Hair analysis is a crucial method in forensic toxicology with potential applications in revealing doping histories in sports. Despite its widespread use, knowledge about detectable substances in hair is limited. This study systematically assessed the detectability of prohibited substances in sports using a multifaceted approach. Initially, an animal model received a subset of 17 model drugs to compare dose dependencies and detection windows across different matrices. Subsequently, hair incorporation data from the animal experiment were extrapolated to all substances on the World Anti-Doping Agency’s List through in-silico prediction. The detectability of substances in hair was further validated in a proof-of-concept human study involving the consumption of diuretics and masking agents. Semi-quantitative analysis of substances in specimens was performed using ultra-performance liquid chromatography–tandem mass spectrometry. Results showed plasma had optimal dose dependencies with limited detection windows, while urine, faeces, and hair exhibited a reasonable relationship with the administered dose. Notably, hair displayed the highest detection probability (14 out of 17) for compounds, including anabolic agents, hormones, and diuretics, with beta-2 agonists undetected. Diuretics such as furosemide, canrenone, and hydrochlorothiazide showed the highest hair incorporation. Authentic human hair confirmed diuretic detectability, and their use duration was determined via segmental analysis. Noteworthy is the first-time reporting of canrenone in human hair. Anabolic agents were expected in hair, whereas undetectable compounds, such as peptide hormones and beta-2 agonists, were likely due to large molecular mass or high polarity. This study enhances understanding of hair analysis in doping investigations, providing insights into substance detectability.

头发分析是法医毒理学的重要方法，在揭示体育运动中的兴奋剂历史方面具有潜在的应用前景。尽管其用途广泛，但对头发中可检测物质的了解仍然有限。这项研究采用多方面的方法系统地评估了体育运动中违禁物质的可检测性。最初，动物模型接受了 17 种模型药物的子集，以比较不同基质的剂量依赖性和检测窗口。随后，通过计算机预测，将动物实验中的毛发掺入数据外推到世界反兴奋剂机构清单上的所有物质。头发中物质的可检测性在一项涉及利尿剂和掩蔽剂消耗的概念验证人体研究中得到了进一步验证。使用超高效液相色谱-串联质谱法对样品中的物质进行半定量分析。结果显示，血浆具有最佳的剂量依赖性，检测窗口有限，而尿液、粪便和头发与给药剂量表现出合理的关系。值得注意的是，头发对合成代谢剂、激素和利尿剂等化合物的检测概率最高（17 中的 14），而 β2 激动剂未检测到。速尿、坎利酮和氢氯噻嗪等利尿剂显示出最高的毛发掺入率。真人头发证实了利尿剂的可检测性，并通过分段分析确定其使用持续时间。值得注意的是人类头发中首次报道了坎利酮。头发中预计含有合成代谢剂，而无法检测到的化合物，如肽激素和 β-2 激动剂，可能是由于大分子质量或高极性造成的。这项研究增强了对兴奋剂调查中头发分析的理解，提供了对物质可检测性的见解。