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  • 1. :Prepared byNajah Al-harbi
  • 2. Dianabol• Dianabol (1 7 alpha-methyl-1 7 beta-hydroxil-androsta-1.4 dien-3-on)• is an anabolic steroid with moderate to high androgenic qualities.• is an orally applicable steroid with a great effect on the protein metabolism.
  • 3. ?What are steroids• Powerful drugs that many people use as a short cut to boost their athletic performance or improve their appearance.• Properly called anabolic-androgenic steroids.
  • 4. What do “anabolic” and “androgenic” mean?Anabolic = ““building body tissueAnabolic-androgenic steroids help to increase muscle mass and body weight, and speed up bone maturationAndrogenic“ = “promoting male characteristicsAnabolic-androgenic steroids promote development of facialhair, deeper voice, balding, and other male characteristicsAll““anabolic” steroids are also “androgenic
  • 5. What? do steroids do• Doctors sometimes prescribe low doses of anabolic- androgenic steroids to treat people with serious medical conditions.• Example: A man’s testosterone level decreases due to a testicular tumor – Dose prescribed is only enough to bring the testosterone level back up to normal – Testosterone levels and side effects are monitored closely by the doctor
  • 6. ?Why do people use steroids• To make muscles bigger and stronger• To be a better athlete• To get an edge over the competition• To look better• Pressure from friends or coach
  • 7. DianabolHistory• Methandrostenolone (Dianabol) is an anabolic steroid originally developed by John Ziegler and released in the US in 1956 by Ciba.• It was used as an aid to muscle growth by bodybuilders until its ban by the FDA under the Controlled Substances Act.• Despite this, methandrostenolone continues to be produced in countries such as Mexico under the trade name Reforvit-b, and is being manufactured in Russia, as well as Thailand, and subsequently is still seen on the United States black market.• Production in most of Western Europe and the United States has ceased.
  • 8. .Cont• Methandrostenolone does not react strongly with the androgen receptor, instead relying on activity not mediated by the receptor for its effects.• These include dramatic increases in protein synthesis, glycogenolysis, and muscle strength over a short space of time.• However, due to its mode of action, it decreases the rate of cell respiration and decreases production of red blood cells.
  • 9. ?How to be use• Dianabol has a short half life of only about 3-5 hours.• As a result, multiple applications of dianabol throughout the day are necessary to achieve a stable blood level.• Dianabol should also be taken with food if possible to decrease possible gastrointestinal pain which may result from its use.• Dianabol reaches the bloodstream within 1-3 hours.
  • 10. .Cont• dianabol is normally used in a mass building stack to jump- start gains in muscle mass and strength as well as to improve pumps.• The norm is to use a dose of 15-40 mg a day for a period of 4-6 weeks for this purpose.• Dianabol should not be used for more than eight weeks at the absolute maximum and only under supervision from a health care professional.• Blood work is important for all enhanced athletes. A full blood panel should be carried out before and after a cycle and assessed by a certified health care practitioner.
  • 11. ?What are the health risks• It´s important to note that steroid abuse is the cause of many side effects, and that the simple use of steroids can not be blamed for this. It´s when the line is crossed from use into abuse that we see all of these effects as being not only possible but probable.• It is metabolized into estradiol by aromatase. This means that without the administration of aromatase inhibitors such as Anastrozole or Aminoglutethimide, estrogenic effects will appear over time in men.
  • 12. Harmful side effects• Normally, if doses are kept low (15-20 mg daily), these side effect are less pronounced and sometimes not seen at all depending on the invidual users response to the drug.• Due to dianabol falling in the class of 17-AA steroids, it is liver toxic and can cause considerable strain on the liver in high doses and over extended periods of time.• Even in doses as small as only 10 mg a day, increases in liver values may be seen, although when such moderate doses are used, liver values tend to return to normal after use of dianabol is halted.
  • 13. Harmful side effects• Since dianabol quickly increases body weight due to both hypertrophy and excess fluid retention, increases in both blood pressure and heart rate can occur, and in extreme cases may require the use of an antihypertensive drug such as catapressin.• Bearing in mind that dianabol readily converts into estrogens, gynecomastia is always a possibility for the dianabol using athlete.• Dianabol may also cause serious acne on the face, neck, chest, back and shoulders since the sebaceous gland is stimulated due to dianabols conversion into dihydrotestosterone.• If a genetic predisposition to hair loss is present in an individual, the use of dianabol may accelerate it.
  • 14. Harmful side effects• The drug causes severe masculinising effects in women even at low doses.• In high dosages of 50 mg+/ day aggressive behavior in the user can occasionally be ebserved.• After dianabol use is discontinued, a loss in strength may be seen since excess fluid retention is reversed.• On the plus side, dianabol is a "feel good" steoroid known to improve: mood, appetite, self confidence, and self esteem.
  • 15. precaution• Water intake should be kept HIGH while using dianabol. This is one of those things that just cant be stressed enough. This will help your organs and especially your liver in the metabolism of dianabol.• As a recommendation, try to take in between 1-2 gallons of water each day while dianabol is used.• For any enhanced athletes, water intake should be kept high.
  • 16. Biological assay• The success of this legislation and/or other interventions assumes the availability of robust screens that can detect the presence of all performance-enhancing drugs.• In the case of anabolic steroids, current approaches utilize sensitive assays such as GC/MS or ELISA to detect the presence of known steroids in biological samples.• These assays suffer from a severe limitation: they require pre- existing knowledge of the precise compounds that are being abused.• An athlete can evade detection by using an anabolic steroid that is not known to authorities .
  • 17. .Cont• Indeed, an emerging clandestine industry has allowed athletes to evade detection via the use of novel “designer steroids”.• Several test-evading designer steroids have been identified in the past 3 years including two existing compounds that were never marketed and a novel chemical entity that was specifically synthesized to evade detection (tetrahydrogestrinone, THG).
  • 18. Reagents and InstrumentsGas chromatography/mass-spectrometry (GC/MS)• A ThermoFinnigan Trace DSQ GC/MS system was used for these studies. Gas chromatographic separation was performed with a Phenomenex ZB-5 (5% phenyl-95% dimethyl-polysiloxane) column (15 meters, 0.25 mm ID, film thickness 0.50 µm). Following data acquisition, total ion spectra found in the samples were compared to known spectra contained in the NIST 98 Library using the Finnigan Xcalibur software.
  • 19. MethodsIsolation and identification of anabolic steroids• A single lot of normal human male serum (Gemini cat#100-512, Lot# H01904Y) and a single lot of normal human female serum (Gemini cat#100-110F, Lot# H00903Y) were used throughout these experiments in order to exclude interindividual differences as a source of variation. Where indicated, serum was spiked with 0.1-10 nM nandrolone (Steraloids cat# E4050-000), 0.1-10 nM methandienone (Steraloids cat# A0130-020) and in some cases female serum was adjusted to contain concentrations of testosterone (T, 15 nM) and dihydrotestosterone (DHT, 3 nM) that are within the range normally present in healthy adult men. For analysis, serum (2.5 ml) was extracted two-times with 2 volumes of ethyl acetate. The two ethyl acetate fractions were pooled, washed twice with water and then dried under nitrogen.
  • 20. .Cont• The steroid-screening assay was performed by incubating the ethyl acetate fraction with agarose- bound androgen receptor ligand binding domain (AR).• To do so, thioredoxin-linked AR (~30 µg total protein; specific activity 2.2 pmol bound ligand/µg, Invitrogen) was incubated with 40 µl Ni-NTA agarose (50% slurry) in a total volume of 300 µl of buffer A (20mM Tris, pH 8, 500mM NaCl, 5mM imidazole) at 4 °C for 1 hr.• Ni-NTA beads were pelleted and washed once with 1 ml of ligand binding buffer (30 mM Tris pH 8.0, 150 mM KCl) then resuspended in 300 µl of ligand binding buffer containing 0.4 mg/ml ovalbumin.
  • 21. .Cont• Immediately prior to assay, the dried ethyl acetate serum fraction was dissolved in 2 ml of ligand binding buffer by incubating for 30 min at room temperature and then incubated with the immobilized AR beads for 1 hr at room temperature followed by 1 hr at 4 °C.• Immobilized AR was then pelleted and washed once with 2 ml ligand binding buffer to remove any unbound ligand.• To dissociate the bound ligands from receptor, immobilized AR was incubated for 30 min. at 65 °C in 400 µl PBS and the released ligands were collected in the supernatant.• This elution was repeated and the two PBS- fractions were pooled, filtered through a 0.22 µm PTFE filter, and extracted two-times with 3-volumes of ethyl acetate. The organic phases were pooled and dried under nitrogen.
  • 22. .Cont• The dried samples were derivatized with 20 µl of N,O -bis (trimethylsilyl) trifluoroacetamide containing 1% trimethylchlorosilane (Pierce) at 55 °C for 6 hr, then 4 µl were analyzed by GC/MS run in scanning mode over a M/Z range of 50-700. At the start of each run, the column temperature was held at 50 °C for 1 minute, increased using a gradient of 25 °C /min up to 300 °C, and held at 300 °C for an additional 8 minutes.