posted April 10, 2000 08:45 AM            

Two male bodybuilders were recently arrested by the French customs in Strasbourg (France) in possession of 2050 tablets and 251 ampoules of various anabolic steroids. It was claimed that the steroids were for personal use and not for trafficking as suggested by the police. Urine and hair specimens were collected from both suspects to clarify the claims. Nandrolone, stanozolol, testosterone and their corresponding metabolites were identified in the urine of both subjects. After decontamination, the hair was hydrolyzed by sodium hydroxide in presence of deuterated internal standards. After extraction with ethyl acetate and silylation, the drugs were identified by GC-MS (gas chromatography-mass spectrometry) in the electron impact mode. Hair from both males were positive for nandrolone, testosterone and stanozolol, clearly indicating steroids abuse.

The standard in drug testing for doping control is gas chromatography/mass spectrometry conducted on a urine sample. For the past 20 years, hair analysis has been proposed for identifying drug abusers in forensic science. Specimens can be collected under close supervision without embarrassment and are not subject to evasive maneuvers. In contrast with urine, hair analysis has a wide window of detection, ranging from weeks to months, depending on the length of the hair shaft, and provides information concerning the pattern of an individual's drug abuse. The International Olympic Committee does not yet recognize hair analysis, although this technology is accepted in most courts of Justice.

In France during a famous bicycle race, the newspapers documented the degree in which doping seemed to be supervised in some teams by managers and doctors. Use of anabolic steroids and other substances was officially banned in the mid-seventies by sports authorities. This policy has been enforced through urine testing before competition. It is well known, however, that a latency period is all that is necessary to defeat these tests. Nevertheless, hair analysis could be a promising tool when testing for periods that are not accessible to urinalysis any more. We have developed different sensitive methods for testing hair for amphetamines, anabolic steroids and their esters and corticosteroids. For anabolic steroids and their esters, 100 mg of powdered hair were treated with methanol in an ultrasonic bath for extraction of esters, and then alkaline digested for an optimum recovery of other drugs. Analysis was conducted by gas chromatography coupled to a triple quadruple mass spectrometer. Thirty cyclists were sampled and tested both in hair and in urine. Anabolic steroids were detected twice (nandrolone 1 case, and testosterone undecanoate 1 case) in hair (out of 25 analyses) compared to none in urine (out of 30 analyses).

The actual anti-doping control rules applied in sports (as established by the International Olympic Committee and the International Sport Federations) state that a positive case is chemically established by the unequivocal detection of a forbidden parent molecule and/or any of its metabolite(s) in urine, no matter the amounts which were administered and when the drug was taken. Screening is accomplished most of the time by using GC-MS (gas chromatography-mass spectrometry) procedures. These have been optimized to detect most if not all of the forbidden compounds, which are put on a list. Recently, attempts have been made on scalp hair to demonstrate the value of this matrix as a possible means for differentiating between therapeutic uses and doping abuse. Naturally occurring molecules, like testosterone and its metabolites, could also be differentiated from their synthetic counterparts. Positive cases are more often challenged in courts and retrospectively in time of the drug(s) intake is becoming an important issue for evaluating the responsibility of the person. This can be based on hair analyses if the drugs have been taken at regular intervals. On the other hand, anabolic agents, when taken to improve the physical performances, follow complex regimens with the mixing of various formulas and dosages. Scalp hair references ranges for these as well as for endogenous substances still wait to be established statistically for competing, well-trained athletes. The incorporation rate into blond or gray hair is poorer than that of dark colored hair, raising the question of individual�s equality against the controls, a very important matter of concern for the sport's governing bodies. The frequency of hair cutting and short haircuts necessary to gain speed in specific sports, like swimming, are other critical factors. So far, a minority of prohibited substances could be detected in scalp hair with the sensitivity and specificity required in the context of the sport's activities. From the above, clear limitations of the usefulness of hair analysis in doping control analysis are obvious until a lot more data relevant to this particular field have been collected.

When current anti-doping programs were developed, the most frequently used doping agents were xenobiotics, such as stimulants and anabolic steroids that are readily detectable in urine with the use of gas chromatography and mass spectrometry. As control of traditional doping agents became effective, some athletes turned to other means to improve performance, including blood doping and the application of recombinant peptide hormones such as erythropoietin and growth hormone. Doping with these agents is not easily detected in urine samples, and therefore new strategies must be developed as a supplement to those already in use. Such strategies will probably include analyzing blood samples, as several of the most promising methods that are able to detect modern doping agents use blood as the analytical matrix. Non-autologous (auto-logous; from the same organism) blood doping results in an admixture of self and foreign red blood cells that can be detected in a blood sample with the methods available. Methods to indicate doping with erythropoietin (Glycoprotein hormone) include the indirect finding of an elevated level of soluble transferrin receptor in serum, or a direct demonstration of a shift from the normal to an abnormal spectrum of erythropoietin isoforms. To indicate doping with growth hormone, a set of serum parameters including insulin growth factors and their binding proteins are under investigation as indirect evidence. A direct method using isotopic differences between "endogenous" (your body's own) and "recombinant" (of or resulting from new combinations of genetic material) growth hormones is being investigated. A similar method has been established to detect the administration of testosterone esters. Several legal and ethical questions must be solved before blood sampling can become a part of routine doping control.

posted April 10, 2000 03:44 PM         

Slim

posted April 10, 2000 04:39 PM            

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even my battle with death could only keep me out of the gym for 35 days