Mareck Ute, Geyer Hans, Guddat Sven, Haenelt Nadine, Koch Anja, Kohler Maxie, Opfermann Georg, Thevis Mario, Schänzer Wilhelm
Institute of Biochemistry, German Sport University Cologne, Carl-Diem-Weg 6, 50933 Cologne, Germany.
Rapid Commun Mass Spectrom. 2006;20(12):1954-62. doi: 10.1002/rcm.2545.
Anastrozole (2,2'-[5-(1H-1,2,4-triazol-1-ylmethyl)-1.3-phenylene]bis(2-methylpropionitrile)) and exemestane (6-methylenandrostan-1,4-diene-3,17-dione) are therapeutically used to treat hormone-sensitive breast cancer in postmenopausal women. For doping purposes they may be used to counteract adverse effects of an extensive abuse of anabolic androgenic steroids (gynaecomastia) and to increase plasma testosterone concentrations. Excretion study urine samples and spot urine samples from women suffering from metastatic breast cancer, being treated with anastrozole or exemestane, were collected and analyzed to develop/optimize a detection system for anastrozole and exemestane to allow the identification of athletes who do not comply with the internationally prohibited use of these cancer drugs. The assay was based on liquid-liquid extraction after enzymatic hydrolysis following liquid chromatography/tandem mass spectrometry (LC/MS/MS). Anastrozole, exemestane and its main metabolite (17-dihydroexemestane) were identified in urine by comparison of mass spectra and retention times with respective reference substances. An assay validation for the analysis of anastrozole and exemestane was performed regarding lower limits of detection (anastrozole: 0.02 ng/mL; exemestane: 3.1 ng/mL; dihydroexemestane: 0.5 ng/mL), interday precisions (6.6-11.1%, 4.9-9.1% and 5.6-8.3% for low [10 ng/mL], medium [50 ng/mL] and high [100 ng/mL] concentration) and recoveries (ranged from 85-97%).
阿那曲唑（2,2'-[5-(1H-1,2,4-三唑-1-基甲基)-1,3-亚苯基]双(2-甲基丙腈)）和依西美坦（6-亚甲基雄甾-1,4-二烯-3,17-二酮）在治疗上用于治疗绝经后妇女的激素敏感性乳腺癌。出于兴奋剂使用目的，它们可能被用于对抗大量滥用合成代谢雄激素类固醇的不良影响（男性乳房发育）并提高血浆睾酮浓度。收集并分析了接受阿那曲唑或依西美坦治疗的转移性乳腺癌女性的排泄研究尿样和即时尿样，以开发/优化阿那曲唑和依西美坦的检测系统，以便识别不符合国际禁用这些癌症药物规定的运动员。该测定基于液相色谱/串联质谱（LC/MS/MS）后酶促水解的液-液萃取。通过将质谱和保留时间与相应参考物质进行比较，在尿液中鉴定出阿那曲唑、依西美坦及其主要代谢物（17-二氢依西美坦）。针对阿那曲唑和依西美坦的分析进行了测定验证，包括检测下限（阿那曲唑：0.02 ng/mL；依西美坦：3.1 ng/mL；二氢依西美坦：0.5 ng/mL）、日间精密度（低浓度[10 ng/mL]为6.6 - 11.1%，中浓度[50 ng/mL]为4.9 - 9.1%，高浓度[100 ng/mL]为5.6 - 8.3%）和回收率（范围为85 - 97%）。
