Inside a Sports Drug Testing Lab

Now that the summer Olympics in London have come to a close, most assume performance-enhancing drug ("PED") testing has as well. However, PED laboratories test year round--during competition and off-season training--employing multiple technologies to keep athletes in check and sports drug free. To this end, tests are done routinely during the year to establish a baseline level for each subject, and to guarantee that all potential Olympians are tested at least once before competition.

In 2010, the last year for which there is officially published data, WADA--the World Anti-Doping Agency--executed testing on 180,584 urine and blood serum samples taken from athletes in Olympic sports alone (compared to 77,683 in non-Olympic sports). Peak sampling occurred in 2008, an Olympic year, with 202,067 individual tests. To handle that volume, WADA has accredited 35 established testing labs spread among 32 countries worldwide. These 35 facilities share the burden of proving whether an athlete has illegally enhance his or her performance using any of WADA's prohibited substances. In fact, every year the organization must update its banned-compound list to keep pace with contemporary use. Now, it is undeniable that performance-enhancing drug testing has evolved into a fully global pharmacology network.

Part of the impetus for such a vast net is the proliferation of prohibited substances use across multiple sports.

"We test for over 250 [prohibited] compounds," said Dr. Anthony Butch, director of the official WADA lab located at UCLA.

To illustrate the challenges of detecting one of so many illicit substances, Butch described the path of one subject's urine sample through his lab. In this example, he explained the processes involved in testing for testosterone and some 60 similar substances classified under anabolic agents on WADA's prohibited list. The complex procedure for one athlete's urine sample--from receiving, signing, unsealing, separating, testing and diagnosis--starts with the mail.

To begin, there is the knock on the door from the delivery service, he said, signaling samples have arrived.

"The first thing we look for is signs of leakage," Butch said. "Each box is sealed with tape, every athlete has A and B samples, and each vial inside has a cap or tape that must be broken once to get in."

Here begins the "chain of command," a sign-in process to document each stage in handling the vials. A laboratory employee signs for the package, another must sign when the seal is broken, and so on throughout every step. This is key because allegations of improper handling can affect the outcome of an athlete's appeal.

"All the appeals I've been to," Butch recounted, "[athletes] debate the chain of command. The science is pretty hard to argue."

After the urine samples are taken and logged, they are portioned into "aliquots," or, "little vials--about 20 for each A and B sample," said Butch. These 20 or so aliquots provide the amount needed for the many sub-categories (such as the 60-plus anabolic agents) of the prohibited substances to be detected. Aliquots are kept in a lock-and-key refrigerator until ready for centrifuge.

"Then, we look at color and pH," Butch described. "If [the urine] is clear, it's probably water. If it's purple, that's not urine. If the pH level is too close to 1.000, that's mostly water."

Following this are the gas and liquid chromatography phases, where individual compounds may be detected. This is a process of separating aliquots into their different components by exposing them to increasing gradients of high heat. First, Butch and his team pass small amounts of urine through a straw. "The garbage passes," he said, "and the steroids stick." Then, they wash the fine tube with solvents to concentrate samples.

The reason to use both gas and liquid chromatography in anabolic screening is because each of the more than 60 possible agents in that sub-category alone reacts uniquely to heat. In either gas or liquid state, the residues are placed in ultra-thin chambers with internal diameters of 0.25 mm wide and gradually subjected to heat in stages from 180 degrees Celsius, then to 230, to 270, and last to 300. At each level, a different compound will stick, then release.

"Those release times depend on the specific gravity [or, exact molecular mass]," Butch said. "We know from that if we have a positive sample."

For Dr. Butch, that's a standard urinalysis that he and his team can accurately complete within 48 hours. Accuracy is paramount, as testing stakes are incredibly high for athletes. One positive test could cause them to lose their Olympic eligibility, their medals, or face a suspension or ban from the sport.

"This is an Olympic athlete's career," said Dr. Butch, "You miss one 'Games' and you could be done."

Such precise testing is done in a place designed for such exacting work. Butch's officially certified WADA facility occupies 20,000 square feet just off the UCLA campus and employs 50 full-time medical professionals. In fact, it is the largest WADA-accredited PED lab on the globe. The United States is one of only three countries to feature two licensed centers. (The others are Germany and Portugal.)

"We test more than anyone else in the world," Butch said, "which is about 50,000 urine samples a year, versus 19,000 in the next highest lab."

That second-busiest processing facility for PED testing is in Salt Lake City, Utah. Dr. Daniel Eichner, a self-described friend of and collaborator with Butch, has run the Salt Lake facility for more than a year, having worked previously for both the Australian and U.S. Anti-Doping Agencies. Eichner said the importance to WADA of operating the second lab in the U.S. is for redundancy, "in the case the L.A. lab couldn't operate," he said.

Often, that lab must work on tight deadlines, such as during competition, Butch said. While industry standard turnaround for samples is 10 business days, during the Olympics it can shorten to 48-to-72 hours from receipt, through full testing, to return. Protocol demands that every Olympian be tested prior to competing, and be subject to random testing; some athletes undergo multiple tests. And, Olympic competition itself cues testing.

"Obviously, we must test the winners," Butch added. He said all medalists undergo testing immediately following their event.

Another lab imperative is when an "A" sample--which supplies the first round of testing--shows positive. Butch and his colleagues must then enact a round of testing for confirmation on the subject's "B" sample, which is the second of the required two batches collected from each subject. This second round is the same scientific process as the first, with added rigor and safeguards: all B samples, unlike A samples, are tested in isolated batches by a single technician so the sample never changes hands.

"A negative [untainted] test is my friend," Butch said. "It takes a lot of work to run the first test, let alone a second."

To advance his urinalysis science, Eichner's drug lab has concentrated on developing tests to identify new substances coming from the renegade black market. He is now focusing on blood analysis. Though serum must be kept cold (which is highly expensive and time-sensitive), popular human growth hormone is best detected through blood tests.

But, the importance of screening is not just to protect fairness in sport or to ensure the health of athletes, said Leslie Henderson, professor of physiology and neurobiology at the Geisel School of Medicine at Dartmouth College. She works with hamsters, mice and rats to detect the sometimes-permanent alternations anabolic steroids can inflict on the nervous system when taken during adolescence. Particularly shocking is her research with animal subjects, which has shown that steroid use during this life stage has lifelong effects including emotional hardships (depression, aggression, and sexual dysfunction), and physical ailments (like cancers and liver and kidney diseases). She hesitates to extrapolate too far, but has worried that these frightening results may prove true among people.

In the Olympic realm, Henderson mentioned, this would apply to young athletes, often teenaged women, in sports like swimming and gymnastics, where androgenic (male characteristic producing) drugs boost muscle mass, strength, and performance.

She hopes that her rodent research, drug detection by labs, and WADA regulation might inform and protect a coming generation of young athletes who may not understand the risk.

"Kids see these athletes with perfect bodies," Henderson. "They think they're healthy, but they're not."

Want to test your knowledge of performance-enhancing drugs? Take WADA's Play True Quiz.

I'll be watching the Olympics again this year whenever I can. I'll watch for the explosive performances and tight finishes on the track and field. I'll look for the photo-finish touch in the pool. I'll be tuning in for gymnastics' superhuman-like shows of power and balance.

And, I'll be watching a new Olympic pastime: seeing who gets caught using performance-enhancing drugs, or PEDs. "Citius, Altius, Fortius"--the Olympic motto--translates to: Faster, Higher, Stronger. Does that sentiment drive some athletes to seek enhanced results?

This post marks the beginning of a blog series covering PEDs and how they have affected Olympic games and athletes. PEDs have profoundly changed modern sporting contests and media coverage of them for decades. In addition to the Major League Baseball steroid scandals and July's renewed accusations of Lance Armstrong's blood doping, there have been instances and investigations of doped Olympians dating back to East German and Soviet-era athletes more than 40 years ago.

Only more recently have governing bodies--namely the World Anti-Doping Agency, "WADA," which tests international contests such as London's 2012 Olympics--attempted to test for and regulate their use and abuse. Along with competitive advantages, PEDs carry severe health risks. They can even kill.

Dr. William Mitchell is an orthopedic surgeon and an expert on this subject, having worked with professional and amateur athletes in greater Boston for more than 25 years and having served as contributing editor to The Encyclopedia of Sports Medicine. Of the many drugs on the black market, Mitchell spotlights "the big three": erythropoietin, or EPO; human growth hormone, also known as hGH; and synthetic testosterone. All are prominently featured on WADA's nine-page list of prohibited substances, which went into effect on January 1 this year. It provides the standard for prohibited chemicals among international athletes and will guide official Olympic testing in coming weeks.

Dr. Mitchell walked me through the first section of WADA on anabolic agents, or synthetic versions of testosterone. At the top of the long list are the lab-made hormones androstenediol and androstenedione. Along with many similar anabolics, these are laboratory-made versions of the body's main strength inducing hormone, testosterone.

Both men and women produce testosterone naturally. It imparts primary sex characteristics in males, like a deep voice, facial hair and sexual organs. In both genders, it metabolizes amino acids from the diet into the proteins that make up muscle fibers. Therefore, the more testosterone in the body, the more muscle building potential.

Athletes take these synthetic forms of testosterone hoping to gain strength and muscle density, decrease recovery time after training, and reduce the incidence of injury during intense workouts.

But side effects of these drugs go beyond the simple risk of being caught using. Rage, depression, severe acne and baldness, in both genders, may be the best-known side effects. Less widely circulated are the more severe repercussions of chronic use, like liver abnormalities and tumors, heart and circulatory impairment, cholesterol risks, and the added danger of contracting infectious diseases, like HIV or hepatitis, from shared needles. Every one of these can be life threatening. Because of these dangers, and the use of anabolic agents across so many athletes in multiple sports, the WADA ranked them at the top of their prohibited list, said Mitchell.

Scrolling down the document, you don't have to go far to find the second and third of Mitchell's "big three," hGH and EPO.

Like the "andro-" drugs, hGH is an anabolic hormone very similar to testosterone. Naturally secreted by the pituitary gland in both sexes, it too increases muscle mass. Mitchell relates it very closely to testosterone in that hGH builds proteins from the food we eat so that bone and muscle can grow in density. Olympians in strength and speed events--sprinting, power lifting, swimming, boxing--may be competing against athletes who have used hGH.

Technically, hGH is available only by doctor's prescription, and it is typically used to help young children with deficiencies leading to inhibited growth. Though it hasn't been studied as a performance-enhancer--the ethical implications of such a study are troubling--baseball fans allegedly saw it in action in Barry Bonds. HGH increased Bonds' muscle mass along with his shoe size and even his skull size, said Mitchell.

"That's human growth hormone," he said. "That's what it does."

HGH brings its own set of risks. Topping the list is cardiomyopathy, an enlarging and thickening of muscles in the heart, that weakens heart function over time. HGH can also impair glucose regulation, leading to type-two diabetes. Over prolonged use, joints, tendons, ligaments and muscles can deteriorate, causing an ironic lack of strength in the aging hGH athlete.

This brings us to the last of the "big three," EPO. The drug, epoietin alpha, is a laboratory version of erythropoietin, a naturally-occurring hormone produced by the kidneys and liver that stimulates red blood cell production by the bone marrow. By helping to increase the number of red blood cells, which contain hemoglobin molecules that transport oxygen from the blood to the muscle, EPO boosts the amount of fuel muscles have to burn for energy. In medical applications, it has been used to treat lack of blood iron, or anemia, in patients with greatly impaired kidney function from diseases like AIDS. It can also be used before surgery, like open-heart procedures, to counter the effects of anticipated blood loss.

But, it has been implicated in the death of at least 18 cyclists during alleged heavy use in the 1990s. These cyclists were victims of bleeding events: stroke, heart attack, and blood clots in the lungs called pulmonary edemas.

With so many grave risks, are the perceived benefits worth it? In fact, none of PEDs' touted performance benefits--taken at high doses acquired on the unregulated black market, and with prolonged use or abuse--have been proven. After all, giving athletes high doses of dangerous drugs for research purposes would be highly unethical.

Mitchell agrees: "Doping increases health risks when doses and amounts of hormonal use is not regulated and can lead to overdosing and catastrophic health risks including death."

Chris Doorley

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