title: High performance liquid chromatography-high resolution mass spectrometry and micropulverized extraction for the quantification of amphetamines, cocaine, opioids, benzodiazepines, antidepressants and hallucinogens in 2.5 mg hair samples.
abstract: A high performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS) method for simultaneous screening and quantification of 28 drugs was developed and validated for 2.5 mg hair samples. Target drugs and their metabolites included amphetamines, cocaine, opioids, benzodiazepines, antidepressants, and hallucinogens. After decontamination, hair samples were extracted with 200 μL of a mixture of water: acetonitrile:1 M trifluoroacetic acid (80:10:10, v/v) using a 5 min simultaneous pulverization/extraction step. The extracts were analysed by HPLC-HRMS in an Orbitrap at a nominal resolution of 60,000, with concomitant in source collisional experiments (in source CID). Gradient elution on an Atlantis T3 column resolved 28 target compounds and 5 internal standards. Total chromatographic run time was 26 min. Calibration was achieved by linear regression analysis utilizing six calibration points; R2 ranged from 0.9964 to 0.9999, the limits of quantification were 0.1 ng/mg for 8 compounds, 0.2 ng/mg for 16 compounds and 0.5 ng/mg for 4 compounds; mean relative errors from -21% to +23% were obtained; relative standard deviation, used to estimate repeatability and intermediate reproducibility at three concentrations, was always less than 20%. Process efficiency and recoveries for all analytes were better than 65 and 73%, respectively, at any concentration. The method was applied to hair samples from forensic investigations that contained a broad assortment of drugs of abuse and pharmaceuticals. The use of concomitant HRMS full scan and CID afforded the possibility of retrospective analysis for discovering untargeted drugs.
authors.affiliation: Department of Public Health and Environmental Medicine, University of Padova, and Forensic Toxicology and Antidoping, University Hospital of Padova, Via Falloppio 50, I-35121 Padova, Italy. donata.favretto@unipd.it
authors.forename: Donata
authors.initials: D
authors.lastname: Favretto
authors.forename: Susanna
authors.initials: S
authors.lastname: Vogliardi
authors.forename: Giulia
authors.initials: G
authors.lastname: Stocchero
authors.forename: Alessandro
authors.initials: A
authors.lastname: Nalesso
authors.forename: Marianna
authors.initials: M
authors.lastname: Tucci
authors.forename: Santo Davide
authors.initials: SD
authors.lastname: Ferrara
mesh_terms: D000662:Amphetamines / Q000032:analysis; D000928:Antidepressive Agents / Q000032:analysis; D001569:Benzodiazepines / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003042:Cocaine / Q000032:analysis; D006197:Hair / Q000737:chemistry*; D006213:Hallucinogens / Q000032:analysis; D006801:Humans; D013287:Illicit Drugs / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000032:analysis*; D052617:Solid Phase Microextraction / Q000379:methods*; D015813:Substance Abuse Detection / Q000379:methods*
chemical_list: D000662:Amphetamines; D000928:Antidepressive Agents; D006213:Hallucinogens; D013287:Illicit Drugs; D004364:Pharmaceutical Preparations; D001569:Benzodiazepines; D003042:Cocaine