Objective The purpose of this preliminary study was to assess the utility of applying a non-targeted comprehensive drug screening using High Resolution Mass Spectrometry to a cohort of patients attending the Emergency Department (ED) due to drug intoxication; and to evaluate the differences in toxicological profile between methamphetamine (MAMP) and amphetamine (AMP) intoxicated patients.

Retrospective observational study of thirty-nine patients, 24 in the group MAMP and 15 in AMP group, which were attended in the ED of public hospitals in the Balearic Islands due to recreational drug poisoning between March 2019 and December 2020. Urine samples (500μL) were extracted by solid phase extraction using reversed-phase cartridges. The extracts were analyzed using an Agilent 1290 Series UPLC system coupled to an Agilent 6550 QTOF-MS system working in ESI positive mode. For chromatographic separation an Acquity UPLC® HSS T3 (2.1×75mm, 1.8μm) column with formic acid 0.1% in water and acetonitrile as mobile phases was used. Data mining was performing employing data independent acquisition mode using MassHunter Qualitative software (version B.08.00) in combination with a PCDL library (ForTox PCDL database, Agilent Technologies). [M+H]+ ions were recorded from 50–1000m/z values using 3 different collision energies (10, 20, 40eV). MS/MS spectra matching was manually checked and for tentative identification at least two product ions had to match those in the library with a score≥70%.

The most frequent classical drugs of abuse detected were cocaine (94.9%), MDMA (51.3%) and ketamine (41.0%), without significant differences between MAMP and AMP patients. MAMP related compounds such as 4-hydroxymethamphetamine (29.2%), 1,2-dimethyl-3-phenylaziridine (37.5%) and cyclopentamine (4.2%) were detected in MAMP users, while 3-hydroxyamphetamine (20%) was the only AMP related compound detected in AMP users. The most frequent new psychoactive substances (NPS) detected were cathinones, including mephedrone (23.1%), methylone (17.9%), metamfepramone (10.3%), 4’-methyl-α-pyrrolidinopropiophenone (2.6%) and 3′4′-methylendioxi-alfa-pyrrolidinopropiophenone (2.6%). Mephedrone were detected only in MAMP patients. Piperazines were detected in MAMP patients (58.3%) but not in AMP patients, specifically benzylpiperazine (33.3%) and N-(2-hydroxyphenyl)piperazine (2.6%). Moreover, 12.5% and 6.7% of the MAMP and AMP patients, respectively, were positive to tryptamines (5.1% to 5 or 6-metoxytryptamine, 5.1% to N,N-diethyltryptamine and 2.6% to etryptramine). Methyl lysergate was detected in 5.1% AMP patients. Regarding medicinal drugs, benzodiazepines were found in 50.0% and 46.7% of the MAMP and AMP patients, respectively, mainly oxazepam and midazolam. In addition, opiates were found in 29.2% and 33.3% of MAMP and AMP users, respectively, antidepressants in 25.0% and 33.3% (mainly clomipramine), and neuroleptics in 16.7% and 26.7% (mainly risperidone). A statistically significant higher prevalence of risperidone was found in AMP patients (P=0.014). Codeine and dextromethorphan were mainly found in AMP patients, although not statistically significant. Other less common medical drugs (frequency between 5–15%) included paracetamol, anticonvulsants, NSAIDs, antihistamines, anesthetics, antiretroviral drugs, beta-blockers, and ranitidine.

Candidates from many classes of drugs of abuse and medicinal drugs were tentatively identified in both MAMP and AMP patients using non-targeted drug screening. MAMP patients showed a higher prevalence of NPS (mephedrone and piperazines) while AMP patients showed a higher prevalence of medicinal drugs (opiates and opioids, antidepressants and neuroleptics).

Conclusion: MAMP and AMP overdosed users represent a high-risk population due to their elevated polyconsumption of drugs. MAMP users drug profile seems to be different with respect AMP users. We were able to identify a combination of classical, medicinal and NPS drugs in both populations using QTOF-MS methodology.