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    Testosterone Abuse

Doping

The instinct for man to compete against his peers dates back to the earliest times, and so does the temptation to gain a competitive advantage by unfair means. Today, this unfair advantage can come in many forms, but enhancing performance by chemical means is one of the most sophisticated and hardest to detect. With a responsibility to the public to ensure fair competition, as well as to maintain the health of the athletes themselves, authorities must use the highest performance cutting-edge technology detect and enforce the principle of clean competition.

Stable isotope analysis provides a unique tool for determining whether an athlete has used performance enhancing drugs or if they simply benefit from an natural capacity for high performance. The World Anti Doping Agency has declared that stable isotope analysis is a mandatory technique for sports testing, but similar principles can also be applied to the detection of steroid abuse in cattle.

Detecting synthetic steroids

The application of synthetic forms of endogenous anabolic androgenic steroids by an athlete is known to boost that athletes performance. This is unfair and so must be detected by means of a urinary analysis. GC/C/IRMS has been approved by the World Anti-Doping Agency (WADA) as the official method for detecting this doping procedure and is laid out in WADA Technical Document TD2016IRMS.

GC/C/IRMS

Gas Chromatography / Combustion / Isotope Ratio Mass Spectrometry requires the very highest sensitivity to be able to detect the smallest levels of steroid abuse whilst offering analytical robustness that ensures that the sample analysis is consistently successful, especially during major sporting events. Our AnthrovisION GC/C/IRMS system provides exceptional levels of performance for the most demanding situations.

ionOS® Software

ionOS provides a number of powerful features for the anti-doping laboratory. All data processing is done within ionOS which means that all data processing can be traced back to the original instrument data. Our patented Peak Mapping algorithm is able to automatically detect isotope standards such as USADA-33 and USADA-34 and automatically apply an isotope calibration. Finally, our Calculated Columns functionality is able to automatically calculate Endogenous Reference Compound (ERC) and Target Compound (TC) combinations and flag them as lying outside the permissible ranges.

Sports doping publications using our instruments

Our customers use our instruments to do some amazing research in the sports doping application field. To show you how they perform their research and how they use our IRMS instruments, we have collected a range of peer-reviewed publications which cite our products. You can find the citations below and then follow the links to the publishing journal should you wish to download the publication.

If you would like to investigate our available citations in more detail, or email the citation list to yourself or your colleagues then take a look at our full citation database.

18 results:

Application of stable carbon isotope analysis to the detection of 17beta-estradiol administration to cattle.
Journal of chromatography. A (2005)
C Buisson, M Hebestreit, a Preiss Weigert, K Heinrich, H Fry, U Flenker, S Banneke, S Prevost, F Andre, W Schaenzer, E Houghton, B Le Bizec

The use of anabolic agents in food producing animals is prohibited within the EU since 1988 (96/22/EC directive). The control of the illegal use of natural steroid hormones in cattle is still an exciting analytical challenge as far as no definitive method and non-ambiguous analytical criteria are available. The ability of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) to demonstrate the administration of 17beta-estradiol to bovine has been investigated in this paper. By comparison of 13C/12C isotopic ratio of main urinary estradiol metabolite, i.e. 17alpha-estradiol, with two endogenous reference compounds (ERCs), i.e. dehydroepiandrosterone (DHEA) and 5-androstene-3beta,17alpha-diol, the differentiation of estradiol metabolite origin, either endogenous or exogenous, has been proved to be achievable. After treatment, the delta(13)C(VPDB)-values of 17alpha-estradiol reached -27 per thousand to -29 per thousand, whereas delta13CVPDB-values of DHEA remained between -13 per thousand and -20 per thousand depending on the diet, maize and grass, respectively. A significant difference of delta13CVPDB between ERCs and 17alpha-estradiol was measurable over a period of 2 weeks after estradiol ester administration to the animal.
Tags: carbon , food , spor , gaschrom

Monitoring Anabolic Steroids in Meat-Producing Animals. Review of Current Hyphenated Mass Spectrometric Techniques
Chromatographia (2004)
B. Le Bizec, P. Marchand, D. Maume, F. Monteau, F. Andre

Monitoring anabolic steroids in meat-producing animals is a challenging task. It implies very specific and sensitive analytical methods able to detect and identify sub-μg kg−1 residue levels in complex biological matrices such as meat, urine, or hair. Gas and liquid chromatography coupled to mass spectrometry are the most efficient means of achieving these objectives. In this paper we review how developments in mass spectrometry have been rapidly applied to this problem, how efficient analysis of anabolic steroids in urine, edible tissue, and hair has been achieved, and, later, how measurement of conjugate steroids and determination of the origin of natural steroid hormones has been achieved. The performance characteristics of different mass spectrometers (quadrupole, ion-trap, electromagnetic, isotope-ratio, tandem, and hybrid instruments), the efficiency of different acquisition techniques (LR-SIM, HR-SIM, MRM), and, finally, sample introduction (gas chromatography and liquid chromatography, with discussion of alternative interfaces) are discussed, with numerous applications
Tags: carbon , food , spor , gaschrom

Urinary 19-norandrosterone purification by immunoaffinity chromatography: application to gas chromatography/combustion/isotope ratio mass spectrometric analysis.
Rapid communications in mass spectrometry : RCM (2002)
M C Desroches, J C Mathurin, Y Richard, P Delahaut, J de Ceaurriz

The detection of exogenous 19-norandrosterone (19-NA) in urines was investigated by using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). 19-NA is, for the first time to our knowledge, isolated from urinary matrix by specific immunoaffinity chromatography (IAC) before analysis. The sample preparation consisted of a preliminary purification of urine by solid-phase extraction after hydrolysis by beta-glucuronidase. Unconjugated 19-NA was thus isolated by IAC and directly analysed by GC/C/IRMS. Optimisation of IAC purification was achieved and the reliability of the technique for anti-doping control is discussed.
Tags: carbon , spor , gaschrom

Gas chromatography-combustion-isotope ratio mass spectrometry analysis of 19-norsteroids: application to the detection of a nandrolone metabolite in urine.
Journal of chromatography. B, Biomedical sciences and applications (2001)
J C Mathurin, V Herrou, E Bourgogne, L Pascaud, J de Ceaurriz

Determination of whether the major metabolite of nandrolone in urine, 19-norandrosterone (19-NA), is exogenous or endogenous in origin is one of the most exciting challenges for antidoping laboratories. Gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) can be used to differentiate these two origins by carbon isotopic ratio analysis. A complete method for purification of 19-NA in urine has been established. Acetylated ketosteroids, and in particular 19-NA, are isolated from the urine matrix before analysis after hydrolysis and purification of urine by reversed-phase and normal solid-phase extraction. The limit of detection for 19-NA was about 60 ng with recoveries of 54-60%. Evidence of exogenous administration of 19-NA may be established from isotope ratio determination from the 13C/12C ratios of several synthetic 19-norsteroids compared to those obtained for endogenous steroids.
Tags: carbon , spor , gaschrom

Gas chromatographycombustionisotope ratio mass spectrometry to control the misuse of androgens in breeding animals new derivatisation method applied to testosterone metabo.pdf
Rapid Communications in Mass Spectrometry (2001)
Nicol T Prévost S1

A new derivatisation reaction applied to the analysis of steroids by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) was studied. The trimethylsilylated steroids were characterised by well-resolved chromatographic signals, no peak tailing, reproducible 13C/12C measurements (0.32 per thousand, n = 28), good signal-to-noise ratio and absolute intensity (5 x 10(-9) A, 20 ng), and a slow degradation of copper oxide pellets in the combustion furnace. In addition, two new metabolites and one precursor of testosterone in bovine have been brought into consideration and used for GC/C/IRMS measurements, namely, 3beta-hydroxy-5alpha-androstan-17-one (epiandrosterone), 3beta,17alpha-dihydroxy-5alpha-androstane, and 3beta,17alpha-dihydroxy-5-androstene. The new findings have been applied to an elimination study in bovine of testosterone metabolites after an intramuscular injection of testosterone enanthate. Significant differences (up to 4 per thousand) between testosterone metabolites and precursor were detectable at least three weeks after administration
Tags: carbon , spor , gaschrom

Characterization of exogenous testosterone in livestock by gas chromatography/combustion/isotope ratio mass spectrometry: influence of feeding and age.
Rapid communications in mass spectrometry : RCM (2000)
V Ferchaud, B Le Bizec, F Monteau, F Andre

The detection of exogenous testosterone in bovine urine was investigated by using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). The carbon isotopic ratio measurement of epitestosterone, etiocholanolone (testosterone metabolite) and DHEA (testosterone precursor) in female bovine urines after testosterone enanthate administration was carried out. An important modification in the 13C/12C ratio of testosterone metabolites was observed, such that significant differences between precursor and metabolites of testosterone occurred until three weeks after intramuscular administration of testosterone enanthate. The factors influencing the 13C/12C of endogenous steroids were studied especially through cattle feeding and age. The DHEA mean delta13C value was found to vary between -25 and -26/1000 when hay and concentrate diet were used for fattening. On the other hand the delta13C value observed when maize silage was used increased to -20/1000. Testosterone metabolites showed the same delta13C increase as their precursor. Moreover, we observed a clear relationship between age and efficiency of misuse determination. Indeed, because of the lower concentration of natural hormones in young animals, the contribution of exogenous molecules increases significantly compared with older subjects. Consequently, demonstration of administration is easier to achieve in calves than in mature animals.
Tags: carbon , food , spor , gaschrom

Detection of exogenous intake of natural corticosteroids by gas chromatography/combustion/isotope ratio mass spectrometry: application to misuse in sport.
Rapid communications in mass spectrometry : RCM (2000)
E Bourgogne, V Herrou, J C Mathurin, M Becchi, J de Ceaurriz

A detailed procedure for the analysis of exogenous hydrocortisone and cortisone in urine by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) is proposed. As urinary levels of hydrocortisone are rather low for GC/C/IRMS analysis, the focus is on the main corticosteroid metabolites, tetrahydrocortisone (THE) and tetrahydrocortisol (THF). Following different solid phase extraction purifications, THE and THF are oxidized to 5beta-androstanetrione before analysis by GC/C/IRMS. Significant differences in delta(13)C per thousand values of synthetic natural corticosteroids and endogenous human corticosteroids have been observed. Therefore, a positive criterion, to detect exogenous administration of synthetic corticosteroids in anti-doping control, is proposed.
Tags: carbon , spor , gaschrom

Analysis of exogenous dehydroepiandrosterone excretion in urine by gas chromatography/combustion/isotope ratio mass spectrometry.
Rapid communications in mass spectrometry : RCM (1999)
M Ueki, M Okano

A detailed procedure for the analysis of exogenous dehydroepiandrosterone (DHEA) in urine by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) has been established for detecting doping with DHEA. The average delta-value (parts per thousand difference of (13)C/(12)C ratio from the isotope ratio standard) of 26 synthetic steroids commercially available was -30.1 +/- 2.6, and was significantly lower than that of human endogenous DHEA in urine of the world class athletes who had participated in the XVIIth Olympic Winter Games (-20.3 +/- 2.1, n = 446). Although large inter-individual variations of urinary DHEA excretion were observed following a single oral administration of 50 mg of DHEA, no significant inter-individual difference was found when the excretion of exogenous DHEA was monitored in terms of delta-values using GC/C/IRMS; the minimum delta-values were observed around 6-8 h after the administration, and the values returned to the base level at over 72 h after the dosing. Thus, the deviations in delta-values of DHEA and its diol metabolites are considered to be conclusive evidence for detecting doping with DHEA. Some successful cases of detection of doping with DHEA from athletes are also reported.
Tags: carbon , spor , gaschrom