Analytical methods to determine the potential misuse of the ghrelin mimetics capromorelin (CP‐424,391), macimorelin (macrilen, EP‐01572) and tabimorelin (NN703) in sports were developed. Therefore, different extraction strategies, i.e. solid‐phase extraction, protein precipitation, as well as a “dilute‐and‐inject” approach, from urine and EDTA–plasma were assessed and comprehensive in vitro/in vivo experiments were conducted, enabling the identification of reliable target analytes by means of high resolution mass spectrometry. The drugs’ biotransformation led to the preliminary identification of 51 metabolites of capromorelin, 12 metabolites of macimorelin and 13 metabolites of tabimorelin. Seven major metabolites detected in rat urine samples collected post‐administration of 0.5–1.0 mg of a single oral dose underwent in‐depth characterization, facilitating their implementation into future confirmatory test methods. In particular, two macimorelin metabolites exhibiting considerable abundances in post‐administration rat urine samples were detected, which might contribute to an improved sensitivity, specificity, and detection window in case of human sports drug testing programs. Further, the intact drugs were implemented into World Anti‐Doping Agency‐compliant initial testing (limits of detection 0.02–0.60 ng/ml) and confirmation procedures (limits of identification 0.18–0.89 ng/ml) for human urine and blood matrices. The obtained results allow extension of the test spectrum of doping agents in multitarget screening assays for growth hormone‐releasing factors from human urine.

中文翻译：

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对生长素释放肽模拟物Capromorelin，Macimorelin和Tabimorelin作为掺杂控制目的潜在标志物代谢物形成的综合见解

开发了用于确定生长激素释放肽模拟物Capmoremore（CP‐424,391），Macimorelin（macrilen，EP‐01572）和Tabimorelin（NN703）在运动中的潜在滥用的分析方法。因此，评估了从尿液和EDTA-血浆中提取不同的提取策略，即固相提取，蛋白质沉淀以及“稀释并注射”的方法，并在体内/体外进行了全面的研究。进行了实验，能够通过高分辨率质谱鉴定可靠的目标分析物。药物的生物转化导致初步鉴定了己莫瑞林的51种代谢物，美莫瑞林的12种代谢物和塔比莫林的13种代谢物。在大鼠尿液样品中检测到的七种主要代谢产物在每次口服0.5-1.0 mg剂量后收集后进行了深入表征，从而有助于将其应用于未来的验证性测试方法中。特别是，在给药后的大鼠尿液样本中检测到了两种具有大量丰富成分的Macimorelin代谢物，这可能有助于提高人类运动药物检测程序的灵敏度，特异性和检测范围。更多，完整的药物已用于符合世界反兴奋剂机构标准的初始测试（检出限为0.02–0.60 ng / ml）和确认程序（鉴定限为0.18–0.89 ng / ml）用于人尿和血液基质。获得的结果可以扩展人尿中生长激素释放因子的多目标筛选测定中掺杂剂的测试范围。