Jihyun Yoon (Doping Control Center, Korea Institute of Science and Technology)

Ki Hun Kim (Doping Control Center, Korea Institute of Science and Technology)

Jihyun Yoon (Doping Control Center, Korea Institute of Science and Technology)

Jiyeong Hong (Doping Control Center, Korea Institute of Science and Technology)

Han Bin Oh (Department of Chemistry, Sogang University)

Ki Hun Kim (Doping Control Center, Korea Institute of Science and Technology)

 Dried blood spots (DBS) have been suggested as an alternative to venous blood samples due to several advantages such as cost-effectiveness, sample stability during transport and storage, and reduced invasiveness for athletes. For this reason, recent sports doping research tends to promote the application of DBS, but it is still challenging. According to previous studies, the analytical test using DBS samples has been performed based on physicochemical properties, so each class is required respective sample preparation. However, the number of available spots per athlete is limited. Therefore, expanding the use of DBS samples requires the development of more practical and efficient testing methods. In this study, the extraction of two testing classes—10 steroid esters and 2 peptides—is carried out simultaneously. Each class was spiked into blank whole blood, spotted 20 μL of spiked blood onto cellulose cards, and dried. Based on liquid-liquid extraction, the DBS samples were extracted methyl tert-butyl ether (MTBE):water (1:1, v:v) mixture, then each layers was separated. Steroid esters were detected in the MTBE layer, and peptides were detected in the aqueous layer. To enhance detection efficiency, chemical derivatization using Girard’s reagent T was applied for steroid esters. The samples had adequate sensitivity to meet World Anti Doping Agency (WADA)’s minimum required performance levels (MRPL) requirements for each target.

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