Evidence map›Paper›PMID 40042693›Full record

ArticleForensic toxicology2025

Investigation of commercially available recombinant and conventional β-glucuronidases to evaluate the hydrolysis efficiencies against O-glucuronides and N-glucuronides in urinary drug screening.

Akira Namera, Takeshi Saito, Masataka Nagao

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Article in Forensic toxicology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

3 authors.

Akira NameraDepartment of Forensic Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-Ku, Hiroshima, 734-8551, Japan. namera@hiroshima-u.ac.jp.
Takeshi SaitoDepartment of Emergency and Critical Care Medicine, Tokai University School of Medicine, 143 Shimokasuya, Kanagawa, 259-1193, Japan.
Masataka NagaoDepartment of Forensic Medicine, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-Ku, Hiroshima, 734-8551, Japan.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

purposeTo achieve the rapid analysis of drug metabolites in urine, we examined the differences in the hydrolysis efficiencies against O-glucuronide and N-glucuronide by two commercially available glucuronidases and three commercially available recombinant ones.

methodsThe metabolites analyzed included oxazepam-O-glucuronide, amitriptyline-N-glucuronide, and diphenhydramine-N-glucuronide. Hydrolysis was performed using commercially available five enzymes at two different temperatures, and the reaction progress was monitored for up to 360 min. The amount of hydrolyzed product was quantified using liquid chromatography-tandem mass spectrometry.

resultsAlthough no enzyme selectivity was observed for the hydrolysis of O-glucuronide, the hydrolysis efficiency against N-glucuronide varied significantly, depending on the enzyme and reaction temperature. Among the enzymes evaluated, IMCSzyme 3S and the enzyme derived from E. coli demonstrated superior hydrolysis of N-glucuronides under optimal conditions. For IMCS RT, good results were also obtained by adding twice the amount of enzyme specified.

conclusionsSuitable enzymes and hydrolysis conditions were determined for the rapid and systematic screening of drug metabolites in human urine. These findings are expected to streamline the analytical workflow and reduce the need for tedious sample preprocessing.

Indexed as

GlucuronidaseGlucuronidesSubstance Abuse DetectionAmitriptylineChromatography, LiquidDiphenhydramineHumansHydrolysisRecombinant ProteinsTandem Mass SpectrometryTemperatureAmitriptylineDiphenhydramineGlucuronidaseGlucuronidesRecombinant ProteinsDrug screeningHydrolysis efficiencyN-GlucuronidesO-GlucuronidesRecombinant β-glucuronidase

Identifiers

PMID40042693
PMCPMC12241241

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