Development and validation of fast UHPLC-MS/MS screening method for 87 NPS and 32 other drugs of abuse in hair and nails: application to real cases

Abstract

Interest on keratinized matrices analysis for clinical and forensic purposes has been recently grown due to the wide temporary detection window for psychotropic and toxic substances entrapped after repeated consumption. In this study, the first UHPLC-MS/MS screening method to quantify 119 molecules among most abused classic drugs and new psychoactive substances was developed and fully validated. An amount of 25 mg hair or nails samples, added with the internal standard mixture, were cut and incubated with 500 µL M3® buffer reagent at controlled temperature. After cooling, 1l supernatant was injected in the chromatographic system equipped with an Oasis HLB column. After the 10 min chromatographic separation through a gradient mobile phase (aqueous ammonium formate, phase A; acetonitrile, phase B), the target compounds were detected in multiple reaction monitoring mode. The method was linear (r 2 always better than 0.99) in a calibration range of 0.01-10 ng compound for mg hair and of 1-200 pg compound per mg nail. Recovery of analytes under investigation was always better than 75% and no significant ion suppression due to matrix effect was observed. Intra-assay and inter-assay precision and accuracy were always better than 15%. The applicability and trueness of the method were examined by analysing real samples of hair and nail from users of psychoactive drugs in recreational contexts. Both classic drugs and new psychoactive substances could be determined as result of repeated use and accumulation in keratin matrices

Hair analysis can support the follow‐up addiction care after acute New Psychoactive Substances intoxication: Illustration by two cases

“…31 Interestingly, DIP hair concentrations in Case #1 are around 40 times lower than that observed in a regular DIP abuser (4400 pg/mg). 32 Thus, our results suggest iterative or sporadic rather than chronic 17 ), stimulant and UR-144 use could be here very carefully considered as moderate, whereas MXE use appears to be rather in the order of high use levels.…”

“…Taking into account that the mean growth rate of human scalp hair is approximately 1 cm/month, 15 hair analysis allows retrospective information on former drug exposure and segmental hair analysis allows to assess the possibility of long‐term repeated exposure (here, each 2‐cm length hair section of Case # 1 or # 2 represents approximately 2 months of consumption) 16 . Accordingly, hair analysis represents an efficient tool for NPS prevalence evaluation on repeated use in various populations of drug consumers 17–26 . Nevertheless, the interpretation of NPS hair concentrations remains challenging because of the lack of reference values to discriminate between chronic and occasional use 25,27,28 .…”

“…Those NPS correspond to four stimulants (three cathinone derivatives: 4‐MEC, NEB, and alpha‐PVP; and one amphetamine derivative: EPH), one ketamine derivative exhibiting hallucinogenic/dissociative properties (MXE), and one synthetic cannabinoid (UR‐144). Despite sparse literature data on NPS hair concentrations, which predominantly concern not well‐documented cases (i.e., hair concentrations are reported from NPS users without details on frequency and level of use) or sometimes only one reported hair concentration (e.g., for UR‐144, 100 pg/mg in a not documented case 17 ), stimulant and UR‐144 use could be here very carefully considered as moderate, whereas MXE use appears to be rather in the order of high use levels.…”

“…16 Accordingly, hair analysis represents an efficient tool for NPS prevalence evaluation on repeated use in various populations of drug consumers. [17][18][19][20][21][22][23][24][25][26] Nevertheless, the interpretation of NPS hair concentrations remains challenging because of the lack of reference values to discriminate between chronic and occasional use. 25,27,28 However, in case of an established NPS acute intoxication, hair analysis can provide the drug abuse history of the victim and, consecutively, is able to support clinicians in post-acute medical management.…”

Medicinal Cannabis and Synthetic Cannabinoid Use

“…The rapid growing popularity of new SC on the darknet and the constant synthesis of new substances have become a challenge for forensic laboratories. This analytical challenge involves not only the large range of compounds and metabolites to detect, but also to discern the type of biological specimens (conventional and non-conventional matrices) to be investigated [ 12 ]. Pellegrini et al proposed a screening method for the quantification of three SC (JWH-122, JWH-210 and UR-144) and their respective metabolites JWH-122 N-(4-hydroxypentyl), JWH-122 N-(5-hydroxypentyl), JWH-210 N-(4-hydroxypentyl), JWH-210 N-(5-hydroxypentyl), UR-144 N-(4-hydroxypentyl) and UR-144 N-(5-hydroxypentyl) in urine using a fast sample extraction and two different analytical techniques: ultra-high-performance liquid chromatography–high-resolution mass spectrometry (UHPLC-HRMS) and high-sensitivity gas chromatography–mass spectrometry (GC-MS) [ 13 ].…”