Testosterone Testing/Flaws in the T/E test

by cyclewife on Jul 27, 2006 3:07 PM EDT Comment 13 comments

To avoid further speculation, I decided to do some research on testosterone testing. I happened upon a great (but very scientific) article entitled, "EVALUATION OF INFLUENTIAL FACTORS ON THE T/E RATIO AS DETERMINED IN DOPING ANALYSIS." article link here. While I am NOT a physiologist, I do have a nursing degree, so I though I would share some of the article's highlights.

below the fold...

The T/E test
1. The most widely used and accepted method of testing for testosterone abuse. From the article, "The ratio of the concentration of testosterone glucuronide to the concentration of
epitestosterone glucuronide (T/E ratio) as determined in urine is the most frequently used
method to prove testosterone abuse by athletes. A T/E ratio higher than 6 has been
considered as proof for abuse in the past; however, cases of naturally occurring higher T/E
ratios have been described."

The test is very accurate (I do not have the actual accuracy percentages, though). However, false positives (meaning the T/E ratio IS elevated, but due to some other cause besides doping) do occur.
Many factors can influence the T/E ratio

Factors influencing T/E ratios

Lab inconsistencies
A collaborative study of international lab testing facilities showed variations both between labs and within the same lab in regards to T/E ratio results. At the time the article was written, there was no standardization of testing methods for the T/E test. I am not sure of how standardized the testing procedures are currently.

Microbial degredation
The article claims, "Bacterial activities in urine may cause significant changes in measured steroid profiles, as was brought forward in the Diane Modahl case in 1994 [48-49]. Because
urine samples are collected under unsterile conditions, bacteria have the chance to
grow when samples are stored too long at a too high temperature." So, storing urine at improper temperatures can change test results. However, it has never been proven that bacterial "activities" have caused an initially negative result to turn into a positive.

Endogenous factors:

Several endocrine disorders can raise a T/E ratio
T (testosterone) levels change throughout the day. T levels are highest in the morning, between 6-8 a.m., and T levels are lowest between 8-10 p.m.
Asians have a lower T/E ratio. Only caucasians were used for the T/E ratio guidelines that the IOC uses. Until Asians are factored into the population used to figure acceptable T/E ratios), an Asian could potentially be taking testosterone and still come out with a negative test result.

**4. As quoted from the article, "The influence of exercise on the T/E ratio is still unclear." Several studies have been done producing conflicting results. However, the point I want to make is that a person's T/E levels remain relatively CONSTANT. A rider's body does not just start cranking out a bunch of excess testosterone (even if it is the penultimate stage). Some studies showed that exercise (performed by conditioned athletes on a tour de france style course) lowered T/E levels, some showed exercise increased the ratio, and some studies showed the ratio remained unchanged. Clearly more research needs to be done.

Exogenous factors

Many medicines/hormones can influence T/E levels. Flagyl (an antifungal) raises T/E ratios, while hCG (a hormone normally secreted by pregnant women) lowers T/E ratios. hCG is commonly used by athletes who are trying to mask a high T/E ratio (due to testosterone abuse).
For those of you wondering about Floyd's beer, ethanol does SLIGHTLY increase the T/E ratio. HOWEVER, it takes a LOT, not just a single drink. The article states, "the increase was observed at dosages higher than 1 g/kg

bodyweight. A dose of 2 g/kg bodyweight resulted in an increase of 30-90%, which was, however, insufficient to reach the IOC criterion of 6." So one beer is not enough to raise Floyd's T/E level to above 6.
3. Steriods (especially anabolic steroids) alter T/E ratios. However, I could not find out any specific information on cortisone (the steroid Floyd was using for his hip) and T levels.

Other tests

T/LH ratio--another test as old as the T/E ratio. Can only be done with males.
GC-combustion-isotope ratio MS (GC-C-IRMS). Very expensive time consuming test. Differentiates between endogenous T (produced by the the body) and exogenous T (from plant sources). Can only be performed by certain certified labs. This is the test is the most widely accepted test for confirming a positive test result. More studies need to be done on the effects of ethnicity and diet on test results using this method.
Hair analysis
High-performance liquid chromatography-mass spectrometry (HPLC-MS)--a new technique. Faster and cheaper than GC-MS.

Final thoughts
I hope and pray that the lab performing Floyd's tests is reliable, accurate, and honest. We must now wait for the test results from sample B before we jump to any conclusions. The number of factors that can influence a test is HUGE--this is a very complicated matter.

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B test vs A

Is the B test from a sample different than the A? If it's taken at the same time and place (i.e. both blood samples) won't the B test logically produce the same results unless the first test result was so grey that little changes mean positive or negative. So luck of the draw. I just don't see how (short of freezing the sample, right, Tyler?) anything will change in the B test.

Good stuff over on the BBC chat site about doubts of testerone testing, etc.

Let's hope it's not true.

by tt the bear on Jul 27, 2006 3:24 PM EDT reply actions 0 recs

Thanks

That's a great article. I would still like to know what FL's baseline hormone levels were over the years and during the last few races. Where they normal, say 1.5:1 or 2:1? I still don't see any obvious reason for FL's levels to jump to twice the "normal" baseline.

Still shocked, depressed, turned off, and trying to withhold judgment before I throw in the towel.

by Rydr1 on Jul 27, 2006 4:09 PM EDT reply actions 0 recs

This concerns me as much as anything else...

I hope and pray that the lab performing Floyd's tests is reliable, accurate, and honest.

There is no way for us to know whether the testing is on the level. No independent verification, no publication of the results, no information about the people performing and supervising the testing.

by socal on Jul 27, 2006 4:13 PM EDT reply actions 0 recs

Hear, hear

Also, how is the chain of custody verified on samples?

Thanks socal for voicing what has been on my mind as well.

by bethie on Jul 27, 2006 4:19 PM EDT up reply actions 0 recs

Not looking good

For hypothesis #3 today ... Questions from an earlier post:

Can anybody explain why Landis popped so early on the climb of Le Mont-Ventoux during this year's Dauphiné Libéré? My theories have ranged from:

1. It got steep and his hip started hurting.

2. Sevilla attacked after about 4 km of climbing and Landis redlined.

3. He was taking a page from Tyler Hamilton's play book (i.e., turn in a completely anonymous performance due to a "stomach ailment" when actually in a blood withdrawal phase), or

4. It was just a "bad day," so turn the rest of the race into a training ride. Go figure.

The team's website noted at the time that it was a "dark day" for Phonak. They had high expectations for Landis and the team. The whole team worked together on the lead out and "recovery" after he blew up. The after-race pictures showed one bummed out Landis.

Not very happy about this whatever the case...

by Rydr1 on Jul 27, 2006 4:14 PM EDT reply actions 0 recs

WADA changed the acceptable ratio recently

It is now 4:1, down from 6:1.

If anyone can tell us why, that'd be nice.

I cynically posit WADA changed it because thay felt like they weren't catching enough dopers.

-Cosmo http://cyclocosm.com

by Cosmo on Jul 27, 2006 4:19 PM EDT reply actions 0 recs

from Velochimp

Floyd Landis' higher than normal testoterone positive drug test could be defended. There are several factors which lead to this positive test. The first is the fact that WADA recently lowered the threshold of testosterone:

The World Anti-Doping Agency has lowered the limit for the maximum T/E level from 6:1 to 4:1. Some athletes have naturally high levels, and can prove this through a series of tests.
-Cyclingnews via Matt Howey on OCC Listserv

The second case is that Landis' Stage 17 breakaway to Morzine could have raised his testoserone levels just because of the type of effort that was involved.

"If drug testing occurs in close proximity to the completion of a
testosterone enhancing workout, testosterone levels and the
testosterone:epitestosterone ratio could be inflated beyond the nanoscopic
tolerances allowed in drug testing."

"The characteristics of a testosterone provoking resistance workout are high
volume, moderate to high intensity, short rest intervals, and the use of
large muscle masses. The protocol should provide frequent bouts of
glycolytic work. This type of workout stimulation is commonly employed by
"natural" body builders."

More info via via Matt Howey on OCC Listserv

The day long breakaway where Landis was pushing huge watts up some tough climbs could have contibuted to increasing Landis' Testosterone output. According to the above site. The reading may be unusually high right after the effort which is when the drug testing would occur.

Compared to other cases of doping or positive drug tests, this might have an easier way out. There are lots of cases that could give a higher than normal reading without using illicit methods.

by ELVISGOAT on Jul 27, 2006 4:45 PM EDT reply actions 0 recs

That T/E steroid test beatable since mid 80's

"...By 1982, the International Olympic Council had developed a test for the detection of excess levels of testosterone in athletes, known as the "Testosterone: Epitestosterone test". In this test, levels of testosterone vs/ epitestosterone are measured, and if the testosterone level is 6x that of the epitestosterone level, it can safely be concluded that some form of testosterone has been used by the athlete. This is because testosterone is commonly no more than 6x the natural level of epitestosterone found naturally in the body. Thus, if there were more than that ratio, it was not naturally occurring, in all probability. The IOC was, as usual, one step behind the athletes. The GDR had already done a study on their athletes using a form of testosterone which would leave the body quickly, and thus they would be ready for the IOC test within three days of their last injection (6). They then developed a protocol to allow their athletes to continue steroid use, ceasing it only long enough to pass the drug test. In addition, the German firm Jenapharm, who had been supplying the government with steroids for their athletes, developed an epitestosterone product to administer to athletes to bring the ratio back to normal without discontinuing steroid use.(5)

Their doping methods were so advanced, however, that they remained undetected for many years, until late 1989 when information was leaked to the western media about a government sponsored program of systematic anabolic steroid administration and concealment....."

http://www.steroid.com/

Are you going to tell me that an athlete competing at the high level Landis did is going be allowed by his training staff to fail a drug test that has been easily beatable since the mid '80s?

I'm not saying Landis wouldn't cheat. I'm saying Landis would not cheat with a drug that was of little benefit to him in a mountain stage, and when there is such an easy way to cover the presence of testosterone in the bloodstream by raising epitestosterone levels.

I'd believe a cheating accusation at this level. I would not believe an accusation that Landis' high-priced training staff screwed up trying to cheat this badly.

by afs on Jul 27, 2006 5:23 PM EDT reply actions 0 recs

beating the test

I totally agree. There are tons of ways to try to beat the test: epitestosterone, hCG, probenecid and bromantan. Of course, these "cover-ups" are detectable too. But at this level of cycling, IF someone was helping a rider dope, I would think that they would at least TRY to cover it up.

I was unaware that the T/E ratio had been lowered to 4. Maybe they decided to figure Asians into the mix (Asians have a much lower normal T/E ratio.) Adding this group in could skew the norm and make it much lower (very unfair to non-asian riders).

PLEASE put a rush on the tests (and I hope Floyd's team demands every testosterone test known to man)!! Studies by INDEPENDENT labs please!! Let's have those lab results published. And I would like to see how Floyd's questionable test stacks up against Floyd's norm (he has been riding for a while--I am sure they have samples from years back--oh, wait--unless they lost or "froze" the old samples). How accurate are these "labs?" I want ANSWERS!! It will be a LONG 1-2 weeks.

by cyclewife on Jul 27, 2006 6:55 PM EDT up reply actions 0 recs

Issues in detecting abuse of anabolic steroids

Here's a theory regarding FL's reported "normal" T levels and "reduced" E levels resulting in the reportedly high T/E ratio of 11. http://www.npr.org/templates/story/story.php?storyId=5587194. I'm not an endocrinologist, but here goes. Probably won't have the "final" answer thru all the appeals for another 8-10 months:

* Chronic administration of testosterone (T) inhibits the body's natural production of T, epitestosterone (E) and luteinizing hormone (LH) as evidenced in urine samples.

* Short half-life formulations of T are available (in oral, gel, or patch form or by injection) which can produce serum concentrations of T within the reference range for normal, but can temporarily result in high urinary ratios of T/E. However, the ratios typically fall to < 6:1 within 4-6 hours (the typical length of a Tour stage).

* For the user who discontinues taking T, the T/E ratio reverts to baseline (typically ~ 1.0) within about 20 weeks. Additionally, male non-users of T have relatively stable T/E ratios, which fluctuate around a mean, with a CV that typically will not exceed +/- 30%. T/E ratios for T users typically fluctuate much more widely. For the extremely rare athlete with a naturally increased T/E ratio, the T/E remains chronically increased.

* When T/E >10:1, additional invasive and non-invasive test are recommended. Numerous such tests have been proposed and appear to be available, including carbon isotope ratio, and urinary T/LH ratio of > 30, a potentially more sensitive marker of steroid use than the urinary T/E ratio, which remains sensitive for twice as long as urinary T/E.

Side issues:

* The overall incidence of urinary T/E > 6 in the general population of healthy males not abusing steroids is rare, < 0.8%, as evaluated by Catlin and Hatton and confirmed by Dehennin ... Moreover, it is common for non-power athletes (e.g., distance runners, swimmers, tennis players, soccer players) to utilize physiological doses of T (i.e., TC 100 mg/week) to counter the catabolic effects of stress and exercise on muscle... http://www.clinchem.org/cgi/content/full/43/5/731

* Some rare cases of physiologically high T/E ratios (between 6 and 12) are encountered; these may be attributed to a dysregulation of the testicular secretions of epitestosterone. http://joe.endocrinology-journals.org/cgi/content/abstract/142/2/353

* Until further details are available, one may prudently consider that large and inebriating doses of ethanol (read beer!) may increase the T/E for several hours after ingestion and that some females may be particularly sensitive to the ethanol effect. Males showed no significant effect of ethanol on T/E at doses of 1.0-1.2 g/kg, whereas the same dose produced large increases in another study. At very high doses of ethanol (2 g/kg), the T/E ratios of males increased, although none exceeded 6:1. http://www.clinchem.org/cgi/content/full/43/7/1280

PS: You know you're a cyclist of you actually read this stuff! Additional links FYI:

Lower levels of Epitestosterone following Testosterone administration. http://www.clinchem.org/cgi/content/full/43/7/1280#R24
Issues in detecting abuse of xenobiotic anabolic steroids. http://www.clinchem.org/cgi/content/full/43/7/1280
Origin of physiologically high ratios of urinary testosterone to epitestosterone. http://joe.endocrinology-journals.org/cgi/content/abstract/142/2/353
T/E before and after administering ethanol. http://www.clinchem.org/cgi/content/full/43/7/1280
Issues in detecting abuse of xenobiotic anabolic steroids and testosterone by analysis of athletes' urine. http://www.clinchem.org/cgi/content/full/43/7/1280
Detection of anabolic steroid administration. . http://www.clinchem.org/cgi/content/full/43/5/731
Masking Drug Use. http://entertainment.howstuffworks.com/athletic-drug-test6.htm

by Rydr1 on Jul 27, 2006 8:55 PM EDT reply actions 0 recs