[rational head]

Removed for revision.

[shag]

Nice job, RH. I'll resume banging the drum I was banging yesterday: where in the process you've laid out comes testing/confirmation that the sample is not contamined? (I promise I'll stop harping on this soon.)

[rational head]

Nice job, RH. I'll resume banging the drum I was banging yesterday: where in the process you've laid out comes testing/confirmation that the sample is not contamined? (I promise I'll stop harping on this soon.)

Look above in step 1. Sample integrity. That's the first time they check for pH - before trucking it to LNDD. Then, each time they open an A or B sample after thawing it, they would check for pH to stay under 7. Also, during one (of the total three) confirmation tests for both A- and B-samples they estimated free Epitestosterone and conjugated Epitestosterone. The ratio was under 5% ( as supposed to) for A-sample but not for B-sample. As I analyzed in the other topic, WADA does not define EXACTLY
how a lab should treat contamination. That's possibly why LNDD did not consider B-sample contaminated. That's my reading.

[Cal]

RH,

Thanks for the outline. I realize I have been looking for a more definitive answer to the scientific back and forth. I probably need to be more patient. From your examination, were the tests done according to International Standards and WADA protocol? Are there any glaring mistakes? You seem to answer these questions by stating the crux of the debate is over 1 vs. 4 metabolites needing to test positive.

Other posts suggest the lab may not have read the T/E ratio correctly in the CIR (post #284 by duckstrap in the Powerpoint Analysis thread). It has also been raised, with very little exploration, "Why did the tests not show FL positive for synthetic cortisone?" I suppose I am waiting for these questions to be further answered and waiting for the scientist's to be at a point where they can say, "The lab is accurate in its assessment or the lab is not accurate." Maybe this will never be possible.

Cal

[rational head]

RH,

Thanks for the outline. I realize I have been looking for a more definitive answer to the scientific back and forth. I probably need to be more patient. From your examination, were the tests done according to International Standards and WADA protocol? Are there any glaring mistakes? You seem to answer these questions by stating the crux of the debate is over 1 vs. 4 metabolites needing to test positive.

Other posts suggest the lab may not have read the T/E ratio correctly in the CIR (post #284 by duckstrap in the Powerpoint Analysis thread). It has also been raised, with very little exploration, "Why did the tests not show FL positive for synthetic cortisone?" I suppose I am waiting for these questions to be further answered and waiting for the scientist's to be at a point where they can say, "The lab is accurate in its assessment or the lab is not accurate." Maybe this will never be possible.

Cal

Cal, I did not put any right/wrong evaluations on my description. Nor did I outline all outstanding issues. That's not the purpose of me starting the thread because there are OTHER threads dealing with it.

My purpose was to provide an easy guidance/platform for understanding of what was supposed to be done vs what was actually done - a sort of an outline of a BIGGER PICTURE.

My thought was that if members understood the WHOLE process/chain in terms of expectation vs. what actually happened it would be easier to follow other technical issues under other threads.

A backgrounder.

Edit: I also thought that understanding how those analyzers work (step 10) would be helpful for everyone
in terms of following all the inaccuracies brought up by many sceptics of the test results.

[Pelotonium]

10. How does GC-MS- work, please keep it short and simple!!?

Try looking at this website and following along with RH's excellent description - it worked for me:
Gas Chromatography Mass Spectrometer

In fact, looking at the mass spec part of it, I'd say it works a lot like a cathode-ray tv tube. It's just a television for nerdy chemists!

Just goes to show that you learn a lot just reading The Daily Peloton Forums ™. Vaughn, you can mail the cheque to my usual account. ;-)

Andy

This post has been edited by Pelotonium: 18 October 2006 - 03:41 PM

[12string]

Rational Head,
Thanks for explaining the technical steps to testing. Is there a part of this testing procedure that is characterized by 'interpretation' of test results, similar to the way a radiologist interprets x-rays?

If there is such a step, does it have the same susceptability to bias that x-rays have? I know from friends that interpretations of a suspicious mammogram can run the gamut from 'no problem' to 'need a biopsy'.
Thanks again,
Art

[Steve in ATL]

I'm going to expand point 1, from an athlete's perspective.

1. What happened first?

a. Soon after (whoever wins whatever finishes) s/he peed in a bottle. It was split into 2 samples, A and B, each at least 20 mL (remember this, dummies, ).

a1). The first thing that happens is that you get a WADA (in my case, USADA) escort, who notifies you that you are to be tested, goes over a form that explains why and how you are being tested, and what the consequences of various actions (such as fleeing the facility before testing) are. The escort stays with you until you report for testing. You have 1 hour after notification to report, and they allow extra time for awards ceremonies, etc. You sign, date, and the escort time-stamps the form.

a2) Once you report to the testing station, you may not leave. You are allowed to have 1 witness / representative with you. No cameras, cell phones, recording devices are allowed in the sample collection area.

a3) Drinks in sealed containers are kept in the collection area to help the process along.

a4) -ADA representatives are not allowed to hand you anything (other than paperwork). You have to get your own drinks.

a5) You select a pee cup from a container of sealed pee-cups. This makes the selection random, and you have to then verify to the sample collection witness that the container is sealed and has not been tampered with.

a6) The -ADA representative must watch you pee in the cup. The whole process must be witnessed. You must be bare from knees to sternum so that you cannot fake the sample.

a7) After peeing in the cup, you must seal it yourself, and then proceed to the sample processing area (after having dressed again, of course). Again, the -ADA representatives cannot handle your sample or hand you anything.

a8) In the sample analysis area, the athlete must choose, again, at random, a tamper-proof sample analysis kit consisting of a styrofoam container, two sample jars with tamper-proof lids (the jars are glass, and have a screw-on, ratcheting top that is spring-loaded so that you have to physically break the top in order to unscrew it). The kit is sealed (which you have to verify) and numbered (this is the famous "Armstrong Number" that L'Equipe used to track down who's sample was who's). The number is recorded on a form, along with information on what drugs you may have taken in the last 3-5 days. We'll call this the Test Form for the rest of this.

a9) The athlete divides his own urine into the two containers. The containers are then sealed by the athlete. The athlete is required to check to make sure that the containers are in fact sealed, unable to be opened, and are not leaking (NPI).

a10) In the sample analysis area, in front of you and your witness, the sample's pH is checked. There was one other check they did also - specific gravity, if memory serves. This testing is done, if I recall, on leftover urine, but it may not have been - my memory is fuzzy. If they did it prior to sample division, they separated out a small quantity of urine ot perform the test on. This is recorded on the test form.

a11) The athlete places the samples into the sample container, and it is sealed. the sample container is then handed over to the -ADA representative. This is the first time that the -ADA handles the sample.

a12) The test form is completed and signed. The athlete gets a copy, the -ADA representative hold the copy with the name and Sample ID number, and the lab gets the copy that has only the sample ID number and the drug information.

a13) The athlete is then advised on turn-time of the testing (4-6 weeks, in my case) and is then free to leave.

On a personal note, I will say that all of the USADA representatives were polite, professional and friendly. Several of the congratulated me quite warmly. It was a very different experience than I thought it would be.



b. Sample integrity was checked BEFORE transporting it.

b1) That's the process (pH / Specific gravity) that I detailed above.


c. They were sealed and shipped to LNDD, (at minus 5 C temperature)

c1) Again, the sealing part is detailed above. I did not see how they stored the sample after I turned it over, but again, the sample container was a heavy (and faily large - 3" x 7" x 7") styrofoam box.

[rational head]

Rational Head,
Thanks for explaining the technical steps to testing. Is there a part of this testing procedure that is characterized by 'interpretation' of test results, similar to the way a radiologist interprets x-rays?

If there is such a step, does it have the same susceptability to bias that x-rays have? I know from friends that interpretations of a suspicious mammogram can run the gamut from 'no problem' to 'need a biopsy'.
Thanks again,
Art

You're welcome, Art.

I am quite familiar with the GC-MS technology but I don't make my living interpreting it every day. There are few members of this forum who appear to have used the instrumentation on a daily basis.

With all these disclaimers in place, I'll answer your question directly. All the stages of Floyd's testing were subject to human interpretation. But in a much different way than a radiologist does. A radiologist looks at an IMAGE or a PICTURE. Interpretation of a simple black-and-white X-ray (you mentioned) requires an experienced reader to carefully compare organ sizes and proportions to those he/she expects to be normal (physiologically speaking). It's an Art based on experience. There are some clear criteria in place on how to read properly but it's still an art. Interpreting an MRI image or a CAT scan image (also a branch of radiography) is arguably less subjective as there is powerful help from computer-generated color-rich images allowing closer look into organ - including full 3-D reconstructions.

Unlike a radiologist any spectrometrist interprets a set of printouts, graphs and tables. His job is closer to a mathematician's skill-set. For example, to identify testosterone or it's metabolites he'll employ his skill of
expecting steep peaks in certain position on a chromatogram (or a spectrogram). These peaks are characteristic of the ionized "whole" T-molecule (called a "dominant diagnostic ion") or its fragments. It's a relatively straight forward job to identify low molecular weight proteins. There are tables and libraries to assist with the task. The computer library contains thousands upon thousands of pre-programmed well identified molecular signatures. They alone can give away most known proteins. But a skillful technician is still very important. A lab (and WADA) requires to have the result verified (and signed) by another tech. Mistakes happen when an unexpected molecular fragment appears or when sensitivity is limited or when
calibration or other instrument performance issues interfere. You may see a thread where RJBora dissected
LNDD plots. It's important to double check.

Still, in my experience, these tests are less prone to human error than it may seem. Particularly when
another human checked the results and the procedure was validated.

I hope I answered your question.
------------
EDIT: I just checked my writeup for spelling errors and in stead noticed that I am a dummy . I should have mentioned in the appropriate places above, that a simple X-ray could be a good analogy for a T/E screening test - quick, inexpensive and accurate enough. If it finds some abnormal "shape" a doc would order an MRI - more accurate but more specialized and expensive. Not much unlike an IRMS test triggered by an abnormal T/E test. Screen for T/E cost (in LA) $150 (along with "cost-free" 200 other markers). IRMS cost $300 and takes twice as long. The machine is around $100,000 (without the GC portion and the "combustor"/interface). Only half of WADA labs have it.

[lakeArrowheadrider]

According to WADA one or more of these CIRs must exceed a specific criteria for an athlete to be declared positive. Floyd argues that ALL FOUR must be over the limit. This is the major point of the case.

Thanks for the great explanation Rational head. Please keep the information flowing for us layman. It really helps and someone may need to write an edition of "Sport Drug Testing for Dummies", to be available at most book stores.

I thought the WADA code was ambiguous with regards to the specific criteria for the a CIR positive. You state WADA says one or more? Has new information araised that I have missed?

[Steve in ATL]

in reference to the above quote, anybody ever get performance anxiety?

One of the other riders who was tested took quite a while to produce - something like 2 hours. The mind does funny things - no one gets that dehydrated track racing!

[MacRoadie]

One of the other riders who was tested took quite a while to produce - something like 2 hours. The mind does funny things - no one gets that dehydrated track racing!

Funny, whenever I'm at the ADT Center, I have to pee all the time. Probably something to do with the ridiculously long walk to the can....

This post has been edited by MacRoadie: 19 October 2006 - 06:29 PM

[rational head]

OMJ, thank you for your answer.

is there any way to distinguish these metabolites once the graph is run or is it just guessing at that point?

If these metabolites are close enough to be shoulders and included in the calculations, how does one quantify the t vs the other, and on any of these graphs, does it look like enough to change a test to close to the borderline of positive or over to negative?

At what point is scientific certainly lost in the test is there is evidence that coelution is happening?

I am sorry guys but this is a science for dummies thread!

Jr, you are asking good questions but I think they are technical enough to be asked in the appropriate thread.
You have 3-4 experts there dissecting every aspect of the graphs in that thread.

http://www.dailypelotonforums.com/main/ind...?showtopic=1530

So, I am asking you (and all like you) to keep THIS thread free of complicated technical discussions because there is a very clear interest expressed here to keep things at a basic level..

If you don't hear me, I'll exercise my moderating privilege and transfer posts like that into appropriate places.

Thank you.

[rational head]

I am moving Jr's questions and OMJ and Duck's answers to this more specific thread.

http://www.dailypelotonforums.com/main/ind...?showtopic=1530

Thanks Jr for your questions and your appreciation of my request.
I also appreciate OMG and Duck's answers to Jr. I PMed all of you with more explanations.
If there is a problem let me know.

Edit:[b] Jr, I just took a shot at your questions in "slides" thread (along with Duck and OMJ) with some links to GC-MS that are slightly above beginner's level. Hope it helps!

[Dumas]

According to WADA one or more of these CIRs must exceed a specific criteria for an athlete to be declared positive. Floyd argues that ALL FOUR must be over the limit. This is the major point of the case.

EDIT: spelling

The WADA Technical Document TD2004EAAS isn´t that clear about it:
“The results will be reported as consistent with the administration of a steroid when the ¹³C/¹²C value measured for the metabolite(s) differs significantly i.e. by 3 delta units or more from that of the urinary reference steroid chosen. In some Samples, the measure of the ¹³C/¹²C value of the urinary reference steroid(s) may not be possible due to their low concentration. The results of such analysis will be reported as “inconclusive” unless the ratio measured for the metabolite(s) is below -28‰ based on non-derivatized steroid.”

In Floyds case no metabolite reach the criteria -28o/oo. Floyds values are, depending on the reference from different researchers, very close or in the reference population.
The term "Metabolite(s)" is not explicit and in doubt, you should use it in the favour of the athlete.

[lakeArrowheadrider]

According to WADA one or more of these CIRs must exceed a specific criteria for an athlete to be declared positive. Floyd argues that ALL FOUR must be over the limit. This is the major point of the case.

Rational Head:

I am sorry to repeat this question. I am curious/confused. Has the WADA code now been confirmed as "one or more" for a positive CIR? Is that because in that code WADA refers to them as Metabolite(s)? This is a huge part of Floyds case. The way I read your post is "one" being over is a positive.

Thanks

[one-mint-julich]

from cyclingnews:

The Landis argument: The Landis team argues that the WADA procedure is vague as to whether all metabolites must be out of range, or just one metabolite. The Chantenay-Malabry lab uses the standard that any one of four metabolites coming up 'significantly different' (ie. positive) than the negative control will confirm the positive result. Landis lawyer, Howard Jacobs, said that the UCLA Olympic Analytical Lab which tests for the US Anti-Doping Agency, checks only two metabolites and requires that both metabolites be positive. Dr. Don Catlin, director of the UCLA lab refused to comment on the issue.

Dr. Christiane Ayotte, the director of Canada's doping control laboratory, said that the WADA technical document is very clear on the question, and in drafting the document, 'metabolite(s)' was used to concisely state that if one or more metabolites is abnormal, this confirms exogenous testosterone. "We never imagined that this would be taken in any other way" said Ayotte.

The analysis: The Chantenay-Malabry lab and the INRS in Quebec both agree that any one metabolite showing a significant difference from the negative control indicates a positive test. Dr. Ayotte said that depending on the time the sample is taken after administration of testosterone, the profile of metabolites showing altered carbon isotope ratios will vary, and claims that the science clearly shows that any one metabolite is sufficient to demonstrate exogenous testosterone. If there are varying interpretations of the technical document among the WADA labs, or different procedures in place for exogenous testosterone analysis, this may be enough of a technicality to call the adverse analytical finding into question.

D, thanks for the article. Unfortunately, I couldn't open it.

[rational head]

We had a lot of discussion on the legal/technical interpretation of the pertinent WADA document in these threads.
http://www.dailypelotonforums.com/main/ind...?showtopic=1525
http://www.dailypelotonforums.com/main/ind...?showtopic=1538
http://www.dailypelotonforums.com/main/ind...?showtopic=1530
http://www.dailypelotonforums.com/main/ind...?showtopic=1569
It says black-on-white "one or more parameters of the urinary steroid profile ", page 1 of 11. So, if you see only "more" and another fan sees only "one", it does not change the text. It's totally appropriate to discuss what is "is" but let's do it in the right places. We've opened dozens of specific threads on every issue related to Floyd's case.

Anyway the discussion here is not about WADA clarity but about what is the technical essence of the case in laymen's terms. I highlighted the number of metabolites NOT to promote WADA, or Floyd's position but to pin up what may become the main issue.

So, tomorrow I'll add more background to the question of WHY it's important to account for testosterone metabolites.

I'll try to keep my explanations "non-partisan", because it helps more members to get their own opinion.

[rational head]

Rational Head:

I am sorry to repeat this question. I am curious/confused. Has the WADA code now been confirmed as "one or more" for a positive CIR? Is that because in that code WADA refers to them as Metabolite(s)? This is a huge part of Floyds case. The way I read your post is "one" being over is a positive.

Thanks

Lake... I NEVER meant to focus on the number or a count of metabolites in this thread. It's irrelevant untill you and others understand the significance of the count. I posted about it few times. Arguments can go on (and will go on) for years after the case is closed either way because it's just a question of how one WANTS to read and see the text. Let's get off word parceling and continue with the background technical discussion. I will post my opinion eventually but it's as irrelevant as any of us here.

[cycling newbee]

According to WADA one or more of these CIRs must exceed a specific criteria for an athlete to be declared positive. Floyd argues that ALL FOUR must be over the limit. This is the major point of the case.

This has probably been addressed elsewhere so I apologize in advance if that is the case. However, because I don't know where to find the answer in all the many posts and threads, could you provide the link to the exact rule (or cite the exact rule) (or steer me to the thread and post number to find the answer if you happen to have that handy)?

I was just now able to go through the "dummy" threads started by Jimmy and Rational Head. I again would like to take the opportunity to thank each of the "scienctists" for their insight and time in breaking it down further for us. Props to you all.


EDIT: I just reviewed Dumas's post above a second time. Is that all that is said or is there other language more in line with that which I quoted above from RH's post?

This post has been edited by cycling newbee: 23 October 2006 - 04:06 AM