Asli Cakir Alptekin of Turkey crosses the finish line during the Women's 1500m Final of the London 2012 Olympic Games

Anti-doping technology to catch the Olympic cheats

Sportsmen and women are not the only ones training hard to get good results in Rio de Janeiro later this year.

In August 2015, Asli Cakir Alptekin of Turkey had to hand back the 1500 metres gold medal she won at the 2012 Olympic Games in London after admitting to blood doping - one of the latest examples of drug-related cheating in sports, a problem that had grown enormously since 1968, when the Swedish pentathlete Hans-Gunnar Liljenwall became the first athlete to be disqualified from the Olympics for a doping offence. He had drunk a few beers to calm his nerves, which was banned under new regulations set by the International Olympic Committee (IOC).

In 2016, when the Games return to Latin America for the first time since those early days of drug control, things will have moved on dramatically in the fight against doping.

With every subsequent edition of the quadrennial competition, more substances have been banned, more athletes have been selected for screening, more sports federations have officially adhered to global anti-doping regulation and more sophisticated performance-enhancing drugs and doping methods have been developed.

The technology designed to expose the cheats has become more advanced, too, with the result that more are being caught. In just one example, Russia as a nation was banned from international track-and-field events in November 2015 for systematic doping.

For every new drug or doping scheme that becomes available, a new test technology or procedure has to be developed, validated and implemented by the World Anti-Doping Agency (WADA), the independent organisation set up in 1999 to standardise and coordinate international policy initiatives in the battle for clean sport.

While the quantity of the job at hand might look as arduous as that done by a software antivirus company that has to chase several new threats a week, the quality of the results produced by the latest biochemical technologies is growing exponentially.

This means that the detection of prohibited drugs, hormones and metabolic modulators at the Rio Olympics will be possible for both lower concentrations and longer time windows than in London 2012.

The Rio anti-doping lab

Every time an athlete competing anywhere in the world is singled out for an anti-doping test, the samples collected (blood or urine) can only be analysed at a WADA-accredited laboratory. There are currently only 34 such laboratories worldwide (there used to be 35 until last November, when the accreditation of the Laboratory Anti-doping Centre Moscow was indefinitely suspended following the ongoing Russian athletics doping scandal).

In Latin America, there are four WADA-accredited labs located in Bogota (Colombia), Habana (Cuba), Mexico City (Mexico) and Rio de Janeiro (Brazil). The latter, the Brazilian Doping Control Laboratory (LBCD), will be in charge of screening the tens of thousands of tests carried out this summer before and during the Games.

The lab is co-located at the Federal University of Rio de Janeiro’s Chemical Institute. Francisco Radler de Aquino Neto works full-time at the building as both head of the LBCD and professor at the university’s Chemical Institute. He says that during the Games he and his team will be mapping over 500 substances in a constant stream of athlete samples arriving at the lab.

In 1968, poor old Liljenwall could have easily argued he wasn’t aware that alcohol consumption was not allowed in his sport and people would not have had much trouble believing him. Today, every young kid practising any sport in a local, regional or national federation is made aware from a very early stage of the need to be familiar with ‘The Prohibited List’, a document updated and published every 1 January by WADA that - as its name implies - leaves nothing to the imagination.

The Prohibited List

‘The List’, as it is known in the world of sport, is the most visited document on WADA’s website. It specifies the names of approximately 200 substances and doping methods banned by the global anti-doping agency. That number prompts a question, though: Why will the Rio anti-doping lab be looking to detect over 500 substances when the 2016 version of The List details fewer than half that number?

“The challenge is that not every substance that is prohibited is specifically listed by WADA,” answers Matt Fedoruk, science director at the US Anti-Doping Agency (USADA, the organisation that in 2012 famously charged former cyclist Lance Armstrong with an anti-doping rule violation). “The labs are responsible for detecting both the listed substances and substances that are pharmacologically or structurally related to the listed substances and their metabolites.”

To help them do just that, accredited labs must keep up with the latest developments in screening instrumentation. Anthony Butch, professor of pathology and laboratory medicine at the UCLA School of Medicine and director of the UCLA Olympic Analytical Laboratory (the world’s largest WADA-accredited lab), says there are certain compounds that have been on the Prohibited List for a number of years, but only now are methods being developed to actually detect them.

“There’s no doubt that the Brazilian lab will be using extremely sensitive instrumentation,” he says. “They will be using gas chromatography triple quads (quadrupole mass spectrometry systems) instead of GC single quad systems, which gives you another layer of sensitivity. We were already identifying a lot of anabolic steroids with the GC single quad. Now we are using a GC triple quad, which is kind of MS/MS [tandem mass spectrometry, a method for structure determination and analysis of complex molecules].

“Sensitivity is much better because you’re looking at a product that comes from the precursor molecule, so your background noise is much lower and it improves your specificity,” Prof Butch explains. “You can detect probably half the concentration that you can do with some of the single-quad instruments. That’s technology that they will have in Rio because they’re buying the best of the best for the Games.”

USADA’s Fedoruk also highlights the constant evolution that mass spectrometry equipment has witnessed over the years and how this has helped anti-doping labs to detect ever-lower levels of substances. “Ten years ago a laboratory maybe was able to detect nanogram-per-millilitre amounts of substances,” he says, “whereas now they are down to detecting picogram levels.”

A forensic approach

Just as important as the increasing sensitivity of the instruments employed at the labs is the development of novel methodological approaches to sample testing and the conduction of research on new drugs, supplements and potential performance-enhancing techniques.

One of the conditions of being a WADA-accredited laboratory is that the lab must invest in research continuously. “A laboratory investing in a certain area of research will typically begin to work on a new method of analysis, which will then be developed and its effectiveness validated,” says the head of the Brazilian anti-doping lab, Professor Radler.

“After that, some time will pass before the rest of the labs are able to begin using the new methodology. For the Olympic Games, the IOC takes a number of initiatives that have not yet been disseminated, but it wishes that the Olympic laboratory begins to implement during the event. We already know of at least five procedures that are not routinely observed by all the WADA-accredited labs which will be implemented in our laboratory for the Games,” he reveals.

It is this relentless push for cutting-edge research that by the mid-2000s helped set in motion what is now the most significant change of paradigm in the fight that biochemical scientists are bringing to doping athletes.

Up until that point, the entire doping-detection movement relied exclusively on the use of toxicological testing. In other words, the use of screening techniques designed to detect specific drugs that had been determined to enhance performance.

Yet experience proved that there was a major shortcoming with that approach. It was discovered that some drugs (particularly the most powerful, such as erythropoietin, commonly known as EPO) were capable of boosting performance in endurance sports for several weeks after administration, yet they only remained in the human body for less than 48 hours.

This created a situation where the only way to get infringing athletes to test positive was by collecting samples literally a few hours after the drug had been administered.

Cheating cyclists, boxers, rowers, marathon runners, mixed martial art fighters, even racehorses, had suddenly found a way to artificially increase the supply of oxygen to their muscles, providing them with an unfair advantage over clean athletes whose muscles would tire earlier in training and competition. And they were all getting away with it.

“So what happened, and this is a really exciting development, is that WADA and the authorities said: Let’s not only go after the drug, but let’s go after the trace that the drug leaves behind,” says Yannis Pitsiladis, professor of sport and exercise science at the University of Brighton, director of the FIMS Reference Collaborating Centres of Sports Medicine for Anti-Doping Research and a member of the IOC’s Medical and Scientific Commission.

He is referring to the introduction of biomarkers to the anti-doping movement, which ultimately led to the establishment of the Athlete Biological Passport (ABP) in 2008. The ABP is a monitoring programme that forces athletes to regularly report for testing so that, over time, an individual longitudinal profile of each athlete is built where fluctuations between biomarker data points may suggest the use of doping.

“We’re moving towards an era that is more forensic-based rather than just pure toxicology-based,” says USADA’s Fedoruk. “In urine and blood samples there are substances that our bodies produce naturally, but that can change with the use of doping products. Those changes last a longer period of time than the prohibited substances themselves when we look at them in the body.”

Growth hormone (GH) biomarkers were applied for the first time in Olympic anti-doping tests in London 2012. For the upcoming Games in Rio, newly developed GH biomarkers will be added to the battery of tests. “Rather than looking for the recombinant growth hormone itself, we are now looking at two biomarkers, IGF-1 and P3MP, which allows us to look at the effects of GH for a much longer period of time,” says Fedoruk.

As part of the efforts to discover more indirect biomarkers of doping, some labs have focused on the identification of metabolites of banned drugs. Recent research around longer-term metabolites of stanozolol, for example, has allowed anti-doping agencies to extend the window of detection for the use of this fairly common anabolic steroid from hours to weeks.

From his prestigious laboratory in Los Angeles, Professor Butch gives me another example of a relatively new biomarker technology. It was developed a couple of years ago by his team. “It’s a method to determine doping with HCG (human chorionic gonadotropin), where we can speciate the different isoforms of HCG to detect doping. The Brazilian anti-doping lab has been in contact with us, and that is going to be available in Rio as well,” he confirms.

Forty-eight years after exposing the first Olympic doping cheat, the Summer Games return to Latin American soil armed with a long list of technological weapons that should be able to detect slightly more sophisticated doping schemes than having a couple of beers ahead of the action.

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