Simple validated method of LC–MS/MS determination of BZ agent in rat plasma samples. Issue 4 (10th January 2020)

• Record Type: Journal Article
• Authors: Herman, David; Dlabkova, Alzbeta; Cechova, Lenka; Vanova, Nela; Misik, Jan; Jun, Daniel; Zdarova Karasova, Jana
• Is Part Of: Drug testing and analysis. Volume 12:Issue 4(2020)
• Page Start: 431
• Page End: 438
• Publication Date: 2020-01-10
• Subjects: 3‐quinuclidinyl benzilate -- agent BZ -- chemical warfare agent -- incapacitating; Drugs -- Analysis -- Periodicals; Drug testing -- Periodicals; Chemistry, Forensic -- Periodicals; 615.1901
• DOI: 10.1002/dta.2742
• Languages: English
• ISSNs: 1942-7603

Abstract

Abstract: Agent BZ (3‐quinuclidinyl benzilate) is a centrally acting synthetic anticholinergic agent, considered as a potential military incapacitating chemical warfare agent. Despite its significance as a model compound in pharmacological research and its potential misuse in chemical attacks, few modern analytical methods for BZ determination in biological samples have been published. The goal of the present work is to develop and validate a sensitive and rapid LC–MS/MS method for the determination of agent BZ in rat plasma. The sample preparation was based on solid‐phase extraction on C‐18 cartridges. The reversed‐phase HPLC coupled with the mass spectrometer with electrospray ionization in the positive ion‐selective reaction monitoring mode was employed in the BZ analysis. Atropine was used as an internal standard. The presented method is selective, accurate, precise, and linear (r 2 = 0.9947) in a concentration range from 0.5 ng/mL to 1 000 ng/mL and sensitive enough (limit of detection 0.2 ng/mL; limit of quantification 0.5 ng/mL) to determine the BZ plasma levels in rats exposed to 2 mg/kg and 10 mg/kg of BZ. The highest level of BZ in plasma was observed 5 minutes after intramuscular administration (154.6 ± 22.3 ng/mL in rats exposed to 2 mg/kg of BZ and 1024 ± 269 ng/mL in rats exposed to 10 mg/kg). After 48 h, no BZ was observed at detectable levels. This new method allows the detection and quantification of BZ in biological samples after exposure of an observed organism and it will be further optimized for other tissues to observe the distribution of BZ in organs. Abstract : An LC–MS/MS method to determine BZ agent (a potential military incapacitating chemical warfare agent) was designed, validated, and compared with other available methods and successfully used for analysis of BZ in rat plasma samples.