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On-line Deconjugation of Glucuronides Using an Immobilized Enzyme Reactor Based Upon Beta-Glucuronidase

On-line Deconjugation of Glucuronides Using an Immobilized Enzyme Reactor Based Upon Beta-Glucuronidase

M P Di Marco, G Felix, V Descorps, M P Ducharme, I W Wainer

J Chromatogr B Biomed Sci Appl. 1998 Sep 18;715(2):379-86.

PMID: 9792524

Abstract:

An immobilized enzyme reactor based upon beta-glucuronidase (BG-IMER) has been developed for the on-line deconjugation of substrates. The activity of the BG-IMER and its applicability to on-line deconjugation was investigated. The BG-IMER was coupled to a reversed-phase column (C8 or C18) and the latter column was used to separate substrates and products eluted from the beta-glucuronidase reactor. The activity of the BG-IMER was followed by measurement of percent deconjugation and the parameters investigated were: substrate concentration, pH (4 to 6), temperature (r.t., 37 degrees C), enzyme-substrate contact time using flow-rates of 0.1 to 1.0 min/min (15-1.5 min). The glucuronides used in the evaluation of the BG-IMER were: 4-methylumbelliferyl-beta-D-glucuronide, p-acetaminophen-beta-D-glucuronide, 3'-azido-3'-deoxythymidine-beta-D-glucuronide, phenyl-beta-D-glucuronide, chloramphenicol-beta-D-glucuronide, estradiol-17-beta-D-glucuronide and morphine-beta-D-glucuronide. The development of on-line HPLC deconjugation of glucuronide substrates using the BG-IMER will facilitate the identification of metabolites and quantification of aglycones in metabolic and pharmacokinetic studies.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP16110104	Paracetamol β-D-glucuronide		16110-10-4	Price
AP17685051	Phenyl-β-D-glucuronide		17685-05-1	Price

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