Development and validation of a fast TEIS-TQMS screening method for the identification of 8 NPS in non-biological...

The number of New Psychoactive Substances (NPS) on the drug market is steadily increasing and there are no signs that this trend will change in the coming years. The constantly evolving drug market necessitates forensic experts to develop new methods for NPS analysis. With the growing number of analytes, the diversity of these compounds also increases, making it more challenging to develop screening methods that enable the analyses of a large number of the drugs simultaneously. It should also be noted that methods enabling quick NPS analysis are important due to constant changes in the popularity of "legal highs", usually from several to a dozen of NPS are popular on the drug market at a given time. Therefore, it was decided to develop a research method for rapid screening determination of compounds from three groups of NPS: cannabinoids, tryptamines, and cathinones. The analyzed substances included 5-MEO-MIPT, N-ethylcathinone, UR-144, MDPBP, 3-MMC, XLR-11, 5F-AKB48, and alpha-PVP. As a result of the conducted work, a method enabling the analysis of samples containing 8 NPS in one minute, was developed. To achieve this, a Thermal Extraction Ion Source technique was applied in conjunction with a Triple Quadrupole Tandem Mass Spectrometry (TEIS-TQMS). TEIS can also be coupled with high-resolution mass spectrometers and ion trap mass analyzers for the analyte identification. The first use of TEIS-TQMS in the field of "legal highs" analysis is reported and its application enables the examination of "designer drugs" with a significant simplification of the sample preparation stage. Sample preparation for real samples involved a simple solid-liquid extraction with methanol and an addition of internal standard. Importantly, the high speed and efficiency of the TEIS-TQMS technique with analysis times as short as one minute per sample have the potential to substantially reduce the time required for toxicological evaluations, expert reporting, and overall case handling. This aspect is particularly valuable in forensic and prosecutorial workflows, where timely and reliable analytical results are crucial for supporting law enforcement investigations and informing prosecutorial decisions. As such, the proposed methodology may play a key role in streamlining the early stages of criminal proceedings, especially in cases involving large volumes of seized materials. The research method was validated in accordance with the ANSI/ASB norm. We have determined potential interferences, selectivity, matrix effects, and the limit of detection. Validation results included calculated limits of detection ranging from 0.01 to 6.47 µg/ml, matrix effects from -97% to 241%, and coefficients of variation from 9 to 84%. The usefulness of the developed method was confirmed by analyzing real samples and the results were subsequently validated using LC-MS/MS.