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J Anal Toxicol. 2016 Nov;40(9):732-737. Epub 2016 Jul 11.

Variations in enzymatic hydrolysis efficiencies for amitriptyline and cyclobenzaprine in urine.

Author information

1
Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208, USA.
2
Integrated MicroChromatography Systems, LLC, 541 Main Street, Suite 117, Columbia, SC 29208, USA.
3
PSO Laboratory, LLC, 2250 South Washington Ave, Suite 202, Lansing, MI 48910, USA.
4
Department of Chemistry and Biochemistry, University of South Carolina, Columbia, SC 29208, USA morgansl@mailbox.sc.edu.

Abstract

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.

PMID:
27405366
DOI:
10.1093/jat/bkw062
[Indexed for MEDLINE]

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