Eun Sook Jeong1,2, So-Hee Kim1,3, Eun-Ju Cha1,3, Kang Mi Lee1, Ho Jun Kim1, Sang-Won Lee3, Oh-Seung Kwon1, Jaeick Lee1. 1. Doping Control Center, Korea Institute of Science and Technology, Hwarang-ro 14-gil 5, Seongbuk-gu, Seoul, 136-791, Korea. 2. Department of Pharmacology and Pharmacogenomics Research Center, School of Medicine, Inje University, 875, Haeun-daero, Haeundae-gu, Busan, Korea. 3. Department of Chemistry, Research Institute for Natural Sciences, Korea University, 145, Anam-ro, Seongbuk-gu, Seoul, 136-701, Korea.
Abstract
RATIONALE: Doping analysis is a two-step process consisting of a screening step for prohibited substances and a confirmation step to verify the presence of specific substances found during the screening. The entire process must be performed within a limited time period, but traditional screening procedures commonly employ separate analytical methods for each class of prohibited substances being screened and thus require a great deal of human resources and instrumentation. A single simple and rapid multiresidue analytical method that could accommodate multiple classes of prohibited substances would be extraordinarily useful in doping analyses. METHODS: Urine samples were extracted via two consecutive liquid-liquid extractions at different pH values following enzymatic hydrolysis. Analyses were performed by ultrafast liquid chromatography/triple-quadrupole mass spectrometry with polarity switching and time-dependent selected reaction monitoring. RESULTS: We developed a rapid multiresidue screening and confirmation method for efficient high-throughput doping analyses. The present method was validated with regard to the limits of detection (0.01-100.0 ng/mL for screening analyses and 0.2-500.0 ng/mL for confirmation assays), matrix effects (48.9-118.9%), recovery (20.6-119.7%) and intra- (0.6-17.6%) and inter-day (4.0-20.0%) precision. CONCLUSIONS: A multiresidue analytical method was developed and validated for screening and confirming the presence of performance-enhancing drugs. A total of 210 substances from diverse classes of prohibited substances were successfully identified with an analytical run time of 10 min.
RATIONALE: Doping analysis is a two-step process consisting of a screening step for prohibited substances and a confirmation step to verify the presence of specific substances found during the screening. The entire process must be performed within a limited time period, but traditional screening procedures commonly employ separate analytical methods for each class of prohibited substances being screened and thus require a great deal of human resources and instrumentation. A single simple and rapid multiresidue analytical method that could accommodate multiple classes of prohibited substances would be extraordinarily useful in doping analyses. METHODS: Urine samples were extracted via two consecutive liquid-liquid extractions at different pH values following enzymatic hydrolysis. Analyses were performed by ultrafast liquid chromatography/triple-quadrupole mass spectrometry with polarity switching and time-dependent selected reaction monitoring. RESULTS: We developed a rapid multiresidue screening and confirmation method for efficient high-throughput doping analyses. The present method was validated with regard to the limits of detection (0.01-100.0 ng/mL for screening analyses and 0.2-500.0 ng/mL for confirmation assays), matrix effects (48.9-118.9%), recovery (20.6-119.7%) and intra- (0.6-17.6%) and inter-day (4.0-20.0%) precision. CONCLUSIONS: A multiresidue analytical method was developed and validated for screening and confirming the presence of performance-enhancing drugs. A total of 210 substances from diverse classes of prohibited substances were successfully identified with an analytical run time of 10 min.
Authors: Eduardo Luiz Rossini; Dmytro S Kulyk; Emelia Ansu-Gyeabourh; Taghi Sahraeian; Helena Redigolo Pezza; Abraham K Badu-Tawiah Journal: J Am Soc Mass Spectrom Date: 2020-05-14 Impact factor: 3.109