Innovative Anti Doping Tests

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Chris Gadsden

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Oct 28, 2019
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The authors claim that by focusing on differences between frozen vs. non-frozen RBCs, with their model, they can detect 100 % of autotransfusers (some for up to 5 weeks after the treatment) who use cryopreservation (-80 degrees C) with no false positives, when the control group has been sea level, high altitude and hypoxic chamber athletes.
That's fantastic. Maybe, just maybe the days of blood doping are drawing to a close.
 
This is breakthrough research, but unfortunately freezing blood has also lost its relative advantage when one can't infuse 4-5 bags collected in October-February during a GT but instead must use a handful of 200-300 ml microinfusions that will likely survive in a non-frozen form.

Interestingly the arguments how Malm et al dismiss the viability of standard +4 C storage is very similar to that of Michael Ashenden in 2002 when -- despite his arguments -- blood dopers were many in number and used mainly their refrigerated (+4 C) blood (e.g. Freiburg+almost all clients of Fuentes+USPS) with a few exceptions.

Wrong in 2002, but hopefully not wrong today.

 
A little bit of wishful thinking at the moment but maybe a few years ago some of us would have said DBS testing was wishful thinking too:

View: https://twitter.com/RobinParisotto1/status/1288431094950293510


Parisotto's Tweet refers to an article titled 'Wearable Electronic Sweat Sensor Detects Health Biomarkers' the key takeaway from which is:
Engineers at Tufts University have shared their research in developing a flexible electronic sensing patch that analyzes sweat for a variety of biological markers, including sodium, ammonium, pH and lactate, directly from sweat and in a band format that can be sewn on to clothing.
The science bit:
The sensor platform is built as a patch where threads are placed on to the fabric gauze of a commercial bandage, the gauze enabling sweat transport from the sensing area on the patch to the reverse side of the patch for evaporation. However, the gauze edges are sealad to prevent sweat evaporating from the sensing area.

Sweat is wicked at the sensor surface for real-time measurement, and the threads connect to wireless readout electronics that permit real-time data acquisition and collection for detection and sensing of the biomarkers.
You can see what this looks like here. Basically, it's a Band Aid.



Change the markers being tested for and you can see where this could go. If it gets over the hurdles still ahead.
The study provides proof of feasibility, but lacks strict standardized measurements and statistically validated data at this point. The authors nod to this in their discussion write-up, and suggest that this will be the next step for development of the sensor: "The sensors can be statistically validated for their effectiveness in real-time continuous measurements of multiplexed biomarkers in the context of underlying clinical trials in lieu of sparse standardized measurements."
You can get your geek on with the full paper here.
 
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Perhaps you could summarise it for us...
  • a very sensitive method with no false positives.
  • WADA has the method under scrutiny with the reports available, but a Swedish anti-doping agency executice is optimistic.
I read like 3-4 articles about the method in the past 24 hours and not one mentioned that it only detects frozen blood, not the refrigerated one -- the latter one being cheaper, more accessible and fully viable for guys who don't need like 5-10 BBs during a season.
 
Thanks. So the usual hype without insight. Funny that WADA aren't on the hype train with it as well, though. I mean, they were promising us that our good friend Yannis Pitsiladis's mythical ground-breaking blood doping breakthrough was to have been ready for the Tokyo Games, which should have started a week ago.
 
So the usual hype without insight. Funny that WADA aren't on the hype train with it as well...
In defence of Malm et al, it at least looks a very good method in what it is doing, actually very good.

And it is "fair" in the sense that it'll make the llfe of former blood freezer-dopers (with a lot of money) - who donated blood during the OFFseason - more miserable when they have to use the same methods as their poor competitors.

Had it been the other way (detecting only refrigerated blood), a part of me would've found the method ethically dubious from frair-play perspective.

...If you are interested in coauthoring a short CN-article on the issue (I can do my part for free), I can write and DM you a memo on the history/application of the both of methods.
 
A little bit of wishful thinking at the moment but maybe a few years ago some of us would have said DBS testing was wishful thinking too:

View: https://twitter.com/RobinParisotto1/status/1288431094950293510


Parisotto's Tweet refers to an article titled 'Wearable Electronic Sweat Sensor Detects Health Biomarkers' the key takeaway from which is:The science bit:You can see what this looks like here. Basically, it's a Band Aid.



Change the markers being tested for and you can see where this could go. If it gets over the hurdles still ahead.You can get your geek on with the full paper here.
I'm really struggling to see how this could have anti-doping applications. Measuring ammonium, sodium etc. concs. in sweat compared to measuring metabolites of PEDs is very, very different. Developing a real time sensor to do this via skin sampling is even more different. There has been methods developed in the past that may be more applicable, this for example:


But even this would need to be tested to see if the metabolites of interest were actually seen, which is a very big if.
 
But even this would need to be tested to see if the metabolites of interest were actually seen, which is a very big if.
The post itself is a big if and is meant to be taken as such. That said, playing hypotheticals, Parisotto said:
Some biological markers have been detected in sweat for years so presumably many drug metabolites which 'do their work' in muscle tissue can in theory be transported (and detected) in the sweat that ensues from the tissue. Of course the research has to be done.
WADA are not exactly supportive of looking at other matrices. This is from an ExCo meeting in 2019:
Dr. Rabin introduced the subject on the advantages and disadvantages of using doping control matrices different from urine and blood. He reminded that at present, only urine and blood were accepted matrices and that any other matrices could be used to support the results of an Adverse Analytical Finding but not to override it. WADA frequently received research proposals to use other matrices like saliva, hair, breath, sweat, and these were almost systematically not considered to be a priority. Examples included detection of clenbuterol in hair, perfluorocarbons in breath. Saliva was discussed by the HMRC in the past but did not have support due to the abundance of confounding factors. Sweat may be problematic due to contamination. Dried blood or urine spots were not considered to be alternative matrices since the biological fluids were the ones approved for doping control analysis. The questions posed to the HMRC was to whether WADA should give more consideration to alternative matrices, at least at the research level.

Several of the HMRC members gave their opinion. Prof. Handelsman believed there could be a place for the use of hair analysis. Prof. Ayotte mentioned that hair would only be useful for chronic administrations and if it was a matter of sensitivity, none of the other matrices would compare to urine or blood. Dr. Vernec noted it could be useful to distinguish the cases where a unique exposure was claimed. Dr. Barroso raised the fact that drugs stuck to hair differently depending on the colour and thickness. Dr. Wan noted that hair analysis is used in horse racing doping control, but even the follicle could become contaminated and there was no consensus on how to clean the hair. Dr. Pascual added that hair was always accessible to tampering. Prof. Ayotte proposed that WADA should convene a working group that should look into the published literature and recommend whether the results would be defendable in court. There was not much support for saliva: it could only be used for certain hormones but it could be easily contaminated with blood, and the collection required that the person didn’t eat for a certain period of time. Breath would be limited to a few substances and could be contaminated with saliva, which in turn could be contaminated with blood.
There have been some studies that looked at sweat, such as this, but the collection issue seems to be the first stumbling block.

Does this have the potential to replace blood and urine? No it does not, for the simple reason that there are fewer markers present in sweat (this paper lists them as " the opiates, buprenorphine, amphetamines, gamma hydroxybutyrates, cocaine, and cannabinoids"). Does that mean there is no potential application here should the Tufts people get this thing across the line? No. The UCI currently uses DBS for pre-race Tramadol testing. Depending on the markers present in sweat maybe it is possible that limited applications could present themselves.

All that said, I'm not betting any money on this developing into anything. It's a whole train of ifs.
 
The post itself is a big if and is meant to be taken as such. That said, playing hypotheticals, Parisotto said:WADA are not exactly supportive of looking at other matrices. This is from an ExCo meeting in 2019:There have been some studies that looked at sweat, such as this, but the collection issue seems to be the first stumbling block.

Does this have the potential to replace blood and urine? No it does not, for the simple reason that there are fewer markers present in sweat (this paper lists them as " the opiates, buprenorphine, amphetamines, gamma hydroxybutyrates, cocaine, and cannabinoids"). Does that mean there is no potential application here should the Tufts people get this thing across the line? No. The UCI currently uses DBS for pre-race Tramadol testing. Depending on the markers present in sweat maybe it is possible that limited applications could present themselves.

All that said, I'm not betting any money on this developing into anything. It's a whole train of ifs.
The post itself is a big if and is meant to be taken as such. That said, playing hypotheticals, Parisotto said:WADA are not exactly supportive of looking at other matrices. This is from an ExCo meeting in 2019:There have been some studies that looked at sweat, such as this, but the collection issue seems to be the first stumbling block.

Does this have the potential to replace blood and urine? No it does not, for the simple reason that there are fewer markers present in sweat (this paper lists them as " the opiates, buprenorphine, amphetamines, gamma hydroxybutyrates, cocaine, and cannabinoids"). Does that mean there is no potential application here should the Tufts people get this thing across the line? No. The UCI currently uses DBS for pre-race Tramadol testing. Depending on the markers present in sweat maybe it is possible that limited applications could present themselves.

All that said, I'm not betting any money on this developing into anything. It's a whole train of ifs.
Both papers you link are reviews, which is a bit annoying as it means reading all the references. Taking the second paper, that one also references another review in the sweat analysis section and one paper that looked at detecting the drugs you list in both saliva and sweat. The results are very mixed and they conclude in the abstract that saliva would be useful. They can't detect most of the drugs in sweat it seems (it's late so I've not signed in to get the full paper). The Tufts device wouldn't work to detect these things. Sweat or saliva might be useful, but it's going to be through sampling and analysing with something like the patch I linked to.
 
Not any anti-doping test but innovative nonetheless (if it works): Pitchside saliva tests could be used to diagnose concussion
“For the first time we have successfully identified that these specific salivary biomarkers can be used to indicate if a player has been concussed,” Professor Antonio Belli, the report’s senior author, said. “We now have a laboratory-based, non-invasive diagnostic test using saliva, which is a real gamechanger, and provides an invaluable tool to help clinicians diagnose concussion more consistently and accurately.

“The test could be used not only in sport, from grassroots to professional levels, but also in healthcare and military settings. In community sports, these biomarkers may provide a diagnostic test that is comparable in accuracy to the level of assessment available in a professional sports setting.”
 
Currently, testing labs analyze samples using tandem mass spectrometry (MS) and gas or liquid chromatography. These approaches break up molecules in the sample and separate the fragments, yielding spectra that can reveal the identity of the original, intact compounds. But it can be tough to differentiate molecules with minor structural differences -- including isomers -- that distinguish endogenous steroids from exogenous ones, such as the synthetic anabolic steroids athletes take to build muscle.

To accentuate those differences, Chouinard pairs MS with ion mobility (IM) spectrometry, a separation technique he learned as a graduate student with Richard Yost, Ph.D., at the University of Florida. Yost's team and others found that the differences between isomers could be made even more apparent if the molecules in a sample were modified prior to IM-mass spec analysis by reacting them with other compounds. After Chouinard set up his own lab in 2018, he applied this technique by reacting steroid samples with ozone or acetone in the presence of ultraviolet light -- reactions already well-established among researchers who study lipid isomers, but new in the anti-doping arena.
source
 
This one, depending on instrument resolution etc. would work pretty well. IM-MS instruments have been around for a while and honestly, I kind of assumed this was being done already.
Lacking a science background this one confused me because - similar to your own reaction - it sounds like something that's been around for yonks.

I wasn't sure if this wasn't just Olympic year puffery. (I'm holding my breath waiting for the quadrennial round of gene-doping stories.)
 
Lacking a science background this one confused me because - similar to your own reaction - it sounds like something that's been around for yonks.

I wasn't sure if this wasn't just Olympic year puffery. (I'm holding my breath waiting for the quadrennial round of gene-doping stories.)
As a standalone technique, IMS has been around since the 50s/60s and gets used for things like chemical weapons detection on battlefields and explosives and narcotics detection at airports. If you've ever had luggage swabbed going through security then it's highly like it was analysed via an IMS system ( probably made by Smiths Detection). It works at ambient pressure and the devices can be made to be really robust. There's one on the ISS I was once told a cool story about.

Paired with mass spectrometry it's been around since, I think, the 90s as a useable research tool and found its way into vendor instruments in the 2000s. They're pretty much standard in most big pharma research labs these days and they're also being turned towards healthcare for disease detection.

They work by separating things out based on their collisional cross-section. The theory is that small conformational changes that don't really affect chromatography and are hard to detect via mass spectrometry, can be separated using IM, that's where the line about endogenous vs exogenous comes in your quote. Usually it's placed between the chromatography column and mass spectrometer (although it can be used as a detector in it's right it would be very unlikely in this case). In this case the article describes reacting the compounds with something, this is done to increase the difference in collisional cross-sections between the analytes, so lower resolution instruments can still separate them.
 
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