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UNCERTAIN
GOLD Drug testing at the Olympics
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very other year, athletes from around the world gather to compete in cities whose names soon become international watchwords. For two weeks or so, the Olympic Games crowd other programming off of television and sweep most other news off the radio. But behind the stories of triple lutzes, bro ken records, professional rivalries, and personal bests is a separate struggle, rarely discussed during the continuous stream of reporter commentary. Few viewers or listeners realize that the final test of a gold or silver medal may actu ally lie in the contents of the specimen bot tles the three top athletes, plus a few oth ers chosen at random, are required to fill after each competition. During the two weeks of competition, the International Olympic Committee (IOC)-accredited laboratory works around the clock to analyze thousands of urine
selves. Competitors in third-century Forensic drug testing Greece were said to have eaten bread soaked with opium during the games to has evolved into an improve their endurance and eliminate pain of overexertion and injury. After important part of the stimulants such as strychnine, ephedrine, amphetamine were introduced in the judging athletes' and 1800s, canal jumpers in Amsterdam and cyclists in America and Europe started to performance experiment with them. samples for prohibited drugs. The staff may have only 24 h, from the time the sam ples are collected to the awards cere mony for that event, to deliver the reports that determine whether the athletes will really receive their medals. At stake are the reputations and careers of the ath letes, their coaches, their doctors, and the countries they represent. Athletic drug abuse, or "sports dop ing," is as old as the Olympic Games them
In the 1930s, the revival of the Olympic Games brought with it increasing abuse of narcotics and stimulants for athletic ad vantage. In contrast to the more active con trol of professional horse and dog races, where sports doping was considered a bla tant form of cheating, no such regulation was imposed on human sporting events in that era. It wasn't until 1960 that thefirstfatality from stimulant drugs was reported offi cially. The death of an Olympic cyclist
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from amphetamine abuse shocked the in ternational community. In 1967, the IOC responded to the growing concern over sports doping and formed the Medical Commission, giving it a mandate to test athletes at the Olympics for evidence of drug abuse. The 1968 Olympics were the first to require systematic drug testing. Since then, the Medical Commission has continued to develop protocols and propose regulations on sports doping. Its mission is twofold: to reduce the inci dence of this form of cheating and to dis suade athletes from using drugs that could weaken their systems over time or bring on heart attacks or fatal seizures.
Testing
Before the sample leaves the collection site, the athlete splits the specimen into two bottles labeled "A" and "B" and wit nesses a complicated labeling and sealing process that is designed to guarantee the forensic chain of custody. Sample A is tested routinely; sample Β is stored for retesting in the presence of the athlete or a representative in the event that sample A is positive. Bowers and Catlin say each sample must go through at least five sep arate protocols for screening, including derivatized and underivatized GC/MS for steroids and stimulants, immunoassays for peptide and glycoprotein hormones, and LC for diuretics. According to IOC rules, all confirmations have to be performed by GC/ MS, even though that method may not be ideal for detecting some of the suspected analytes.
assay cocktail that catches everything," he explains. Another advantage is that com mon reducing agents such as vitamin C and acetaminophen interfere with the peroxidase-based color generation schemes of immunoassays but not with GC/MS. Classes of prohibited drugs
Incidents of abuse continue to be reported for narcotics and the older stimulant drugs, even strychnine (rat poison). Most athletes shun strychnine of their own ac cord because with a single dose, the drug can go very quickly from acting as a stim ulant to causing fatal seizures. Catlin says that laboratory staff were surprised when one urine test at the recent games in Bar Building up the labs celona turned up positive for it. Testing at Olympic events takes place at an IOC-accredited laboratory set up at a Since the 1970s, however, new classes nearby university or research institu of drugs such as anabolic androgenic ste tion in or near the city where the games roids and growth hormone have become are being held. Setting up is no small un more prominent. One likely reason is the dertaking; the laboratory, which may have pharmaceutical industry's increasing use 12-20 GC/MS instruments working of recombinant DNA technology to massaround the clock during the games, usu produce hormones that are nearly indistin ally takes at least six months to equip, guishable from the native human ones. staff, and validate before the games be Not all drugs are tested for in all Olym gin. The national sporting body of the pic events. Currently, say Bowers and Cat country hosting the games is responsible lin, there arefiveclasses of prohibited for collection and testing and for funding drugs; two specified types of prohibited the laboratory operation. blood doping methods; and several other classes of drugs that may be restricted or In addition to the temporary labs that allowed, depending on the sport, the dose, must be set up for the Olympics each time, or a legitimate medical reason for use. there are 24 permanent IOC-accredited Each GC/MS system may be opti Prohibited drug classes include anabolic labs throughout the world that perform mized for the detection of a different type drug tests on athletes for qualifying heats, of drug and its metabolites. "Almost none agents, stimulants, narcotics, diuretics (potentially abused by boxers and other non-Olympic competitions, and other na of the compounds are excreted as the weight-classed athletes), and peptide and tional-level amateur and professional parent drug," Bowers says. "Sometimes sporting events. Don Catlin of the Univer the only way to determine whether a drug glycoprotein growth hormones and their analogues. sity of California, Los Angeles, and Larry was used is to piece together information Bowers of Indiana University serve as di from characteristic metabolites. There Anabolic steroids, used to build mus rectors of the two permanent IOC- accred fore, the hardest part of setting up the cle mass and to hasten recovery after ited drug-testing laboratories in the lab is establishing a library of multiple me strenuous activity, are the most com United States. Catlin, who built the labora tabolites for each drug. If youfindthree monly abused drugs among male athletes tory that served the 1984 Summer Olym metabolites and you have MS data on all of both in and out of the Olympics. Female pics in Los Angeles, also serves as a mem them, that's usually pretty conclusive." athletes sometimes use them as well, says ber of the IOC's Medical Commission. Detection limits for most analytes are in Catlin, and may get a greater effect from the ng/mL range. a much smaller dose, which can be too low Catlin says it took almost three years to detect easily. Last November, he says, to build the facility at UCLA. He recalls Bowers says one major difference be 11 Chinese athletes, at least one of whom that when the IOCfirstapproached him to tween IOC testing and standard work had broken a number of records in world do it in 1981, he was reluctant. "I hadn't place drug testing is that the IOC uses competitions in the past two years, tested heard of half the drugs on their list, and GC/MS rather than immunoassays for positive for dihydrotestosterone. they were talking about doing 150 sam screening. The reason, he says, is the ples a day. In those days, that kind of work large number of drugs and excreted me Among the steroids, testosterone load was extraordinary. These days the tabolites that must be assayed for in a sin abuse in men poses the most problems for instruments are better and more stable." gle sample. "No one's got an immuno detection, Catlin says. So far, no method
Each GC/MS system may be optimized for a different type of drug or metabolite
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has successfully distinguished between natural and synthetic testosterone, and natural levels fluctuate, but generally the ratio of testosterone to epitestosterone is fairly constant and ratios > 6:1 are considered suspect. Forbidden doping methods include "blood doping," where the athlete tries to enrich his or her blood in the weeks before competition. Formerly this was done by taking a pint or more of blood for storage and retransfusion after several weeks of training and recovery. The more modern method, which requires no transfusions, is to administer the hormone erythropoietin, which stimulates the production of red blood cells. The drawback for investigators, says Catlin, is that synthetic erythropoietin, like the peptide and glycoprotein hormones, is potent enough to use at very low doses. GC/MS, the official confirmation method of the IOC regulations, isn't sensitive enough to detect the resulting traces in urine and can't distinguish them from natural hormones. The other doping method involves the pharmaceutical, chemical, or physical manipulation of the urine sample itself. Physical or chemical manipulation is usually overcome by the IOC regulations, which require sample collection to be supervised by a trained collector who tests the samples for specific gravity and pH at the collection site. If the samples are too dilute or too alkaline, the athlete must stay and give another specimen at least 30 min later. Most recently, the IOC investigators have become concerned over the increased use of "masking" drugs. Athletes sometimes use these drugs in an attempt to block the normal excretion of prohibited drugs such as steroids in the urine, thereby preventing their detection, but the masking drugs themselves are detectable indicators of drug abuse. Restricted drugs include alcohol, caffeine, marijuana (which isn't illegal in all countries and isn't tested for unless an individual sport requires it), local anesthetics, corticosteroids, and beta blockers. Allowances and acceptable doses depend on the sport and on the event judges' discretion. Some of the thresholds are fairly straightforward—for competition purposes, unacceptable caffeine levels are ~ 10-15 times the level a normal coffee
drinker might have. For other drugs, how- as erythropoietin that aren't excreted at ever, the question of what constitutes detectable concentrations or easily identifitherapeutic use and what is sports doping able forms in urine. can be a real dilemma. For instance, antiBut requiring blood samples is the subhypertensives such as beta blockers that ject of a complicated and long-standing inslow down the heart rate are often vital for ternational debate. Unlike urine testing, heart patients, but they are forbidden in which is acceptable to all the member archery and pistol-shooting events, where countries of the IOC, Catlin explains, contestants are trained to shoot between "Blood testing has traditionally been much heartbeats so as to avoid involuntary hand more controversial. Some cultures conor body movements caused by the pulse. sider it to be very invasive." Blood testing also brings up the issues of AIDS, the need for trained phlebotomists to collect The future—blood testing? samples, the need for universal precauIOC drug testing methods are still evolving, say Bowers and Catlin. The games at tions, and the fact that athletes don't like Lillehammer (Norway) gave the Medical getting stuck with needles repeatedly. "We're all being very careful to respect Commission its first opportunity to collect blood samples from athletes in a small the athlete," Bowers says. However, Catlin remains optimistic that blood testing number of events to determine the possible added value of testing blood. Blood will eventually be allowed at the Olympics. If so, IOC-accredited laboratories will testing, which has never been part of the get the chance to test for analytes that are IOC protocols despite its established use in clinical and forensic settings, could difficult to determine by currently approved methods. Deborah Noble open the door to testing for analytes such
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