Phytochemicals: Extraction, Purification & Analysis. Introduction, Some of the medicinal plants & uses, different processes of extraction & purification. Analysis requirement & Analytical technique.

1. Dr. A. Amsavel

2. Content
 Source Of Drugs
 History Of Medicines Used From Plants
 Recent Development & Technology
 Pharmacology Of Photochemical
 Plants, Medicinal Chemistry & Treatments
 Type Of Extractions
 Isolation And Purification
 Analysis of Drug
 Job Opportunities In Pharma Industries
 References

3. Sources Of Drugs
1. Natural substances:
From plants, animals, microorganisms, etc.
2. Semi synthetic substances:
These are drugs that are manufactured by partial
synthesis.
3. Synthetic substances:
These are drugs which are manufactured by synthetic
process

4. Medicines from plant: Early evidence
 60,000 years ago plants were buried with the
Neanderthal man – plants found to be medicinal value
 4,000 year old Sumerian clay tablet recorded numerous
plant remedies
 3500 yrs ago Egyptian civilization had information on
medicinal plants
 4500 years ago in China ~1600 recorded herbal
medicine standard (pharmacopeia).
 Herbal medicines were in use around 3000 years in
India.

5. Path to Modern Medicine
 Many herbs are still in use since several centuries
 Some have been lost, due to lack of document
 Many herbal remedies have sound scientific basis
 Hippocrates (460-377 B.C.), known as the Father of
Medicine, who used various herbal medicine
 William Withering was the first to scientifically
investigate a folk remedy
 His studies (1775-1785) of foxglove to treat heart
failure set standard for pharmaceutical chemistry.

6. Development in 19th & 20th Centuries
 Scientists began Extracting & purifying the active
molecule from medicinal plants
 Breakthrough: Serturner isolated morphine from opium
poppy in 1806
 Today huge number of drugs are isolated from plant or
semi-synthetic
 About 80% of the rural population in developing nations
using herbal medicine [WHO-2005]
 25% of prescriptions drugs written in the U.S. is from
plants
 More than 60% of anticancer & anti-infective drugs are
natural products.

7. Medicine in India
• Ayurvedic medicines were referred in Rig-Veda
• Ancient Medical scholars: Atreya, Agnivesa Charaka &
Shusruta
• 800BC – Shushruta’s written treatise on surgery is the oldest
record.
• 80% population uses Folk & documented systems of medicine
• Ayurveda, Unani, Siddha & Homoeopathy (AYUSH)
• > 7000 plants used for medicinal purpose
• The domestic demand of medicinal plants has been estimated
around 2 lakhs MT
• Turnover of this industry is about 10000 Cr
•

8. Active ingredients in plants
Active contents used for medicine in the plants are;
 Alkaloids
 Glycosides
 Polyphenols
 Antioxidants
 Tannin and etc..

9. Physiological effects of alkaloids
 Alkaloids used as Anesthetics,
Stimulants, Hallucinogens &
Depressants
 > 12,000-alkaloids are known to exist in
~ 20% of plant species
 But only few are exploited as medicinal
purposes.
 Analgesics : morphine and codeine,
 Muscle relaxant: Thiocolchicoside
 Antibiotics: berberine,
 Aanticancer: vinblastine,
 Sedative : scopolamine.
 Addictive stimulants-caffeine, nicotine,
cocaine, & ephedrine

10. Glycosides
Sugar /glucose molecule (glyco-) is attached to the
active molecule & Steroids
Glycosides are used to treatment of ;
 Appetite.
 Heart dieses
 Purgative
 Skin dieses
 Hemolytic anemia
 Anticancer: Amarogentin, ailanthone & polygalin
 Also used to synthesis hormones (progesterone &
cortisone

11. Plants and Medicinal purpose
Plant Dieses Drugs
1. Foxglove - heart disease –digitalis
2. Willow bark - pain, fever - aspirin
3. Fever Bark Tree - malaria - quinine
4. Snakeroot - hypertension - reserpine
5. Vinca - leukemia - vincristine
6. Taxus - ovarian&breast cancer – taxol
7. Gloriosa Superba - Gout -colcicine
8. Vocagine bark -De-addiction -Ibogaine

12. Pharmacological Action:
Treatment Plants
 Anticancer Vinca
 Anti Spasmodic Datura, belladonna
 Anti asthamatics Ephedra
 Anthelmintic Kurchi
 Anti amoebic Ipecac
 Anti diabetic Gymnema
 Cardio tonics Digitalis, squill
 Carminatives Cinnamon, clove,
 Expectorant Vasaka
 Laxatives Senna, Aloe

13. Plant material
 Plant parts contain chemical components that are
medically active;
 Leaves, flowers,
 Roots, rhizomes,
 Stems, barks,
 fruits,
 grains & seeds, or whole
 Collected, processed (dried), and preserved
.

14. Aspirin: Willow Bark
 Bark of willow trees (Salix spp.) used for reducing fever
and relieving pain – used in the form of a tea
 In 1828 Salicin was first extracted as chemical.
 In 1898 Felix Hoffman (Bayer) synthesized
acetylsalicylic acid
 Aspirin:
 antipyretic (fever reducing)
 anti-inflammatory
 analgesic (pain relieving)
 Also used for heart attacks, strokes, gout, and colon
cancer

15. Malaria and Fever bark tree
 Malaria is still the world's most prevalent disease
 2 to 3 million people die each year from malaria
 Fever bark tree (Quina-quina) used for treatment of
malaria
 In 1820 French scientists isolated the alkaloid
quinine
 36 alkaloids found in Cinchona bark and 4 have anti-
malarial properties

16. Snakeroot for hypertension
Sarpaganti-Chivan amelpodi (Rauwolfia
serpentina)
 For over 4000 years, Snakeroot used
for the treatment of snakebites, insect
stings, and even mental illness
 In 1952 the alkaloid reserpine was
isolated from the roots from dozens of
alkaloids
 Reserpine – widely used for treatment
for hypertension

17. Cancer therapy
 Cancer is uncontrolled growth of abnormal cells.
 In the late 1950's National Cancer Institute and
USFDA began search for plants with anti-cancer
properties
 Thousands of plants have been scientifically
screened, and several have become standard
chemotherapy for different forms of cancers

18. Vinblastine and Vincristine
Periwinkle / Vinca rosea :
 Contains > 95 alkaloids in range of 0.2-1%
 Used by traditional healers as treatment for diabetes
 In 1950's, University of Western Ontario (Canada ) and Eli
Lilly Pharmaceuticals found no evidence of usefulness in
treating diabetes
 But Isolated alkaloids from the leaves were effective
against leukemia cells
 Vincristine: Treatments for leukemia (99%
reduction) lymphomas
 Vinblastine- is used for leukemia Hodgkin's disease
 Both alkaloids also used for other types of cancer

19. Vinca Rosea Plant

20. Gloriosa superba seed
 Tuber is used in Ayurvedic
medicine- abortifacient,
antiarthritic, antihaemorrhoid,
antileprotic, purgative , ulcer
piles etc ….
 Soup made from leaf or tuber –
for women ‘s ailment.
 Seed contain: Colchicine &
Colchicoside and several alkaloids
Isolated used in alophahty
medicine
 Anti gout, anti inflammatory,
anticancer etc

21. Pacific yew trees:Taxol
 Taxol is isolated from the bark of the
Pacific yew, Taxus brevifolia
 National Cancer Institute discovered
the Anti-tumor properties
 Clinical trials showed taxol
especially promising in treating
ovarian and breast cancer
 Taxol - Recently approved by USFDA

22.  Raw opium is the air-dried latex from opium poppy (Papaver
somniferum L)
 It contains several alkaloids: -> 10.0% morphine, >2.0% codeine,
Noscapin (~ 6%)
 Morphine-a powerful analgesic
 Codeine (Methyl morphine) & Noscapin - is an antitussive agent
 Several Morphine derivatives used as medicine
Opium

23. Phytochemical Screening & Use
1- Selection of promising plant materials.
2- Proper collection & authentication of selected plants.
3- Drying of plant materials.
4- Grinding of the dried plants.
5 - Packing, storage and preservation
6 – Extraction of constituents.
7 - Methods of separation and purification.
8- Methods of identification of isolated compounds
9-Method of testing for quality
10-Establish safety & Efficacy of drugs

24. Extraction
 Extraction: is the separation of medicinally active
portion of plants by using selective solvent and
suitable methods of extraction.
 Non polar solvents (petroleum ether and hexane) will
dissolve non-polar compounds (fats and waxes).
 Polar solvents (Alcohols, and water) will dissolve polar
compound (alkaloid salts and sugars).
 like dissolve like
 Single or mixture of solvents used based on based on
solubility

25. Methods of extraction :
1. Infusion
2. Decoction
3. Digestion
4. Maceration
5. Continues hot extraction- Soxhlet
6. Supercritical fluid extraction
7. Counter current extraction
8. Ultrasonication-Assisted Extraction
9. Microwave assisted extraction

26. Method of Extraction
Extract the readily soluble
constituents of crude drugs from
plant into solvent medium.
Infusion :
 Macerating for a short period
of time with cold or boiling
water
Decoction :
 Boiled in a specified volume
of water for a defined Time

27. Method of Extraction
Digestion :
 Heat gently during with solvent for
extraction. Extraction of Morphine
Maceration :
 Solid ingredients are placed in a
closed container with solvent and
allowed to stand for at least 3 days
(3 - 7 days) with frequent agitation

28. Soxhlet Extraction
•Continuous extraction of a component from a solid .
•Large amount of drug can be extracted with small
quantity of solvent.
•Efficient, Faster & Economic

29. Supercritical Fluid Extraction
 Extraction using SCF under
controlled pressure and/or
temperature, Eg Carbon dioxide
 Non-toxic solvents
 No harmful residue.
 Thermally labile compounds
can be extracted at low
temperatures

30. Counter-Current Extraction
 A liquid-liquid extraction process
 Solvent and product flow in opposite
directions.
 Crude drug is extracted with much
smaller volume of solvent
 It is done at room temperature or
warm temperature based on the
nature ie thermo labile molecule

31. Ultrasonication-Assisted Extraction:
 Extract under ultrasound waves -
frequencies >20 kHz
 Solid and liquid particles are
vibrated & accelerated , So solute
quickly diffuses into solvent
 Less expensive, simple and efficient
 Increase of extraction yield and
faster
 For thermolabile compounds.
 Eg.Saponin Glycoside from ginseng,

32. Microwave-assisted Extraction :
 Microwaves Extraction operate at
2.45 GHz frequency
 Microwaves possess electric &
magnetic fields which enhance the
diffusion efficiency
 Less solvent consumption,
 shorter operational time,
 high recoveries and reproducibility
 Eg. Taxanes from Taxus brevifolia needles,
 Anti-oxidative phenolic compounds from
tomato

33. Solvents used for Extraction
 Petroleum ether :- Fixed oils, Phytoglycerols.
 Benzene:- Fixed oils, Phytosterols.
 Chloroform;- Alkaloids
 Acetone:- Phytosterols
 Ethanol:- Carbohydrates, Glycosides
 Saponin:- Phenolics, Tannins, proteins, Amino
 acids
 Water:- Proteins, Amino acids, Glycosides,
Gums,
 Mucilages, Carbohydrates

34. Selection of Solvents

35. Factors influence the Extraction
• Length of the extraction time
• Solvent used,
• The solvent-to-sample ratio.
• pH of the solvent,
• Temperature,
• Particle size of the plant parts,

36. Concentration
The extract contains huge
solvent quantity and small
amount of active molecule.
Concentration:
Evaporate the solvent and
enrich the active content
 Concentration under vacuum,
Thin film evaporator, Freeze

37. Separation And Isolation
The most difficult issue is isolation and purification
of active content from other molecules;
 Column chromatography
 fractional crystallisation,
 Fractional distillation,
 fractional liberation.
 Chemical reaction or method is based on groups or
moieties

38. Purification
Crystallisation
 Formation of crystals at the point of super saturation
in the solvent by concentration, slow evaporation,
cooling etc
Fractional distillation:
 Fractionation based on boiling point
Sublimation
Chromatography separation :
 Column chromatography : separation based on the
solubility /partition coefficient

39. Industrial level
Structured approach
R&D:
 Process development, Process Optimization, lab
validation and Scale up
Production:
 Selection of suitable Equipment
 Unit operation
 Process validation and manufacturing
Analysis:
 Analysis, Analytical technique, Pharmacopeia, &
compliance

40. Research & Development
 Extraction & Purification
 Process chemistry
 Thermo chemistry
 Kinetics
 Scale up
 Technology transfer
 Structure elucidation
 Patent search
 New analytical techniques
 Simple & easy test methods
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41. Manufacturing/production:
 Process Chemistry
 Technology transfer
 Chemical reaction
 Handling of Chemicals
 Unit operations
 Reaction, distillation,
 filtration, drying, blending
 Formulation, etc
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42. Standardization parameter for plant drugs

43. Method Development & Analysis of drug
 Identification and confirmation of molecule & structure
 Method development & method validation
 Qualitative and quantitative Analytical technique
 HPLC-high performance liquid chromatography
 HPTLC-high performance thin layer chromatography
 GC-gas chromatography
 UV –Ultraviolet spectrometry
 FTIR –FT Infrared spectrometry,
 MS-Mass spectrometry

44. Analytical technique
Ultraviolet-Visible &Infrared Spectrometry
Functional group identification and quantitative estimation
Nuclear Magnetic Resonance Spectroscopy (NMR)
The number and nature of chemical entities in a molecule
Mass Spectroscopy : Mass of the molecule
TGA/ DTA: Thermal property
Melting, Decomposition, hydration/ solvation, polymorph of
maolecule,
XRD: crystal structure

45. Quality Control
Pharmacopeia & in-house standard:
 Physico-chemical properties
 Identification
 Chiral /Optical rotation
 Assay/ Purity
 Residue on Ignotion
 Heavy metals
 Other tests:LOD/ water / pH
 Particle size etc.
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46. Analytical technique
 Chromatography
 Spectrophotometry
 UV- Vis & FTIR
 Impurities: Inorganic &
Organic & Residual solvents
 Polymorph /Crystal structure-
XRD
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47. Chromatography
Chromatography used
extensively for testing of
purity and impurity in
complex molecule;
 TLC
 Gas Chromatography
 HPLC
 Gel permeation
 Ion chromatography
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48. D
The Following are ensured for effectiveness of
treatment;
Quality,
Safety,
Efficacy &
Strength
Final Medicinal form

49. References
1. Mukharji Pulok K. 2002,Quality Control of Herbal Drugs,1st
edition Published by Business Horizon, p.p 380-421.
2. Mehta R.M. 2010, Introduction to Pharmaceutics ,5th edition
Published by Vallabh Prakashan, p.p150-167.
3. Rangari v.d. 2002 ,Pharmacognosy and Phytochemistry ,1st
edition ,volume 1st Published by Career Publication, p.p.95-98
4. Cooper J.W, Gunn`s Colin 2010-2011, Register of General
Pharmacy p.p.308-393

50. References
1. S. S. Handa. 2008.-An Overview of extraction Techniques for
Medicinal and Aromatic Plants.
2. Phytochemicals: Extraction Methods, Basic Structures and
Mode of Action as Potential Chemotherapeutic Agents
James Hamuel Doughari 2012
3. Barnes, J.; Anderson, L.A. & Phillipson, J.D. (2007). Herbal
Medicines. A guide for Healthcare Professionals, third ed.
4. Philipson, J.D. (2007). Phytochemistry and
pharamacognosy. 68:2960-2972.
5. http://www.dbtindia.nic.in/therapeutics/
6. http://www.medicinalplants.in/
7. http://www.nmpb.nic.in/

52. THANK YOU