In vitro cleavage of paracetamol glucuronide by human liver and kidney beta-glucuronidase: determination of paracetamol by capillary electrophoresis.
A capillary electrophoresis (CE) method was developed using paracetamol glucuronide as a novel probe for human beta-glucuronidase activity. Using UV detection without prior sample clean-up procedures, fast and reliable quantitation of the released paracetamol was possible. The method showed good precision, accuracy and sensitivity with a limit of detection of 0.25 microM (38 ng/ml) and a limit of quantitation of 1 microM (151 ng/ml). The suitability of the method has been shown for enzyme kinetic studies using different liver and kidney homogenates, respectively. Our data clearly demonstrate that paracetamol glucuronide is cleaved by human beta-glucuronidase thereby releasing paracetamol. The CE method presented is not only a valuable tool for measuring human beta-glucuronidase activity, but also allows investigation of the contribution of deglucuronidation of paracetamol glucuronide to the disposition of paracetamol.
Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, Germany.
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Bohnenstengel
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HK
Kroemer
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B
Sperker
D000082:Acetaminophen / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D018712:Analgesics, Non-Narcotic / Q000032:analysis* / Q000378:metabolism; D019075:Electrophoresis, Capillary; D005966:Glucuronidase / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D018712:Analgesics, Non-Narcotic; D000082:Acetaminophen; C037386:acetaminophen glucuronide; D005966:Glucuronidase