Evaluation of abalone β-glucuronidase substitution in current urine hydrolysis procedures.
This study examined the potential of abalone β-glucuronidase as a viable and cost effective alternative to current hydrolysis procedures using acid, Helix pomatia β-glucuronidase and Escherichia coli β-glucuronidase. Abalone β-glucuronidase successfully hydrolyzed oxazepam-glucuronide and lorazepam-glucuronide within 5% of the spiked control concentration. Benzodiazepines present in authentic urine specimens were within 20% of the concentrations obtained with the current hydrolysis procedure using H. pomatia β-glucuronidase. JWH 018 N-(5-hydroxypentyl) β-d-glucuronide was hydrolyzed within 10% of the control concentration. Authentic urine specimens showed improved glucuronide cleavage using abalone β-glucuronidase with up to an 85% increase of drug concentration, compared with the results obtained using E. coli β-glucuronidase. The JWH 018 and JWH 073 carboxylic acid metabolites also showed increased drug concentrations of up to 24%. Abalone β-glucuronidase was able to completely hydrolyze a morphine-3-glucuronide control, but only 82% of total morphine was hydrolyzed in authentic urine specimens compared with acid hydrolysis results. Hydrolysis of codeine and hydromorphone varied between specimens, suggesting that abalone β-glucuronidase may not be as efficient in hydrolyzing the glucuronide linkages in opioid compounds compared with acid hydrolysis. Abalone β-glucuronidase demonstrates effectiveness as a low cost option for enzyme hydrolysis of benzodiazepines and synthetic cannabinoids.
Division of Forensic Toxicology, Armed Forces Medical Examiner System, Dover, DE, USA.
Brittany
B
Malik-Wolf
Shawn
S
Vorce
Justin
J
Holler
Thomas
T
Bosy
D000701:Analgesics, Opioid / Q000652:urine; D000818:Animals; D001569:Benzodiazepines / Q000652:urine; D002186:Cannabinoids / Q000652:urine; D003061:Codeine / Q000378:metabolism; D003362:Cost-Benefit Analysis; D004926:Escherichia coli / Q000201:enzymology; D049851:Gastropoda / Q000201:enzymology*; D005966:Glucuronidase / Q000378:metabolism*; D006372:Helix, Snails / Q000201:enzymology; D006868:Hydrolysis; D008140:Lorazepam / Q000031:analogs & derivatives / Q000378:metabolism; D009022:Morphine Derivatives / Q000378:metabolism; D010076:Oxazepam / Q000031:analogs & derivatives / Q000378:metabolism; D013048:Specimen Handling; D016482:Urinalysis / Q000379:methods*
D000701:Analgesics, Opioid; D002186:Cannabinoids; D009022:Morphine Derivatives; D001569:Benzodiazepines; C026256:oxazepam glucuronide; D010076:Oxazepam; D005966:Glucuronidase; D008140:Lorazepam; C018108:morphine-3-glucuronide; C031290:lorazepam glucuronide; D003061:Codeine