Thomas Andreas, Kohler Maxie, Schänzer Wilhelm, Kamber Matthias, Delahaut Philippe, Thevis Mario
Center for Preventive Doping Research and Institute of Biochemistry, German Sport University Cologne, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany.
Rapid Commun Mass Spectrom. 2009 Sep;23(17):2669-74. doi: 10.1002/rcm.4176.
Doping control analysis of performance-enhancing peptides in urine represents a challenging requirement in modern sports drug testing. Low dosing, effective metabolism and short half-life lead to target concentrations in the low fmol/mL range in urine. Synthetic adrenocorticotropic hormone (1-24, Syn-ACTH-en) shares all these characteristics and improved analytical performance is required for its sufficient determination by means of liquid chromatography/tandem mass spectrometry (LC/MS/MS). The desired effects for cheating sportsmen are mainly due to enhanced release of corticosteroids as well as androgenic steroids into the circulation after systemic administration of the drug. Immunoaffinity purification with coated magnetic beads and subsequent liquid chromatography with nano-ultra-performance liquid chromatography (UPLC) coupled to tandem mass spectrometry (high resolution/high mass accuracy) of Synacthen from urinary specimens is described in the present study. The general proof of principle was obtained by analysis of excretion study urine samples and validation was performed with focus on the limit of detection (3 pg/mL), linearity, precision (<20%), recovery ( approximately 30%), robustness, specificity and stability. For all experiments, the ACTH fragment 1-17 was used as the internal standard.
尿液中促性能提升肽的兴奋剂控制分析是现代运动药物检测中一项具有挑战性的要求。低剂量、高效代谢和短半衰期导致尿液中的目标浓度处于低飞摩尔/毫升范围内。合成促肾上腺皮质激素（1-24，Syn-ACTH-en）具有所有这些特征，通过液相色谱/串联质谱法（LC/MS/MS）对其进行充分测定需要提高分析性能。对作弊运动员的预期效果主要是由于在全身给药后，皮质类固醇以及雄激素类固醇向循环系统中的释放增加。本研究描述了用包被磁珠进行免疫亲和纯化，随后对尿样中的合成促皮质素进行液相色谱分析，该液相色谱采用纳米超高效液相色谱（UPLC）与串联质谱联用（高分辨率/高质量准确度）。通过对排泄研究尿样的分析获得了一般原理证明，并重点针对检测限（3 pg/mL）、线性、精密度（<20%）、回收率（约30%）、稳健性、特异性和稳定性进行了验证。对于所有实验，促肾上腺皮质激素片段1-17用作内标。
