T/E

www.doping.chuv.ch
Les$perspec(ves$an(,dopage$$
pour$Sochi$2014$
Mar(al$Saugy$$
$
Laboratoire$Suisse$d’Analyse$du$Dopage$
$
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Summary:!!
!
!
) Detec.on!of!EPO!Doping!:!Stairways!to!Heaven!
!!
) !Testosterone!!always!!at!the!Front!!
!
) !!Where!doping!analyses!can!help!the!athletes!to!detect!cancer!
) !Perspec.ves!
!!
!
www.doping.chuv.ch
Blood$parameters
!!
!
Gareau,!Audran,!Baynes,!Flowers,!!
Duvallet,!Sénécal!&!Brisson!
Nature,(1996(,380,(113(
!
Erythropoie.n!abuse!in!athletes!
!
Ra.o!S)transferin!receptors/Ferri.n!!
$
$
EPO
Ferritin
$$
Tour$de$Suisse$1996$
Hct
sTFR
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Blood$Doping$with$EPO$
Bjarne Riis
Tour de France
1996
Mr.$60%$
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1997:$No$Start$Rule$
$
1997$in$cycling:$$
NO$START$Rule:$$50%$
$
$
$
Later$based$on$Hgb$$:$170$g/l$
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Mean Hct
R,$EPO$Clinical$Study$53
Mean Reti
3.7
51
Hct!
Hct (%)
49
2.9
2.5
47
2.1
45
Reti (%)
On
&
Off
3.3
1.7
43
Ret!
R,EPO$
41
1.3
0.9
39
0.5
0
5
10 15 20 25 30 35 40 45 50 55 60 65 70 75
Days
Haematologica.!2002!Jun;87(6)!
The$interpreta(on$of$secondary$blood$markers$can$get$hazardous$in$case$of$a$discon(nuous$rhEPO$treatment.$
Robinson!N,!Saugy!M,!Buclin!T,!Gremion!G,!Mangin!P.!
!
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R,$EPO$Detec(on$:$Direct$approach$
Lasne$&$de$Ceaurriz,$Nature,$2000$
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Test$IEF$(iso,electro$focalisa(on)$
!
!Based!on!differences!in!the!glycan!chains!between!endogenous!!
!and!!recombinant!from!1st,!2nd!et!3rd!genera.on)!
!
!
3rd!genera.on!
C.E.R.A.!
1st!genera.on!
rEPO!
Endogenous!EPO!
2nd!genera.on!
NESP!
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Separa(on$of$the$molecules$based$on$their$electric$charge$$
$
C.E.R.A.!
rEPO!
Endogen.!
NESP!
NESP!
Endogen.!EPO!
Endogen.!
rEPO!
C.E.R.A.!
rEPO!(1st!gen)!
rEPO!(1st)!
C.E.R.A.!(3rd)!
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NESP!!
(2nd)!
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Belgian!Classics!
2001!
OG!2008!Beijing!
OG!!Salt!Lake!City!2002!
Tour!de!France!2008!
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R,$EPO$Detec(on$:$ $Indirect$approach$
Hgb$
Large between-rider variance!
Hgb!
!
Population mean value is 149 g/L for males and
133 g/L for females
Between-rider variance = 57.2g/L
Within-rider variance = 28.2g/L
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R,$EPO$Detec(on$:$ $Indirect$approach$
Haemoglobin:
from population to athlete-specific
values
Male population mean = 149
Population limit = 170
False positives
False negatives
No Start
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Hgb:$from$popula(on$to$athlete,specific$values$
maximal &
minimal values
expected values
differentiation based on
previous test history
athlete with naturally low hgb
athlete with medium hgb
athlete with naturally high hgb
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$
Main$principles$of$the$athlete$passport$$
$$
$
)  1$:$$Individual$
$
)  2$:$$Longitudinal$
$
)  3$:$$Based$on$an$adap(ve$model$$
$
,$ $4$:$$Defines$normal$individual$
$
$$limits$for$biological$parameters$
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Normal$heamtological$passport$
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Examples$of$Blood$passports$
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Prevalence$of$blood$manipula(on$
IAAF!Blood!tests:!
2001!)!2009!
Blood!doping!!Biomarkers:!!
!
!
Tool!to!measure!the!
prevalence!of!blood!doping!
!
www.doping.chuv.ch
Prevalence$of$blood$manipula(on$
Blood!tests!2001)2009!
Mean:$12%$
Country$A:$78%$
Country$B:$$1%$
Prevalence M2:
Assumption of rhEPO microdoses
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!
!
) !Detec.on!of!EPO!Doping!:!Stairways!to!Heaven!
!!
) !Testosterone!!always!!at!the!Front!!
) !Where!doping!analyses!can!help!the!athletes!to!detect!cancer!
) !Perspec.ves!
!!
!
www.doping.chuv.ch
Testosterone$
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Testosterone:$endogenous$or$exogenous?$
T/E$Ra(o$in$urine$$
(Testosterone$/$Epitestosterone)$
Testosterone$
Epitestosterone$
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T/E$in$athlete$Popula(on$
2004
1982
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Testosterone$applica(on:$ $40$mg$oral$$$
!
!
!
!
!
!
!
!
!
!24
!
!
!!
22.2
18
T/E
12
6.9
6
1.1
0
!!
0
2
4
2.7
3.1
6
8
Time in h
40!mg!testosterone!
1.0
10
0.9 0.9
12
24
4.0
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1000.00
S1
S3
S4
Mean (S2,S5,S6,S7)
100.00
10.00
T/E
Inter,variability$of$$
metabolism$
1.00
0.10
0.01
0
5
10
Time (h)
15
20
25
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Isotopes$in$Humans$
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13C/12C$in$nature$
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Comparison$between$T/E$and$$
δ13C‰$of$androsterone$aber$intake$testosterone$
δ13C‰
-30
25
22.2
-28.1
-28
3
-27.9
-27.3 -27.1
-26.9
-26
-24.4
-24.7
6.9
-24.0
-24
2.7
1.1
-22
0
2
4
6
3.1
1.0
8
Time (h)
10
0.9
12
0.9
24
0
T/E
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IRMS
Inter,variability$of$metabolism$
3!selected!profiles!
T/E
Temps /heures
T/E
S4
δ13C values
T/E
S3
δ13C values
T
values
δ13C
δ13C /‰
T/E
S1
Androstérone (!)
Etiocholanolone (")
Androsténol (!)
Temps /heures
Temps /heures
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Clinical study | Baume et al. Eur. J. Applied Physiol. (2006) 98:329-340
Healthy
volunteers
Placebo
9
300 mg mannitol
!!!
!!!
Testosterone
8
testosterone undecanoate
(80 mg + 115 mg mannitol)
3 times a week for 4 weeks
Steroidomics
6 samples per volunteer
t00 | before the first pill intake
t01-04 | on day 24 at 0, 4, 8, and 24 h after pill intake
t05 |13 days after the last pill intake
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Database
Steroid metabolism
All peaks
m/z 95-1000
Putative steroid
metabolites
N = 234
N > 5000
20
15
U03
LV2
10
U13
5
U20
U06
0
U11
U25
U14
U01
-5
U18
U09
-10
U17
U05
U19
U07
U10
Steroidomics
U24
-15
-20
-20
-10
0
LV1
10
20
J. Boccard et al., Forensic Sci. Int., 213 (2011) 85-94
www.doping.chuv.ch
!
!
) !The!first!steps!
!
) !Detec.on!of!EPO!Doping!:!Stairways!to!Heaven!
!!
) !Testosterone!!always!!at!the!Front!!
) !Where!doping!analyses!can!help!the!athletes!to!detect!cancer!
) !Perspec.ves!
!!
!
www.doping.chuv.ch
www.doping.chuv.ch
Case$Report:$HCG$
13.09.2011$
Immulite!:!!
!ELISA!:! !
!21!mIU/mL
!10!mIU/mL
02.10.2011$
Immulite!:!!
ELISA!:! !
!49!mIU/mL!
!25!mIU/mL
!Serum!ELISA:!135!!mIU/mL!!
11.10.2011$
Contact!with!the!athlete,!
!inves.ga.on!and!immediate!opera.on!for!tes.s!cancer.!
Prevalence!in!WADA!labs:
!
2010:! !12.7/100’000
2011:! !19.3/100’000!
!
!
!Prevalence!in!US!pop.!(same!age)!!
!
!
!1980!–!95:!
!16.7/100’000!
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!
!
) !The!first!steps!
!
) !Detec.on!of!EPO!Doping!:!Stairways!to!Heaven!
!!
) !Testosterone!!always!!at!the!Front!!
) !Where!doping!analyses!can!help!the!athletes!to!detect!cancer!
) !Perspec.ves!
!!
!
www.doping.chuv.ch
www.doping.chuv.ch
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WELCOME !!!