title: Introduction of solid-phase microextraction as a high-throughput sample preparation tool in laboratory analysis of prohibited substances.
abstract: A fully automated, high-throughput method based on thin-film solid-phase microextraction (SPME) and liquid chromatography-mass spectrometry was developed for simultaneous quantitative analysis of 110 doping compounds, selected from ten classes and varying in physical and chemical properties. Among four tested extraction phases, C18 blades were chosen, as they provided optimum recoveries and the lowest carryover effect. The SPME method was optimized in terms of extraction pH, ionic strength of the sample, washing solution, extraction and desorption times for analysis of urine samples. Chromatographic separation was obtained in reversed-phase model; for detection, two mass spectrometers were used: triple quadrupole and full scan orbitrap. These combinations allowed for selective analysis of targeted compounds, as well as a retrospective study for known and unknown compounds. The developed method was validated according to the Food and Drug Administration (FDA) criteria, taking into account Minimum Required Performance Level (MRPL) values required by the World Anti-Doping Agency (WADA). In addition to analysis of free substances, it was also shown that the proposed method is able to extract the glucuronated forms of the compounds. The developed assay offers fast and reliable analysis of various prohibited substances, an attractive alternative to the standard methods that are currently used in anti-doping laboratories.
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada N2L 3G1.
authors.forename: Ezel
authors.initials: E
authors.lastname: Boyacı
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada N2L 3G1.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Gorynski
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada N2L 3G1.
authors.forename: Angel
authors.initials: A
authors.lastname: Rodriguez-Lafuente
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada N2L 3G1.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bojko
authors.affiliation: Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada N2L 3G1. Electronic address: janusz@uwaterloo.ca.
authors.forename: Janusz
authors.initials: J
authors.lastname: Pawliszyn
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid; D004300:Doping in Sports; D005260:Female; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D009930:Organic Chemicals / Q000652:urine*; D052617:Solid Phase Microextraction / Q000295:instrumentation; D015813:Substance Abuse Detection / Q000295:instrumentation / Q000379:methods*
chemical_list: D009930:Organic Chemicals
keywords: Drugs; Multiwell high throughput system; Quantitative determinations; Solid phase microextraction