WEBINARS - IMCS

ON-DEMAND WEBINAR:

Evaluation of two β-Glucuronidases for Urine Drug Testing

Dr. Nguyen Nguyen earned his PhD in Chemistry from UC-Davis and his clinical chemistry fellowship from Baylor Scott & White. Dr. Nguyen began his career in biopharmaceutical and transitioned to clinical research at UC-SF. He has setup a number of small clinics and a reference lab. He is currently the clinical chemist/toxicologist and safety officer at the Baylor Scott and White Medical Center. With over 10 years of experience, he’s here to share with you one of his recent works in drug testing.

Full Abstract

OBJECTIVES
Liquid chromatography tandem mass spectrometry (LC-MS/MS) has been the gold standard method for urine drug testing (UDT). Analytical sensitivity and specificity of drugs have increased dramatically due to mass spectrometry detection. In addition, the beta-glucuronidase in the sample workup step also helps recover parent drug compounds by hydrolyzing glucuronides on metabolites, which were formed through normal metabolism. There are many types and generations of beta-glucuronidases available, and here, we are evaluating two of the current modern products for their deconjugation performance on commonly prescribed opioids and benzodiazepines at room temperature (20-25°C).

METHODS
Urine remnants of specimens, previously confirmed positive, were pooled together to include many opioids and benzodiazepines of interest. Optimization study was carried out by varying each product amount from 0 to 100uL. The samples were incubated for 15min at room temperature along with deuterated internal standards. A diluent of 43% methanol in water was added prior to MS/MS detection of 12 opioids and 7 benzodiazepines. Results were normalized to the highest amount for graphical evaluation. We define performance adequacy if conjugation achieved of at least 80% of highest normalized value.

RESULTS & CONCLUSION
For benzodiazepines, the recovery was best achieved by using 20uL of our current enzyme or IMCSzyme RT. For opioids, the recovery was best achieved by using 80uL and 20uL of the current enzyme/buffer product and IMCSzyme RT, respectively. Both products offered similar performance in extracting parent opioids and benzodiazepines compounds from urine at room temperature.

β-glucuronidases recover drug compounds by hydrolyzing glucuronides on metabolites formed by normal metabolism. In this talk, Dr. Nguyen Nguyen, Clinical Chemist/Toxicologist and Safety Officer at Baylor Scott & White, presents his group's evaluation of two novel β-glucuronidase products, IMCSzyme RT and "CE" (their "current enzyme"), for recovery of common opioids and benzodiazepines at room temperature.

At the end of this webinar, participants will:

Learn about the phases of drug metabolism and why β-glucuronidase enzymes is necessary in urine drug testing,
Recognize the advantages of novel room-temperature β-glucuronidases
Discuss the possible mechanisms behind the enzyme- and patient-dependent differences in deconjugation performance between the two enzyme products.

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Caleb R. Schlachter, Ph.D.

Principal Scientist

Caleb R. Schlachter, Ph.D., as the Principal Scientist at IMCS, leads and provides guidance for several research and development projects that involve proteins, including enzymes for glycan hydrolysis and glycan synthesis. He has co-authored multiple patents, posters, and peer-reviewed articles on β-glucuronidases and sulfatases.

Gray D. Amick, Ph.D.

Director of Operations

Gray D. Amick, Ph.D., is the Director of Operations at IMCS with over 26 years of experience in forensic DNA analysis and toxicology. Prior to joining IMCS, he led forensic DNA testing for the Richland County Sheriff’s Department as technical leader and lab director. He has been court-qualified as an expert over 100 times and has authored and co-authored multiple posters and peer-reviewed articles.

Amanda C. McGee

Research Scientist

Amanda C. McGee is a Research Scientist at IMCS involved with enzyme characterizations, new analytical method developments, and advanced technical support. She joined IMCS with several years of experience in analytical testing for active pharmaceutical ingredients as per cGMP, USP and ICH guidelines. She has co-authored peer reviewed articles in the Journal of Analytical Toxicology and presented research at national and international conferences.

L. Andrew Lee, Ph.D.

Co-Founder and Chief Scientific Officer

L. Andrew Lee, Ph.D. co-founded IMCS and leads research and development efforts in enzyme engineering and automated micro-chromatography workflows. He directs new market efforts in glycan synthesis, supported by three NIH Fast-Track awards.