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Steroid Precursors and Prohibited Substances. Athletic Drug Testing: What Is It?.

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Athletic Drug Testing: What Is It?

  • “Anti-doping programs seek to preserve what is intrinsically valuable about sport. This intrinsic value is often referred to as ‘the spirit of sport,’ it is the essence of Olympism; it is how we play true. The spirit of sport is the celebration of the human spirit, body and mind, and is characterized by the following values

    • Ethics, fair play and honesty

    • Health

    • Excellence in performance

    • Character and education

    • Fun and joy

    • Teamwork

    • Dedication and commitment

    • Respect for rules and laws

    • Respect for self and other participants

    • Courage

    • Community and solidarity

  • Doping is fundamentally contrary to the spirit of sport”

World Anti-Doping Agency. The World Anti-Doping Code. Accessed; 12/15/2010. Available at:

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Athletic Drug Testing: Who Monitors?

  • World Anti-Doping Agency (WADA)1

    • Established in 1999 as an international independent agency composed and funded equally by the sport movement and governments of the world

    • Key activities include scientific research, education, development of anti-doping capacities, and monitoring of the World Anti-Doping Code, which harmonizes anti-doping policies across all sports and countries    

  • United States Anti-Doping Association (USADA)2

    • Established in 2000 as the national anti-doping organization for the Olympic movement in the United States

    • The US Congress recognized USADA as "the official anti-doping agency for Olympic, Pan American and Paralympic sport in the United States.“

  • National Collegiate Athletic Association (NCAA)3

    • When Proposal No. 30 was approved at the 1986 NCAA Convention and Proposal Nos. 52-54 were approved at the January 1990 Convention, NCAA institutions reaffirmed their dedication to the ideal of fair and equitable competition at their championships and postseason certified events




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Athletic Drug Testing: How Do They Monitor?

  • Athletes are selected randomly and may be tested “in season” or “out of season”

  • A urine sample is given under close scrutiny of a testing officer, and volume, pH, temperature, and in some cases specific gravity of the sample are immediately tested to ensure that there has been no attempt to alter the sample

    • At least 75 mL must be given under close scrutiny

    • The urine is split into 2 portions (bottles "A" and "B“)

  • Only the athlete handles the urine and collection containers until sealed with tamper-proof strips, overwrapped with tamper-evident seals, and coded

  • In some cases, blood samples are taken and undergo a similar rigorous collection and analysis process

  • The independent official observing the sample procedure records the information and ensures that a chain-of-custody record is continuous

    • This guarantees that the sample content is protected and that the sample tested is from the correct athlete

  • The anonymous samples are analyzed by an independent, accredited laboratory for presence of substances banned for a given sport

    • In many cases, the unused sample may be stored for future testing

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Athletic Drug Testing: Where Does Liability Lie?

  • Athletic commissions maintain a “strict liability” policy regarding drug testing

    • Athletes are responsible for any prohibited substance, metabolites, or markers in their body, whether intentional or accidental

  • Some brands of dietary supplements can contain unknown contaminants that may contain trace amounts of banned substances

    • A 2007 study of dietary supplements found that 12.5% contained banned substances not declared on the label

  • NSF International certification program

    • Provides assurance to athletes that the supplements they use will not contain a banned substance

Abbreviation: NFL, National Football League.

Martello S, et al. Food Addit Contam. 2007;24(3):258-365.

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Permissible and Nonpermissible Substances

  • Agents prohibited at all times

    • Anabolic agents

      • Steroids, steroid precursors

    • Hormone antagonists and modulators

      • Erythropoeitin (EPO)

    • Chemical and physical manipulation

      • Intravenous fluids or blood products, unless exempted

    • Specified substances

      • Beta blockers, masking agents, or diuretics

  • Major sources of information on prohibited substances

    • NCAA Banned Substance List

    • World Antidoping Agency (WADA)

    • United States Antidoping Association (USADA)

Abbreviation: NCAA, National Collegiate Athletic Association.

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NCAA Rules Regarding Nutritional Supplements (1)

  • Colleges and coaches are limited in what nutritional supplements can be provided to athletes

    • No muscle-building supplements and products with unproven or potentially unsafe ingredients can be provided

    • Institutional staff cannot sell supplements to athletes

    • Over-the-counter and prescription products may also be prohibited1

      • Decongestants and weight loss products that contain phenylpropanolamine (a stimulant)

      • Beta-2 agonists (asthma medications)

      • Ephedrine

      • Alcohol

      • Caffeine (urinary levels ≥ 15 μg/mL are not allowed)

        • ~5 regular cups of coffee within a few hours of testing for a healthy and average-sized man2

Abbreviation: NCAA, National Collegiate Athletic Association.

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NCAA Rules Regarding Nutritional Supplements (2)

  • Permissible to provide to athletes

    • Vitamins/minerals

    • Energy bars

      • May contain no more than 30% of kcals from protein

    • Calorie replacement drinks

      • Shake-type protein drinks with no more than 30% of kcals from protein

    • Electrolyte replacement drinks

  • Not permissible to provide to athletes

    • Amino acids (including amino acid chelates)

    • Chondroitin

    • Chrysin

    • CLA

    • Creatine/creatine-containing compounds

    • Garcinia cambogia (hydroxycitric acid)

    • Ginkgo biloba

    • Ginseng

    • Glucosamine

    • Glycerol

    • Green tea

    • HMB (hydroxy-methylbutyrate)

    • Melatonin

    • MSM (methyl sulfonyl methane)

    • Protein powders

    • St. John’s Wort

    • Tribulus

    • Weight gainers

    • Yohimbe

Adapted from NCAA bylaw 16.5.2.g. Available at:

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Substance Classes of Concern

  • Testosterone and its structural analogues (anabolic steroids)

  • Precursors of testosterone (prohormones)

  • Human growth hormone (HGH)/insulin-like growth factor 1 (IGF-1)

  • Human chorionic gonadotropin (HCG)

  • Clenbuterol

  • Aromatase inhibitors

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Testosterone and Anabolic Steroids

  • Testosterone and its analogues work mainly by interacting with androgen receptors

    • 2 main sites of androgen receptors are muscle myonuclei and satellite cells

  • Once testosterone or its analogues dock with the receptors, processes are set in motion that can increase muscle protein synthesis

  • Effectiveness of anabolic steroids was debated and questioned for a long time

    • Many studies used dosages much lower than typically used by athletes

    • To discourage steroid use, the efficacy/activity of steroids was not communicated

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Benefit/Risk of Anabolic Steroid Use


  • Increased muscle mass and strength

  • Improved recovery after workouts

  • Increased aggressiveness, which may intensify training


  • Cardiovascular disease

    • Dramatic decreases in HDL

    • LDL may increase

  • Liver damage

    • Peliosis hepatitis, esophageal varices resulting from portal hypertension

  • Sexual side effects

    • In males: gynecomastia, testicular atrophy, and decreased sperm count

    • In females: masculinization

  • Reduced tendon and ligament flexibility (more tears)

  • Aggressiveness (‘roid rage) and withdrawal symptoms

  • Acne

The American College of Sports Medicine issued a position statement in 1984 deploring the use of anabolic steroids by athletes (based on health risks and ethics)1

1. American College of Sports Medicine. Med Sci Sports Exerc. 1987;19(5):534-539.

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Testosterone Increases Muscle Size and Strength With or Without Exercise

  • Subjects

  • 43 healthy young men across 4 groups

  • Treatment

  • 600 mg testosterone or placebo weekly for 10 weeks

  • Exercisers lifted weights 3 times/week

aP < .05 for the comparison between the change indicated and that in either no-exercise group.

b P < .05 for the comparison between the change indicated and that in the group assigned to placebo with no exercise.

cP < .05 for the comparison between the change indicated and the changes in all 3 other groups.

Reprinted from Bhasin S, et al. N Engl J Med. 1996;335(1):1-7.

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Testosterone Precursors/Prohormones

  • Androstenedione/androstenediol

  • 19-Norandrostenedione/19-norandrostendiol

  • Dehydroepiandrosterone (DHEA)

  • Others

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Summary of Prohormones

  • Reasons for not using prohormone supplements

    • Poor quality control

      • Study of 16 DHEA products found that only 7 were within 90% to 110% of stated label claim

      • 3 had no DHEA at all, 1 was 150% of label claim

    • Uncertainty regarding optimal doses

    • Potential safety concerns

      • Possible stimulation of the growth of cancers or hypertrophy of tissues such as the prostate

      • Androgenic effects (eg, facial hair in females)

    • Research on possible health benefits of prohormones (eg, bone density, cognition) is sketchy at best

    • Similar lack of efficacy for the nor- versions of androstenediol/androstenedione at 156 mg/day (combination) for 8 weeks in resistance-trained men

Abbreviation: DHEA, 5-dehydroepiandrosterone.

Parasrampuria J, et al. JAMA. 1998;280(18):1565; Van Gammeren D, et al. Eur J Appl Physiol 2001;84:426.

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Summary of Androstenedione (Andro)

  • One biochemical step away from testosterone

  • Best study was performed by King et al

    • 20 weight-training subjects

    • 8 weeks of weight training

    • 100 mg andro 3 times/day for 3 different 2-week periods during training (rice flour placebo)

  • Beyond effect of exercise itself, andro:

    • Did not increase muscle mass or strength

    • Did not increase free or total testosterone

    • Did not decrease body fat

    • Decreased HDL cholesterol

    • Increased estradiol and estrone

Abbreviation: HDL, high-density lipoprotein.

King DS, et al. JAMA. 1999;281(21):2020-2028.

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Summary of 5-Dehydroepiandrosterone (DHEA)

  • DHEA and its sulfated conjugate DHEA-S are the steroids of greatest abundance in the blood

  • DHEA can be converted to androstenedione, which then can be made into testosterone

  • Levels of DHEA typically decrease after age 25

  • Supplementation of weight lifters with 150 mg DHEA/d for 8 weeks had no effects on testosterone, body composition, or strength beyond the effect of training

  • Other studies have also generally not shown efficacy for strength or muscle mass in older and non-exercising populations

Brown GA, et al. J Appl Physiol. 1999;87(6):2274-2283.

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Human Growth Hormone (HGH)

  • HGH is normally produced by the pituitary gland

    • Not a steroid, but nonspecifically promotes growth of various tissues (eg, bone, organs)

    • HGH levels naturally decrease with age

    • Many of the biologic effects of HGH are from increased production of insulin-like growth factor 1 (IGF-1)

    • Major effects include reduction in body fat and increased lean mass

    • Effects are most pronounced in elderly and untrained subjects

Blackman MR, et al. JAMA. 2002;288(18):2282-2292; Rudman D, et al. N Engl J Med. 1990;323(1):1-6.

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Human Growth Hormone (HGH)

  • Even though there is evidence for altered body composition, there are a number of concerns associated with HGH

    • No evidence that HGH actually improves muscle function, strength, or performance in young adults or trained subjects

      • Inconsistent results for body composition changes

    • Potential for a wide variety of side effects

      • Acromegaly/gigantism, organomegaly

      • Carpal tunnel syndrome

      • Swollen feet and ankles

      • Joint pain

      • Glucose intolerance

  • Supplementation of HGH is now more prevalent than ever because of easy availability of recombinant HGH

    • Banned by both WADA and NCAA, and there is now a blood test

    • May cost ≥ $1,000 per month

Abbreviations: NCAA, National Collegiate Athletic Association; WADA, World Anti-Doping Agency.

Crist DM, et al. J Appl Physiol. 1988;65(2):579-584; Deyssig R, et al. Acta Endocrinol (Copenh). 1993;128(4):313-318; Yarasheski KE, et al. Am J Physiol. 1992;262(3):E261-E267; Yarasheski KE, et al. J Appl Physiol. 1993;74(6):3073-3076.

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Human Chorionic Gonadotropin (HCG)

  • Peptide hormone that arises in the placenta during pregnancy

    • Similar in structure to luteinizing hormone

  • Male steroid users employ HCG after a cycle of steroids to stimulate the testes to restart production of testosterone

  • May have some anabolic effects

  • Banned in males by both NCAA and WADA

Abbreviations: NCAA, National Collegiate Athletic Association; WADA, World Anti-Doping Agency .

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  • Not a steroid, but rather a 2-adrenergic agonist

  • Human use is illegal in the United States

    • Some parts of Europe use inhaled (not oral) clenbuterol for humans as a bronchodilator

    • Clenbuterol is approved for some veterinary applications in the US (eg, horses with lung obstruction)

  • Popular with bodybuilders because of fat-burning and muscle mass-building properties (no studies of these properties in humans)

  • Side effects have occurred in people eating the meat of animals that received clenbuterol

    • Skeletal muscle tremors, agitation, palpitations, dizziness, nausea, muscle cramps, rapid heart rate, and headache

  • Clenbuterol use carries the risk of toxicity, is illegal in the US, and it is on the WADA and NCAA banned substance lists

Abbreviations: NCAA, National Collegiate Athletic Association; WADA, World Anti-Doping Agency.

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  • WADA, USADA, and NCAA are the primary agencies responsible for establishing requirements for athletic supplements

  • It is the athlete’s responsibility to ensure that no banned substances are in his/her blood

  • Although some banned substances may contribute to increased strength, speed, or athletic ability

    • They are associated with serious adverse effects

    • They have inconsistent efficacy (if any)

    • Their detection can result in elimination from sport participation

  • Nutritional supplements may also be considered prohibited or restricted and may contain traces of banned substances

    • For supplements, look for certification of the absence of banned substances and other unsafe contaminants by NSF International or other organizations

Abbreviations: NCAA, National Collegiate Athletic Association; USADA, United States Anti-Doping Association ; WADA, World Anti-Doping Agency.