Technical note: Development of a UHPLC-MS/MS method for the analysis of hCG and IGF-I from dried blood spots: A...

Two of the most used hormones in sport, included in the WADA list of banned molecules, are human chorionic gonadotropin (hCG) and insulin-like growth factor (IGF-I). Male athletes may use pharmaceutical preparations of hCG to stimulate the production of testosterone before competition and IGF-I to stimulate muscle protein synthesis, promote glycogen storage, and improve lipolysis. Dry matrix microsampling and dried blood spots (DBS) are very promising strategies to improve current doping control activities, as they can lead to a significant improvement in compound stability, the simplification of sample collection and pre-analytic phases, the facilitation of their automation and a reduction of the risks of sample contamination and tampering. In this study, a protein extraction protocol from DBS, specifically hCG and IGF-I, was developed. Through the testing of various extraction solvents and subsequent analysis by Qubit fluorometry and nanoLC-MS/MS, we established an effective protein extraction procedure from DBS, specifically targeting hCG and IGF-I. The nanoLC-MS/MS bottom-up proteomic study reported a good protein extraction protocol from DBS, demonstrating good reproducibility and abundance related to the peptide sequences of hCG and IGF-I detected.