As hypoxia‐inducible factor stabilizers (HIFs) can artificially enhance an athlete's erythropoiesis, the World Anti‐Doping Agency prohibits their use at all times. Every urine sample for doping control analysis has to be evaluated for the presence of HIFs and therefore sensitive methods that allow high sample throughput are needed. Samples suspicious for the presence of HIFs need to be confirmed following the identification criteria established by the World Anti‐Doping Agency. Previous work has shown the advantages of using turbulent flow online solid‐phase extraction (SPE) procedures to reduce matrix effects and retention time shifts. Furthermore, the use of online SPE allows for automation and high sample throughput. Both an initial testing procedure (ITP) and a confirmation method were developed and validated, using online SPE liquid chromatography–tandem mass spectrometry (LC–MS/MS), with limits of detection between 0.1 ng/ml (or possibly lower) and 4 ng/ml (or higher for GSK360a) and limits of identification between 0.1 ng/ml (or possibly lower) and 1.17 ng/ml. The ITP only takes 6.5 min per sample. To the best of our knowledge, these are the first ITP and confirmation methods that include more than three HIFs without the need for manual sample preparation.

中文翻译：

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用于检测和确认尿液中缺氧诱导因子稳定剂的自动化样品制备

由于缺氧诱导因子稳定剂 (HIF) 可以人为地增强运动员的红细胞生成，因此世界反兴奋剂机构始终禁止使用它们。必须评估用于兴奋剂控制分析的每个尿液样本是否存在 HIF，因此需要允许高样本通量的敏感方法。需要按照世界反兴奋剂机构制定的识别标准来确认怀疑存在 HIF 的样本。以前的工作已经显示了使用湍流在线固相萃取 (SPE) 程序减少基质效应和保留时间偏移的优势。此外，使用在线 SPE 可以实现自动化和高样品通量。开发并验证了初始测试程序 (ITP) 和确认方法，使用在线 SPE 液相色谱-串联质谱 (LC-MS/MS)，检测限在 0.1 ng/ml（或可能更低）和 4 ng/ml（GSK360a 或更高）之间，鉴定限在 0.1 ng/ml ml（或可能更低）和 1.17 ng/ml。ITP 每个样本仅需 6.5 分钟。据我们所知，这些是第一个包含三个以上 HIF 且无需手动样品制备的 ITP 和确认方法。