Institut de médecine légale, 11 rue Humann, Strasbourg 67000, France.
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J Anal Toxicol. 2023 Nov 1;47(8):703-708. doi: 10.1093/jat/bkad050.
Clomiphene or clomifene is a selective estrogen receptor modulator used to treat female fertility in case of ovulatory dysfunction. In sport, clomiphene is prohibited at all times for use by athletes and is listed in the section S4.2 (hormone and metabolic modulators) by the World Anti-Doping Agency. Indeed, clomiphene can indirectly increase testosterone levels in the body and can mitigate some side effects of synthetic steroid abuse. Despite its prescription to millions of subjects, its detection in human hair or nail clippings has never been reported. The aim of this study was to develop a specific method to identify clomiphene in hair and nail clippings by liquid chromatography-quadrupole tandem mass spectrometry. The procedure was then applied in a case of challenged doping results. The method involves sonication/incubation for 1 h of 30 mg of pulverized material in 1 mL of methanol in the presence of 2 ng diazepam-d5 used as internal standard. The chromatographic separation was performed using a HSS C18 column with a 15 min gradient elution. After spiking blank hair and nail with the corresponding amounts of clomiphene, linearity was verified from 1 to 500 pg/mg (r2 = 0.9994 and 0.9995 for hair and nail, respectively). The limit of detection was estimated at 0.3 pg/mg for both matrices. No interference was noted from endogenous compounds, particularly steroids. Clomiphene was identified at 85 and 20 pg/mg in the pubic hair and the fingernail clippings, respectively, of a male athlete challenging an adverse analytical finding.
克罗米酚或氯米芬是一种选择性雌激素受体调节剂，用于治疗排卵功能障碍的女性生育能力。在运动中，运动员在任何时候都被禁止使用克罗米酚，并且被世界反兴奋剂机构列入 S4.2 节（激素和代谢调节剂）。事实上，克罗米酚可以间接增加体内的睾丸激素水平，并减轻一些合成类固醇滥用的副作用。尽管它被开给了数百万患者，但从未有人报道过在人体毛发或指甲屑中检测到克罗米酚。本研究的目的是开发一种通过液相色谱-串联质谱法鉴定毛发和指甲屑中克罗米酚的特定方法。然后将该方法应用于一个有争议的兴奋剂检测结果案例中。该方法涉及在 1ml 甲醇中加入 30mg 粉碎材料，在 2ng 地西泮-d5 作为内标存在的情况下进行 1 小时的超声/孵育。采用 HSS C18 柱，在 15 分钟梯度洗脱下进行色谱分离。用相应量的克罗米酚对空白毛发和指甲进行加标后，验证了线性范围为 1 至 500pg/mg（毛发和指甲的 r2 值分别为 0.9994 和 0.9995）。两种基质的检测限均估计为 0.3pg/mg。未发现内源性化合物（特别是类固醇）的干扰。在一位挑战阳性分析结果的男性运动员的阴毛和指甲屑中，分别鉴定出 85 和 20pg/mg 的克罗米酚。
