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Wednesday 15 June 2011
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Drug Discovery Drugs in sport:
from Nature Jonathan Knight
Jonathan Knight is a contributing correspondent for Nature . Don Catlin's lab has struck a major blow against drug abuse in
athletics, by developing a test for a shadowy 'designer steroid'.
Jonathan Knight visits the scientists who are striving to keep
sport clean. Nature
Nature Structural &
Neuropsychopharmacology Their lives in his hands: athletes' reputations can
be shattered if Don Catlin finds a banned drug in
a sample. Molecular Psychiatry JOHN HAYES Signaling Gateway
Therapeutics When you arrive at the Olympic
Analytical Laboratory in Los
Angeles, it's clear that this isn't
an operation that goes out of its
way to make visitors feel
welcome. Nestled among carbody shops, its narrow façade
shoulders up against chain-link
fences topped with razor wire.
The mirrored-glass front door of
this drug-testing lab is locked,
and will only be opened if you are
meant to be there. Numerous
television crews have been turned
away in recent weeks, and a sign
inside the door reminds staff not
to admit an athlete who filed a
complaint against the lab after it
found steroids in her urine. Although the fences actually belong to the neighbouring car-repair yards,
this defensive appearance is fitting for a lab that is at war with the
suppliers of performance-enhancing drugs. After a summer of intense
work on the residue from an anonymously supplied syringe, the lab
rocked the world of sport last month by revealing that an undisclosed
number of athletes had tested positive for a designer steroid, known as
tetrahydrogestrinone or THG, which had evaded standard tests (see
Nature 425 , 752; 2003).
Keeping things under wraps is an essential part of the game, says lab
director Don Catlin. Indeed, most of the lab's employees were in the http://www.nature.com/drugdisc/news/articles/426114a.html Page 1 of 4 drug [email protected] 6/15/11 9:09 AM dark about THG until after the news became public. "If what we were
working on had gotten out, people would have stopped using it before
we had a test," Catlin says.
Drug testers and drug-using athletes have been locked in battle for
more than three decades. When the International Olympic Committee
(IOC) ran its first drug tests during the 1968 games in Mexico City, no
one was found using anything other than alcohol. But heavier testing in
Munich in 1972 caught seven athletes using banned drugs. Since then,
testers and dopers have been engaged in an arms race — with the users
of illicit drugs generally being perceived to have the upper hand.
An unlikely hero
Catlin did not expect to become involved in this struggle. In 1982, he
was an established endocrinologist at the University of California, Los
Angeles (UCLA), when the IOC came to ask if he would run the testing
for the 1984 Los Angeles games. "They showed me a list of banned
substances," Catlin recalls, "and I said, 'I don't think we can do this'."
But when the IOC made it clear that it was offering to greatly expand
his lab and hire the staff he would need, he rose to the challenge.
Today Catlin's lab, which is still part of UCLA, has three dozen
employees. Its biggest customers, apart from the US Anti-Doping Agency
(USADA), are the National Collegiate Athletic Association and the
National Football League. Most of the work involves the routine
processing of number-coded urine samples that arrive by courier almost
daily. These can be analysed for several dozen hormones, stimulants,
diuretics and other banned substances, depending on what the customer
asks for. Most of the tests involve a combination of gas chromatography
and mass spectrometry. The desktop machines that run these
procedures occupy almost every available surface in the lab.
Besides this routine, there is a continuous effort to improve the tests. In
the EPO lab, where technicians run antibody tests to detect the hormone
erythropoietin, which boosts red-blood-cell counts so that more oxygen
gets to the muscles, group leader Andreas Breidbach is looking for ways
to see further back in time. Right now, evidence of EPO use is hard to
find after about a week. But the benefits last longer. Knowing this, many
athletes stop doping ahead of an event. Breidbach hopes to extend the
window of detection by tweaking each step of the test to improve its
sensitivity. "We are always improving because the athletes are always
improving," he says.
One of the lab's research achievements is a
method for spotting testosterone abuse. Unlike
synthetic steroids, testosterone and its
breakdown products are naturally present in
the urine of dopers and non-dopers, whether
they are male or female. The standard test for
testosterone doping looks for an elevated ratio
of the hormone relative to its cousin
epitestosterone. These are normally present at
equal levels in the urine, and the IOC has set a
ratio of 6:1 as the cut-off for declaring a
positive test. This is bad news for the rare
athlete with a naturally elevated ratio. And the
test is not hard to beat: athletes can, for
instance, take epitestosterone as well as
JOHN HAYES http://www.nature.com/drugdisc/news/articles/426114a.html Page 2 of 4 drug [email protected] 6/15/11 9:09 AM Catlin's group hit on the idea of using carbon
isotope ratios to distinguish between natural testosterone and its labmade counterpart. Synthetic testosterone contains less of the heavy
isotope carbon-13 compared with the natural compound. This is because
the hormone's manufacturers begin their synthesis with an extract from
yams, which happens to contain a compound with the same four-ring
structure as steroids. Plants incorporate carbon-13 — present at low
levels in the atmosphere — much less efficiently than do animals.
Although the isotope test is expensive and not always used, it has made
a difference. Once athletes became aware of the test in the late 1990s,
many stopped using testosterone, Catlin says.
But the battle continues. Just how far ahead are drug-using athletes and
their suppliers? No one knows for sure, although part of the answer may
lie in a slim binder tucked away on a shelf in the spectral analysis room
at Catlin's lab.
In this room, Yuliya Kucherova pores over gas-chromatography traces
and mass spectrograms, looking for the signatures of steroids. In some
cases, the steroid shows up as a distinct peak by gas chromatography,
which separates compounds based on their volatility. In others,
confirmation by mass spectrometry is essential. This involves breaking
molecules into fragments and determining their molecular weights.
Kucherova, after reading dozens of spectra
a day for years, has no trouble spotting
banned compounds. But occasionally a new
peak shows up that doesn't correspond to
any known drug, prescription or otherwise.
Is it a new synthetic steroid? A Chinese
herb? A rare prescription medication?
These mystery peaks wind up in the special
binder, and several dozen are awaiting
further investigation. By the numbers: identified only by a
code, rows of athletes' urine
samples are screened for a wide
range of banned substances.
JOHN HAYES Once in a while, Catlin and his colleagues
get lucky. Kucherova last year spotted a
peak she had actually seen previously. It
was the synthetic steroid norbolethone, first
made in 1966 as a possible treatment for
short stature. But it proved too toxic and
was scrapped by its maker, the
pharmaceutical company Wyeth. It has not
been made commercially since. Although only one athlete tested positive for norbolethone, that result
was a turning point for the lab. It helped Catlin to convince the sporting
authorities that the underground market for steroids was being served
by clandestine chemists and manufacturers. He made the case that to
keep ahead of the game, the testers needed to work in secret on what
the next underground drug might be. Earlier this year, the USADA
awarded Catlin's lab a grant to probe such questions.
The group was preparing to launch into this work when the anonymously
posted syringe came through the door. A team of three, which rapidly
rose in number to eight, was immediately assigned to the task of
identifying the residue inside. People worked late, sometimes all night.
Details of the investigation were shared internally on a 'need-to-know'
basis, but the sense of urgency and excitement was pervasive. "We've http://www.nature.com/drugdisc/news/articles/426114a.html Page 3 of 4 drug [email protected] 6/15/11 9:09 AM never had a project of this magnitude," enthuses Michael Sekera, the
lab's scientific director. "It was an amazing experience."
It was clear right away that normal tests would never have spotted
THG. Before analysis, samples are chemically treated to make steroids
show up as single sharp peaks in the gas chromatograph. But this
process sees THG produce 25 little peaks that don't look like a steroid at
The researchers turned to mass spectrometry to get a clean signature.
Then they worked backwards to guess at the compound's structure. They
confirmed their hunch by making the compound from scratch and
showing that it produced the same signature pattern. Then they found a
primate laboratory that was willing to give THG to a baboon, so that
they could have some idea how it would come out in the urine. Catlin
and his colleagues devised a new test, and began running it on athletes'
urine samples. The whole process took two months.
The story isn't over, Catlin says, as he anticipates that there may be
legal challenges. These have become more frequent as the stakes in
sports have risen. When Catlin first started in the drug-testing business,
athletes often reacted to being caught with resignation, even contrition.
"You caught me doc, I messed up," Catlin recalls one athlete saying.
Now, athletes' lawyers send chemists into the lab in the hope of finding
flaws in its testing procedures.
So Catlin has to be very careful. He won't discuss the results of the THG
tests as long as the federal investigation into the scandal continues. Not
that he knows the names of the accused athletes. All samples are coded
before they arrive. Even an athlete's attorneys, who are entitled to
observe the testing, say only which number they represent, and not
In any case, Catlin and Sekera say that they are more interested in
where to go from here. To keep ahead of the people making drugs you
have to start making them yourself, they argue. You need to design
them, find out how they break down in the body, and develop a test for
Whether the lab will get the funds to continue with this approach is a
matter for the sports authorities to decide. The bulk of the USADA grant
was consumed in figuring out how to test for THG. But if Catlin and his
colleagues do resume this work, don't expect them to breathe a word of
what they're up to. "We are very good at keeping things to ourselves,"
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