Division of Forensic Toxicology, Armed Forces Medical Examiner System, Dover, DE, USA.
J Anal Toxicol. 2014 Apr;38(3):171-6. doi: 10.1093/jat/bku003. Epub 2014 Jan 31.
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.
本研究考察了鲍鱼β-葡糖苷酸酶作为一种可行且具有成本效益的替代方法的潜力，以替代当前使用酸、赫氏粪产碱杆菌β-葡糖苷酸酶和大肠杆菌β-葡糖苷酸酶的水解方法。鲍鱼β-葡糖苷酸酶成功地将奥沙西泮-葡糖苷酸和劳拉西泮-葡糖苷酸水解至加标对照浓度的 5%以内。真实尿液标本中的苯二氮䓬类药物与当前使用赫氏粪产碱杆菌β-葡糖苷酸酶的水解方法获得的浓度相差 20%以内。JWH 018 N-(5-羟基戊基)β-d-葡糖苷酸的水解率与对照浓度相差 10%以内。与使用大肠杆菌β-葡糖苷酸酶相比，真实尿液标本中使用鲍鱼β-葡糖苷酸酶可提高 85%的药物浓度，从而改善葡糖苷酸的裂解。JWH 018 和 JWH 073 羧酸代谢物的药物浓度也增加了 24%。鲍鱼β-葡糖苷酸酶能够完全水解吗啡-3-葡糖苷酸对照物，但与酸水解结果相比，真实尿液标本中只有 82%的总吗啡被水解。可待因和氢吗啡酮的水解在不同标本之间存在差异，表明与酸水解相比，鲍鱼β-葡糖苷酸酶在水解阿片类化合物的葡糖苷酸键方面可能效率较低。鲍鱼β-葡糖苷酸酶作为一种低成本的酶水解苯二氮䓬类药物和合成大麻素的选择方法，具有有效性。
