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18Feb 2015
Feb 18, 2015

A Time-course Study Reveals a 93% Hydrolysis of Buprenorphine & Norbuprenorphine

ABSTRACT OVERVIEW

Ameritox conducted a time-course study with IMCSzyme® for buprenorhpine urinalysis.  Buprenorphine is a commonly prescribes analgesic for pain management and treatment of opioid dependence.  In conventional analysis of buprenorphine by GC/MS, hydrolysis is required to cleave the glucuronides.  Hydrolysis by acid or base is ineffective for buprenhorphine, but is achievable with beta-glucuronidase enzymes. Enzyme hydrolysis, though, has traditionally been costly and time-consuming with reported incubation times of up to 16 hours depending on the source1,2.

Ameritox tested the effectiveness of IMCSzyme relative to beta-glucuronidase from abalone.  Data showed that the recovery of norbuprenorphine from the glucuronide control improved with heat activation and longer incubation times while the maximum hydrolysis of buprenorphine glucuronide controls at room temperature were equivalent to the measurements with heated incubation.

Total buprenorphine and norbuprenorphine in patient samples compared well between the enzyme treatment and total measurements from monitoring the intact glucuronides.  Overall, the percent hydrolysis exceeded 93% for all target analytes in patient samples in 30 minutes or less. 

References:

  1. M. A. El Sohly, W. Gul, S. Feng and T. P. Murphy. Hydrolysis of conjugated metabolites of buprenorphine II. the quantitative enzymatic hydrolysis of norbuprenorphine- 3-β-d glucuronide in human urine. J. Anal. Toxicol. 29: 570-573 (2005).
  2. S. Feng, M. A. El Sohly and Duckworth, D. T. Hydrolysis of conjugated metabolites of buprenorphine. I. the quantitative enzymatic hydrolysis of buprenorphine-3- β-d-glucuronide in human urine. J. Anal. Toxicol. 25: 589-593 (2001).

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IMCSzyme product information: www.imcszyme.com

Information summarized by IMCS from “Buprenorphine Hydrolysis Using a Novel Recombinant beta-glucuronidase for Urine Drug Testing” – presented by Ameritox, Ltd. at SOFT 2014

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