Centre for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Germany.
Drug Test Anal. 2011 Nov-Dec;3(11-12):791-7. doi: 10.1002/dta.364. Epub 2011 Oct 13.
The use of growth factors for accelerated healing of sports injuries is restricted under the terms of the World Anti-Doping Agency (WADA) anti-doping code. Cheating athletes have used the black market as a source of performance-enhancing substances. Drugs that currently undergo clinical trials are frequently offered--despite the unknown health risks associated with the administration of unapproved pharmaceuticals. Recently, a new growth factor (referred to as fibroblast growth factor 1/FGF-1) with known effects on the repair and regeneration of damaged tissue was detected in an unlabelled black market product confiscated by the German customs. The identification of the protein was achieved by one- and two-dimensional polyacrylamide gel electrophoresis (SDS-PAGE and 2D-PAGE), different proteolytic digestions, immunological methods and nano-liquid chromatography high-resolution/high-accuracy Orbitrap mass spectrometry. The SDS-PAGE analysis revealed slight differences concerning the molecular weight of recombinant human and black market FGF-1. Using in-gel proteolysis, a truncation or modification located at the N-terminus of the protein was suggested. These findings demonstrate that drug candidates without clinical approval can be readily obtained from the black market, regardless of potential dangerous consequences for the consumer, which corroborates the necessity of proactive and preventive doping control approaches. In that regard, physiological concentrations of blood and urine specimens collected from healthy individuals were analyzed and were found to range below 28 pg/ml in urine, while there was no detectable FGF-1 in plasma.
世界反兴奋剂机构(WADA)反兴奋剂规则限制了生长因子在运动损伤加速愈合中的使用。作弊运动员将黑市作为获取性能增强物质的来源。目前正在进行临床试验的药物经常被提供——尽管与未经批准的药物给药相关的未知健康风险。最近,在德国海关没收的未贴标签的黑市产品中发现了一种新的生长因子(称为成纤维细胞生长因子 1/FGF-1),它对受损组织的修复和再生有已知的作用。通过一维和二维聚丙烯酰胺凝胶电泳(SDS-PAGE 和 2D-PAGE)、不同的蛋白水解消化、免疫方法和纳升液相色谱高分辨率/高精度轨道阱质谱法鉴定了该蛋白。SDS-PAGE 分析显示重组人源和黑市 FGF-1 的分子量略有差异。通过胶内蛋白水解,提示蛋白的 N 端存在截断或修饰。这些发现表明,即使对消费者存在潜在的危险后果,未经临床批准的候选药物也可以轻易从黑市获得,这证实了积极主动和预防性兴奋剂控制方法的必要性。在这方面,分析了来自健康个体的血液和尿液标本的生理浓度,发现尿液中的浓度低于 28pg/ml,而血浆中没有可检测到的 FGF-1。