Center for Preventive Doping Research and Institute of Biochemistry, German Sport University Cologne, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany.
J Chromatogr A. 2012 Oct 12;1259:251-7. doi: 10.1016/j.chroma.2012.07.022. Epub 2012 Jul 28.
In the present study, a screening assay was developed comprising 11 prohibited peptides (<1.5 kDa) that are sufficiently purified from urine using weak cation exchange with subsequent determination of all substances by means of nanoUHPLC separation coupled to high resolution tandem mass spectrometry. These peptides included Gonadorelin (LH-RH), Desmopressin and 9 growth hormone releasing peptides (GHRP-1, -2, -4, -5, -6, Hexarelin, Alexamorelin, Ipamorelin and a GHRP-2 metabolite); however, the procedure is expandable to further target analytes or metabolites. The method was validated with a main focus on qualitative result interpretation considering the parameters specificity, linearity (0-500 pg/mL), recovery (45-95%), precision (<20% at 100 pg/mL), limits of detection (2-10 pg/mL), robustnesss and ion suppression. The proof-of-principle was shown by analysing excretion study urine samples for LHRH, Desmopressin and GHRP-2.
在本研究中，开发了一种筛选分析方法，该方法包括从尿液中使用弱阳离子交换充分纯化的 11 种禁用肽（<1.5 kDa），然后通过纳米 UHPLC 分离与高分辨率串联质谱联用的方法测定所有物质。这些肽包括促性腺激素释放激素（LH-RH）、去氨加压素和 9 种生长激素释放肽（GHRP-1、-2、-4、-5、-6、Hexarelin、Alexamorelin、Ipamorelin 和 GHRP-2 代谢物）；然而，该程序可以扩展到进一步的目标分析物或代谢物。该方法经过验证，主要侧重于定性结果解释，考虑了特异性、线性（0-500 pg/mL）、回收率（45-95%）、精密度（<20%在 100 pg/mL 时）、检测限（2-10 pg/mL）、稳健性和离子抑制等参数。通过分析 LHRH、Desmopressin 和 GHRP-2 的排泄研究尿液样本，证明了原理的可行性。
