Abstract
PurposeWe developed and validated a method for quantitative analysis of 50 psychoactive substances and metabolites (antidepressants, benzodiazepines and opioids) in oral fluid samples using simple liquid–liquid extraction procedure followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS).MethodOral fluid samples were collected using Quantisal™ device and extracted by liquid–liquid extraction with 1.0 mL of methyl tert-butyl ether and then analyzed using LC–MS/MS.ResultsThe method attended method validation criteria, with limits of quantification as low as 0.5 and 1.0 ng/mL, and linearity between 0.5–50.0 ng/mL for antidepressants, 0.5–25.0 ng/mL for benzodiazepines and 1.0–50.0 ng/mL to opioids. During method validation, bias and imprecision values were not greater than 16 and 20%, respectively. Ionization suppression/enhancement bias results were not greater than 25%. No evidence of carryover was observed. Sample stability studies showed that almost all analytes were stable at 25 °C for 3 days and at 4 °C for 7 days. Freeze–thaw cycles stability showed that most antidepressants and opioids were stable under these conditions. Autosampler stability study showed that all analytes were stable for 24 h, except for nitrazepam and 7-aminoclonazepam. Thirty-eight authentic oral fluid samples were analyzed; 36.8% of the samples were positive for 2 drugs. Citalopram was the most common drug found, followed by venlafaxine.ConclusionsThe method was validated according to international recommendations for the 50 analytes, showing low limits of quantification, good imprecision and bias values, using simple liquid–liquid extraction, and was successfully applied to authentic oral fluid samples analysis.
Highlights
Oral fluid is used as an alternative matrix for diagnostic in clinical and workplace applications; drug testing under driving, drug monitoring and criminal justice settings have been increasing over the last 20 years [1,2,3,4,5,6]
The aim of this work was to develop and validate an analytical method for simultaneous and quantitative analysis of 50 psychoactive drugs of antidepressants, benzodiazepines and opioids, which are widely circulating in the world, in oral fluid samples, using simple liquid–liquid extraction and LC–MS/MS
methyl tert-butyl ether (MTBE) and hexane were evaluated individually as binary and ternary mixtures, using analytes’ peak areas as the measure of response
Summary
Oral fluid is used as an alternative matrix for diagnostic in clinical and workplace applications; drug testing under driving, drug monitoring and criminal justice settings have been increasing over the last 20 years [1,2,3,4,5,6]. The Roadside Testing Assessment (ROSITA) study, the objective of which was assessing drug and alcohol driving problems in six countries in Europe and four American states, highly recommended the start of random drug testing for government officials [10]; oral fluid was considered the most relevant biological matrix applied for roadside testing situations [2]. This matrix is useful to field sample collection, such as at parties and music festivals; one of the main objectives of this study is to establish solid patterns into drug consumption using oral fluid [11]. Mohr et al [12] evaluated the use of synthetic stimulants and hallucinogens in an electronic dance music festival, and concluded that paired blood, urine and oral fluid sampling, was the best choice for monitoring these populations
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