Simple extraction of toxicologically relevant psychotropic compounds and metabolites from whole blood using mini-QuEChERS followed by UPLC-MS/MS analysis.

The determination of psychotropic drugs and metabolites in blood is relevant in the context of both therapeutic drug monitoring and clinical and forensic toxicology. LC-MS/MS is the preferred method for these assays. However, LC-MS/MS is particularly susceptible to matrix ionization effects and appropriate sample preparation is required to minimize these effects. In this study, a simple, single-step, mini-QuEchERS extraction procedure, coupled to UPLC-MS/MS, was developed and validated for the determination of 15 toxicologically relevant compounds in whole blood, including psychoactive drugs and some metabolites. The assay was linear in the range of 25-1,000 ng ml-1 , fulfilling criteria for accuracy and precision. Extraction yields (71.9-87.7%) and matrix effects (-3.3 to +4.4%, with the exception of codeine, which had matrix effects of -35.36 to -28.14%) were acceptable for the majority of the evaluated compounds, using a single internal standard. The assay was applied to 238 clinical specimens from patients admitted to an emergency service, with 22 samples presenting quantifiable concentrations of 11 different compounds. The developed assay is a simple and efficient strategy for determination of target psychotropic drugs and metabolites in forensic and clinical toxicology.

Determination of cannabinoids in plasma using salting-out-assisted liquid-liquid extraction followed by LC-MS/MS analysis.

The detection of the markers of Cannabis consumption in biological specimens is an important task for drug testing laboratories in varous contexts. A simple assay combining salting-out assisted liquid-liquid extraction sample preparation and LC-MS/MS analysis was applied to the measurement of Δ9 -tetrahydrocannabinol, 11-nor-9-carboxy-Δ9 -tetrahydrocannabinol (THC-COOH), 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in 100 µl plasma specimens. The assay had linearity of 1-100 ng ml-1 for THC-COOH and 0.5-50 ng ml-1 for the other tested cannabinoids. Assay validation criteria were fulfilled. Extraction yields (88.7-97.3%) and internal-standard correct matrix effects (-9.6 to +5.4%) were acceptable. The assay was applied to 238 clinical specimens from trauma patients, with 19 samples presenting quantifiable concentrations of at least one of the target compounds. The developed assay is a simple and efficient strategy for simultaneous measurement of Δ9 -tetrahydrocannabinol, THC-COOH, 11-hydroxy-Δ9 -tetrahydrocannabinol, cannabinol and cannabidiol concentrations in plasma specimens.

Blood phosphatidyl ethanol levels as a tool to detect alcohol misuse in trauma patients.

INTRODUCTION: There is a strong need for a reliable marker of harmful alcohol consumption to identify injured patients that can benefit from alcohol interventions, and blood phosphatidyl ethanol (PEth) has not previously been tested on this population. This study aims to compare the performance of blood PEth concentration, blood alcohol concentration (BAC) and the Alcohol Use Disorders Identification Test Consumption (AUDIT-C) for the screening of alcohol misuse in trauma patients. METHODS: Prospective cross-sectional study of 238 adult patients presenting in the emergency department with any type of trauma. PEth concentration was determined in whole blood by high-performance liquid chromatography with tandem mass spectrometry. Consent, AUDIT-C score and demographic data were obtained. RESULTS: The sample consisted of majority male (67.6%), single (46.2%) and employed (66%) patients. The most common type of trauma was traffic collision (63.9%). The mean age was 41.7 years. We found a significant correlation between PEth levels with AUDIT-C score (Spearman's r = 0.654; p < .0001). PEth had an area under the ROC curve of 0.885 to detect hazardous alcohol consumption (AUDIT-C score ≥ 6) and PEth ≥23.9 ng/mL cutoff point provided 91.2% of sensitivity and 78.4% of specificity. Twelve patients reported alcohol abstinence, but had quantifiable levels of PEth. CONCLUSIONS: PEth levels and AUDIT-C score had a moderate correlation in our population. PEth was useful to identify 12 cases of underreporting of alcohol consumption habits. PEth shows promising results, but more research is needed to identify the best screening tool for alcohol misuse in trauma patients.

Validation of an analytical method for the simultaneous determination of 16 drugs and metabolites in hair in the context of driving license granting.

The use of psychoactive substances has been associated with increased risk for traffic accidents. Hair testing has become a routine practice in clinical and forensic toxicological laboratories, with a unique perspective in the investigation of drug consumption. The study aimed to develop and validate a UHPLC-MS/MS method for the determination of multiple drugs in hair, to be used for toxicological examination in driving license granting. Sample preparation was a one-step liquid extraction of milled hair with methanol, which was incubated for 15h at 50°C. The chromatographic separation was performed in a reversed phase column, with a run time of 2.2min. Measured compounds were cocaine, benzoylecgonine, norcocaine, anhydroecgonine methyl ester, cocaethylene, amphetamine, methamphetamine, methylenedioxyamphetamine, methylenedioxymethamphetamine, fenproporex, amfepramone, mazindol, codeine, morphine, 6-monoacetylmorphine, and tetrahydrocannabinol. The assay was linear for all substances (r>0.99), accurate (86.63-105.87 %), and precise, with a cv ranging from 1.9-13.5 % for intra-assay and 3.3-14.3 % for inter-assay. There was no significant carry over effect and the internal standard corrected matrix effect was minimal. The relative uncertainty percentages were below 9% for all the substances at cut-off values. The method was successfully applied to 50 hair samples from injured drivers, with 12% of positivity, including cocaine, MDMA and THC.