Pharmacological characteristics of anabolic steroids, including nandrolone Testosterone is the ancestor and the most potent androgen of the group that incorporates the hormone nor-testosterone, which is better known in sports under the old trade name “Nandrolone” and which is characterised by anabolic effects (muscle building).
Associated with the weight-increasing activity of anabolic steroids upon skeletal muscles has been the enhancement in glycogen muscular content. The continuous administration of anabolic steroids will not, however, result in a progressive increase of glycogen deposition but may induce a gradual decrease. Androgens act on osteoid matrix, stimulate the osteoblastic function and promote the linear growth of the skeleton by interference on epiphyses.
During puberty, however, androgens may hasten epiphyseal closure, markedly interfering with the tallness of young athletes.
The effect on muscle protein anabolism stimulated the research and development of many synthetic compounds (e.g. nandrolone, mesterolone, oxandrolone, oxymetholone, stanozolol, stebolone, etc.) in the hope to divorce the anabolic effect from androgenic activity. Although some dissociation has been realised, all the synthetic compounds - including nandrolone - still possess androgenic action on testes, prostate, penis, seminal fluid, and so on (Benzi e Bellotti, 1990).
The rate of absorption of androgens is related to the way of administration.
Testosterone is almost completely inactivated when ingested per os because of the passage through the liver, while some chemical modifications result in a considerable protection against metabolic hepatic degradation.
The use of androgens in form of sublingual preparations appears to be an appropriate method, even if much of the hormone is swallowed with saliva because of the need for stay in contact with the buccal mucosa for an extended period of time.
The effectiveness of parenteral administration (subcutaneous, intramuscular, etc.) of solutions of steroid in oil or aqueous suspensions is related to the delay in the absorption from the site of injection.
Nandrolone phenpropionate (Durabolin) is a parenteral preparation in oil whose activity is prolonged because of esterification with proprionate. Injections of nandrolone phenpropionate at weekly intervals are suitable to maintain good anabolic effects. Furthermore, another ester of nandrolone, namely nandrolone decanoate (Deca-Durabolin), is characterised by a longer duration of anabolic effects when injected once every 3 or 4 weeks. Adverse reactions by anabolic steroids, including nandrolone Androgens may promote both sodium and potassium retention, and tissue oedema.
Furthermore, the administration of synthetic compounds, including nandrolone, results in testicular atrophy or inhibition of testicular growth and function (hypospermia, hypozoospermia, azoospermia), in gynecomastia, and in increase in prostate glandular secretion and size. Intermittent discontinuation of anabolic steroid use by athletes probably allows some side effects to return to normal. A typical adverse reaction by anabolic steroids on the skin is represented by enhancement of secretion rate of sebaceous glands, increasing skin oiliness and materials blocking the ducts of sebaceous and sweat glands.
The final result is the development of acne.
The primary adverse reaction of anabolic steroids on the liver is represented by the accumulation of bile into the biliary collecting system, with jaundice and increase in serum bilirubin, alkaline phophatase and transaminase levels.
A debated problem is represented by the possibility to generate hepatic tumours. Anabolic steroids may induce the peliosis hepatis characterised by small, blood-filled cystic spaces disseminated randomly in the liver parenchyma.
The evolution of peliosis hepatis to benign or malignant tumours is suspected. Anabolic steroid adverse reactions in female are furthermore characterised also by alopecia, hirsutism, deepening of the voice, enlargement of clitoris, amenorrhea, and increase or (less commonly) decrease in libido, vaginal mucosa atrophy, breast size decrease, appetite increase, etc. (Benzi, 1993).
Analytical methodology to detect nandrolone in athletes Over the past years, screening for abuse and/or misuse of nandrolone by athletes has been a subject of controversy and discussion, as data have been reported on the presence of norandrogens of endogenous origin in several mammalian species, including humans.
At present, high performance analytical techniques are suitable to detect trace amounts of steroids in blood and urine. In that respect, gas-chromatography/mass spectroscopy (GC-MS) is considered the gold standard for the quantitative analysis of nandrolone in blood and its two major metabolites 19-nor-androsterone and 19-nor-etiocholanolone in urine (Björkhem and Ek, 1982; Masse et al, 1985; Ozer and Temizer, 1987; De Boer et al, 1990; Dehennin et al, 1996; Meier-Augenstein, 1999; Dehennin et al, 1999; Müller et al, 1999). In that case, 19-nor-androsterone in urine may be considered as a specific marker metabolite of nandrolone. As a rule, the presence in urine of 19-nor-androsterone in a concentration greater than 2 ng/ml for males (or 5 ng/ml for females) is regarded as proof that nandrolone was assumed by athletes for improving psychophysical performance.
This cut-off value is the result of a consensus decision of several authorities including the IOC sub-commission for doping and biochemistry in sport. It has been confirmed by the FIFA itself. This means that the presence of 2 or more ng/ml of 19-nor-androsterone in urine from football players will give a doping “positive” result.
Genesis of the nandrolone-marker metabolite 19-nor-androsterone We need to clarify whether a relation might exist between raised levels of the doping-marker metabolite 19-nor-androsterone in urine samples and events possibly related to use of nandrolone or its pro-hormones by football players.
Endogenous production of nandrolone or 19-nor-androsterone by athletes Nandrolone has been found in human ovarian fluid, possibly as intermediate in enzymatic conversion of androgens to oestrogens (Dehennin et al, 1987), in urine from women in their 6th or 14th week of pregnancy (Van Eenoo et al, 1999; Mareck-Engelke et al, 1999), in healthy volunteers by collection of urine before and after prolonged intense effort (Le Bizec et al, 1999), and in volunteers trying moderate recreational sport activities (Reznik et al, 2001).
On the basis of a study on the endogenous production of 19-nor-androsterone in 137 amateurs and 358 football players from the first and second divisions of the Swiss football national league the conclusion is that none of the football players were excreting any nandrolone metabolites before effort. After the match, the spontaneous production of 19-nor-androsterone by the body of the players was an exceptional event. In fact, only 27 out of 495 football players (5.4 %) showed traces of nandrolone metabolites in the urine (urine concentrations < 2 ng/ml: IOC cut-off for males)), and only 3 out of 495 football players (0.6 %) showed urine concentrations of 19-nor-androsterone included between 2 and 3 ng/ml (Saugy et al, 1999; Robinson et al, 2001).
The quoted study was performed without previous recommendation to the players to avoid use of medicinal products, food supplements, and so on.
Thus, the urine concentrations of the quoted 3 players are possibly the result of use of preparation(s) incorporating nandrolone or pro-hormones, and so on. In any case, this research confirms that the presence of 19-nor-androsterone in a concentration greater than 2 ng/ml may be the proof that nandrolone and/or pro-hormones were assumed by players.
Consumption of meals containing nandrolone Various types of meat (pork, beef, boar, veal, lamb, chicken, fish, etc.) can contain nandrolone and, in theory, may contribute to the increased levels of 19-nor-androsterone in urines in a concentration greater than 2 ng/ml. As things stand, two possibilities can be distinguished: presence of nandrolone in meat due to illegal nandrolone treatment of the animals, or natural presence of nandrolone in meat.
The level of nandrolone in meat from illegal treated animals is too low to show up - after the ingestion by the athletes - a raised level of 19-nor-androsterone in a concentration greater than 2 ng/ml in urine samples taken one, two or several days after. In the same way, we consider the consumption of an ‘injection spot’ containing nandrolone as absolutely unlikely (the ‘injection spot’ is the location where the anabolic steroids are illegally injected in the animals in question).
Before the ingestion, the meat is instinctively subjected by the athlete to a simple, visual inspection and an incorporated disgusting cyst of non-assimilated nandrolone in meat (e.g. steak, cutlet, fillet, etc.) is easy to see and impossible to eat.
The hypothesis of unintentional intake of nandrolone through consumption of non-castrated male pig and wild boar meat (La Bizec et al, 2000) can be ruled out because in the European Union only very small quantities of non-castrated male pig and wild boar meat are eaten.
In any case, these quantities are too low to show up a raised level of 19-nor-androsterone in a concentration greater than 2 ng/ml in urine samples taken one, two or several days after. Offal, such as livers and kidneys, may have quite higher nandrolone content but very few non-castrated male pigs and wild boars are slaughtered in the European Union both as the European rules on offal use are very restrictive and the consumers do not like the queer taste of meat that is clearly distinguishable from other kinds of pork. Consumption of food supplements incorporating nandrolone or pro-hormones On the basis of the composition of food supplements manufactured according to European rules and circulating in the European Union, the conclusion is that they cannot result in the absorption of nandrolone, androstenedione, pro-hormones or metabolites suitable to be converted by the player body into 19-nor-androsterone. A different situation is represented by the use of food supplements manufactured in the United States by a dietary supplement industry that generates annual sales estimated at US $ 12 billion.
According to the “Dietary Supplement Health and Education Act of 1994”, in the US it is possible to prepare products containing anabolic steroids, more or less incorporated in a ‘melange’ of amino acids, carbohydrates, vitamins, and so on. Several anabolic steroids are sold over-the-counter (OTC: i.e., no medical prescription is required) because the quoted Act enables companies to market steroids out of the medical goal to diagnose, prevent, or cure diseases.
In which case, companies escape US Food and Drug Administration (FDA) regulation, and content and purity of ingredients are left to manufacturers (Angell and Kassirer, 1998). In the quoted products from US the presence of anabolic steroids is sometimes indicated on the packaging, while otherwise there is only general information.
Often the package labels do not warn that they may contain substances that are banned by sports organisations and may cause the detection of steroids or their metabolite(s) in urine. Therefore, also the unintentional use of US supplements added with IOC banned steroids can cause positive urine test results for nandrolone.
The European legislation defines as “medicinal products” the preparations containing anabolic steroids and, therefore, the food supplements from the US incorporating steroids are illegal in the European area. However, the products from US should be obtained by firms illegally importing the products or should be illegally purchased by Internet. In US, the steroids androstenedione and 4-androstene-3,17-dione are frequently present in over-the-counter (OTC) capsules. The nandrolone-marker metabolite 19-nor-androsterone may be found in urine samples obtained after oral administration of OTC capsules containing androstenedione.
The urinary concentrations are small but, if these samples are from football players, practically all subjects would have tested positive for the nandrolone-marker 19-nor-androsterone (Catlin et al, 2000). Possibly, in OTC capsules containing androstenedione, the 19-nor-androstenedione could be present as contaminant released, e.g. during manufacturing.
Thus, the presence of the nandrolone marker metabolite in urine samples may be due to the metabolic conversion by the player body of the contaminant 19-nor-androstenedione to the doping-marker 19-nor-androsterone. We cannot, however, exclude the conversion by the athlete body of androstenedione itself to 19-nor-androstenedione, because aromatisable androgens may be converted to estrogens in humans, and this conversion is associated with C-19 demethylation to 19-nor-steroids.
Use of homeopathic products containing sterols and steroids Vegetable or animal sterols and steroids may be present in homeopathic products.
In accordance with the European and the Italian legislation, the homeopathic products should be characterised by a concentration of active component equal to or less than 1/100 of the lower dosage of active molecule in allopathic preparations. In that case, 1/100 of the lower dosage of nandrolone phenpropionate (25 mg/week, equivalent to 3.6 mg/day) corresponds to a concentration of 36 mg active molecule in homeopathic mother tincture that, in turn, is gradually diluted (potentised: D1 = 1:10; D2 = 1:100; D3 = 1:1,000; etc.).) to a relative high degree. Thus, the unintentional increase of 19-nor-androsterone in urine samples by use of homeopathic products can be ruled out because of the very small quantities of steroids administered.
On the other hand, does not seem to be feasible the biochemical conversion by the body of vegetable or animal sterols and steroids into the doping-marker
19-nor-androsterone.
Use of nandrolone as medicinal product We consider the use of intramuscular injections of medicinal products containing nandrolone to be unlikely because the injection of nandrolone phenpropionate, or decanoate, or undecylate, and so on, supports the presence of the hormone in the athlete body for about one, or two, or three weeks, respectively.
Thus, for long time it is easy to found the 19-nor-androsterone in urines by doping controls. This does not seem the case of oral ingestion of tablets, powder, etc. of anabolic steroids or pro-hormones during the post-match recovery period (the sporting activity came to a standstill) or just at the first day of the “three, four or five day break” between two football matches.
The presence in the body of the doping-marker metabolite may be limited to one or two days after the ingestion, increasing the effect of the training performed during this break and maintaining in the football players the induced ‘fight and flight’ characteristics during the next match. During the match the “tail” of psychophysical effects induced by nandrolone may be still present, even if nandrolone itself and its metabolites were eliminated by the body one or two days before, possibly avoiding the presence of 19-nor-androsterone in urine during an eventual doping control.
Thus, a drug action “tail” will be consistent with the drug molecule “ghost”. Of course, this doping protocol by alternate anabolic steroid oral administration is quite intricate and an error in the programme of administration allows finding 19-nor-androsterone in urine samples. Conclusions We consider the presence in urine samples of the doping-marker metabolite 19-nor-androsterone in a concentration greater than 2 ng/ml for males (or 5 ng/ml for females) as the result of use of nandrolone and/or its pro-hormones.
Particularly, the quoted hormones may be incorporated in food supplements illegally imported from the US or other extra-European area. Raised concentrations of nandrolone metabolite(s) were lately found in the urine samples of many football players.
The recent “nandrolone lesson” confirms that the inclusion of drugs in the IOC lists of forbidden substances does not seem suitable to prevent drug abuse and misuse in sports if the institutional bodies are unwilling, or are turning a blind eye on doping events, or are taking a soft stand against the doping offenders, and so on.
A forty-year-old ambiguous policy was unreliable to block the drug abuse and misuse in sports. As the saying goes, the bad workman blames his tools. We must, however, consider the question from all angles. Sports are activities by industrial and commercial concerns, sometimes with shares quoted on the Stock Exchange.
The football players and, generally speaking, the athletes may be more or less well-paid machine tools to make money and/or power. The market requires increasing productivity of human machine tools. Drug abuse and misuse are intended to be a way.
To break the ‘sports connection’ we need an international strong action - mainly based on transparent information - also to avoid the increasing dissemination of drug abuse and misuse in amateur and young population.
The kind of action is unlikely because puts a spoke in interested parties’ wheel. On the other hand, as the saying goes, we can’t make an omelette without breaking the eggs. Otherwise, it still remains to give the very expensive antidoping action as a bad job and to give free rein to drug abuse and misuse in sports.

Gianni Benzi
Dipartimento di Scienze Fisiologiche-Farmacologiche – Università degli Studi di Pavia – Italia European Medicines Evaluation Agency (EMEA) - London - UK