A high-sensitivity ultra-high performance liquid chromatography/high-resolution time-of-flight mass spectrometry (UHPLC-HR-TOFMS) method for screening synthetic cannabinoids and other drugs of abuse in urine

Abstract

The continuing emergence of designer drugs imposes high demands on the scope and sensitivity of toxicological drug screening procedures. An ultra-high performance liquid chromatography/high-resolution time-of-flight mass spectrometry (UHPLC-HR-TOFMS) method was developed for screening and simultaneous confirmation of both designer drugs and other drugs of abuse in urine samples in a single run. The method covered selected synthetic cannabinoids and cathinones, amphetamines, natural cannabinoids, opioids, cocaine and other important drugs of abuse, together with their main urinary metabolites. The database consisted of 277 compounds with molecular formula and exact monoisotopic mass; retention time was included for 192 compounds, and primary and secondary qualifier ion exact mass for 191 and 95 compounds, respectively. Following a solid-phase extraction, separation was performed by UHPLC and mass analysis by HR-TOFMS. MS, and broad-band collision-induced dissociation data were acquired at m/z range 50-700. Compound identification was based on a reverse database search with acceptance criteria for retention time, precursor ion mass accuracy, isotopic pattern and abundance of qualifier ions. Mass resolving power in spiked urine samples was on average FWHM 23,500 and mass accuracy 0.3 mDa. The mean and median cut-off concentrations determined for 75 compounds were 4.2 and 1 ng/mL, respectively. The range of cut-off concentrations for synthetic cannabinoids was 0.2-60 ng/mL and for cathinones 0.7-15 ng/mL. The method proved to combine high sensitivity and a wide scope in a manner not previously reported in drugs of abuse screening. The method's feasibility was demonstrated with 50 authentic urine samples.

Bioanalytical methods for the determination of synthetic cannabinoids and metabolites in biological specimens

“…Extraction of SCs using SPE has been used in blood and serum [91], urine [92][93][94][95][96] and oral fluid [45,97,98]. Different types of SPE columns can be used depending on the cost, availability and the nature of the analytes of interest.…”

“…The main challenge of non-targeted LC-HRMS screening methods is to achieve sufficient selectivity to identify low concentrations of SC [19]. Several non-targeted methods using HRMS technique for screening of drugs of abuse including SCs in biological matrices were published [89,94,114,115]. These methods are enabling forensic laboratories to obtain accurate data of MS and MS/MS without pre-selecting substances.…”

Current applications of high-resolution mass spectrometry for the analysis of new psychoactive substances: a critical review

“…Enzymatic hydrolysis with β-glucuronidase has been performed in a limited number of cases prior to SPE. [42,43].…”

“…Sundström et al [43] developed a NPS screening method in urine which included various hydroxylated and carboxylated synthetic cannabinoids metabolites which were positively detected in authentic urine samples.…”

Target and Non-target Techniques for the Quantitation of Drugs of Abuse, Identification of Transformation Products, and Characterization of Contaminants of Emergent Concern by High Resolution Mass Spectrometry

“…Nowadays, highresolution instruments such as the quadrupole time of flight (Q-TOF) or the Orbitrap are routinely used in urine analysis as screening tools for doping (Li et al 2013;Musenga and Cowan 2013;Sundström et al 2013). However, few methods have been implemented for the analysis of illicit drugs in wastewaters Fedorova et al 2013) despite clear advantages offered by full-scan HRMS such as enabling the screening of previously-acquired data for drugs of abuse that are identified after initial data processing has occurred.…”