University of Bucharest, Faculty of Chemistry, Department of Analytical and Physical Chemistry, Sos. Panduri, No. 90, Sect 5, Bucharest, 050663, Romania; Romanian Doping Control Laboratory, Bldv. Basarabia, No. 37-39, Sect 2, Bucharest, 022103, Romania.
Romanian Doping Control Laboratory, Bldv. Basarabia, No. 37-39, Sect 2, Bucharest, 022103, Romania.
Anal Biochem. 2023 Dec 1;682:115336. doi: 10.1016/j.ab.2023.115336. Epub 2023 Oct 6.
The use of growth hormone-releasing hormones (GHRHs) is prohibited in sports according to the regulations of the World Anti-Doping Agency (WADA). Considering the complexity of urine samples and the low concentrations at which these analytes should be detected, analyzing GHRHs is a challenging task. In most of the studies, GHRHs are analyzed using UHPLC-HRMS with an orbitrap. The present developed and validated method for some GHRHs (tesamorelin, CJC-1295, sermorelin (GRF 1-29), sermorelin (3-29)-NH, somatorelin) is based on the triple quadrupole UHPLC/MS-MS method with solid phase extraction (SPE) with weak cation exchange and is able to detect concentrations as low as 0.2 ng/mL (LOD), a limit of quantification (LOQ) at 0.6 ng/mL, and linearity across the range of 0.1 ng/mL to 1.2 ng/mL. The present method developed by our doping control laboratory was validated according to WADA technical documents for selectivity, limit of detection (LOD), carryover, reliability of detection, stability and recovery. The results show that the method has adequate recoveries and sensitivity, hence, it can be employed for routine screening in anti-doping laboratories.
根据世界反兴奋剂机构（WADA）的规定，生长激素释放激素（GHRHs）在运动中被禁止使用。考虑到尿液样本的复杂性和这些分析物应该被检测到的低浓度，分析 GHRHs 是一项具有挑战性的任务。在大多数研究中，使用超高效液相色谱-高分辨质谱联用仪（UHPLC-HRMS）和轨道阱分析 GHRHs。本研究开发并验证了一些 GHRHs（特莫瑞林、CJC-1295、生长激素释放肽（GRF 1-29）、生长激素释放肽（3-29）-NH、索玛肽）的方法，该方法基于带有固相萃取（SPE）的三重四极杆 UHPLC/MS-MS 方法，采用弱阳离子交换，能够检测低至 0.2ng/mL（LOD）的浓度，定量限（LOQ）为 0.6ng/mL，线性范围为 0.1ng/mL 至 1.2ng/mL。本实验室开发的方法按照 WADA 技术文件进行了选择性、检测限（LOD）、交叉污染、检测可靠性、稳定性和回收率的验证。结果表明，该方法具有足够的回收率和灵敏度，因此可用于反兴奋剂实验室的常规筛选。
