Optimal enzymatic hydrolysis of urinary benzodiazepine conjugates.
Conditions for the enzymatic hydrolysis of benzodiazepine conjugates were systematically examined. Optimal recovery of the free drugs occurs when 1 mL of urine buffered to pH 4.5 is incubated with 5000 U of Helix pomatia beta-glucuronidase at 56 degrees C for 2 h. Urine specimens containing conjugates of temazepam, oxazepam, lorazepam, alpha-hydroxyalprazolam, 2-hydroxyethylflurazepam, and N-desalkyl-3-hydroxyflurazepam as targeted benzodiazepines were used. The freed drugs were quantitated by gas chromatography-mass spectrometry.
Meatherall
R
R
Department of Biochemistry, St. Boniface General Hospital, Winnipeg, Manitoba, Canada.
D000818:Animals; D001569:Benzodiazepines / Q000652:urine*; D002417:Cattle; D004926:Escherichia coli / Q000201:enzymology; D008401:Gas Chromatography-Mass Spectrometry; D005966:Glucuronidase / Q000378:metabolism*; D006372:Helix, Snails / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D008099:Liver / Q000201:enzymology
D001569:Benzodiazepines; D005966:Glucuronidase