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Xavier de la Torre
Application of GC-C-IRMS in the ABP
Athlete Biological Passport Symposium
5-7 November 2018Rome, Italy
• What we know so far
• Previous steps on IRMS application in doping analysis
• Longitudinal application
• Application to real passports
• Conclusions
Outline
Athlete Biological Passport Symposium - November 2018 2
What we know…..
• ABP needs to accumulate data before being effectiveDifficulties on the Results Management – Interaction with APMUsDelay on the final decisionNot fully understood by Athletes and Media
• ABP evaluation is not always effectiveDHEAT transdermalFemale athletesMicro dosing
• The inclusion of additional parameters is not easyHarmonization of new markers detection
Hydroxylated steroidsSulphated steroids
Athlete Biological Passport Symposium - November 2018 3
Previous steps on IRMS application in doping analysis
• IRMS Steroids data are more stable than the corresponding concentrations• Delta values are stable in a short, mid and long term period for a given individual• Delta and delta-delta values are normally distributed in the population and
individuals• The individual variability is much lower than the population one• Longitudinal evaluation of the delta values extends the detection window and an
equivalent approach to steroid module of the ABP is possible
Athlete Biological Passport Symposium - November 2018 4
Variability Concentrations vs Delta values (Male-M1)
SD=0,2
SD=0,3
SD=0,4
CV%=23
CV%=27
CV%=30
-29,0
-28,0
-27,0
-26,0
-25,0
-24,0
-23,0
-22,0
-21,0
-20,0
-19,0
17-Mar-13 6-May-13 25-Jun-13 14-Aug-13 3-Oct-13 22-Nov-13 11-Jan-14 2-Mar-14 21-Apr-14
δ13
C ( ‰
)
Time
5bdiol
M-3DS POP
M+3DS POP
µ-3DS LONG
µ+3DS LONG
5bdiol
Athlete Biological Passport Symposium - November 2018 5
Mean Mean ± 2SD Mean ± 3Sd Outlier
d T_M1 d T_M6 d T_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d A_M1 d A_M6 d A_M12-26
-25
-24
-23
-22
-21
-20δ
13C
(‰)
Mean Mean ± 2SD Mean ± 3SD Outlier
d Et_M1 d Et_M6 d Et_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d 5adiol_M1d 5adiol_M6
d 5adiol_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d 5bdiol_M1d 5bdiol_M6
d 5bdiol_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d E_M1 d E_M6 d E_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
δ 13C (‰) A δ 13C (‰) Et δ 13C (‰) 5adiol
δ 13C (‰) 5bdiol δ 13C (‰) T δ 13C (‰) E
Variability of δ13C (‰) values (TCs)Short, medium an long term period Vol #1
Athlete Biological Passport Symposium - November 2018 6
Variability of δ13C (‰) values (TCs)Short, mid an long term period
Variability of δ13C (‰) values (ERCs)Short, mid an long term period
Mean Mean ± 2SD Mean ± 3SD Outlierd 11bOHA_M1
d 11bOHA_M6d 11bOHA_M12
-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d 11KetoE_M1d 11KetoE_M6
d 11KetoE_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d PD_M1 d PD_M6 d PD_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
Mean Mean ± 2SD Mean ± 3SD Outlier
d PT_M1 d PT_M6 d PT_M12-26
-25
-24
-23
-22
-21
-20
δ 13
C (‰
)
δ 13C (‰) 11OHA δ 13C (‰) 11KetoE
δ 13C (‰) PD δ 13C (‰) PT
Vol #1
Athlete Biological Passport Symposium - November 2018 7
Variability of ∆δ values Short, mid an long term period
Mean Mean ± 2SD Mean ± 3SD Outlierdd (PD-A)_M1
dd (PD-A)_M6dd (PD-A)_M12
-2
-1
0
1
2
3∆
δ
Mean Mean ± 2SD Mean ± 3SD Outlierdd (PD-Et)_M1
dd (PD-Et)_M6dd (PD-Et)_M12
-2
-1
0
1
2
3
∆δ
Mean Mean ± 2SD Mean ± 3 SD Outlierdd (PD-5adiol)_M1
dd (PD-5adiol)_M6dd (PD-5adiol)_M12
-2
-1
0
1
2
3
∆δ
Mean Mean ± 2SD Mean ± 3SD Outlierdd (PD-5bdiol)_M1
dd (PD-5bdiol)_M6dd (PD-5bdiol)_M12
-2
-1
0
1
2
3
∆δ
Mean Mean ± 2SD Mean ± 3SD Outlierdd (PD-T)_M1
dd (PD-T)_M6dd (PD-T)_M12
-2
-1
0
1
2
3
∆δ
Mean Mean ± 2SD Mean ± 3SD Outlierdd (PD-E)_M1
dd (PD-E)_M6dd (PD-E)_M12
-2
-1
0
1
2
3
∆δ
∆δ (PD-A) ∆δ (PD-Et) ∆δ (PD-5adiol)
∆δ (PD-5bdiol) ∆δ (PD-T) ∆δ (PD-E)
Vol #1
Athlete Biological Passport Symposium - November 2018 8
Normale Attesa
-25.0 -24.5 -24.0 -23.5 -23.0 -22.5 -22.0 -21.5 -21.0 -20.5X <= Soglia Categoria
0
5
10
15
20
25
30
N. o
sser
vazi
oni
-24.5 -24.0 -23.5 -23.0 -22.5 -22.0 -21.5 -21.0 -20.5Valore
-3
-2
-1
0
1
2
3
Valo
ri N
orm
ali A
ttesi
Normale Attesa
-26.0 -25.5 -25.0 -24.5 -24.0 -23.5 -23.0 -22.5 -22.0 -21.5X <= Soglia Categoria
0
2
4
6
8
10
12
14
16
18
20
22
N. o
sser
vazi
oni
-26.0 -25.5 -25.0 -24.5 -24.0 -23.5 -23.0 -22.5 -22.0Valore
-3
-2
-1
0
1
2
3
Val
ori N
orm
ali A
ttesi
Normale Attesa
-27.0 -26.5 -26.0 -25.5 -25.0 -24.5 -24.0 -23.5 -23.0 -22.5 -22.0 -21.5
X <= Soglia Categoria
0
2
4
6
8
10
12
14
16
N. o
sser
vazi
oni
Distribution of δ13C (‰) in the Population Normal distribution and P-P plots
A (n=76)
Et (n=73)
5bdiol (n=71)
Etiocolanolone
-25.5 -25.0 -24.5 -24.0 -23.5 -23.0 -22.5 -22.0 -21.5Valore
-3
-2
-1
0
1
2
3
Valo
ri N
orm
ali A
ttesi
Athlete Biological Passport Symposium - November 2018 9
-1.0 -0.5 0.0 0.5 1.0 1.5 2.0 2.5Valore
-3
-2
-1
0
1
2
3
Valo
ri N
orm
ali A
ttesi
Normale Attesa
-1.5 -1.0 -0.5 0.0 0.5 1.0 1.5 2.0X <= Soglia Categoria
0
5
10
15
20
25
30
35
N. o
sser
vazi
oni
Normale Attesa
-1.0 -0.5 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0X <= Soglia Categoria
0
2
4
6
8
10
12
14
16
N. o
sser
vazi
oni
-1.0 -0.5 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5Valore
-3
-2
-1
0
1
2
3
Valo
ri N
orm
ali A
ttesi
Distribution of ∆δ in the Population Normal distribution and P-P plots
PD-E (n=62)
PD-Et (n=71)
Athlete Biological Passport Symposium - November 2018 10
-23.1-22.9
-22.7-22.5
-22.3-22.1
d A
0
2
4
6
8
10
12
N. o
sser
vazi
oni
d ANDRO: SW-W = 0.9558, p = 0.2760
A
-24.2-24.0
-23.8-23.6
-23.4-23.2
d Et
0
2
4
6
8
10
12
N. o
sser
vazi
oni
d ETIO: SW-W = 0.9533, p = 0.2398
Et
-24.30-24.04
-23.78-23.52
-23.26-23.00
d 5adiol
0
1
2
3
4
5
6
7
8
N. o
sser
vazi
oni
d 5a3aD: SW-W = 0.9462, p = 0.1729
5adiol
-24.30-24.08
-23.86-23.64
-23.42-23.20
d 5bdiol
0
2
4
6
8
10
12
14
N. o
sser
vazi
oni
d 5b3aD: SW-W = 0.8984, p = 0.0123
5bdiol
-24.50-24.16
-23.82-23.48
-23.14-22.80
d T
0
1
2
3
4
5
6
7
8
9
10
N. o
sser
vazi
oni
d TESTO: SW-W = 0.9569, p = 0.3344
T
-25.10-24.82
-24.54-24.26
-23.98-23.70
d E
0
2
4
6
8
10
12
N. o
sser
vazi
oni
d Epi: SW-W = 0.9633, p = 0.4371
E
-24.20-23.94
-23.68-23.42
-23.16-22.90
d 11OHA
0
1
2
3
4
5
6
7
8
N. o
sser
vazi
oni
d 11bOHAndro: SW-W = 0.9657, p = 0.5631
11OHA
-23.50-23.24
-22.98-22.72
-22.46-22.20
d 11KetoE
0
2
4
6
8
10
12
N. o
sser
vazi
oni
d 11KetoEtio: SW-W = 0.9621, p = 0.3903
11KetoE
-23.80-23.42
-23.04-22.66
-22.28-21.90
d PD
0
2
4
6
8
10
12
N. o
sser
vazi
oni
d Pdiolo: SW-W = 0.9683, p = 0.5797
PD
-23.3-23.1
-22.9-22.7
-22.5-22.3
d PT
0
1
2
3
4
5
6
7
8
9
N. o
sser
vazi
oni
d Ptriolo: SW-W = 0.9738, p = 0.7430
PT
Athlete Biological Passport Symposium - November 2018 11
Distribution of δ 13C (‰) in Vol #1 (n=28) (Shapiro Wilk Normality test; P>0,05)
-1.10-0.82
-0.54-0.26
0.020.30
Dd PD-A
0
2
4
6
8
10
12
N. o
sser
vazi
oni
?d (PD-A): SW-W = 0.9677, p = 0.5885
∆δ (PD-A)
0.10 0.44 0.78 1.12 1.46 1.80
Dd PD-Et
0
2
4
6
8
10
12
N. o
sser
vazi
oni
?d (PD-Et): SW-W = 0.9381, p = 0.1336
∆δ (PD-Et)
0.00 0.28 0.56 0.84 1.12 1.40
Dd PD-5adiol
0
1
2
3
4
5
6
7
8
9
N. o
sser
vazi
oni
?d (PD-5a3aD): SW-W = 0.9655, p = 0.5336
∆δ (PD-5adiol)
-0.30 0.24 0.78 1.32 1.86 2.40
Dd PD-5bdiol
0
2
4
6
8
10
12
14
N. o
sser
vazi
oni
?d (PD-5b3aD): SW-W = 0.9043, p = 0.0228
∆δ (PD-5bdiol)
0.10 0.46 0.82 1.18 1.54 1.90
Dd PD-T
0
1
2
3
4
5
6
7
8
9
N. o
sser
vazi
oni
?d (PD-T): SW-W = 0.9363, p = 0.1347
∆δ (PD-T)
1.2 1.4 1.6 1.8 2.0 2.2
Dd PD-E
0
1
2
3
4
5
6
7
8
9
10
N. o
sser
vazi
oni
?d (PD-EPI): SW-W = 0.9236, p = 0.0620
∆δ (PD-E)
Athlete Biological Passport Symposium - November 2018 12
Distribution of ∆δ (PD-TC) Vol #1 (n=28)
Reference Population vs. Pharmaceutical Preparations
δ Ref Pop (A) δ Drugs-32.8 -31.6 -30.5 -29.3 -28.1 -26.9 -25.8 -24.6 -23.4 -22.3 -21.1
0
2
4
6
8
10
12
14
16
18
N. O
bser
vatio
ns
n=40
N=105
[1] Ueki M. et al. Rapid Commun. Mass Spectrom., 1999, 13, 2237–2243.[2] de la Torre X. et al. J. Pharm. Biomed. Anal., 2001, 24, 645–650.[3] Cawley A. et al. Drug Test. Anal., 2010, 2, 557–567.[4] Forsdahl G. Drug Test. Anal., 2011, 3, 814–819.[5] Brooker L. Drug Test. Anal., 2014, 6, 996–1001. [6] Pharmaceutical seized by the Carabienieri (NAS) 2014
Athlete Biological Passport Symposium - November 2018 13
Individual Reference vs. Pharmaceutical Preparations & Population
δ Ref Pop (A) δ Drugs δ A Vol#1-32.8 -31.6 -30.5 -29.3 -28.1 -26.9 -25.8 -24.6 -23.4 -22.3 -21.1
0
2
4
6
8
10
12
14
16
18
N. O
bser
vatio
ns
n=28
Athlete Biological Passport Symposium - November 2018 14
Individual Reference vs. Pharmaceutical Preparations & Population
δ Ref Pop (A) δ Drugs δ A Vol #1 δ A Vol #4-32.8 -31.6 -30.5 -29.3 -28.1 -26.9 -25.8 -24.6 -23.4 -22.3 -21.1
0
2
4
6
8
10
12
14
16
18
N. O
bser
vatio
ns
n=28
Athlete Biological Passport Symposium - November 2018 15
Longitudinal application
Athlete Biological Passport Symposium - November 2018 16
To test the sensitivity of the IRMS Bayesian Model where theABP Steroid module and the positive criteria for IRMS basedon population references fail.
Objective
δ (‰ )
Time
Androstenedione orTestosterone administration
Detection capacity using WADA criteria
Athlete Biological Passport Symposium - November 2018 17
Experimental and Methods
SP IRMSEnzymatic hydrolysis, liquid-liquid extraction with TMS derivate and measure in multi-target screenings GC/MS/MS
Enzymatic hydrolysis, liquid-liquid extraction, HPLC purification and GC-C-IRMS detection
Mazzarino, M. et al. Anal. Chim. Acta 683, 221–6 (2011).de la Torre, X., et al. Anal. Chim. Acta 756, 23–29 (2012).
3 samples/week for 1 month2 Samples/month for 6 months1 Sample/month for 1 year
28 samples/volunteer
Urine samples collection, longitudinal study
Athlete Biological Passport Symposium - November 2018 18
3 Male Caucasian volunteers (41 ± 8 yrs)100 mg androstenedione (AED) p.o. (ASN Androstene 100 TM, Hood River, OR, USA)Urine samples collected before and for 5 days after the administrationThe androstenedione capsule composition was verified and the δ13C (‰) value determined (-30.5 ± 0.3; n=5)
Administration studies
1 Male Caucasian volunteer (43 yrs) Gel (Testogel ®, Schering) , 50 mg/day (eq. to 5 mg T), every 24 h for 4 daysUrine samples collected before and for 36 h after the last administration
The testosterone δ13C (‰) value was determined (-29.6 ± 0.3; n=5)
Oral Androstenedione
Transdermal testosterone
Athlete Biological Passport Symposium - November 2018 19
Oral Androstenedione
Athlete Biological Passport Symposium - November 2018 20
AED metabolism and marker
O
O
OH
O
OH
OH
O OH
O
O
H
O
OH
H
OH
OH
HO
O
OHO
O
OH
O
OH
OHO
OH
OH
AED
7α-OH-AED
7α-OH-T
T
6α/β-OH-AED
6α/β-OH-T
4-OH-AED
4-OH-T
5α/β-DHT
5ααβ -Adiol5αββ-Adiol5βαβ-Adiol
5α/β-androstane-3,17-dione
Androsterone (5α-androstane-3α-ol-17 one)
Epiandrosterone (5α-androstane-3β-ol-17 one)
Etiocholanolone (5β-androstane-3α-ol-17 one)
Epietiocholanolone (5β-androstane-3β-ol-17 one)
5βββ-Adiol
OH
O
H
O
OO
O
OH
O
OH
OH
2α/β-OH-AED
2α/β-OH-T
Athlete Biological Passport Symposium - November 2018 21
IRMS Results
Athlete Biological Passport Symposium - November 2018 22
SP ABP moduleCase #1 (Sample at 11.5 h)
Athlete Biological Passport Symposium - November 2018 23
Athlete Biological Passport Symposium - November 2018 24
SP ABP moduleCase #1 (Sample at 21.5 h)
IRMS population-based: ∆δERC-TC(‰) vs Time (h)
-1012345678
0 12 24 36Pd-A Pd-Etio Pd-TPd-E Pd-5aAdiol Pd-5bAdiol
-1012345678
0 12 24 36Pt-A Pt-Etio Pt-TPt-E Pt-5aAdiol Pt-5bAdiol
-1012345678
0 12 24 3611KE-A 11KE-Etio 11KE-T
11KE-E 11KE-5aAdiol 11KE-5bAdiol
-1012345678
0 12 24 36
11OHA-A 11OHA-Etio 11OHA-T11OHA-E 11OHA-5aAdiol 11OHA-5bAdiol
33.5 h
30 h33.5 h
33.5 h
Case #1
Athlete Biological Passport Symposium - November 2018 25
IRMS population-based
Inconclusive sample
Case #1 (Sample at 38.5 h)
Athlete Biological Passport Symposium - November 2018 26
-26.079
-28
-27
-26
-25
-24
-23
-22
-21
-201 3 5 7 9 11 13 15 17 19 21 23 25 27
d5a3aD 0.50% 99.50%
-26.72
-28
-27
-26
-25
-24
-23
-22
-21
-201 3 5 7 9 11 13 15 17 19 21 23 25
d5b3aD 0.50% 99.50%
-25.406
-28
-27
-26
-25
-24
-23
-22
-21
-201 3 5 7 9 11 13 15 17 19 21 23 25 27
dEtio 0.50% 99.50%
-24.087
-28
-27
-26
-25
-24
-23
-22
-21
-201 3 5 7 9 11 13 15 17 19 21 23 25 27 29
dAnd 0.50% 99.50%
IRMS Bayesian approach δTC(‰) Case #1 (Sample at 38.5 h)
δ TC(
‰)
Athlete Biological Passport Symposium - November 2018 27
-23.198
-28
-27
-26
-25
-24
-23
-22
-21
-201 3 5 7 9 11 13 15 17 19 21 23 25 27 29
dPd 0.50% 99.50%
-23.555
-28
-27
-26
-25
-24
-23
-22
-21
-201 3 5 7 9 11 13 15 17 19 21 23 25
d11bOHAnd 0.50% 99.50%
Case #1 (Sample at 38.5 h)
δPd(‰) δ11bOHAnd(‰)
Athlete Biological Passport Symposium - November 2018 28
IRMS Bayesian approach δERC(‰)
IRMS Bayesian approach ∆δERC-TC(‰)
2.881
-2
-1
0
1
2
3
4
1 3 5 7 9 11 13 15 17 19 21 23 25 27
dPd.5a3aD 0.50% 99.50%
3.522
-2
-1
0
1
2
3
4
1 3 5 7 9 11 13 15 17 19 21 23 25 27
dPd.5b3aD 0.50% 99.50%
0.889
-2
-1
0
1
2
3
4
1 3 5 7 9 11 13 15 17 19 21 23 25 27
dPd.And 0.50% 99.50%
2.208
-2
-1
0
1
2
3
4
1 3 5 7 9 11 13 15 17 19 21 23 25 27
dPd.Eti 0.50% 99.50%
Case #1 (Sample at 38.5 h)
Athlete Biological Passport Symposium - November 2018 29
∆δ E
RC-T
C(‰
)
Transdermal Testosterone
Athlete Biological Passport Symposium - November 2018 30
Transdermal T administration
Gel (Testogel ®, Schering)
50 mg/ day (eq. to 5mg T)
Every 24 h x 4 days
Athlete Biological Passport Symposium - November 2018 31
Campione n° T-Gel 14 CRITERI DI VALUTAZIONE
11bOH-Andro 11KetoEtio Pregnantriolo Pregnandiolo POSITIVE INCONCLUSIVE
Andro 0,2 -0,6 0,1 0,8 ANDRO INCONCLUSIVE
Etio 1,0 0,2 0,9 1,6 ETIO INCONCLUSIVE
5a3aA 3,6 2,9 3,6 4,3 a DIOLO INCONCLUSIVEINCONCLUSIVE!5b3aA 1,2 0,5 1,2 1,9 ENTRAMBI I DIOLITesto 2,1 1,4 2,1 2,8 UN DIOLO E IL TESTO INCONCLUSIVE
Epitesto 0,9 0,1 0,9 1,6 EPITESTODHEA -0,2 -1,0 -0,3 0,5
δ 13C/12C uc Ampl 44 (mV) Range di linerità (mV)
Frazione 1-7-8Etio -23,8 0,15 3719 350-7000Andro -23,0 0,09 3302 350-700011bOH-Andro -22,8 0,23 1259 350-7000
Frazione 2-65a3aA -26,5 0,29 2847 350-700011KetoEtio -23,6 0,09 1711 350-7000
Frazione 5-95b3aA -24,1 0,24 4335 350-7000Pregnandiolo -22,2 0,21 7143 350-7000
Frazione 3-6ptTesto -25,0 0,03 1832 350-7000Pregnantriolo -22,9 0,31 5158 350-7000
Frazione 4DHEA -22,6 0,11 751 350-7000
Frazione 4EpiEpitesto -23,8 0,11 1716 340-7000
Athlete Biological Passport Symposium - November 2018 32
IRMS Bayesian approach δERC(‰) (Sample at 12 h last adm)
Athlete Biological Passport Symposium - November 2018 33
δ ERC
(‰)
PPV = 99,9999991 % PPV = 99,999998 %
IRMS Bayesian approach ∆δERC-TC(‰) (Sample at 12 h last adm)
Athlete Biological Passport Symposium - November 2018 34
∆δ E
RC-T
C(‰
)
Methods comparison
Methods
Case WADALab ABP SP module WADA
TD2016IRMS IRMS Bayesian
Androstenedione100 mg p.o.
1 25.5 11.5 33.5 38.5
2 23.75 13.5 28.7 37.5
3 32 15 54.0 59.5
T gel 4 x 50 mg
(every 24 h) 1 0 72 (??) 96 > 104
It is evident that IRMS values evaluated with a Bayesian approach, increase the sensitivity of the technique
Time window of detection (h)
Athlete Biological Passport Symposium - November 2018 35
Athlete Biological Passport Symposium - November 2018 36
Steroid Profile ITP
Steroid Module ABP
IRMSConfirmation Method
IRMS ITP
Isotopic Module ABP
IRMSConfirmation Method
Run time 15’ / sample
One GC/C/IRMS analysis per SamplePositive QC
GC: Agilent 6890;Column : HP-5MS : 5% PhenylmethylsiliconeInjector T: 280°COven program: 150°C (1’)- @25°C/min- 260°C (3’)-@ 40°C/min-310°C (2.7 min)
Injection: 2 μL Splitless
GC/C/IRMSThermo Delta Advantage
E
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Analysis Sequence
Confirmation Screening
25
PQCNQC
PQCNQC
5
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Increasing workload capacity
Step h
3-12 mL Urine
3Free fraction removal
Hydrolysis + L/L
HPLC / sample 0,71,7
4-5 Injections / sample 1
Total time for 30 samples 54
Injections per sequence > 120
Step h
6 mL Urine
2No Free fraction removal
Hydrolysis + L/L
HPLC / sample 0,30,6
1 Injection / sample 0,3
Total time for 30 samples 19,4
Injections per sequence < 35
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Real passports
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Steroid Module
Samples collected 2015-2018
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-28.0
-27.0
-26.0
-25.0
-24.0
-23.0
-22.0
-21.0
-20.0
26-Nov-16 6-Mar-17 14-Jun-17 22-Sep-17 31-Dec-17 10-Apr-18 19-Jul-18
13C δ (‰)
T A E 5a 5b PD
ca. 3 years
-1.0
-0.5
0.0
0.5
1.0
1.5
2.0
2.5
3.0
26-Nov-16 6-Mar-17 14-Jun-17 22-Sep-17 31-Dec-17 10-Apr-18 19-Jul-18
∆δ
PD-T PD-A PD-E PD-5a PD-5b
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Isotopic Module
SD < 0.7 ‰
IRMS data are stable
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Isotopic Module
A Et T
5aAdiol 5bAdiol PD
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Steroid Module
2 Laboratories involved
2 Laboratories involved
-28.0
-27.0
-26.0
-25.0
-24.0
-23.0
-22.0
-21.0
-20.0
-19.0
23-Oct-15 18-Aug-16 14-Jun-17 10-Apr-18 4-Feb-19
13C δ(‰)
T A E 5a 5b PD
-26.0
-24.0
-22.0
-20.0
-18.0
-16.0
16-Dec-16 6-Mar-17 25-May-17 13-Aug-17 1-Nov-17
13C δ (‰)
T A E 5a 5b PD
(ca. 1 year) (ca. 2 years)
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Isotopic Module
Athlete Biological Passport Symposium - November 2018 46
Steroid Module
-2.0
-1.0
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
31-Jan-16 18-Aug-16 6-Mar-17 22-Sep-17 10-Apr-18 27-Oct-18
∆δPD-T PD-A PD-E PD-5a PD-5b
-31.0
-29.0
-27.0
-25.0
-23.0
-21.0
31-Jan-16 18-Aug-16 6-Mar-17 22-Sep-17 10-Apr-18 27-Oct-18
13C δ (‰)
T A E 5a 5b PD
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Isotopic Module
Athlete Biological Passport Symposium - November 2018 48
Isotopic Module
A Et T
5aAdiol 5bAdiol PD
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Isotopic Module
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……. But the steroid module of the ABP has some additional information
Conclusions and Future Perspectives
IRMS Bayesian model:
Improves the sensitivity for the detection of doping with
pseudo endogenous steroids
The simplicity and the same approach used for the Steroid
Profile Adaptive Model should allow an easy inclusion in the
Athlete Biological Passport
The use of IRMS should be extended not only to
confirmations
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Future
Greater harmonization among Laboratories is needed
Study of potential confounding factors needed
Acknowledgments
My IRMS colleagues @ the Laboratorio Antidoping FMSI
Italian Ministry of Health (“Ministero della Salute,Commissione per la vigilanza sul doping e sulla tutelasanitaria delle attività sportive”) for the financialsupport of the initial part of the project
World Antidoping Agency (WADA) for financial support
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