The document presents methods for the estimation and utilization of various phytoconstituents, including forskolin, digoxin, vinca alkaloids, and podophyllotoxin. It details their biological sources, extraction processes, identification tests, and utilization in treatment for conditions such as cardiovascular issues and cancer. Each compound's specific methods of cultivation and chemical properties are also outlined, alongside their therapeutic applications in traditional and modern medicine.

1. Industrial Production ,
Estimation and Utilization of
the following
Phytoconstituents
Presented By-
Mrs. Poonam Nilesh Chougule
Assistant Professor
AMCP- Peth Vadgaon.

2.  Forskolin,
 Sennoside,
 Artemisinin,
 Diosgenin,
 Digoxin,
 Atropine,
 Podophyllotoxin,
 Caffeine,
 Taxol,
 Vincristine and
 Vinblastine.

3. Forskolin
 Synonym- Forskohlii, Plectranthus
barbalus
 Biological Source- Forskolin is a
diterpenes is obtained from the dried
roots of Coleus forskohlii
 Family- Labiatae
 Method of cultivation- Vegetative
method-stem cutting.
 Harvesting – In the month of Autumn.

4. Chemical constituents-
 Forskolin is a diterpenes.
 It contains various diterpenoid
derivatives.
 The leaves give Coleon E
(Methylenequinine), barbatusin and
Coleon F.
 The root contains Coleonol B,
Coleonol C, Deoxycoleonol.

5. Leaves and
flowering top
Roots

6. Extraction and Isolation
 Dried roots of Coleus forskohlii
 Extract with Chloroform or Ethyl acetate (1 :
10) using soxhlet apparatus
 Collect the organic extract
 Evaporate to dryness to yield crude extract of
forskolin.
 Saparation done by Column chromatography
and by using activated charcoal.
 The residue obtained from the eluate is
purified and crystalized by usnig organic
solvent
 Pure form of Forskolin obtained.

7. Identification test-
 TLC Method-
 Stationary phase- Absorbent- pre-
coated Silica gel
 Mobile phase- Benzene: Ethyl Acetate
(85: 15)
 Sample – Root extract
 Detecting agent- Anisaldehyde
sulphuric acid.
 Observation- Violet to Purple color
spot.

8. Estimation
 Forskolin is estimated by two methods,
 I . Method 1 - HPLC
 II. Method 2 – HPTLC
 1. HPLC-
 Chromatographic System- HPLC equipped with 1) LC 8A
Pump and II) Photo array detector in combination with class
LC10A Software.
 Chromatographic condition-
 Column- ODS (Octodecylsilane) C ₁₈, 5 micrometer 250*
4.6mm
 Mobile phase- Acetonitrile: Water (50: 50)
 Wavelength- 220 nm
 Flow rate- 1.6ml/ min
 Injection volume- 20 microlitre.

9.  Standard Preparation:
Dissolve 10mg of forskolin with 15ml of
acetonitrile in 25ml of volumetric flask and
makeup the volume up to 25ml with
acetonitrile.
 Sample Preparation:
Dissolve 250mg of sample with 25ml of
acetonitrile. Filter the solution & makeup
the volume up to 100ml with actonitrile.
 Procedure: Inject the standard and sample
preparation (10μl) & record the
chromatogram. Now calculate the
percentage of Forskolin content from the

10. Utilization:
 Forskolin has Vasodilator and cardio
stimulant effects. Drug is used for
treatment of glaucoma, CHF and
asthma.
 In Ayurvedic traditional medicine it
prefered for heart diseases and
abdominal colic.
 It also used in the treatment of
hypotension, spasmolytic and platelet
aggregation inhibitory activity.

11. Digoxin
 Synonym- Foxglove leaves, digitalis.
 Biological source-
Digoxin is Cardiac glycoside obtained
from the dried leaves of plant Digitalis
purpurea and Digitalis lanata.
 Family- Scrophulariaceae
 Method of cultivation- Seed
propagation method.
 Harvesting- Hand picking method.

12. Digitalis leaves

13. Extraction and Isolation
 Coarsely powdered leaves extracted with 50%
Ethanol at low temp. and filter,
 To the filtrate add Lead acetate solution, impurities
ppt out
 Centrifuge to remove the ppt
 Cardiac glycosides present in the supernatant
liquid
 Extract with chloroform and collect chloroform
extract.
 Evaporate under vacuum
 The residue of Cardiac glycoside obtained
 Purify it by chromatography.
 Get pure Digoxin.

14. Identification test-
 Chemical test- By Keller- Killani test,
Legal test and Baljet test.
 TLC method-
Stationary phase- Absorbent Pre-coated
Silica gel
Mobile phase- Benzene: Ethanol (7:3)
Sample- plant extract (leaves)
Detecting agent- p-Anisaldehyde,
perchloric acid
Observation- UV 350nm, Blue spot.

15. Estimation-
 Colorimetric estimation-
Digoxin extract is treated with 3,5-
Dinitro benzoic acid and Benzyl
trimethyl ammonium hydroxide.
Bluish Red color is formed.
Calorimetrically measured at 550nm.

16.  Assay of Digoxin: IP 66
Weigh about 40mg of digoxin, dissolve in
95% ethanol and makeup volume to 50
ml.
Pipette out 5ml from it and makeup
volume to 100ml again with 95% ethanol.
Again pipette out 5ml from above solution
and add 3ml alkaline picric acid.
Allow to stand for 30 mins.
Measure the absorbance at 495nm.
 Utilization- Mainly used in the treatment of
CHF.

17. Vinca alkaloids
 Synonym- Periwinkle leaf
 Biological source- Vincristine and
Vinblastine are Dimeric indole alkaloid
obtained from the whole plants of
Catharanthus roseus.
 Family- Apocynaceae
 Chemical constituents- about 90
alkaloids isolated from catharanthus
plant. Important alkaloids are dimer
indole alkaloids Vincristine and
Vinblastine which posses Anti-cancer
activity.

18. Extraction and Isolation-
 Vinca leaves and herbs are extracted with
aqueous alcoholic acetic acid solution.
 After concentration of aqueous extract the
residue further extracted with 2% HCL.
 NaOH is used to adjust the pH of extract to 4.
 This is again extracted with benzene.
 The pH of solution again raised to 7.
 Again extracted with benzene.
 pH 7 benzene extract are concentrated, dried
and dissolved in benzene- methylene
chloride (65:35) solution.

19.  This is separated by chromatography
on neutral alumina column with
benzene- methylene chloride solution
as eluent technique.
 Evapouration of the vinblastine- rich
fractions gives vinblastine sulphate
which is recrystalised from alcohol.
 Further elution of the column gives
vincristine which is dried and
crystallized from alcohol.

20. Identification test
 By TLC method-
Stationary phase- Absorbant pre coated
silica gel
Mobile phase- ACN : Benzene (30:70)
Sample- 1mg in 0.1ml of 25% H2o in
methanol.
Detecting agent- 1% solution of Cerric
ammonia sulfate in 85% phosphoric
acid.
Rf value- 0.39

21. Estimation-
 HPLC method
 Utilisation-
 Vinblastine sulphate is used for the treatment
of Hodgkins disease.
 Vincristine sulphate is used for acute
haemolytic leukaemia. Highly active in
childhood leukaemia. And also used in breast
cancer.
 Both vincristine and vinblastine are resulting
in tumour cell death during replication of
DNA.
 They are used as intravenously.

22. Podophyllotoxin
 Synonym- Indian podophyllum,
Himalayan may-apple.
 Biological source- Podophyllotoxin is
resin obtained from the rhizomes and
roots of Podophyllum hexandrum or
podophyllum emodi.
 Family- Berberidaceae
 It grown in forest region of Himalyas
from kashmir to sikkim in Himachal
pradesh.

23. Chemical Constituents-
 Podophyllum contains 3.5 to 6% of resin
whose active principles are Lignans, its
biosynthethically flavonoids.
 The most important ones present in the
podophyllumresin, podophyllitoxin, α-
peltatin and β-peltatin.
 Indian Podophyllum is derived of α and
β- peltatin.
 Also presence of
dimethylpodophyllotoxin,
dehydropodophyllotoxin and quercetin-a-
tetra-hydroxy flavonol.

24. Extraction and Isolation
 Podophyllum resin extracted from
dried rhizomes and roots using an
alcoholic percolation method.
 It contains not less than 40% of
podophyllum resin.

25.  The powdered and dried rhizomes and roots
of podophyllum are extracted with 90%
ethanol in soxhlet extractor.
 Alcohol is removed by distillation and the
alcoholic extract as concentrated till it
become syrup-like consistancy.
 A mixture of water containing 2% HCL is
poured into this syrup at 5ᵒC with continuous
stirring.
 Mixture allow stand for 2hrs below 5ᵒC and
then dissolved in sufficient quantity of 90%
hot alcohol.
 This solution is filtered and alcohol distilled
off.

26. Utilization-
 It is used as a drastic but slow acting
purgative.
 Podophyllotoxin possesses Anti-
tumour properties and may be used in
the treatment of Cancer.
 It is invariable prescribed with other
purgatives, henbane or belladonna to
prevent gripping action in infants.
 It is used in the semi-solid synthetic
production of Etoposide.

27. Thank You