang, Yanyan Liu, Bin Guo, Bo Chen

ABSTRACT

UDP-glucuronosyltransferase (UGT) is a polymorphic family of conjugating enzymes responsible for the elimination of a myriad of xenobiotics and endogenous compounds. The precise reaction phenotyping of this multi-isoform superfamily is hampered by a lack of fast generic methods for directly measuring the diverse glucuronoconjugate metabolites for comprehensive profiling of UGT isoform-specific glucuronidations. We report here a single-shot liquid chromatography-tandem mass spectrometry (LC-MS/MS) method enabling the simultaneous direct measurement of nine intact glucuronides from hepatic microsomal glucuronidations mediated by a battery of isoforms (1A1, 1A3, 1A4, 1A6, 1A9, 2B7, 2B10, 2B15, and 2B17), which represent the majority of human UGTs in drug metabolism. This new method is based on post-incubation pooling of the individual probe reaction samples for nine-in-one cassette analysis with polarity switching multiple reaction monitoring (MRM) of all the marker glucuronides within a single LC-MS/MS injection. The pooled sample strategy overcomes the cross-interferences among the cocktail substrates and also increases the throughput. The periodic polarity switching of the LC-MRM acquisition expands the glucuronide profiling coverage using a generic single-run analysis. The source-induced dissociation of the glucuronoconjugates was evaluated as a generic alternative for their quantitation as their free aglycones, but a significant bias occurs against the traditional assumption that the parent substrates could be used as the surrogates for quantifying their glucuronide metabolites without authentic standards. After collective validations for analyte quantitation and enzyme kinetics, this single-shot cassette quantitative profiling approach may prove useful in large-scale phenotyping of human glucuronidations and rapid screening for UGT inhibitors in natural products. Graphical abstract Multi-reaction monitoring of intact conjugate metabolites for quantitative profiling of human major glucuronidations. More... »

PAGES

8117-8132

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00216-016-9915-z

DOI

http://dx.doi.org/10.1007/s00216-016-9915-z

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1044219237

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27604270


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