Abstract：
In a doping case, a top athlete challenged an anti-doping rule violation, involving molidustat. Molidustat is a stabilizing agent of the hypoxia-inducible factor (HIF) recently developed. It is currently undergoing clinical trials for anemia associated with chronic kidney disease. HIF stabilizers are banned at all times by the World Anti-Doping Agency (class S2). Because of their pharmacological proprieties, these new drugs can enhance athletic performance. The athlete\'s defense wanted to analyze multiple keratinized matrices as they allow long-term investigations. Requests concerning HIF stabilizers are constantly growing. We have therefore developed a liquid chromatography coupled with tandem mass spectrometry method to identify and quantify three molecules of this class: molidustat, vadadustat, and roxadustat. Thirty milligrams of keratinized matrices were incubated in 1 mL of pH 8.4 diammonium hydrogen phosphate buffer for 16 h at 40°C with 1 ng of testosterone-D3, used as internal standard. After extraction with ethyl acetate/diethyl ether (80/20), the organic phase was evaporated, and the dry residue was reconstituted in 30 μL of initial phase. The method was linear from 5 to 1000 pg/mg for the three analytes. Limits of quantification were 2, 0.5, and 5 pg/mg for molidustat, roxadustat, and vadadustat, respectively. The analysis of the athlete\'s head hair (collected 1 month after the urine test) showed a concentration of molidustat of 135 pg/mg, and his beard hair and his fingernails clippings contained 55 and 40 pg/mg, respectively.
摘要：
在掺杂的情况下，一名顶级运动员挑战违反反兴奋剂规则的行为，牵涉到的是Molidustat.Molidustat是最近开发的缺氧诱导因子（HIF）的稳定剂。目前正在进行与慢性肾脏疾病相关的贫血的临床试验。世界反兴奋剂机构（S2级）始终禁止使用HIF稳定剂。由于它们的药理特性，这些新药可以提高运动成绩。运动员的辩护希望分析多个角化基质，因为它们允许长期调查。有关HIF稳定剂的要求不断增长。因此，我们开发了一种液相色谱与串联质谱法相结合的方法来鉴定和定量这类三种分子：moleidustat，vadadustat,和roxadustat.将30毫克角化基质在1mLpH8.4磷酸氢二铵缓冲液中在40°C下与1ng睾酮-D3孵育16小时，用作内标。用乙酸乙酯/乙醚（80/20）萃取后，蒸发有机相,并且将干燥的残余物在30μL的初始相中重构。对于三种分析物,该方法从5至1000pg/mg是线性的。molidustat的定量限值为2、0.5和5pg/mg，罗沙达,和vadadustat,分别。对运动员头毛的分析（在尿液测试后1个月收集）显示moidustat的浓度为135pg/mg，他的胡须头发和手指甲剪含有55和40皮克/毫克，分别。
