Download growth – dedicated call – 7/99

GROWTH – DEDICATED CALL – 1/00
TOPIC III.21
Drugs in Sport – Isotopic Procedures for the Detection of Doping with
Endogenous Compounds
1. CONFORMITY WITH THE WORK PROGRAMME
This topic falls under the Competitive and Sustainable Growth Programme, generic
activity Measurement and Testing. Specifically, it is related to Objective GROW-20006.2.2 Measurement and Testing anti-fraud methodologies for which expressions of
interest have been called.
2. KEYWORDS
Doping, isotopic techniques, hormones, erythropoietin, growth hormone, isotope ratio
mass spectrometry
3. SUMMARY OF OBJECTIVES AND JUSTIFICATION
In the fight against drug abuse in sport one of the most difficult areas to control is the
illegal administration of compounds that are produced naturally by the athlete or racing
animal. The existing approach is to establish the normal level amongst a population and
to apply statistical means to calculate a level above which it is highly improbable that a
finding could occur naturally. However “normal levels” can vary greatly and make this
method unreliable. An alternative approach is to determine the presence of the drug
administration, or doping, by the use of isotopic techniques, such as isotope ratio mass
spectrometry. These techniques allow the detection of the doping substance in the
presence of the “natural” analogue and have the crucial advantage of not being reliant
upon the determination of normal levels within populations and individuals. Such
isotopic techniques are well established in the determination of the authenticity of food
and offer the potential for both screening and the unequivocal confirmation of doping in
sport.
4. BACKGROUND
In the fight against drug abuse in sport one of the most difficult areas to control is the
administration of compounds that are produced naturally by the athlete or racing animal.
These drugs produced synthetically or through biotechnological routes are essentially
identical to the endogenous material. Sporting authorities attempt to control these drugs
in a variety of ways. One approach is to establish the normal level among the
population and to apply statistical means to calculate a level above which it is highly
improbable that a finding could occur naturally. Two approaches have been adopted,
firstly the determination of an absolute threshold for the analyte of interest, and
secondly the determination of the ratio of the analyte of interest to a second structurally
related endogenous compound, the concentration of which is not increased by the
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administration. For example the International Olympic Committee (IOC) accepts that a
ratio greater than 6 for testosterone / epitestosterone (T/E) indicates testosterone misuse.
All of the approaches suffer one drawback or another. “Normal” levels can vary greatly
and can be influenced by stress, controlled release formulations such as skin patches
may not produce urine or blood concentrations high enough to result in a positive call.
Ratios such as T/E can be manipulated by for example doping with a mixture of
testosterone and epitestosterone and it is necessary to be able to detect the “added”
hormones over those naturally present. The use of isotopic ratios, especially carbon 12:
carbon 13 (C12/C13) to indicate an administration is gaining acceptance by the IOC.
Peptide and glycoprotein hormones and their analogues pose a far greater analytical
challenge. Substances such as chorionic gonadotrophin (hCG – a human chronic
gonadotrophin), corticotrophin (ACTH), growth hormone (hGH, somatotrophin) and all
the respective releasing factors for such substances and erythropoietin (EPO) are highly
potent, high molecular weight substances believed to be among the primary drugs
misused in sport today. There is also potential for misuse of these substances in nonhuman athletes, e.g. greyhounds and racehorses.
There are at present no accepted methods for the detection of growth hormone or
erythropoietin and the IOC has recently (1) classified as urgent the need for the
development of such methods capable of detecting these compounds, along with the
quality control tools for the implementation of the new methods. Recent incidents
support this need and support claims of misuse by athletes. Vials of growth hormone
were found in the luggage of a Chinese swimmer entered for the World Championships
and the Tour de France in 1998 was seriously disrupted when French police discovered
team managers with supplies of erythropoietin.
The current approach to Drug Control in sport depends on the concept of “confirmatory
analysis” which is deemed to be the unequivocal identification of the presence of the
banned substance usually based upon mass spectrometric data. With the extended use
of isotopic techniques rapid screening tests and confirmation based on mass
spectrometric data will be possible.
5. ECONOMIC AND SOCIAL BENEFITS
The following economic and social benefits will result from this research:
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The provision of screening and confirmatory methodology using instruments
already in IOC laboratories or under development. Improved methods of detection
will lead to an increase in drug testing.
The provision of techniques that can be applied to other endogenous drugs (or
precursors).
The prevention of deaths of athletes and racing animals.
The maintenance and boosting of reputation by Europe and the drug enforcement
authorities being seen to develop and implement new techniques.
The enhancement of Europe’s ability to attract major sporting fixtures to Member
States, because of its superior drug surveillance capabilities, and the consequent
financial, employment, and entertainment benefits to European citizens
The ability of Europe to defend its own sports people falsely accused of doping.
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It has already been said that the detection of endogenous compound is analytically one
of the most demanding and no simple tests using basic technology currently exist. Only
through the application of the most sophisticated isotopic techniques can a solution be
found to the problem of doping with endogenous compounds. Isotopic techniques, e.g.
Isotope ratio mass spectrometry (IRMS), require capital expenditure of ~£100k per
instrument but there are no alternatives. Further IRMS is already in place in some IOC
laboratories, e.g. Germany and Japan, and it is anticipated that it will replace the t/e
ratio as the tool of control for testosterone doping. Experience in food analysis has
shown that once the first isotopic method has been established, methods for other foods
and food components are soon established thereafter.
The adoption of new analytical procedures to detect doping in sport must be achieved
by international agreement (IOC Lausanne Declaration on Doping in Sport (4 February
1999) given at the World Conference on Doping in Sport) and this will be facilitated by
international co-operation in developing the new techniques.
Additionally the specialisms involved in this research are located in a small number of
expert groups around Europe and it is unlikely to be sufficient expertise within a single
member state. A pan-European approach (including the IOC) will also facilitate the
member states of Europe achieve the harmonisation in doping controls sought by the
IOC and substantiate Europe as the leading player in the global fight against doping.
6. SCIENTIFIC AND TECHNOLOGICAL OBJECTIVES
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To develop isotopic techniques (2H, 13C, 15N, 18O,) that will be appropriate for
routine use in drug testing facilities both as rapid screening procedures and as
confirmatory tools. Emphasis must be given to 15N and 18O to complement ongoing
EU funded research on 13C and 2H.
To focus development on isotopic procedures to detect the illegal presence of
peptide and glycoprotein hormones in humans and animals.
To produce for these techniques quality control procedures commensurate with
ISO17025.
7. TIME SCALE
It is anticipated that additional isotopic procedures and their associated quality
procedures should be available for routine use in doping control laboratories within 24
months.
1. Harmonisation of methods and measures in the fight against doping in sport. 1999.
European Commission Final Project Report SMT4-CT98-6530.