Performance-enhancing drugs continue to threaten the integrity of sporting events and the health and well-being of participants. Drug testing therefore has become a routine practice at major sporting events such as the 1998 FIFA Soccer World Cup (WC '98). This year, the World Cup involves more teams than ever, and the organizers are committed to catching any players misusing drugs.

The Laboratoire National de Despistage du Dopage in Paris, France, one of only 23 laboratories in the world accredited by the International Olympic Committee (IOC), will handle all drug testing for WC '98. HP analytical instruments such as the HP GC/MS system will play a critical role in the analysis of urine samples from players participating in the event.

250-300 Banned Substances

Each urine sample sent to the laboratory must be tested for 250-300 different substances and metabolites banned by FIFA -- no small task even with today's highly sophisticated analytical instruments. Banned substances fall into five main categories:

• Stimulants. This category includes common drugs such as caffeine as well as cocaine and amphetamines

• Analgesics. Morphine and other narcotics allow athletes to continue to train or compete when injured, often causing serious damage and posing the threat of addiction.

• Anabolic Steroids. These muscle-building drugs include naturally occurring hormones, such as testosterone and their derivatives.

• Diuretics. They cause increased urine excretion and are used to mask the use of other banned substances.

• Peptide Hormones. Possibly the most difficult group to detect reliably, they include growth hormone and corticotrophin, which is used to increase the levels of corticosteroids and induce euphoria.

Quantitation Important for Some

In most cases, tests for these substances are qualitative. Any use of a banned substance, in any quantity, contravenes FIFA's rules, and even tiny traces of a banned substance or its metabolites indicate a positive result. Some of the banned compounds occur in the body naturally, such as testosterone, while others are prohibited only in quantities considered abusive, caffeine, for example. In these cases, the substances must be quantitated, and only levels above the designated threshold are considered positive.

Consistent Data Handling Essential

Athletes' careers and reputations depend on the laboratory results; standard operating procedures are therefore put in place for each type of testing. The laboratories operate under Good Laboratory Practice (GLP) and regularly undergo a stringent review by the IOC.

The Paris laboratory has a series of HP GC/MS systems that are linked by an HP ChemLAN laboratory network -- ensuring that methods and analytical data are handled and recorded consistently. In the event of a legal challenge, the system provides evidence that the correct operation has taken place and eliminates the possibility of sample misidentification.

The Techniques Used

Good sample preparation is essential. Five different procedures are used, which include passing the sample through columns that bind specific types of compounds. Contaminating substances can be washed away and molecules of interest concentrated. The concentrated sample undergoes a number of additional steps, including hydrolysis, extraction, and concentration, depending on which substances it is being tested for. Sample preparation involves a mixture of manual and automated processes and takes about two hours.

Gas chromatography combined with mass spectroscopy is the main technique used by the laboratory to detect banned substances. Together, GC and MS provide a "fingerprint" for each compound. The laboratory checks the mass spectra for compounds with molecular weights characteristic of banned substances. Then, at the touch of a button, the laboratory can check the retention time on the GC. Only if both retention time and mass are consistent with a banned substance or a known metabolite is the sample considered positive. Analgesics, anabolic steroids, stimulants, and diuretics can all be identified using this method. GC/MS is also used to quantitate levels of testosterone, caffeine, and ephedrine.

Banned Drugs Detectable 99%

To detect and quantitate corticosteroids, the laboratories use HPLC in conjunction with mass spectroscopy. HPLC is also used to test for the presence of diuretics, while immunoassays are used to detect peptide hormones. Current analytical techniques can detect 99% of the compounds banned by FIFA.

To ensure that compounds are not misidentified, standards and controls are essential. Developing a standard for only the banned substances would not be difficult for the laboratory -- a standard sample of the drug could be added to a urine sample, for example. But laboratories must be able to detect all metabolites of a banned substance, and these can be assessed accurately only by letting a volunteer take the banned drug and using the urine as the positive standard. The negative standard is the urine from a volunteer who has not taken any banned substances. All urine samples tested must be run with the appropriate positive and negative standards.

Quality and accuracy requirements are extremely strict, and laboratories regularly receive faked samples to test their procedures. A single false positive result is sufficient for a laboratory to lose its IOC accreditation. In the event of a false negative result, the accreditation will be suspended until the reason is found and corrective action has been taken.

HP makes the ideal partner for such drug monitoring missions; first by offering a complete range of high-quality analytical instrumentation, then by providing the necessary applications and maintenance support.