Mastrianni Kaylee R, Lee L Andrew, Brewer William E, Dongari Nagaraju, Barna Michael, Morgan Stephen L
Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208, USA.
Integrated MicroChromatography Systems, LLC, 541 Main Street, Suite 117, Columbia, SC 29208, USA.
J Anal Toxicol. 2016 Nov;40(9):732-737. doi: 10.1093/jat/bkw062. Epub 2016 Jul 11.
A collaborative study was conducted to investigate discrepancies in recoveries of two commonly prescribed compounds, amitriptyline and cyclobenzaprine, in patient urine samples when hydrolyzed with different enzymes from different sources. A 2- to 10-fold increase in analyte recoveries was seen for patient samples hydrolyzed using a recombinant β-glucuronidase (IMCSzyme™) over samples hydrolyzed with β-glucuronidase from Haliotis rufescens We report outcomes from four commercially available β-glucuronidase enzymes (IMCSzyme™, Patella vulgata, Helix pomatia and H. rufescens) on patient samples that tested positive for amitriptyline and cyclobenzaprine. Our results confirm reduced hydrolysis of glucuronides by β-glucuronidases isolated from mollusks, but near complete conversion when using the recombinant enzyme. Our premise is that systematic differences in hydrolysis efficiencies due to varying substrate affinity among enzyme subtypes potentially impacts accuracy and reliability of measurements.
开展了一项合作研究，以调查两种常用处方药阿米替林和环苯扎林在患者尿液样本中，使用不同来源的不同酶进行水解时回收率的差异。与使用来自红鲍的β-葡萄糖醛酸酶水解的样本相比，使用重组β-葡萄糖醛酸酶（IMCSzyme™）水解的患者样本中，分析物回收率提高了2至10倍。我们报告了四种市售β-葡萄糖醛酸酶（IMCSzyme™、欧洲笠贝、罗马蜗牛和红鲍）对阿米替林和环苯扎林检测呈阳性的患者样本的检测结果。我们的结果证实，从软体动物中分离出的β-葡萄糖醛酸酶对葡萄糖醛酸苷的水解作用减弱，但使用重组酶时几乎能完全转化。我们的前提是，由于酶亚型之间底物亲和力不同，水解效率存在系统性差异，这可能会影响测量的准确性和可靠性。
