Institute of Biochemistry, Center for Preventive Doping Research, German Sport University Cologne, Am Sportpark Müngersdorf 6, 50933, Cologne, Germany.
CER Groupe, Marloie, Belgium.
Anal Bioanal Chem. 2023 Feb;415(4):669-681. doi: 10.1007/s00216-022-04456-y. Epub 2022 Nov 28.
Potential scenarios as to the origin of minute amounts of banned substances detected in doping control samples have been a much-discussed problem in anti-doping analysis in recent years. One such debated scenario has been the contamination of female athletes' urine with ejaculate containing doping agents and/or their metabolites. The aim of this work was to obtain complementary information on whether relevant concentration ranges of doping substances are excreted into the ejaculate and which metabolites can be detected in the seminal fluid (sf) and corresponding blood plasma (bp) samples. A method was established to study the concentration and metabolite profiles of stanozolol and LGD-4033-substances listed under anabolic substances (S1) on the World Anti-Doping Agency's Prohibited List-in bp and sf using liquid chromatography high-resolution mass spectrometry (LC-HRMS). For sf and bp, methods for detecting minute amounts of these substances were developed and tested for specificity, recovery, linearity, precision, and reliability. Subsequently, sf and bp samples from an animal administration study, where a boar orally received stanozolol at 0.33 mg/kg and LGD-4033 at 0.11 mg/kg, were measured. The developed assays proved appropriate for the detection of the target substances in both matrices with detection limits between 10 and 40 pg/mL for the unmetabolized drugs in sf and bp, allowing to estimate the concentration of stanozolol in bp (0.02-0.40 ng/mL) and in sf (0.01-0.25 ng/mL) as well as of LGD-4033 in bp (0.21-2.00 ng/mL) and in sf (0.03-0.68 ng/mL) post-administration. In addition, metabolites resulting from different metabolic pathways were identified in sf and bp, with sf resembling a composite of the metabolic profile of bp and urine.
近年来，在反兴奋剂分析中，检测到的兴奋剂控制样本中痕量违禁物质的来源的潜在情况一直是一个备受讨论的问题。其中一个有争议的情况是，女性运动员的尿液被含有兴奋剂及其代谢物的精液污染。本研究的目的是获得补充信息，了解相关浓度范围的兴奋剂物质是否排泄到精液中，以及可以在精液（sf）和相应的血浆（bp）样本中检测到哪些代谢物。建立了一种使用液相色谱高分辨率质谱（LC-HRMS）在 bp 和 sf 中研究司坦唑醇和 LGD-4033-世界反兴奋剂机构禁用清单上列出的合成代谢物质（S1）物质的浓度和代谢物谱的方法。针对这些物质的痕量检测，开发并测试了 sf 和 bp 方法的特异性、回收率、线性、精密度和可靠性。随后，对猪经口给予司坦唑醇 0.33 mg/kg 和 LGD-4033 0.11 mg/kg 的动物给药研究的 sf 和 bp 样本进行了测量。所开发的检测方法证明适用于两种基质中目标物质的检测，sf 和 bp 中未代谢药物的检测限分别为 10 和 40 pg/mL，允许估计 bp 中司坦唑醇的浓度（0.02-0.40 ng/mL）和 sf（0.01-0.25 ng/mL）以及 bp 中 LGD-4033 的浓度（0.21-2.00 ng/mL）和 sf（0.03-0.68 ng/mL）给药后。此外，还在 sf 和 bp 中鉴定了来自不同代谢途径的代谢物，sf 类似于 bp 和尿液代谢谱的组合。
