Autologous Doping with Cryopreserved Red Blood Cells – Effects on Physical Performance and Detection by Multivariate Statistics

Abstract

The discovery of erythropoietin (EPO) simplified blood doping in sports, but improved detection methods, for EPO has forced cheating athletes to return to blood transfusion. Autologous blood transfusion with cryopreserved red blood cells (RBCs) is the method of choice, because no valid method exists to accurately detect such event. In endurance sports, it can be estimated that elite athletes improve performance by up to 3% with blood doping, regardless of method. Valid detection methods for autologous blood doping is important to maintain credibility of athletic performances. Recreational male (N = 27) and female (N = 11) athletes served as Transfusion (N = 28) and Control (N = 10) subjects in two different transfusion settings. Hematological variables and physical performance were measured before donation of 450 or 900 mL whole blood, and until four weeks after re-infusion of the cryopreserved RBC fraction. Blood was analyzed for transferrin, iron, Hb, EVF, MCV, MCHC, reticulocytes, leucocytes and EPO. Repeated measures multivariate analysis of variance (MANOVA) and pattern recognition using Principal Component Analysis (PCA) and Orthogonal Projections of Latent Structures (OPLS) discriminant analysis (DA) investigated differences between Control and Transfusion groups over time. Significant increase in performance (15 ± 8%) and VO2max (17 ± 10%) (mean ± SD) could be measured 48 h after RBC re-infusion, and remained increased for up to four weeks in some subjects. In total, 533 blood samples were included in the study (Clean = 220, Transfused = 313). In response to blood transfusion, the largest change in hematological variables occurred 48 h after blood donation, when Control and Transfused groups could be separated with OPLS-DA (R2 = 0.76/Q2 = 0.59). RBC re-infusion resulted in the best model (R2 = 0.40/Q2 = 0.10) at the first sampling point (48 h), predicting one false positive and one false negative. Over all, a 25% and 86% false positives ratio was achieved in two separate trials. In conclusions, autologous re-infusion of RBCs increased VO2max and performance as hypothesized, but hematological profiling by multivariate statistics could not reach the WADA stipulated false positive ratio of <0.001% at any time point investigated. A majority of samples remained within limits of normal individual variation at all times.

Prevalence Estimate of Blood Doping in Elite Track and Field Athletes During Two Major International Events

“…While the discovery of erythropoietin (EPO) suddenly made blood doping simpler, many cheaters returned to blood transfusions upon significant development of detection methods (Schumacher et al, 2012;Salamin et al, 2017). Definitely, autologous blood transfusion is the method of choice with no valid method to date to accurately directly detect such intervention (Malm et al, 2016). Monitoring variations in blood parameters in athletes as implemented with the ABP may, however, be observed and would allow to flag abnormal alterations.…”

Prevalence estimate of blood doping in elite track and field at the introduction of the Athlete Biological Passport

“…While the discovery of erythropoietin (EPO) suddenly made blood doping simpler, many cheaters returned to blood transfusions upon significant development of detection methods (Salamin, Kuuranne, Saugy andLeuenberger 2017, Schumacher et al 2012). Definitely, autologous blood transfusion is the method of choice with no valid method to date to accurately directly detect such intervention (Malm et al 2016).…”

Annual banned‐substance review: analytical approaches in human sports drug testing

“…The performance enhancing effect of autologous blood doping was recently quantified in a setting consisting of two donations of 450 mL of whole blood each, which were cryopreserved as red blood cell concentrates for up to 16 weeks prior to re‐infusion. Recreational athletes underwent a time‐to‐exhaustion treadmill running tests before and after blood withdrawal as well as 48 h and up to 4 weeks post‐infusion, demonstrating a performance increase 48 h after blood transfusion of 15 ± 8% . The relevance of appropriate anti‐doping measures concerning blood doping has been known for many years, and the haematological module of the ABP has proven its utility and robustness in this context in the past .…”

“…Recreational athletes underwent a time-to-exhaustion treadmill running tests before and after blood withdrawal as well as 48 h and up to 4 weeks post-infusion, demonstrating a performance increase 48 h after blood transfusion of 15 AE 8%. [150] The relevance of appropriate anti-doping measures concerning blood doping has been known for many years, and the haematological module of the ABP has proven its utility and robustness in this context in the past. [151,152] Interpreting blood parameters of the peculiar population of elite athletes necessitates substantial and specific expertise, taking into consideration the occasionally exceptional circumstances and conditions acting on sport professionals.…”