Development and Validation of an Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for the Determination of MAM-2201 Metabolites in Human Urine
Keywords:
MAM-2201 N-(4-hydroxypentyl); MAM-2201 N-pentanoic acid; Synthetic cannabinoids; Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry (UHPLC-MS/MS).Abstract
Background. Although synthetic cannabinoids (SCs) are controlled and banned in many countries, they remain prevalent and continue to pose a serious public health problem due to severe intoxications and fatal cases. Their analytical detection is challenging because SCs undergo extensive metabolism, and urine rarely contains parent compounds. Therefore, forensic laboratories need analytical methods to identify and quantify synthetic cannabinoid metabolites.
Objectives. This study aimed to develop and validate a targeted UHPLC-MS/MS method to determine MAM-2201 metabolites in human urine.
Methods. Urine samples were hydrolyzed with β-glucuronidase and extracted via solid-phase extraction using Bond Elut Plexa PCX cartridges. Analysis was performed using a UHPLC–MS/MS system in positive electrospray ionization (ESI) mode with MRM detection on a C18 column.
Results. The method demonstrated good linearity (0.5-45 ng/mL, 0.75-45 ng/mL; r²≥0.996), minimal matrix effects, high recovery, and acceptable accuracy and precision (<15% RSD). LODs were 0.25 and 0.5 ng/mL, and LLOQs were 0.5 and 0.75 ng/mL, respectively. No carryover was observed, and analytes were stable under the tested conditions.
Conclusions. The developed UHPLC-MS/MS method provides a sensitive, simple, and rapid approach for reliable determination of MAM-2201 metabolites in human urine. It is suitable for forensic and toxicological applications.
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