Integrative Pharmacology and Systems Neuroscience Research Group, IMIM (Hospital del Mar Medical Research Institute), Barcelona, Spain.
Department of Diabetes, Endocrinology, Nutritional Medicine and Metabolism, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland.
Drug Test Anal. 2021 Mar;13(3):510-522. doi: 10.1002/dta.2975. Epub 2020 Nov 25.
Dried blood spots (DBSs) sampling is gaining support by the antidoping community because of simplicity and cost-effective characteristics, especially in collection, transport, and storage. Nevertheless, DBS applicability demands specific studies for each of the analytes proposed for testing. Here, GHRP-2 has been selected as a representing member of the growth hormone-releasing peptides (GHRPs) family to provide further evidence of DBS suitability for GHRPs abuse detection in sport testing. An analytical procedure to extract GHRP-2 and its main metabolite (AA-3) from DBS and to detect them by liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed. The method has been validated for the detection of GHRP-2. Specificity and identification capabilities have been assessed in agreement with antidoping guidelines. The low AA-3 levels found in DBS samples prevented its effective application for the determination of this metabolite. The limit of detection (LoD) for GHRP-2 has been established at 50 pg/ml. Long-term stability (>2 years) has been confirmed. The procedure has been successfully applied to actual DBS samples from an administration study with a single intravenous dose of GHRP-2 (100 μg) being detected up to 4 h after drug injection. GHRP-2 concentrations have been higher in venous blood DBS than in capillary blood DBS. Despite the observed differences, a similar detection window has been achieved independently of the type of blood used. In summary, this study provides specific evidence supporting DBS usefulness to detect GHRP-2, and potentially other GHRPs family members, for antidoping tests.
干血斑（DBS）采样因其简单、经济高效的特点，尤其在采集、运输和储存方面，得到了反兴奋剂界的支持。然而，DBS 的适用性要求针对拟检测的每个分析物进行具体研究。在这里，GHRP-2 被选为生长激素释放肽（GHRP）家族的代表性成员，以提供进一步的证据证明 DBS 适用于运动测试中的 GHRP 滥用检测。已经开发了一种从 DBS 中提取 GHRP-2 及其主要代谢物（AA-3）并通过液相色谱-串联质谱（LC-MS/MS）检测它们的分析程序。该方法已针对 GHRP-2 的检测进行了验证。特异性和识别能力已根据反兴奋剂准则进行了评估。DBS 样本中发现的 AA-3 水平很低，阻止了其有效应用于该代谢物的测定。GHRP-2 的检测限（LoD）已确定为 50 pg/ml。已确认长期稳定性（>2 年）。该程序已成功应用于单次静脉注射 GHRP-2（100 μg）给药研究的实际 DBS 样本，在药物注射后 4 小时即可检测到 GHRP-2。静脉血 DBS 中的 GHRP-2 浓度高于毛细血管血 DBS。尽管观察到了差异，但无论使用哪种类型的血液，都实现了类似的检测窗口。总之，这项研究提供了具体的证据，支持 DBS 用于检测 GHRP-2，以及可能用于检测其他 GHRP 家族成员，用于反兴奋剂测试。
