Abstract
Glycerol was identified and isolated from endogenous interferences during analysis of human urine using high-resolution mass spectrometry (HRMS) for doping control. Urinary sample preparation was simple; the samples were diluted with an organic solvent and then analyzed using a liquid chromatography-mass spectrometry (“dilute and shoot” method). Although the interfering ion peaks were observed at the similar retention time of glycerol, the inference could be identified by isolation with HRMS and further investigation. Thus, creatinine was identified as the endogenous interference for glycerol analysis and it also caused ion suppression resulting in the decrease of glycerol signal. This study reports the first identification and efficient isolation of endogenous interferences in human urine for “dilute and shoot” method. The information about ion suppression could be novel to prevent overestimation or a false result for antidoping analysis.
Highlights
The World Anti-Doping Agency (WADA) regulates the list of prohibited substances and requires that an accredited lab should analyze substances with appropriate methods
This study reports the first identification and efficient isolation of endogenous interferences in human urine for “dilute and shoot” method
The sodium adduct of glycerol ([M+Na]+) was only detected and it did not produce product ions through collision-induced dissociation (CID); glycerol analysis was performed via MS
Summary
The World Anti-Doping Agency (WADA) regulates the list of prohibited substances and requires that an accredited lab should analyze substances with appropriate methods. Various prohibited substances were classified to 11 classes including anabolic steroids, beta-2 agoinsts, diuretics, stimulants, masking agent, etc.[1] Glycerol known to regulate osmosis in human body can induce increment of plasma by osmosis.[2] it can lower count of erythrocyte or hemoglobin concentration, which provides a role of ‘masking agent’ to interfere detection of drugs for hematogenesis such as erythropoietin. The oral dosing or intravenous injection of glycerol was prohibited by WADA from 2010 and 4.3 mg/mL of urinary concentration of. Under this license, authors reserve the copyright for their content; they permit anyone to unrestrictedly use, distribute, and reproduce the content in any medium as far as the original authors and source are cited. Redistribution, or reproduction of a work, users must clarify the license terms under which the work was produced
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