Abstract
New psychoactive substances (NPS) represent an important focus nowadays and are continually produced with minimal structural modifications in order to circumvent the law and increase the difficulty of identifying them. Moreover, since there are a high number of different compounds, it is arduous to develop analytical screening and/or confirmation methods that allow the identification and quantification of these compounds. The aim of this work is to develop and validate a bioanalytical method for detecting new synthetic drugs in biological samples, specifically oral fluid, using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS) with minimal sample pretreatment. Oral fluid samples were simply centrifuged and denaturized with different rapid procedures before injection into the LC-MS/MS system. Calibration curves covered a linear concentration range from LOQ to 100 ng/mL. Validation parameters such as linearity, precision, accuracy, selectivity, matrix effect and thermal stability were evaluated and showed satisfactory results, in accordance with US Food & Drug Administration guidelines. The inter-day analytical bias and imprecision at two levels of quality control (QC) were within ±15% for most compounds. This method was able to identify and calculate the concentration of 10 NPS validated in this biological sample, even in the presence of matrix effect.
Highlights
The drugs of abuse scenario is constantly changing with the rapid development of unregulated synthetic and ad-hoc-designed compounds (Anzillotti et al, 2019b; UNODC Early Warning Advisory on New Psychoactive Substances, 2020)
Mass Spectrometer Parameters A mixture of analytes at a concentration of 0.2 μg/mL was infused in the mass spectrometer both for optimizing physical parameters such as declustering potential (DP) and collision energy (CE) and for detecting characteristic analyte transitions
Relative matrix effects were examined by using the approach proposed by Matuszewski: slope CV (%), the slope difference (%), and the assay CV range (%) (Matuszewski, 2006; Food Drug Administration (FDA), 2018) were obtained from five different lots of biological fluid spiked with the same calibration levels
Summary
The drugs of abuse scenario is constantly changing with the rapid development of unregulated synthetic and ad-hoc-designed compounds (Anzillotti et al, 2019b; UNODC Early Warning Advisory on New Psychoactive Substances, 2020). NPS are continually synthesized in illegal laboratories, and their consumption produces significant dangerous effects on human health that are still under investigation, such as agitation, aggression, and acute psychosis, as well as the potential development of dependence or cardiovascular effects (Anzillotti et al, 2019a; UNODC Early Warning Advisory on New Psychoactive Substances, 2020). In this context, the development and validation of analytical methods able to rapidly identify and correctly quantify such compounds are highly encouraged from a clinical and scientific point of view. A recent development has been the discovery of synthetic cannabinoids in liquids for electronic cigarettes, very popular among young people (European Monitoring Centre for Drugs Drug Addiction, 2017)
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