Use of ultra-high pressure liquid chromatography coupled to high resolution mass spectrometry for fast screening in high throughput doping control

• Published in: Journal of Chromatography A 2013-05, Vol.1288, p.82-95
• Main Authors: Musenga, Alessandro, Cowan, David A.
• Format: Article
• Language: English
• Subjects: Analysis; Analytical chemistry; beta-adrenergic antagonists; Bioanalysis; Biological and medical sciences; Chromatography; Cost analysis; dissociation; Diuretics; Doping control; enzymatic hydrolysis; Fast screening; General pharmacology; glucocorticoids; High resolution; High resolution mass spectrometry; hormone antagonists; humans; ion exchange; ionization; ions; Joining; Liquid chromatography; Mass spectrometry; Medical sciences; narcotics; Pharmacology. Drug treatments; pretreatment; rapid methods; Retrospective analysis; Screening; solid phase extraction; sports; urine
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2013.03.006

•Fast simple discriminating screen for almost 200 substances prohibited in sport.•Comprehensive data capture permits detection of new misused substances.•High resolution mass spectrometry at 5ppm mass accuracy simplifies data review.•Data obtained is maximised by use of positive, negative ionisation and CID.•First in the world for use at an Olympic Games evidencing reliability of method. We describe a sensitive, comprehensive and fast screening method based on liquid chromatography–high resolution mass spectrometry for the detection of a large number of analytes in sports samples. UHPLC coupled to high resolution mass spectrometry with polarity switching capability is applied for the rapid screening of a large number of analytes in human urine samples. Full scan data are acquired alternating both positive and negative ionisation. Collision-induced dissociation with positive ionisation is also performed to produce fragment ions to improve selectivity for some analytes. Data are reviewed as extracted ion chromatograms based on narrow mass/charge windows (±5ppm). A simple sample preparation method was developed, using direct enzymatic hydrolysis of glucuronide conjugates, followed by solid phase extraction with mixed mode ion-exchange cartridges. Within a 10min run time (including re-equilibration) the method presented allows for the analysis of a large number of analytes from most of the classes in the World Anti-Doping Agency (WADA) Prohibited List, including anabolic agents, β2-agonists, hormone antagonists and modulators, diuretics, stimulants, narcotics, glucocorticoids and β-blockers, and does so while meeting the WADA sensitivity requirements. The high throughput of the method and the fast sample pre-treatment reduces analysis cost and increases productivity. The method presented has been used for the analysis of over 5000 samples in about one month and proved to be reliable.