Phytochemical Methods: Lecture Notes I

Lecture notes I
Phytochemical methods
R.Venkataraman, Reader in chemistry,

Sri Paramakalyani College,
Alwarkurichi-627412, Tamil Nadu, India
E-mail: rvraman3@rediffmail.com
Introduction
Plants alone synthesise their own food by using chemistry in photosynthesis.
The photosynthetic products are essential for the plant growth and development. They are called
primary metabolites. Plants synthesise a further group of compounds called secondary
metabolites, mostly stored in xylem and phloem region of the cells. The secondary metabolites
are known as phytochemicals.
The secondary metabolites are non-essential to plant life but their contribution to fitness and
survival of the plants are unquestionable.
By instinct or intuition or trial and error, many natural materials have been used
by the mankind as foodstuffs, colouring matters aroma substances, perfumes and medicines for
combating human ailments only because of secondary metabolites.Phytochemistry is that branch of
science which deals with isolation, purification and charecterisation of secondary metabolites of the
plants. Phytochemistry, though ancient, gains significance today because more than 75 pure
compounds derived from higher plants find place in modern medicine.
General Isolation procedure
The plant materials are collected and dried under shade to a constant weight.
It is then extracted in various solvents of increasing polarity from petroleum ether, benzene,
chloroform, ethanol and water (graded extraction).
1. Solvent extraction Alcohol is good all purpose solvent for cold extraction.
Chloroform is a good medium polar solvent for hot extraction
2. Fractional extraction
3. Separation by precipitation
4. Extraction of polar compounds from biological materials
Chromatographic methods
The crude extracts can be subjected to various chromatographic methods
for separation of compounds.
1. Thin layer chromatography TLC
2. Paper chromatography PC
3. High performance liquid chromatography HPLC
4. Gas chromatography GC
Thin layer chromatography (TLC) is superior over paper
chromatography(PC).
HPLC is also superior because of resolution, recovery, speed of operation
in
the range of 0.1mg scale.
Various phytochemical tests reveal the particular
class of compounds. Elemental analysis can guide us to finalise the
probable empirical formula. A single spot in three different solvent systems
in TLC confirms the purity of the sample.
General characterisation procedure

The isolated pure compounds can be charecterised by
the following spectroscopic methods:
1. Ultra violet spectroscopy (UV)
2. Infra red spectroscopy (FTIR)
3. Nuclear magnetic resonance spectroscopy (NMR)
2-D NMR techniques COSY, NOESY, DEPT, INADEQUATE
4. Mass spectrometry(MS) EIMS, FABMS

5. X-ray diffraction studies (XRD)
6. Optical spectroscopy (ORD & CD)
The structure of the compounds can be elucidated
using spectroscopic studies and confirmed by single crystal X-ray
diffraction studies
Conclusion
Phytochemistry finds many applications now-a-days in plant
systemics,
Chemo taxonomical markers, biopharmaceutical chemistry, plant genetics, plant ecology,
plant pathology, agriculture, biopesticides etc... Due to the enormous medical and
economic impact of natural product pharmaceuticals and their demand, special attention
is shown towards the improvement of existing compounds and to discover the new
hitherto unreported secondary metabolites.
Suggestions for further reading
J.B.Harborne, Phytochemical methods, 2nd Ed, Chapman Hall, New York
(1973)
Questions
1. Secondary metabolites are mostly stored in
a) Epidermis b) Pith c) Xylem d) Endodermis
2. Primary metabolites are essentially------------
a) Alkaloids b) Carbohydrates c) Steroids d) Flavonoids
3. A good all purpose solvent for plant extraction is
a) Benzene b) Chloroform c) Ether d) Alcohol
4. The finger print region in IR spectra is
a) below 1200 per cm b) 1500-1650 per cm
c) above 3500 per cm d) 4000-700 per cm
5. The shift of a peak in ultraviolet -visible spectroscopy towards
longer wavelength is called
a) Hypsochromic shift b) Bathochromic shift
c) Isomeric shift d) isotopic shift
6. Expand the following:
a) FTIR b) NOESY c) DEPT d) COSY
Keys to the questions
1. c 2. b 3. d 4. a 5. b
6. a. Fourier Transformation Infra Red
b. Nuclear Overhauser Effect Spectroscopy
c. Distortionless Enhancement of Polarisation Transfer
d. Correlation Spectroscopy
Lecture notes II
Traditional and modern drugs
R.Venkataraman, Reader in chemistry,

Sri Paramakalyani College,
Alwarkurichi-627412, Tamil Nadu, India
E-mail: rvraman3@rediffmail.com
Introduction
Health is not the absence of illness only but a feeling of wholeness. Health care
professionals do use crude herbal drugs earlier from the time immemorial. But in modern
medicine several synthetic chemical compounds are being used. The western allopathic
treatment coupled with eastern traditional healing arts are now-a-days popular in
developed countries like USA. This is known as ‘greening of medicine’. The modern
drugs give side effects whereas the traditional drugs give permanent cure even for chronic
diseases without side effects.
A drug is any substance that is used to modify the physiological system
for the benefit of the recipient—according to WHO. The various sources of drugs include
minerals, plant products, animal products, synthetic products, microorganism etc…
Mechanism of drug action
Drugs normally bind with receptors. The site of drug action is called
receptors. It is also known as binding site. A receptor is a protein molecule embedded
within cell membrane with a part of its structure facing outside cell. The various drug
receptor theories include Occupancy theory, Rate theory, Induced fit theory,
Macromolecular perturbation theory and Activation-Aggregation theory.
Standardisation of drugs
Modern drugs are mostly synthesized with extra care given to its purity.
For example, one isomer of phthalidomide (a hallucinating drug often prescribed for
pregnant mother) is a poison but another isomer is a drug. For estimation of a potency of
a drug, chemical, biological and immunological assays are possible.
Traditional drugs essentially supported by medicinal plants. Sometimes
due to the confusion in vernacular names and trade names, similar morphological
characters etc..., misidentification may be possible. This may pave the way for
adulteration which affects the quality of a crude drug. The quality of a crude drug refers
to the intrinsic value of a drug. Hence these confusions can be rectified by studying their
pharmacognostic characters of medicinal plants such as macroscopic character,
microscopic character, ash value, and extractive value, preliminary phytochemical
screening, fluorescence character, TLC, micro chemical studies histochemical studies and
cytological studies systematically.
Suggestions for further reading
R.S.Satoskar and S.D. Bhandarkar, “Pharmacology and Pharmacotherapeutics”

Popular prakasam, Bombay, Vol.I&II, 10th ed. (1986)
In: George Edward Trease and William Charles Evans (Eds.), “Pharmacognosy”
12th ed. Bailldere, Tindall, London (1983)
Questions
1. The receptor of a drug essentially
a. alkaloid b. protein c. hemoglobin d. fat
2. The least polar solvent is
a. benzene b. water c. n-hexane d. ethyl acetate
3. Radioimmunoassay is mostly used to estimate
a. histamine b. protein c. insulin d. urea
4. Organoleptic methods are based on
a. microscopic characters b. macroscopic characters
c. pharmacological characters d. immunological characters
5. Libermann-burchard test is used to detect
a. alkaloid b. steroid c. saponin d. xanthoprotein
6. The botanical name of NITHYA KALYANI is
a. Vinca rosea b. Ocimum sanctum c. Phyllanthus amarus d.
Artemissia annua
Keys to the questions
1.b 2.c 3.c 4.b 5.b 6.a

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Lecture notes I

R.Venkataraman, Reader in chemistry,

General characterisation procedure

4. Mass spectrometry(MS) EIMS, FABMS

Keys to the questions

6. a. Fourier Transformation Infra Red

b. Nuclear Overhauser Effect Spectroscopy

c. Distortionless Enhancement of Polarisation Transfer

R.Venkataraman, Reader in chemistry,

Mechanism of drug action

Suggestions for further reading

R.S.Satoskar and S.D. Bhandarkar, “Pharmacology and Pharmacotherapeutics”

Keys to the questions

1.b 2.c 3.c 4.b 5.b 6.a

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