Developing Temperature-induced Marker to improve the Detection Window for Small Peptide Hormones in Urine

Abstract

Among the prohibited substances by the World Anti-Doping Agency (WADA), peptides hormones are one of the substances that face challenges in detection. Peptide molecules are susceptible to external factors, such as temperature and time, and known to have short half-life and low concentration in the body. Therefore, the peptide molecules have challenges in detection. The urine sample collected are transported and stored by following the World Anti-Doping Agency (WADA)-provided guidelines for urine sample transportation. However, there is a possibility of unforeseen changes in temperature during the transportation and it can pose challenges in detection. In this study, the behavior of peptide molecules under high temperature was observed. GHRP-2, GHRP-6 and Hexarelin were selected from prohibited peptide molecules and incubated for detection after 0, 1, 2, 5, 7, and 14 days at 60 degrees Celsius. GHRP-2 was present in the Adverse Analytical Findings (AAFs) of the 2021 Anti-Doping Testing figures and was selected for further research. This study utilized Q-Exactive mass spectrometry to observe the peptide molecules. The peak area of selected peptide molecules and its metabolites were observed to decrease in heated condition. Moreover, a possible altered product of peptide molecule was found from incubated samples. Further findings of this study can suggest a method to enhance the detection window of small peptide hormones in urine.