1. :Prepared by
Najah 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 tissue
Anabolic-androgenic steroids help to increase muscle mass
and body weight, and speed up bone maturation
Androgenic“ = “promoting male characteristics
Anabolic-androgenic steroids promote development of facial
hair, deeper voice, balding, and other male characteristics
All““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 user's
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 dianabol's 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 can't 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 Instruments
Gas 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.

20. Methods
Isolation 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.

21. .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.

22. .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.

23. .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.