Semenistaya Ekaterina, Zvereva Irina, Thomas Andreas, Thevis Mario, Krotov Grigory, Rodchenkov Grigory
Anti-Doping Center, Elizavetinskiy per., 10/1, Moscow, 105005, Russian Federation.
Institute of Biochemistry and Center for Preventive Doping Research, German Sport University Cologne, Germany.
Drug Test Anal. 2015 Oct;7(10):919-25. doi: 10.1002/dta.1787. Epub 2015 Apr 13.
Growth hormone releasing peptides (GHRPs) stimulate secretion of endogenous growth hormone and are listed on the World Anti-Doping Agency (WADA) Prohibited List. To develop an effective method for GHRPs anti-doping control we have investigated metabolites of GHRP-1, GHRP-2, GHRP-6, Hexarelin, and Ipamorelin in urine after nasal administration. Each compound was administrated to one volunteer. Samples were collected for 2 days after administration, processed by solid-phase extraction on weak cation exchange cartridges and analyzed by means of nano-liquid chromatography - high resolution mass spectrometry. Six metabolites of GHRP-1 were identified. GHRP-1 in the parent form was not detected. GHRP-1 (2-4) free acid was detected in urine up to 27 h. GHRP-2, GHRP-2 free acid and GHRP-2 (1-3) free acid were detected in urine up to 47 h after administration. GHRP-6 was mostly excreted unchanged and detected in urine 23 h after administration, its metabolites were detectable for 12 h only. Hexarelin and Ipamorelin metabolized intensively and were excreted as a set of parent compounds with metabolites. Hexarelin (1-3) free acid and Ipamorelin (1-4) free acid were detected in urine samples after complete withdrawal of parent substances. GHRPs and their most prominent metabolites were included into routine ultra-pressure liquid chromatography-tandem mass spectrometry procedure. The method was fully validated, calibration curves of targeted analytes were obtained and excretion curves of GHRPs and their metabolites were plotted. Our results confirm that the detection window after GHRPs administration depends on individual metabolism, drug preparation form and the way of administration.
生长激素释放肽（GHRPs）可刺激内源性生长激素的分泌，且被列入世界反兴奋剂机构（WADA）的禁用清单。为开发一种有效的GHRPs反兴奋剂控制方法，我们研究了鼻腔给药后GHRP-1、GHRP-2、GHRP-6、六肽生长激素释放肽（Hexarelin）和依帕莫瑞林（Ipamorelin）在尿液中的代谢产物。每种化合物分别给予一名志愿者。给药后收集2天的样本，通过弱阳离子交换柱进行固相萃取处理，并采用纳升液相色谱 - 高分辨率质谱法进行分析。鉴定出了GHRP-1的6种代谢产物。未检测到母体形式的GHRP-1。在尿液中可检测到GHRP-1（2-4）游离酸长达27小时。给药后47小时内尿液中可检测到GHRP-2、GHRP-2游离酸和GHRP-2（1-3）游离酸。GHRP-6大多以原形排泄，给药后23小时在尿液中可检测到，其代谢产物仅在12小时内可检测到。六肽生长激素释放肽和依帕莫瑞林代谢活跃，以一组母体化合物及其代谢产物的形式排泄。在母体物质完全排出后，尿液样本中检测到了六肽生长激素释放肽（1-3）游离酸和依帕莫瑞林（1-4）游离酸。GHRPs及其最主要的代谢产物被纳入常规超高效液相色谱 - 串联质谱程序。该方法经过了全面验证，获得了目标分析物的校准曲线，并绘制了GHRPs及其代谢产物的排泄曲线。我们的结果证实，GHRPs给药后的检测窗口期取决于个体代谢、药物制剂形式和给药方式。
