Exogenous testosterone's effect on epitestosterone?

[JonnySak]

MMA fighter Chael Sonnen recently tested positive for an elevated t/e ratio (16.9). At his hearing today, his defense was that he was on trt. My question is, how would trt effect the t/e ratio? Does exogenous testosterone also shutdown epitestosterone production creating a scenario where someone can have a high ratio due to exogenous test, and suppressed epitestosterone production, while sill having "normal" testosterone levels?


Thanks for the help!

JS

 

[Millard]

MMA fighter Chael Sonnen recently tested positive for an elevated t/e ratio (16.9). At his hearing today, his defense was that he was on trt. My question is, how would trt effect the t/e ratio? Does exogenous testosterone also shutdown epitestosterone production creating a scenario where someone can have a high ratio due to exogenous test, and suppressed epitestosterone production, while sill having "normal" testosterone levels?


Thanks for the help!

JS

The medical/scientific ignorance of the CSAC was exposed during Sonnen's suspension hearings broadcast live yesterday.

Yes, TRT can cause an athlete to fail testosterone: epitestosterone (T:E) ratio doping test. And TRT (since it involves exogenous testosterone) will obviously cause an athlete to fail the carbon isotope ratio (CIR) test (which is specifically designed to detect exogenous testosterone).

These are the two tests administered to Sonnen to determine his "doping" guilt. So, even if Sonnen had followed the proper procedure in disclosing his medically-diagnosed hypogonadism, he still could have been banned under CSAC's ass-backward policy.

Yet, the CSAC states that an athlete IS PERMITTED testosterone replacement therapy YET they still CAN NOT fail the T:E ratio or CIR test.

This makes no senses.

From what I understand, Sonnen's blood levels of testosterone were within the physiological (normal) range. The fact that he failed the T:E ratio test doesn't necessarily prove that he used SUPRAphysiological levels of steroids (testosterone). It it only suggestive of steroid use.

See Dr. Scally's comments about the T:E ratio from an older article:

"It is not possible to accurately determine the testosterone dose or serum testosterone level from the testosterone/epitestosterone ratio or the urine testosterone level [...]

At best, the T:E ratio can offer putative evidence of exogenous testosterone usage.

[Ratio greater than 4] ...clearly indicates that exogenous testosterone was administered. However, any attempt to extrapolate this ratio to an amount administered is futile."

Read more from this MESO-Rx article at: Did David Jacobs Autopsy Actually Reveal Elevated Testosterone Levels?

 

[JonnySak]

Thanks for the response. The only thing I'd question is this:

"From what I understand, Sonnen's blood levels of testosterone were within the physiological (normal) range."

From what I saw during the meeting, his actual pre fight blood levels were not tested for and unknown (is that possible though? To get a ratio, don't you need to know the level?). Although there were numerous blood tests from the prior two years showing normal to low test levels.

 

[Michael Scally MD]

You have to take into consideration the source of the Carbon detected within T. The Carbon found in exogenous administration will have a different source (ratio) of Carbon isotopes. The following helps to explain some of the background on testing.


Strahm E, Emery C, Saugy M, Dvorak J, Saudan C. Detection of testosterone administration based on the carbon isotope ratio profiling of endogenous steroids: international reference populations of professional soccer players. British Journal of Sports Medicine 2009;43(13):1041-4. Detection of testosterone administration based on the carbon isotope ratio profiling of endogenous steroids: international reference populations of professional soccer players -- Strahm et al. 43 (13): 1041 -- British Journal of Sports Medicine

Background and objectives: The determination of the carbon isotope ratio in androgen metabolites has been previously shown to be a reliable, direct method to detect testosterone misuse in the context of antidoping testing. Here, the variability in the 13C/12C ratios in urinary steroids in a widely heterogeneous cohort of professional soccer players residing in different countries (Argentina, Italy, Japan, South Africa, Switzerland and Uganda) is examined.

Methods: Carbon isotope ratios of selected androgens in urine specimens were determined using gas chromatography/combustion/isotope ratio mass spectrometry (GC-C-IRMS).

Results: Urinary steroids in Italian and Swiss populations were found to be enriched in 13C relative to other groups, reflecting higher consumption of C3 plants in these two countries. Importantly, detection criteria based on the difference in the carbon isotope ratio of androsterone and pregnanediol for each population were found to be well below the established threshold value for positive cases.

Conclusions: The results obtained with the tested diet groups highlight the importance of adapting the criteria if one wishes to increase the sensitivity of exogenous testosterone detection. In addition, confirmatory tests might be rendered more efficient by combining isotope ratio mass spectrometry with refined interpretation criteria for positivity and subject-based profiling of steroids.

 

[JonnySak]

So since exogenous testosterone has different carbon isotopes than natural testosterone, it will always result in an elevated t/e ratio?

Meaning a t/e ratio test is useless for an athlete on trt?

Also didn't know cholesterol was the origin of testosterone.

 

[Bill Roberts]

No, the T/E and isotopic tests are completely different.

If a high percentage of an individual's testosterone is from injection, the isotope test will be failed because the steroid molecules are detectable as being ultimately of plant origin rather than animal origin.

(Synthetic steroids use diosgenin, extracted from the wild yam, as the starting material. Plants incorporate carbon isotopes into the steroid skeleton in a slightly different ratio than animals do.)

But also if a high percentage of an individual's testosterone is from injection, the T/E ratio will be failed, because when T is produced in the body epi-T comes along with it, whereas injected testosterone doesn't include epi-T. If the ratio is too high, it is unlikely or impossible that that much T could have been produced by the body with that little epi-T.

Even if the amount of T is completely reasonable and well within the normal range, or even potentially if low-normal.

As Millard was saying, they are claiming that they allow testosterone replacement therapy, but then fail athletes for the inevitable outcomes of testosterone replacement therapy (different isotope ratio, increased T/E ratio.)

 

[JonnySak]

Thanks for the clarification on the two tests.

But also if a high percentage of an individual's testosterone is from injection, the T/E ratio will be failed, because when T is produced in the body epi-T comes along with it, whereas injected testosterone doesn't include epi-T. If the ratio is too high, it is unlikely or impossible that that much T could have been produced by the body with that little epi-T.

This is what I was getting at in my original question. I understand in the body's natural production, test and epitest are produced at similar rates. But from what you write, it seems that if the body is no longer producing natural T (due to shutdown) it will no longer produce epi-T either. Because if that wasn't true, I would think a low-normal amount of exogenous Test would not fail the t/e ratio test.

 

[JonnySak]

I just realized something, if a person suffers from hypogonadism and produces very low testosterone, he's probably producing very low epi-testosterone also. So regardless of whether exogenous testosterone slows down epi-testosterone production, the boost to normal testosterone levels would throw the ratio out of whack.

 

[Michael Scally MD]

I just realized something, if a person suffers from hypogonadism and produces very low testosterone, he's probably producing very low epi-testosterone also. So regardless of whether exogenous testosterone slows down epi-testosterone production, the boost to normal testosterone levels would throw the ratio out of whack.

Others can probably comment better than I can, since I have not kept up with this area of science. From the news, at least, it seems the area of masking and detection has been an active arena.

The other setting is that of androgen abusers seeking to avoid detection of synthetic androgens or exogenous testosterone by stimulating endogenous testosterone production. Preliminary information suggests that the testosterone-to-epitestosterone (T/E) ratio is unaffected by hCG treatment, consistent with its stimulation of endogenous testosterone production by Leydig cells, producing a characteristic testosterone to estrogen ratio for that individual, which is no different from natural endogenous testosterone production.

Handelsman DJ. The Rationale for Banning Human Chorionic Gonadotropin and Estrogen Blockers in Sport. J Clin Endocrinol Metab 2006;91(5):1646-53. The Rationale for Banning Human Chorionic Gonadotropin and Estrogen Blockers in Sport -- Handelsman 91 (5): 1646 -- Journal of Clinical Endocrinology & Metabolism

Context: The objective of the study was to review the rationale underlying the banning of human chorionic gonadotropin (hCG) and estrogen blockers (antiestrogens, specific estrogen receptor modulators, aromatase inhibitors) in sports for male and female athletes in the light of gender differences in regulation of reproductive physiology.

Evidence Acquisition: We reviewed well-controlled clinical studies of exogenous testosterone effects on human muscle size and strength in men and all available evidence relevant to the effects of hCG and estrogen blockers on blood testosterone in men and women.

Evidence Synthesis: Well-designed placebo-controlled clinical studies in men with suppressed pituitary-testicular axis establish a strong case that, across a wide range from sub- to supraphysiological doses, muscle growth and strength is proportional to exogenous testosterone dose and resulting blood testosterone concentrations. In men, there is unequivocal evidence that hCG and estrogen blockers cause consistent and sustained rise in blood testosterone concentrations. In women, although there has been no direct testing of ergogenic or myotrophic properties of exogenous testosterone in healthy women, either hCG or estrogen blockers do not produce any consistent or biologically significant increase blood testosterone concentrations.

Conclusions: In men undergoing potential stimulation of endogenous blood testosterone concentrations, blood testosterone concentration is a reasonable surrogate measure for muscle growth and increased strength in men. Because hCG and estrogen blockers produce marked increase in blood testosterone concentration in men, this provides strong evidence to support the banning of hCG and estrogen blockers in men. In women, however, the negligible effect on blood testosterone suggests that drug-induced performance enhancement by hCG or estrogen blockers is highly unlikely. Furthermore, routine urinary hCG testing in young women risks invasion of privacy by detecting unrecognized pregnancy. These considerations suggest that prohibition of hCG and estrogen blockers should be restricted to men in which they are well justified.


Basaria S. Androgen Abuse in Athletes: Detection and Consequences. J Clin Endocrinol Metab 2010;95(4):1533-43.

Context: Doping with anabolic androgenic steroids (AAS) both in sports (especially power sports) and among specific subsets of the population is rampant. With increasing availability of designer androgens, significant efforts are needed by antidoping authorities to develop sensitive methods to detect their use.

Evidence Acquisition: The PubMed and Google Scholar search engines were used to identify publications addressing various forms of doping, methods employed in their detection, and adverse effects associated with their use.

Evidence Synthesis: The list of drugs prohibited by the World Anti-Doping Agency (WADA) has grown in the last decade. The newer entries into this list include gonadotropins, estrogen antagonists, aromatase inhibitors, androgen precursors, and selective androgen receptor modulators. The use of mass spectrometry has revolutionized the detection of various compounds; however, challenges remain in identifying newer designer androgens because their chemical signature is unknown. Development of high throughput bioassays may be an answer to this problem. It appears that the use of AAS continues to be associated with premature mortality (especially cardiovascular) in addition to suppressed spermatogenesis, gynecomastia, and virilization.

Conclusion: The attention that androgen abuse has received lately should be used as an opportunity to educate both athletes and the general population regarding their adverse effects. The development of sensitive detection techniques may help discourage (at least to some extent) the abuse of these compounds. Investigations are needed to identify ways to hasten the recovery of the gonadal axis in AAS users and to determine the mechanism of cardiac damage by these compounds.

 

[Bill Roberts]

Thanks for the clarification on the two tests.



This is what I was getting at in my original question. I understand in the body's natural production, test and epitest are produced at similar rates. But from what you write, it seems that if the body is no longer producing natural T (due to shutdown) it will no longer produce epi-T either. Because if that wasn't true, I would think a low-normal amount of exogenous Test would not fail the t/e ratio test.

Yes, epi-T production for a given individual is in relation to their natural T production. So when that that amount is low, so is the epi-T production.

If the only T present is naturally producted, the T/epi-T ratio remains normal regardless that T is low. (Both numbers in the equation are low, so the ratio remains normal.)

But if T is increased by injection while epi-T has no such increase, the ratio increases.

 

[sir_winstrol]

Yes, epi-T production for a given individual is in relation to their natural T production. So when that that amount is low, so is the epi-T production.

If the only T present is naturally producted, the T/epi-T ratio remains normal regardless that T is low. (Both numbers in the equation are low, so the ratio remains normal.)

But if T is increased by injection while epi-T has no such increase, the ratio increases.

Hello Bill & MIchael, I hope you guys (as experts on this subject) can help me out here.

So according to this suspicious T/E ratio can only be detected during the halflife of testosterone injected?

I have a possible in-(strength)competition steroid testing done in exactly 4 months from now, I just quit injecting Testosterone Enanthate. The usual detection time for Test E is 3 months, so should I be worrying about this?

I understand Wada and National anti-Doping organizations test for both Testosterone/Epitestosterone ratio and the Testosterone Enanthate itself, so T/E ratio should be back to normal in around 15 days (halflife of Test E)? 3 months is the detection time for Test Enanthate steroid ester itself right? Are there any additional tests for testosterone?

Am I worrying to much here?

 

[Bill Roberts]

The time duration isn't the half-life, but rather however long it will be in the particular instance before sufficient natural T and epi-T levels are being produced so that the further T remaining from injection doesn't push the T/E ratio over the limit.

This will depend not only on the half-life, but also on the amount of testosterone injected (if the usage was twice as much per week, then it will take twice as long to clear, for example), on the rate of natural recovery, and on individual differences.

 

[sir_winstrol]

The time duration isn't the half-life, but rather however long it will be in the particular instance before sufficient natural T and epi-T levels are being produced so that the further T remaining from injection doesn't push the T/E ratio over the limit.

This will depend not only on the half-life, but also on the amount of testosterone injected (if the usage was twice as much per week, then it will take twice as long to clear, for example), on the rate of natural recovery, and on individual differences.

Thank you for clarifying this. I took a very modest set of 7 x 250 mg Test Enanthate (purchased from an European pharmacy, so should be legit) injected every 5 days, I am 29 years of age, weight 215 lbs, Bf ~7%. I understand it is hard to give an exact estimate how long it takes for T/E ratio to back to normal but would it be correct to assume my T/E/ ratio _should_ be to normal region within 4 months?

I understand WADA and antidoping agencies also use Carbon-Isotope ratio mass spectrometry to distinguish synthetic testosterone from natural test? Is the 3 month detection time given for Test E for this specific test? Sorry for any spelling mistakes.

 

[Michael Scally MD]

Testosterone doping cannot be detected by simply measuring the levels of endogenous hormones such as androgens in biological fluids. The variability in human metabolism of this compound is simply too large. In the antidoping field, evidence of testosterone administration relies on a confirmatory procedure that uses isotope ratio mass spectrometry (IRMS). A technical document established by WADA in 2004, stipulates that a urine sample must be analysed by IRMS to determine the carbon isotope ratios of androgens if the peak area ratio of testosterone/epitestosterone equivalent to the glucuronide is equal to or greater than 4.0 or if altered steroid profiles are determined. Endogenous testosterone is produced in the human body via cholesterol metabolism.

Detection of testosterone doping relies on the general observation that endogenous testosterone has a different 13C content compared to hemisynthetic testosterone used in pharmaceutical preparations. Such detection is possible because the carbon atoms in steroid molecules originate primarily in atmospheric CO2, which is fixed through photosynthesis. The most important photosynthetic pathways used by plants are the so-called C3 and C4 pathways. Significant isotope fractionation occurs during photosynthetic carbon fixation, depending on the mode of CO2 fixation. Thus, the key enzymes that fix CO2 in C3 plants discriminate more strongly against 13CO2 than their analogues in C4 plants. As a result, the two types of plants differ by about 14‰ in the isotopic composition of their tissues. The natural abundance carbon isotopic ratio is expressed as a ? value relative to an international standard.

The practical outcome is the fact that the distribution of carbon isotopes in an animal reflects the relative abundance of food in the diet that originates directly or indirectly from C4 and C3 plants (either plants or animals that are lower in the food chain). Controlled diet studies have shown that the isotopic composition of the whole body of an animal is enriched by about 1‰ as a function of the isotopic composition of its diet.

Although the diet composition of an athlete has a predominant influence on the carbon isotope ratio of the steroids excreted in urine, there have been no published comparisons of this diagnostic parameter in any cohort of elite athletes in a specific sports category. In this study, the range of the carbon isotope ratio of the steroids relevant to antidoping analysis was investigated in urine specimens obtained from top-level soccer player populations residing in six countries, namely Argentina, Italy, Japan, Republic of South Africa, Switzerland and Uganda. The determination of threshold values specific for a given diet and athlete metabolism is expected to significantly improve the detection of testosterone misuse by means of stable isotope methodology.


Strahm E, Emery C, Saugy M, Dvorak J, Saudan C. Detection of testosterone administration based on the carbon isotope ratio profiling of endogenous steroids: international reference populations of professional soccer players. Br J Sports Med 2009;43(13):1041-4. http://bjsm.bmj.com/content/43/13/1041.full.pdf

BACKGROUND AND OBJECTIVES: The determination of the carbon isotope ratio in androgen metabolites has been previously shown to be a reliable, direct method to detect testosterone misuse in the context of antidoping testing. Here, the variability in the 13C/12C ratios in urinary steroids in a widely heterogeneous cohort of professional soccer players residing in different countries (Argentina, Italy, Japan, South Africa, Switzerland and Uganda) is examined.

METHODS: Carbon isotope ratios of selected androgens in urine specimens were determined using gas chromatography/combustion/isotope ratio mass spectrometry (GC-C-IRMS).

RESULTS: Urinary steroids in Italian and Swiss populations were found to be enriched in 13C relative to other groups, reflecting higher consumption of C3 plants in these two countries. Importantly, detection criteria based on the difference in the carbon isotope ratio of androsterone and pregnanediol for each population were found to be well below the established threshold value for positive cases.

CONCLUSIONS: The results obtained with the tested diet groups highlight the importance of adapting the criteria if one wishes to increase the sensitivity of exogenous testosterone detection. In addition, confirmatory tests might be rendered more efficient by combining isotope ratio mass spectrometry with refined interpretation criteria for positivity and subject-based profiling of steroids.