WADA ...

Discussion in 'Steroid Forum' started by Michael Scally MD, May 1, 2011.

  1. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Subject Based Steroid Profiling

    [Also, see: https://thinksteroids.com/community...sterones-effect-on-epitestosterone.134299923/ ]

    Naturally occurring steroids have hypertrophic capacities for muscle tissue and can accordingly increase the athlete’s strength. Therefore, these performance-enhancing substances are listed as prohibited compounds by the World Anti-Doping Agency (WADA). Their detection has subsequently been implemented in routine procedures of doping control laboratories. Nowadays, endogenous steroids are believed to be among the most widespread doping substances misused in sports. In 2008, more than 40% of the total adverse analytical findings and atypical findings reported by WADA-accredited testing laboratories were attributed to endogenous steroids.

    The detection of misuse with naturally occurring steroids remains challenging since analytical screening methods cannot differentiate between synthetic and natural compounds. The costly and laborious confirmation analysis with gas chromatography/ combustion/isotope ratio mass spectrometry has been applied since the end of the 1990s to differentiate exogenous from endogenous steroids in suspicious urine samples, but today availability of synthetic steroids with the same C13/C12 ratio as endogenous ones can seriously limit this approach.

    Supplementation with endogenous steroids alters basal steroid concentrations. In the early 1990s, population-based reference ranges were established for testosterone (T), epitestosterone (E), androsterone (Andro), etiocholanolone (Etio), 5?-androstane-3?, 17?-diol (5???-Adiol), 5?-androstane-3?,17?-diol (5???-Adiol) and a number of steroid ratios i.e. T/E, Andro/Etio, Andro/T. These reference values were applied to screen for outlying values characteristic of doping with steroids. At that period, the studies mainly focused on the administration of T.

    To defeat doping control tests in the early 1990s, athletes started to use new endogenous steroids which alter the steroid profile in different ways than T. 5?-dihydrotestosterone (DHT or androstanolone), a direct metabolite of T, was suited for this purpose as it leaves the T/E ratio unchanged just after administration. Indeed, the enzymatic 5?-reductase conversion of T to DHT is considered as an irreversible process. Moreover, DHT was known to be a 3-times more potent androgen than its precursor as it shows greater affinity for the androgen receptor complex. As DHT misuse was investigated and detection criteria were established in the mid 1990s, the endogenous 5?-androstane-3?,17?-diol was marketed as a prohormone of DHT to obtain the same effects and was sold as a nutritional supplement.

    From 1996 onwards, DHEA has been marketed as a freely available food supplement. Together with androstenedione (Adion) and andostenediol, DHEA became an over-the-counter product in the United States. Elsewhere in the world, DHEA could be obtained via the Internet or as pharmaceutical preparations from local drug stores with a medical prescription. DHEA is known to be the most abundant androgen circulating in the body and its sulfate ester acts as a reservoir or buffer for steroid genesis of androgens. As the high concentrations decline during lifespan, it was believed that DHEA had anti-ageing effects and was marketed accordingly. Although DHEA is generally claimed to be a weak androgen, the anabolic nature of DHEA has been proven using DNA micro arrays to compare its genomic expression profile with that of DHT. DHEA metabolises via Adion to T and is therefore considered a prohormone of T. Both DHT and DHEA have been misused by athletes. Consequently, several groups have reported on the detection of both DHT and DHEA misuse and proposed decision criteria based upon population statistics.

    Although it was already known in the 1990s that subject based reference ranges are more reliable than population-based reference ranges for androgens and their ratios, it is only in 2007 that an evaluation method was proposed for this purpose. In that model, Bayesian inference techniques are used to switch the focus from comparison with a population to the determination of individual reference values as the number of individual records grows. Today, following the Athlete Biological Passport (ABP) paradigm, individual records comprising results from blood and/or urine analyses acquired in different doping control laboratories are stored over time to reveal the biological response of doping or effects of a medical condition requiring a closer examination. The Athlete Steroidal Passport (ASP) is the endocrine module of the ABP that collects steroid profiling data obtained from urine samples. The ASP records some steroids and metabolites which are routinely monitored in steroid profiling for doping control purposes. The ASP currently includes T, E, Andro, Etio, 5???-Adiol, 5???-Adiol, DHEA, and DHT. In addition to the T/E ratio, relevant steroid ratios such as Andro/Etio, 5???-Adiol/5???-Adiol, 5???- Adiol/Etio, and Andro/T are also evaluated. In its present form, the ASP remains mainly operational for the alteration of the steroid profile induced by different routes of T administration.

    Minor steroid metabolites are important markers for specific determination of the misuse in sports with other naturally occurring steroids, for example, 7-keto-dehydroepiandrostenendione, DHEA and Adion which can be easily purchased as food supplements. Together with traditionally monitored steroids, these hydroxylated steroid metabolites provide important information on how the steroid profile is altered after misuse of particular endogenous steroids. Hence, additional markers may provide useful information regarding the administration of other steroids like DHT and DHEA. This work aims to identify novel biomarkers for DHT and DHEA administration in sports, with a selection among a large set of potential markers based on the results obtained by the Bayesian framework of the ASP.


    Van Renterghem P, Van Eenoo P, Sottas P-E, Saugy M, Delbeke F. Subject-based steroid profiling and the determination of novel biomarkers for DHT and DHEA misuse in sports. Drug Testing and Analysis 2010;2(11-12):582-8. Subject-based steroid profiling and the determination of novel biomarkers for DHT and DHEA misuse in sports - Van Renterghem - 2010 - Drug Testing and Analysis - Wiley Online Library

    Doping with natural steroids can be detected by evaluating the urinary concentrations and ratios of several endogenous steroids. Since these biomarkers of steroid doping are known to present large inter-individual variations, monitoring of individual steroid profiles over time allows switching from population-based towards subject-based reference ranges for improved detection. In an Athlete Biological Passport (ABP), biomarkers data are collated throughout the athlete's sporting career and individual thresholds defined adaptively. For now, this approach has been validated on a limited number of markers of steroid doping, such as the testosterone (T) over epitestosterone (E) ratio to detect T misuse in athletes. Additional markers are required for other endogenous steroids like dihydrotestosterone (DHT) and dehydroepiandrosterone (DHEA). By combining comprehensive steroid profiles composed of 24 steroid concentrations with Bayesian inference techniques for longitudinal profiling, a selection was made for the detection of DHT and DHEA misuse.

    The biomarkers found were rated according to relative response, parameter stability, discriminative power, and maximal detection time. This analysis revealed DHT/E, DHT/5?-androstane-3?,17?-diol and 5?-androstane-3?,17?-diol/5?-androstane-3?,17?-diol as best biomarkers for DHT administration and DHEA/E, 16?-hydroxydehydroepiandrosterone/E, 7?-hydroxydehydroepiandrosterone/E and 5?-androstane-3?,17?-diol/5?-androstane-3?,17?-diol for DHEA. The selected biomarkers were found suitable for individual referencing. A drastic overall increase in sensitivity was obtained. The use of multiple markers as formalized in an Athlete Steroidal Passport (ASP) can provide firm evidence of doping with endogenous steroids.
     
    Last edited: May 1, 2011
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  2. heavyiron

    heavyiron Member

    Interesting reading, thank you for posting this!
     
  3. Millard Baker

    Millard Baker Member

    Does anyone know what the actual costs of Athlete Biological Passport (ABP) / Athlete Steroidal Passport (ASP)? This seems prohibitively expensive for widescale implementation. Either this will mean more taxpayer funds going to anti-doping or sports league having greater control over the anti-doping agencies that police them (since anti-doping will become increasingly dependent on their financial support).
     
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  4. markstent

    markstent Junior Member

    This looks very very interesting
     
  5. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    CASE SCENARIO

    You are the head team physician for a major international professional cycling team. A 30-year-old elite professional cyclist presents with concerns regarding his performance. He has been finishing in the top 10 of major international stage races, but he has struggled to break into the top 5. If he were able to improve his performance by as little as 3%, he could potentially win these races. He has worked with a host of well-known coaches during the past 3 years to modify his training regimen, yet his results have not improved, and he expresses significant frustration with this lack of improvement. He states that the best years of his career are probably going to take place during the next 5 years and that he needs to maximize his athletic abilities to increase his income potential and enhance the recognition he receives from his peers and the public. In addition, he believes that his competitors are using performance-enhancing drugs (PEDs) and that this is the reason he is unable to compete with them.

    He would like to know what you think about athletes using PEDs, what the risks and benefits are related to their use, and whether you would consider prescribing him PEDs. His inquiry is serendipitous because the management of the cycling team recently approached you to revisit the team’s antidoping policies. They are concerned with the bad press surrounding athletes who have been suspended for using PEDs and the public perception that many of the athletes within their sport are “cheaters.” Please advise the athlete and cycling team accordingly.


    Finnoff JT, Chimes GP, Murray TH. Performance-Enhancing Drugs. PM&R 2010;2(4):285-93. ScienceDirect - PM&R : Performance-Enhancing Drugs
     

    Attached Files:

  6. heavyiron

    heavyiron Member

    That case scenerio has red flags all over it but Im pro PED use for sure.
     
  7. ergomaniac

    ergomaniac Member

    oh yeah babe. i gotta response.
    my head. exploding. it hurts.
    got more links?.....
     
  8. ergomaniac

    ergomaniac Member

    really. the whole situation is so silly. and reeks of politics. and misguided morality.
     
  9. Millard Baker

    Millard Baker Member

    Dr. Chimes raises some excellent points...

    The anti-doping movement isn't primarily concerned with athletes' health.

    An athlete using steroids may often be healthier than an athlete who doesn't use steroids.

    The medical community could learn a lot from athletes who use steroids and performance-enhancing drugs. (The medical community owes the bodybuilding community, whether it wants to admit it or not, an enormous debt for the extensive data/information collected from bodybuilders' self-experimentation with anabolic steroids and other bodybuilding drugs as this ultimately led to androgen therapies for wasting, hypogonadism, testosterone replacement therapy etc.)

     
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  10. Nemesis RR

    Nemesis RR Member

    Can you post or send me the full article please.
     
  11. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    The full-text article that is quoted in MB's post is an attachment to my post above.
     
  12. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    A Pilot Study On Subject Based Comprehensive Steroid Profiling

    The detection of misuse with naturally produced steroids has always been challenging because of the chemical similarity of these substances with those produced endogenously in the body. Threshold values were established to detect unusually high levels of androgens and altered steroid concentrations caused by illicit steroid supplementation. However, the discrimination between elevated levels caused by cheating and naturally elevated production/ excretion could not always be unambiguously established because natural levels of steroid markers in biological fluids show potentially big inter-individual variances. For many years, reference limits inferred from a large number of measurements in a healthy population provided decision criteria to prove an athlete’s guilt.

    In the early 1980s, the gas chromatography/mass spectrometry measurement of testosterone (T) over epitestosterone (E) ratio and corresponding laboratory reporting threshold of 6 were introduced by the International Olympic Committee. Today, the T/E remains the most important marker of T abuse. As the application of endogenous steroids induces an alteration of several steroid metabolites constituting the steroid profile, the threshold values of additional steroid metabolites, e.g. androsterone (Andro), etiocholanolone (Etio), dihydrotestosterone (DHT) and dehydroepiandrosterone (DHEA), were assessed. These markers can also be used to detect the misuse of other endogenous steroids which became available on the market during the 1990s. In 2004, the World Anti-Doping Agency (WADA) adopted these thresholds in its technical document on endogenous steroids and lowered the T/E threshold from 6 to 4.7 In the mean time, endocrine studies of putative doping cases revealed individuals whose basal steroid profile values could exceed the available decision limits.

    To confirm the findings of an atypical profile, a sample suspicious for endogenous steroid doping has to be submitted to a carbon isotope ratio analysis, which can in some circumstances distinguish the pharmaceutical from natural steroids, by determination of the 13C/12C ratio. However, measurement of the distinct stable carbon isotope signature of a steroid is expensive and labor intensive method, which makes this technique not suitable for screening purposes. As IRMS relies on a distinct 13C/12C in synthetic T preparations, problems arise for formulations of which the difference between synthetic and natural carbon isotope ratios becomes smaller. Therefore, steroid profiling still remains a very important technique to detect the misuse of endogenous anabolic steroids.

    Because the assessment of population-based reference ranges for androgenic steroids in urine in 1992, researchers continued their work on steroid profiling and questioned the appropriateness of population-based referencing. For the T/E ratio, it was proven that subject-based reference ranges are more appropriate than the upper limits obtained from population statistics owing to large inter-individual variations. Moreover, T/E is claimed to be a relatively stable parameter with an intra-individual coefficient of variance (CV) of <30% in men and <60% in women.

    These findings inspired anti-doping organizations to perform longitudinal studies of athletes who once delivered suspicious doping samples. A first strategy for the systematic evaluation of all athletes’ individual variation of their own homeostatic values was suggested in 2002 and first referred to a ‘haematological passport’ to deter blood doping. Since 2007, researchers provided a new tool in the fight against doping with endogenous agents by developing a Bayesian model that proposes reference ranges which combine population-based and individual-based information. For each new record, the adaptive model recalculates the expected reference ranges and thereby progressively evolves from population to subject bases as the number of individual test results grows.

    This Bayesian inferential technique was implemented in the Athlete’s Biological Passport (ABP), which was adopted by WADA. The ABP can be considered as an electronic document that stores individual biomarker records comprising results from blood and urine analyses acquired in doping control laboratories. In particular, the Athlete Steroidal Passport (ASP) is the endocrine module of the ABP that collects steroid profiling data obtained from urine samples.

    Currently, the ASP comprises a small number of steroids, prohormones and main metabolites that are now routinely monitored in steroid profiling for doping control purposes. These steroid profiles usually include T, E, Andro, Etio, 5a-androstane-3a,17b-diol (5aab-Adiol), 5b-androstane-3a,17bdiol (5bab-Adiol), DHEA and DHT. The T/E ratio, on which the Bayesian model was initially validated, is complemented with relevant steroid ratios such as Andro/Etio, 5aab-Adiol/5bab-Adiol, 5aab-Adiol/Etio and Andro/T.

    In contrary to classical metabolic studies which screen for known metabolites, this study aims to find novel biomarkers to improve the detection of T abuse in sports. In a previous work, the usefulness of widening the screening window to minor steroid metabolites was demonstrated. Monitoring other steroid metabolites and evaluation of additional steroid ratios as potential biomarkers can therefore increase the efficiency of detecting T misuse among athletes. In addition to search for new biomarkers, this study reports the usefulness of subject-based referencing for the selected markers within the Bayesian framework of the ABP.


    Van Renterghem P, Van Eenoo P, Sottas P-E, Saugy M, Delbeke F. A pilot study on subject-based comprehensive steroid profiling: novel biomarkers to detect testosterone misuse in sports. Clinical Endocrinology 2011;75(1):134-40. A pilot study on subject-based comprehensive steroid profiling: novel biomarkers to detect testosterone misuse in sports - Van Renterghem - 2011 - Clinical Endocrinology - Wiley Online Library

    Context Until now, the testosterone/epitestosterone (T/E) ratio is the main marker for the detection of testosterone (T) misuse in athletes. As this marker can be influenced by a number of confounding factors, additional steroid profile parameters indicating T misuse can provide substantiating evidence of doping with endogenous steroids. The evaluation of a steroid profile is currently based upon population statistics. As large inter-individual variations exist, a paradigm shift towards subject-based references is ongoing in doping analysis.

    Objective Proposition of new biomarkers for the detection of testosterone in sports using extensive steroid profiling and an adaptive model based upon Bayesian inference.

    Subjects Six healthy male volunteers were administered with testosterone undecanoate. Population statistics were performed upon steroid profiles from 2014 male Caucasian athletes participating in official sport competition.

    Design An extended search for new biomarkers in a comprehensive steroid profile combined with Bayesian inference techniques as used in the athlete biological passport resulted in a selection of additional biomarkers that may improve detection of testosterone misuse in sports.

    Results Apart from T/E, 4 other steroid ratios (6?-OH-androstenedione/16?-OH-dehydroepiandrostenedione, 4-OH-androstenedione/16?-OH-androstenedione, 7?-OH-testosterone/7?-OH-dehydro-epiandrostenedione and dihydrotestosterone/5?-androstane-3?,17?-diol) were identified as sensitive urinary biomarkers for T misuse. These new biomarkers were rated according to relative response, parameter stability, detection time and discriminative power.

    Conclusion Newly selected biomarkers were found suitable for individual referencing within the concept of the Athlete’s Biological Passport. The parameters showed improved detection time and discriminative power compared to the T/E ratio. Such biomarkers can support the evidence of doping with small oral doses of testosterone.
     
  13. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Detecting Anabolic Androgenic Steroids in Human Fingernail Clippings

    A study was designed with the aim to develop a method that would be efficient and sensitive enough to be able to extract and detect AAS that are abused in the practice of doping in sport from nail clipping samples. For an initial study, three AAS with widely ranging chemical properties were chosen to be measured: namely testosterone, testosterone propionate and stanozolol (from Sigma- Aldrich, Poole, UK).

    Samples were collected from male volunteers, who had been taking AAS over the last six months, as well as comparative samples from volunteers without any history of AAS use. An extraction method and a high-performance liquid chromatographic (HPLC) method were developed, analyzed for performance and efficiency, and subsequently LC-ESI-ion-trap-MS (HPLC coupled to an electrospray ionisation mass spectrometry detector) was used to analyse some samples.

    The methods investigated in this preliminary study did not have acceptable sensitivity levels required for the determination of the concentrations expected in the sample type (although they did meet the minimum WADA standard detection capabilities of 2 ng/mL). However, the ESI-ion-trap MS detection did show qualitatively the presence of stanozolol and testosterone although at unquantifiable concentrations. These results provide evidence that AAS, both endogenous and exogenous, do get taken up into the nail plate after administration. This evidence demonstrates that nail clipping analysis warrants further investigation. The method developed in this study can be refined, in terms both of extraction and detection. More sensitive mass spectrometers are becoming available and with improved sample clean-up steps it is expected that AAS could be quantified in such samples.

    With such improvements, nail samples could be a useful additional sample type to combat doping in sport. With a longer detection window covering out of competition periods and nail samples being difficult to adulterate they offer considerable advantages in policing antidoping policies.


    Brown HG, Perrett D. Detection of Doping in Sport: Detecting Anabolic-androgenic Steroids in Human Fingernail Clippings. Med Leg J 2011;79(2):67-9. Detection of Doping in Sport: Detecting Anabolic-a... [Med Leg J. 2011] - PubMed result
     
  14. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    While this study is done in cattle, the implications are clear.

    In this study, the feasibility of desorption electrospray ionization (DESI) multistage tandem mass spectrometry (MSn) for the rapid screening of anabolic steroid esters in bovine hair was studied. To this end a simplified extraction and preconcentration procedure was developed and results obtained were critically compared with state-of-the-art ultra high performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) operated in the multiple reaction monitoring (MRM) mode.


    Nielen MW, Nijrolder AW, Hooijerink H, Stolker AA. Feasibility of desorption electrospray ionization mass spectrometry for rapid screening of anabolic steroid esters in hair. Anal Chim Acta 2011;700(1-2):63-9. ScienceDirect - Analytica Chimica Acta : Feasibility of desorption electrospray ionization mass spectrometry for rapid screening of anabolic steroid esters in hair

    Hormone and veterinary drug screening and forensics can benefit from the recent developments in desorption electrospray ionization (DESI) mass spectrometry (MS). In this work the feasibility of DESI application for the rapid screening of intact esters of anabolic steroids in bovine hair has been studied. Using a linear ion trap both full scan and data-dependent collision induced dissociation MS(n) spectra were acquired in minutes for testosterone cypionate, testosterone decanoate and estradiol benzoate standard solutions deposited on a glass or PTFE surface. However direct analysis of incurred hair failed due to inefficient desorption ionization and the minute quantities of steroid esters present. Therefore a simplified ultrasonic liquid extraction procedure was developed, allowing rapid DESI analysis of a few microliters of the concentrate and a total analysis time of 2-4h per batch instead of 3 days. The potential of this DESI approach is clearly demonstrated by MS(3) data from hair samples incurred with high levels (300-800mugkg(-1)) of steroid esters, levels which do occur in samples from controlled- and illegally treated animals. For much lower levels state-of-the-art ultra high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) screening methods remain the method of choice and might benefit from the proposed simplified extraction as well.
     
  15. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Influence Of Human Chorionic Gonadotropin On Endogenous Steroids Concentrations

    The results of this study were in accordance with previously published papers investigating hcg use in sports. Direct measurement of hCG, testosterone concentration, and T/LH ratio in both urine and blood matrices have been shown to be relevant biomarkers for hCG doping detection.

    [Note: The increase of serum T after hCG injection has been largely documented. This stimulation of T production inhibits the LH synthesis by the pituitary gland through a negative feedback control. These hormones level changes cause a significant increase of the T/LH ratio in serum that was clearly perceived in this study.]


    Strahm E, Marques-Vidal P, Pralong F, Dvorak J, Saugy M, Baume N. Influence of multiple injections of human chorionic gonadotropin (hCG) on urine and serum endogenous steroids concentrations. Forensic Sci Int. Influence of multiple injections of human chorioni... [Forensic Sci Int. 2011] - PubMed result

    Since it is established that human chorionic gonadotropin (hCG) affects testosterone production and release in the human body, the use of this hormone as a performance enhancing drug has been prohibited by the World Anti-Doping Agency. Nowadays, the only validated biomarker of a hCG doping is its direct quantification in urine. However, this specific parameter is subjected to large inter-individual variability and its determination is directly dependent on the reliability of hCG immunoassays used. In order to counteract these weaknesses, new biomarkers need to be evidenced.

    To address this issue, a pilot clinical study was performed on 10 volunteers submitted to 3 subsequent hCG injections. Blood and urine samples were collected during two weeks in order to follow the physiological effects on related compounds such as the steroid profile or hormones involved in the hypothalamo-pituitary axis. The hCG pharmacokinetic observed in all subjects was, as expected, prone to important inter-individual variations. Using ROC plots, level of testosterone and testosterone on luteinizing hormone ratio in both blood and urine were found to be the most relevant biomarker of a hCG abuse, regardless of inter-individual variations.

    In conclusion, this study showed the crucial importance of reliable quantification methods to assess low differences in hormonal patterns. In regard to these results and to anti-doping requirements and constraints, blood together with urine matrix should be included in the anti-doping testing program. Together with a longitudinal follow-up approach it could constitute a new strategy to detect a hCG abuse, applicable to further forms of steroid or other forbidden drug manipulation.
     
  16. Bill Roberts

    Bill Roberts Steroid Forum Leader

    Personally I think the answer, for me anyway, is to just not care about the Olympics.

    For each individual sport, I just don't think the Olympics represent the pinnacle.

    Except perhaps in sports that no one cares about in the first place, that really have no other major competition.

    E.g., we just don't need the Olympics to find who is the fastest sprinter in the world right now, who is the highest-totalling PL'er, or who is the best soccer team, etc.

    Is it possible that part of the extreme doping-obsession isn't just the sanctity, sports-worship aspect combined with the complexes resulted from being stuffed in lockers by the jocks in high school, but also that "only the Olympics" will do this extreme testing so it's your one place to go for "pure" sports? In contrast to say powerlifting competitions or sprint meets or soccer games run by other organizations.

    As to the researchers, they are of course trying to get money to fund their careers.

    When having to go to these obsessive extremes, which seem to be driven more by distorted psychology, attempts to manipulate the public, and personal gain than anything else, my interest becomes lost.

    Similarly, for example even if I never used anabolic steroids, hcg, or GH, if some employer wanted to run 50 different "biochemical profile" tests on me that I had to pass, complete with urine, hair, fingernail clipping, skin cell, and who knows perhaps stool samples my answer would be that I will go elsewhere. Not for me. Take your competition and shove it, basically. Or, as the case may be, we can sit out the season, etc.

    The MLB players succeeded a long time with that basic approach but unfortunately didn't manage to hold the line :(

    And then besides all the above, I want my baseball players, sprinters, etc to be juiced anyway. I don't care to spend my time watching sub-par performances :)
     
    Last edited: Aug 1, 2011
  17. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Forsdahl G, Östreicher C, Koller M, Gmeiner G. Carbon isotope ratio determination and investigation of seized testosterone preparations. Drug Testing and Analysis. http://onlinelibrary.wiley.com/doi/10.1002/dta.373/abstract

    In the present study, the content of a number of black market testosterone products collected in Austria has been analyzed. Additionally, 13?C/12?C ratios were measured for testosterone in the products after cleavage of the testosterone ester. The aim was to determine whether some of these products had similar 13?C/12?C ratios to those normally found for endogenous testosterone, which could prevent a positive isotopic ratio mass spectrometric (IRMS) finding in doping control. Moreover, it was investigated to what extent the preparations contained the masking agent epitestosterone, in order to lower the testosterone/epitestosterone (T/E) ratio in urinary steroid profiles. Out of 30 analyzed products, the declared ingredients differed from the actual content in 10 cases. Epitestosterone, however, could not be found in any of the products. The products displayed ?13CVPDB values between ?23.6 and ?29.4‰. For more than half of these products, the values were within a range reported for endogenous urinary steroids.
     
  18. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Jing J, Yang S, Zhou X, et al. Detection of doping with rhGH: Excretion study with WADA-approved kits. Drug Testing and Analysis. Detection of doping with rhGH: Excretion study with WADA-approved kits - Jing - 2011 - Drug Testing and Analysis - Wiley Online Library

    The detection of recombinant human growth hormone (rhGH) doping using the World Anti-Doping Agency (WADA) approved kits is reported in this research. Twenty-five young male students were selected and divided randomly into two groups with six belonging to the placebo and nineteen to the administration group.

    Thirteen volunteers in one group were administered with a Chinese preparation of rhGH while six volunteers included in the other group were given rhGH made in Switzerland. Both preparations were administered at a dose of 0.1?IU/kg body weight, one injection per day for 14 consecutive days. Blood samples were collected using WADA guidelines and all blood samples were analyzed with WADA-approved Kits 1 and 2. The time window for detection of rhGH doping using WADA-approved kits and criteria are discussed.

    Based on the comparison of the data obtained from this excretion study and from our routine (Chinese population as reference), consideration of the recent WADA criteria for rhGH AAF (Analytical Adverse Findings) is reported statistically. A comparison of data obtained from the two sample groups administered with pharmaceutical preparations, one Chinese rhGH (GenHeal®, S19990019, 1.6?mg (4?IU), Shanghai, China) obtained from prokaryotic cells and the other (Saizen®, S20080036, 1.33?mg (4?IU), Laboratoires Serone S.A., Switzerland) from eukaryotic cells is reported and did not show any significant difference for the detection of doping with rhGH.
     
  19. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Teale P, Scarth J, Hudson S. Impact of the emergence of designer drugs upon sports doping testing. Bioanalysis 2012;4(1):71-88. Impact of the emergence of designer drugs upon s... [Bioanalysis. 2012] - PubMed - NCBI

    Historically, dope-testing methods have been developed to target specific and known threats to the integrity of sport. Traditionally, the source of new analytical targets for which testing was required were derived almost exclusively from the pharmaceutical industry. More recently, the emergence of designer drugs, such as tetrahydrogestrinone that are specifically intended to evade detection, or novel chemicals intended to circumvent laws controlling the sale and distribution of recreational drugs, such as anabolic steroids, stimulants and cannabinoids, have become a significant issue. In this review, we shall consider the emergence of designer drugs and the response of dope-testing laboratories to these new threats, in particular developments in analytical methods, instrumentation and research intended to detect their abuse, and we consider the likely future impact of these approaches.
     
  20. Michael Scally MD

    Michael Scally MD Doctor of Medicine

    Tort N, Salvador JP, Marco MP. Multiplexed immunoassay to detect anabolic androgenic steroids in human serum. Anal Bioanal Chem 2012;403(5):1361-71. Analytical and Bioanalytical Chemistry, Volume 403, Number 5 - SpringerLink

    A multianalyte enzyme-linked immunosorbent assay (ELISA) has been developed for the simultaneous detection of anabolic androgenic steroids (AAS) in human serum. The multiplexed method was developed according to a planar strategy in which the analytes are identified by their location in the microtiter plate. In the immunochemical procedure established here, human serum samples are mixed with a cocktail of antibodies and added to the distinct sections of a microplate biofunctionalized with different haptenized biomolecules. The cocktail of antibodies consists of a mixture of polyclonal antibodies raised against stanozolol (ST), boldenone (B), and tetrahydrogestrinone (THG). The whole immunochemical analytical procedure takes around 2 h including sample preparation, and many samples can be processed simultaneously to screen for the presence of the three AAS in a single run. Using this ELISA, ST, B, and THG can be detected and quantified individually. When used as a screening method, due to the cross-reactivity profiles of the immunoreagents used, the presence of up to 11 AAS can be detected simultaneously. The detectabilities achieved by this method in human serum are below the MRPLs (minimum required performance limits) proposed by WADA (World Anti-Doping Agency) and reference laboratories of the European Community.