A novel screening method for 64 new psychoactive substances and 5 amphetamines in blood by LC–MS/MS and application to real cases

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 129; pp. 441 - 449
• Main Authors: Vaiano, Fabio, Busardò, Francesco P., Palumbo, Diego, Kyriakou, Chrystalla, Fioravanti, Alessia, Catalani, Valeria, Mari, Francesco, Bertol, Elisabetta
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 10.09.2016
• Subjects: Adult; Amphetamine - blood; Amphetamine - chemistry; Amphetamines; Blood; Blood Chemical Analysis - methods; Blood screening; Chromatography, Liquid - methods; Female; Forensic Toxicology - methods; Humans; LC–MS/MS; Male; NPS; NPS intoxication cases; Psychotropic Drugs - chemistry; Sensitivity and Specificity; Street Drugs - chemistry; Tandem Mass Spectrometry - methods; Young Adult; NPS; Amphetamines; Blood screening; LC–MS/MS; NPS intoxication cases
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2016.07.009

[Display omitted] •Blood LC–MS/MS screening for 69 NPS and amphetamines was developed and validated.•Extraction procedure is simple and fast through deproteinization with acetonitrile.•High identification efficiency and sensitivity were allowed by dynamic MRM mode.•LLOQ was 0.1ng/mL for synthetic cannabinoids and 0.3ng/mL for cathinones.•The procedure was successfully applied to three real cases. Identification and quantification of new psychoactive substances (NPS), both in biological and non-biological samples, represent a hard challenge for forensic toxicologists. NPS are increasingly emerging on illegal drug market. Many cases of co-consumption of NPS and other substances have also been reported. Hence, the development of analytical methods aiming at the detection of a broad-spectrum of compounds (NPS and “traditional” drugs) could be helpful. In this paper, a fully validated screening method in blood for the simultaneous detection of 69 substances, including 64 NPS (28 synthetic cannabinoids, 19 synthetic cathinones, 5 phenethylamines, 3 indanes, 2 piperazines, 2 tryptamines, 2 phencyclidine, methoxetamine, ketamine and its metabolite) and 5 amphetamines (amphetamine, methamphetamine, MDMA, MDA, 3,4-methylenedioxy-N-ethylamphetamine − MDEA-) by a dynamic multiple reaction monitoring analysis through liquid chromatography − tandem mass spectrometry (LC–MS/MS) is described. This method is very fast, easy to perform and cheap as it only requires the deproteinization of 200μL of blood sample with acetonitrile. The chromatographic separation is achieved with a C18 column. The analysis is very sensitive, with limits of quantification ranging from 0.1 to 0.5ng/mL. The method is linear from 1 to 100ng/mL and the coefficient of determination (R2) was always above 0.9900. Precision and accuracy were acceptable at any quality control level and recovery efficiency range was 72–110%. Matrix effects did not negatively affect the analytical sensitivity. This method was successfully applied to three real cases, allowing identification and quantification of: mephedrone and methamphetamine (post-mortem); ketamine, MDMA and MDA (post-mortem); AB-FUBINACA (ante-mortem).