Novel microsampling approach using fabric-phase sorptive extraction (FPSE) for cannabinoid analysis in blood

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• FPSE was applied for the first time to HPLC-MS/MS cannabinoid analysis in blood. • Blood microsamples were obtained on three different FPSE patch chemistries. • Sampling and extraction procedures were optimised. • Extraction yields of the seven cannabinoids was always > 85 % • The complete analytical workflow was validated according to international guidelines. • The HPLC-MS/MS method was highly sensitive; LOD ≤ 0.5 ng/mL, LOQ ≤ 1.5 ng/mL. The most important bioactive special metabolites of Cannabis sp. include cannabinoids, namely tetrahydrocannabinol (THC), cannabidiol (CBD) and their analogues. They are promising compounds with many biological activities, although THC is also known as an illicit compound in several countries. Due to their widespread occurrence and biological activity, cannabinoids are frequently investigated in terms of detection and quantification in various biological matrices. The increasing introduction of products based on plant derivatives and their compounds in the recreational, industrial, pharmaceutical, and illicit markets demands the constant development and refinement of rapid and reliable methods for the determination of these analytes in biological samples. Blood microsampling has emerged as a pivotal innovation in bioanalytical science, offering minimally invasive collection, reduced solvent consumption, and broad reliability and applicability across modern workflows. In this study, an innovative dried microsampling technology based on fabric phase sorptive extraction (FPSE) has been exploited for whole blood sampling, using polymeric fabric patches coated with three different sorbent chemistries: polyethylene glycol (CW20M), polydimethylsiloxane (PDMS) and polytetrahydrofuran (PTHF), for the analysis of two main active cannabinoids and five of their metabolites. An efficient HPLC-MS/MS method was developed, obtaining r 2 ≥ 0.9991, LOD ≤ 0.5 ng/mL and LOQ ≤ 1.5 ng/mL for all analytes. Just 20 µL of whole blood were needed to obtain a satisfactory dried FPSE spot. The sorbent activation procedures were optimised for different chemistries, ensuring compatibility with the analytes and high extraction efficiency. Dried sample pretreatment steps by solvent extraction have been optimised before HPLC-MS/MS analysis. Analytical results were satisfactory for all the analytes, with recovery always higher than 85 %, blood matrix effect lower than 14 % and FPSE matrix effect lower than 9 %. For intraday precision, the RSD value was consistently below 9.3 %, while it was lower than 9.8 % for interday precision. This is the first study where FPSE has been applied to the determination of THC, CBD and their main endogenous metabolites as a novel blood microsampling based approach.

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