Journal Article

Liquid Chromatography-Tandem Mass Spectrometry Detection of the Quaternary Ammonium Compound Mebezonium as an Active Ingredient in T61<sup>®</sup>

Katrin M. Kirschbaum, Wolfgang Grellner, Gertrud Rochholz, Frank Musshoff and Burkhard Madea

in Journal of Analytical Toxicology

Volume 35, issue 2, pages 124-128
Published in print March 2011 | ISSN: 0146-4760
Published online March 2011 | e-ISSN: 1945-2403 | DOI: http://dx.doi.org/10.1093/anatox/35.2.124
Liquid Chromatography-Tandem Mass Spectrometry Detection of the Quaternary Ammonium Compound Mebezonium as an Active Ingredient in T61®

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Quaternary ammonium compounds pose an analytical challenge. Mebezonium, a muscle-relaxing agent contained in veterinary euthanasia solution T61, was analyzed in body fluids, organs, and injection sites of a veterinarian by liquid chromatography-tandem mass spectrometry (LC-MS-MS) method. Additionally, embutramide and tetracaine, which are two other active ingredients contained in T61, methadone, xylazine, and analgesics were detected by LC-MS-MS and high-performance liquid chromatography-ultraviolet detection methods. For detection of mebezonium a solid-phase extraction (SPE) combined with ion-pairing reagent heptafluorobutyric acid was developed. Separation was achieved on Phenomenex Synergi Hydro RP C18 column combined with ammonium formate buffer and acetonitrile (pH 3.5). To enrich other drugs, liquid-liquid extraction procedures were used. Most of these drugs were separated on a Restek Allure PFP Propyl column using the mentioned mobile phase. Mebezonium and embutramide were detected in femoral vein serum in concentrations of 10.9 and 2.0 mg/L, respectively. The concentration of xylazine and methadone in serum was 2.0 and 0.4 mg/L, respectively. The LC-MS-MS method with SPE combined with an ion-pairing reagent allowed the quantitation of mebezonium. Methadone was detected in toxic concentrations and was, in combination with xylazine and T61, considered to be the cause of death.

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Subjects: Medical Toxicology ; Toxicology (Non-medical)

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