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.

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Chris Doorley

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