A Comprehensive HPLC–MS-MS Screening Method for 77 New Psychoactive Substances, 24 Classic Drugs and 18 Related Metabolites in Blood, Urine and Oral Fluid

Abstract

To date, more than 800 molecules are classified as New Psychoactive Substances (NPS), and it is reported that this number increases every year. Whereas several cases of polydrug consumption which led to acute intoxication and death are reported, a lack of effective analytical screening method to detect NPS and classical drug of abuse in human matrices affects the prompt identification of the probable cause of intoxication in emergency department of hospitals. In this concern, a fast, simple and comprehensive high-performance chromatography–tandem mass spectrometry (HPLC–MS-MS) screening method to detect and quantify 77 NPS, 24 classic drugs and 18 related metabolites has been successfully developed and validated in blood, urine and oral fluid. A small volume (100 µL) of whole blood samples spiked with internal standard deuterated mixture was added to 70 µL of M3® buffer and after precipitation of blood proteins, the supernatant was evaporated to dryness and reconstituted in 1 mL of mobile phase. Same volume (100 µL) of urine and oral fluid samples spiked with internal standard deuterated mix were only diluted with with 500 µL of M3 ® reagent. One microliter samples of each matrix was injected into HPLC–MS-MS equipment. The run time lasted 10 min with a gradient mobile phase. Mass spectrometric analysis was performed in positive ion MRM mode. The method was linear for all analytes under investigation with a determination coefficient always better than 0.99. The calibration range for blood and oral fluid was from limits of quantification (LOQs) to 200 ng/mL, whereas that for urine was LOQs to 1000 ng/mL. Recovery and matrix effect were always higher than 80%, whereas intra-assay and inter-assay precision was always better than 19% and accuracy was always within 19% of target in every matrix. Applicability of the method was verified by analysis of samples from real cases.

Simultaneous Quantification of 25 Fentanyl Derivatives and Metabolites in Oral Fluid by Means of Microextraction on Packed Sorbent and LC–HRMS/MS Analysis

“…For what concerns the determination of fentanyl and/or its analogues in this matrix, a few methods have already been published. These include two recent publications from Palmquist et al [3,9] and a number of papers comprising fentanyls in combination with other opioids [10] and new psychoactive substances (NPS) [11][12][13]. In all of these published methods, traditional sample preparation techniques such as dilution, SPE or LLE were used.…”

New Psychoactive Substances: Which Biological Matrix Is the Best for Clinical Toxicology Screening?

“…25 Furthermore, OF only provides evidence of recent drug use with a shorter detection window than urine. 25 Trana et al 26 developed a screening method for 77 NPSs and 10 metabolites of different classes in OF, blood, and urine. da Cunha et al 25 presented a screening method for 87 NPSs of different classes, and Rocchi et al 27 described a method for simultaneous screening and quantification of 31 NPSs of different classes in OF.…”

Development and validation of a rapid LC‐MS/MS method for the detection of 182 novel psychoactive substances in whole blood

“…The major challenge in the analysis of NPS resides in the diversity of structures and physicochemical properties among different NPS classes and within a single NPS class. 11 , 16 Several methods are already available in the literature to detect and/or quantify NPS in the main biological matrices 20 , 21 , 22 , 23 , 24 , 25 , 26 and particularly in whole blood, 27 , 28 , 29 , 30 , 31 though methods including a high number of compounds pertaining to different classes are still scarce. 22 In the present work, a LC‐MS/MS screening method for the rapid determination of 182 NPS in blood, including a wide‐panel of SCs and very recently emerged compounds, for example, 4F‐MDMB‐BINACA, 32 , 33 as well as multiple drug classes has been developed.…”