The document discusses diosgenin, a precursor used in pharmaceuticals like oral contraceptives. It is commonly extracted from Dioscorea zingiberensis tubers using acid hydrolysis. The document outlines methods to extract and isolate diosgenin from plant materials like seeds and hairy roots, such as refluxing with sulfuric acid and eluting with hexane. Analytical methods to analyze diosgenin are also presented, including thin layer chromatography using antimony trichloride as a detecting agent. Diosgenin is useful as it can be used in progesterone semi-synthesis and produces estrogenic activity.

1. NADEEM SIDDIQUI
1

2. NADEEM SIDDIQUI 2
 Diosgenin, is a precursor for partial
synthesis of oral contraceptives, sex
hormones, and other steroids, is widely
used in pharmaceutical industry.
Nowadays, many researchers found that
diosgenin has antiproliferative and
proapoptotic effects on cancer cells or on
rheumatoid arthritis synoviocytes.

3. It also shows pharmacological activities such as
antilipoperoxidative and antiskin aging effect.
The main raw material used in industry is Dioscorea
zingiberensis C. H. Wright (DZW)because of the high
content of diosgenin in its tubers.
NADEEM SIDDIQUI 3

4. Plant material:
Seeds were surface sterilization with a sodium hypochlorite
solution (1.5% W/V available chlorine) supplemented with
two drop triton X- 100 for 6 min and rinsed 3 time with
sterile distilled water
They are lift for germination on agar plates containing 0.5%
sucrose at 25°C in the light with a daily 16-hours
photoperiod.
NADEEM SIDDIQUI 4

5. The contents of flasks are harvested, and
fresh and dry weight, after lyophilization,
will be determination individually.
The hairy roots are powdered, extracted
and hydrolyzed by refluxing for 5 h with 1
ML sulfuric acid in 70% isopropanol.
 the extract is adsorbed on an extrelut
column and diosgenin was eluted with n-
hexane.
The organic solvent is evaporated to
dryness. Root tissues of normal 8-month
old plants grown in the field are extracted.
After filtering off cell debris, the liquid
media are lyophilized and extracted using
the same procedure.
NADEEM SIDDIQUI 5

6. Diosgenin can be isolated by the following analytical
methods
 By TLC method: The TLC profile is given:
a) Mixture of chloroform : ethanol (95:5) or chloroform
acetone (3:1)is the solvent system.
b) Silica gel plates are used as the stationary phase.
c) Antimony trichloride (SbCl3) in chloroform is the
detecting agent used.
 By HPTLC method: A stock solution of diosgenin (100µ
g/ml concentration) is prepared by dissolving 1 ml of
diosgenin in 1 ml of chloroform. A calibration curve 1000-
6000 ng/spot is prepared and analysed for
reproducibility, linearity, and validating the proposed
method.
NADEEM SIDDIQUI 6

7. The correlation coefficient (r), coefficient of variance
(CV%), and the linearity of results are also measured.
To perform TLC diosgenin, silica gel 60 F254 pro-
coated plates (10 × 10 cm) are used with hexane : ethyl
acetate.
By using linomat 5 spotter, the pre-coated TLC plates
are developed up to 10cm in the ascending mode. After
this,the plates are sprayed with 20% antimony chloride
in chloroform, and dried in a chromatographic oven at
105°C for 10 minutes. The diosgenin concentration
in the given sample is estimated by comparing the
area of chromatogram of standard diosgenin.
NADEEM SIDDIQUI 7

8. Diosgenin is used due to the following properties:
1) It is used in the semi synthesis of progesteron.
2) Is is frequently used as the main ingredient in
production of early oral contraceptive agents
3) It acts as an unmodified steroid which shows
estrogenic activity.
4) It helps in reducing the serum cholesterol level
NADEEM SIDDIQUI 8

9. NADEEM SIDDIQUI 9