Capillary blood as a complementary matrix for doping control purposes. Application to the definition of the individual longitudinal profile of IGF-1

Abstract

We present a novel procedure to monitor the fluctuations of the levels of IGF-1 in capillary blood in the framework of doping control analysis. Being an endogenous hormone, direct methods are not applicable, so the most effective way to detect the intake of the exogenous hormone would be based on the longitudinal monitoring of the athlete. We have therefore followed the individual variability, in four subjects (two males and two females), of the levels of IGF-1 in capillary blood samples collected three times per day for five days, then once a week for at least two months. Analyses were performed by liquid chromatography coupled to tandem mass spectrometry following a bottom-up approach. The whole protocol, from the sample collection to the instrumental analysis, was validated according to the World Anti-Doping Agency's guidelines and ISO17025. The analytical protocol showed to be fit for purpose in terms of sensitivity (LOD 25 ng/mL and LOI 35 ng/mL), selectivity (no interferences were detected at the retention time of IGF-1 and the internal standard), and repeatability (CV<10%). The linearity was confirmed in the range of 50–1000 ng/mL (correlation coefficient R2 >0.995, with a % relative bias of the experimental concentration of the different calibrators used for the estimation of the linearity lower than 20% for the lowest level and than 15% for the other levels). Stability studies were also performed, also to establish the optimal conditions for transport and storage: samples were stable at 4 °C for up to 72 h and at −20 °C and −80 °C for up to three months. Our preliminary results indicate that, in all subjects, the levels of IGF-1 did not present significant circadian fluctuations and remained stable during the entire period of the study (2–3 months, depending on the subject). The stability over time of IGF-1 levels in capillary blood indicates the possibility of detecting the intake of the non-endogenous hormone based on a longitudinal approach, as it is modeled in the framework of the endocrinological module of the athlete biological passport.