Beyond Analytical Results

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WADA / Science and Investigations Symposium - Istanbul Beyond Analytical Results Martial Saugy, Norbert Baume, Neil Robinson, Nicolas Jan, François Marclay Swiss Laboratory for Doping Analyses Centre Hospitalier Vaudois & University of Lausanne www.doping.chuv.ch

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping /Clean sport Pharmacokinetic / Re-analyses Where do we go?

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping /Clean sport Pharmacokinetic / Re-analyses Where do we go?

Already a long experience The Laboratory interacts with ADO Prof. Donike ologne Laboratory Prof. Ljungqvist IAAF

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping /Clean sport Pharmacokinetic / Re-analyses Where do we go?

The forensic approach of anti-doping 2003-2004: After investigations on blood parameters in the population of professional cyclists, the prevalence of low reticulocytes showed an obvious re-apparition of blood transfusion in the field 10 9 8 7 6 5 4 3 2 1 0 2001 2002 2003 2004 2005 2006 2007 2008 2009 <.4% > 2.0 % Zorzoli & Rossi, 2010, 2012

The forensic approach of anti-doping After the Hamilton case, the Landis case and the Puerto affair, WADA promotes the forensic approach in the fight against doping. The first step was the introduction of the Athlete Biological Passport based on a collaboration between WADA, UCI and the Lausanne Laboratory (LAD) At this stage, the LAD set up a collaboration with the University Institute of Scientific Police & Criminal Sciences. 2 specialists have been hired to explore the Anti-doping intelligence in close collaboration with the Laboratory.

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping /Clean sport Pharmacokinetic / Re-analyses Where do we go?

How DNA can help In 2007, medical equipment was found in a dustbin behind a team hotel during an international rowing competition (Rotsee, Switzerland). The LAD received the mandate by FISA to cooperate for an investigation. More than 30 bottles of drugs and several syringes found in a dustbin. No forbidden substances. 13 infusion systems. Jan & al, Forensic Sci Int. 2011;213:109-13.

How DNA can help DNA analyses on red residues from 10 infusion systems Circumstantial information Dustbin location Inscriptions on packaging

How DNA can help Material found in a dustbin Pre-evaluation Fingermarks? Biological traces?... Contextual information Exploitation Biological traces - Sanction to coaches & officials Analyses Interpretation DNA profiles Source Inference Reference DNA profiles - Leads to no needle policy 8 traces DNA profiles (5 men and 3 women) were corresponding with 8 reference profiles of athletes.

How DNA can help In 2007-2008, IAAF used DNA to control potential manipulation of the athlete s urine sample Based on differences in steroid profiles, IAAF suspected that several athletes were not providing their own urine during out of competition controls. DNA comparison between IC and OOC urine were performed. Urines from 7 athletes were not corresponding, after comparison also with reference blood samples They were charged because of urine substitution and have been suspended for a period of 2 years and 9 months. CAS 2008/A/1718 to CAS 2008/A/1724 http://www.tas-cas.org/d2wfiles/document/3767/5048/0/20091118165643673.pdf (last visited May 2, 2010).

How DNA can help Legitimacy and constraints of DNA tests in Anti-Doping Implementation of DNA profiling is described in 2015 WADA code, BUT : 1. Need to harmonize the profiling kits (loci/markers) 2. Compatible with data protection? 3. DNA profiling doesn t provide information about suspicious result, but only on the identity of the donor. To our point of view, there is a necessity to develop a standardized model of DNA profiling trough a DNA testing policy with guidelines or technical document. Inclusion of DNA profiling in the Athlete's passport? Would be a progress

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping /Clean sport Pharmacokinetic / Re-analyses Where do we go?

Beyond the passport The passport, by the individual and longitudinal examination of the biological pattern from the athlete is one of the best example of the link between Science and Investigation The technical application of the passport is using a body of evidences to assess the normality or abnormality of a biological profile, then possibly to declare an adverse passport finding.

Drug Test Anal. 2012 Nov;4(11):846-53. Schumacher YO 1, Saugy M, Pottgiesser T, Robinson N. Beyond the passport

Drug Test Anal. 2012 Nov;4(11):846-53. Schumacher YO 1, Saugy M, Pottgiesser T, Robinson N. Beyond the passport

Beyond the passport - Competition schedule - Training schedule - Altitude training - Whereabouts - Curve of performances - Lab APMU Expert s investigations - Targeting system Improve the global efficiency of the passport

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping / Clean sport Pharmacokinetic / Re-analyses Where do we go?

The Prevalence of doping / Clean sport IAAF Blood tests: 2001-2009 Tool to measure the prevalence of blood doping Clin Chem. 2011 May;57(5):762-9. Prevalence of blood doping in samples collected from elite track and field athletes. Sottas PE 1, Robinson N, Fischetto G, Dollé G, Alonso JM, Saugy M.

The Prevalence of doping / Clean sport Blood tests 2001-2009 Mean: 12% Country A: 78% Country B: 1% Prevalence M2: Micro-doses of EPO Clin Chem. 2011 May;57(5):762-9. Prevalence of blood doping in samples collected from elite track and field athletes. Sottas PE 1, Robinson N, Fischetto G, Dollé G, Alonso JM, Saugy M.

The Prevalence of doping / Clean sport Neil Robinson 1, Gabriel Dollé 2, Pierre-Yves Garnier 2 & Martial Saugy *1 Bioanalysis 2012, Vol. 4, No. 13, Pages 1633-1643 WADA-IAAF- LAD Collaboration

The Prevalence of doping / Clean sport

The Prevalence of doping / Clean sport Evaluation of longitudinal steroid profiles from football players in UEFA competitions between 2008 and 2012 A Collaborative and retrospective study between 12 European WADA accredited Laboratories

The Prevalence of doping / Clean sport EURO 2008 Austria (OOC) and Switzerland (IC) Harmonization of the steroid profile quantification Solid basis for data comparison EURO 2012 Poland (OOC and IC) Close collaboration with austrian and swiss antidoping labs Possibility to compare results between both competitions Other european competitions UEFA EURO Qualifiers, UEFA Champions Leagues, UEFA Europa League Significant database to valorize!

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping / Clean sport Pharmacokinetic / Re-analyses Where do we go?

Pharmacokinetic / Re-analyses Example of Cocaine case: a rider was found positive with 181 ng/ml BZE and 45 ng/ml MeE. Rider explanation: Mate de coca from his Mother in law in Columbia Urine test: 28 hours after intake Plausible? Pharmacokinetic data can help

Pharmacokinetic / Re-analyses Excretion study with a volunteer RIDER 181 Be 45 Me Volunteer 250 Be 30 Me 1.3 mg of cocaine

Pharmacokinetic / Re-analyses Why should we re-analyze the stored samples: Because the new methods are more sensitive and new metabolites are detected New method Previous method Example of Oxandrolone Guddat et al, Anal Bioanal Chem. 2013 Oct;405(25):8285-94

Pharmacokinetic / Re-analyses Why should we re-analyze the stored samples: Because the new methods are more sensitive and new metabolites are detected Yes, but: When? Do we know if there is a deterrent effect of the re-analyses and is that any relation with the time after the test. Yes, but: How? We need a specific technical document for the re-analyses to harmonize, to target, to decide Yes, but: How much? What is the real ratio cost-benefit of the process long term storage/re-analyses

Where Science meets Investigations Already a long experience The forensic approach of anti-doping How DNA can help Beyond the passport The Prevalence of doping / Clean sport Pharmacokinetic / Re-analyses Where do we go?

Where do we go? Ways for improving the effectiveness of Anti-Doping Code s latitude Facts related to anti-doping rule violations may be established by any reliable means. Forensic context Use of science in service of the law Multidisciplinary research Deeper & broader understanding of doping Scientific innovation Proactive detection & resolution of doping Anti-Doping Intelligence

Acknowledgements To the Turkish Olympic Committee and to WADA And to the LAD Team