Transcript Analytical procedures for medicinal plant products

ANALYTICAL
PROCEDURES FOR
MEDICINAL PLANT
PRODUCTS
(SAMPLING, SAMPLE
PREPARATION AND ANALYSIS)
1
 A LECTURE TOPIC DELIVERED
 AT THE
 13TH MANDATORY WORKSHOP
 ORGANIZED BY
 THE INSTITUTE OF THE PUBLIC
ANALYSTS OF NIGERIA
 ON 3RD -4TH MAY, 2011

AT IKEJA AIRPORT HOTEL
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BY
OSADEBE, PATIENCE
OGOAMAKA
(MPSN, REG.PHARM, WMGA, MIPAN, FIIA)
•
•
•
PROFESSOR OF PHARMACEUTICAL
& MEDICINAL CHEMISTRY & DEAN,
FACULTY OF PHARMACEUTICAL SCIENCES,
UNIVERSITY OF NIGERIA NSUKKA
A MEMBER OF COUNCIL, INSTITUTE OF
PUBLIC ANALYST OF NIGERIA
3
INTRODUCTION
There
is a resurgence of interest in
medicines sourced from nature or
biodiversity.
This resurgence has been precipitated
by the recent increased prevalence of
incidence of drug resistance and
multiple drug resistance and
sometimes unaffordable cost of some
important conventional or orthodox
drugs.
4
INTRODUCTION CONTINUES
The
long
existing
lack
of
chemotherapeutic solution to the
health menace of some essential
diseases (HIV, cancer etc) has also
played a part in the current paradigm
shift to natural sources in search of
Novel medicinal agents.
New lead compounds are developed to
serve as impetus for continuous search
of drugs which could help to proffer
medicinal solutions to some intractable
disease.
5
INTRODUCTION CONTINUES
Advent
of biotechnology has widened
the scope of research and discovery that
are possible with Herbs and plants in
general.
In
India and UK, and some developing
countries, review of herbal remedies
indicated that herbal remedies available
were being marketed even before the
inauguration of medicines Acts(In
India, before the Drugs and cosmetic
Acts of 1948
6
DEFINITIONS OF RELEVANT TERMS
The definitions of terms that will be encountered in the discussion
are given
 Quality : Conformance to set standards or specification
 Quality : to be what something or someone should be, integer,
whole, pertaining to integrity or wholeness, uninterfered with.
 Quality control : This represents the totality of the efforts or
processes that are set up or made to ensure that a drug product
has and /or retains expected stipulations or requirement with
regards to
 purity,
•
 safety ,
 efficacy
 identity and
 content
7
 From
the point of assemblage of starting
material throughout production to the
point of marketing such that the product
delivered to the consumer will exert the
medicinal effect for which is consumed (to
exert the designed efficacy).
 Quality
is controlled so that at the end it
can be assured.
 Quality is
assured when it has been properly
controlled at critical stages in the sourcing
of raw materials, in the production, in the
labeling and packing of a medicinal
product.
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CHARACTERISTIC CONSTITUENTS

These
are
chemically
defined
substances or groups of substances
which
are generally accepted to
contribute substantially to the
therapeutic activity of a herbal
substance, a herbal preparation or one
or more such herbal substances that
are in combination with one or more
such herbal preparations.
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CONSTITUENTS WITH KNOWN
THERAPEUTIC ACTIVITY
 Are
chemically defined substances or group of
substances which are generally accepted to
contribute substantially to the therapeutic activity of
herbal substance , a herbal preparation or a herbal
medicinal product.
 Herbal
medicinal products(HMP) = Medicinal
Plant product(MPP).
• This is defined as any medicinal product, exclusively
containing as active substances one or more substances
or one or more herbal preparations or one or more such
herbal substances in combination with one more such
herbal preparations
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DEFINITION CONTINUED
 Herbal preparations
◦ are preparations obtained by subjecting herbal substances to
treatments such as extraction, distillation, expression,
fractionation, purification, concentration or fermentation.
These include comminuted or powdered herbal substances,
tinctures, extracts, essential oils expressed juices and processed
exudates.
 Herbal substances
-All mainly whole, fractioned or cut plants, plant parts, algae,
fungi, lichen, in an unprocessed usually dried form but
sometimes fresh.
-Certain exudates that have not been subjected to a specific
treatment are also considered to be herbal substances.
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DEFINITION CONTINUED
Herbal
substances are precisely defined by the
plant part used and the botanical name according
to the binomial system(genus, species, variety and
author).
Markers: are chemically defined constituents or
groups of constituents of a herbal substance , a
herbal preparation or a herbal medicinal product
which are of interest for control purposes,
independent of whether they have any therapeutic
activity.
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DEFINITION CONTINUED
Markers serve to calculate the quantity of herbal
substance(s) or herbal preparation(s) in the HMP
if the marker has been quantitatively determined
in the herbal substance or herbal preparations.
Active marker: are constituents or groups of
constituents that serve for analytical purposes
Analytical Marker: are constituents or group
of constituents that serve for analytical purposes.
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DEFINITION CONTINUED
 Quantification:
Adjusting the herbal preparation to a
defined range of constituents, exclusively achieved by
blending different batches of herbal substances and/or
herbal preparations(eg. Quantified extract)
 Specifications: A list of tests, references to analytical
procedures, and appropriate
criteria which are
numerical limits, ranges, or other criteria for the tests
described. It establishes the set of criteria to which a
herbal
substance/herbal preparation or herbal
medicinal product should conform to be considered for
its intended use.
 Conformance to specifications means that the
herbal preparation/ herbal substance and /or herbal
medicinal product when tested according to listed
analytical procedures will meet the listed analytical
criteria.
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DEFINITION CONTINUED
 Specifications
are binding analytical standards that are
agreed to between the appropriate governmental regulatory
agency and the applicant.
 Standardisation:
means adjusting the herbal substance /
herbal preparation to a defined content of a consistuent or a
group of constituents with known therapeutic activity
respectively either by adding excipients or by blending
batches of the herbal substance and/or herbal preparation
(e.g. standardized extracts).
 Traditional
herbal medicinal products: Medicinal
products for human use that fulfill the conditions laid down in
article 16a(1) of Directive 2001/83/EC, as amended.
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DEFINITION CONTINUED
 Solvent:
An inorganic or an organic liquid used for
the preparation of solutions or suspensions in the
manufacture of a herbal preparation or the
manufacture of a herbal medicinal product.
 Decoction: A herbal drug preparation obtained by
placing the material in cold water, bringing it to boil
and simmering for about 15 min or longer(up to
1hour and then allowing the mixture to stand for a
further 15 min
It is a type of aquoeus extract
Not to be confused with concoction which means
a preparation made usually from many
ingredients
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DEFINITION CONTINUED
Maceration:
this is a preparation obtained by
placing the plant material with the whole of the
extraction liquid in a closed vessel and allowing
to stand for 7 days and shaking occasionally
Infusion: This is a type of aqueous extract or
preparation obtained by pouring boiling water on a
specific quantity of plant material and allowing the
mixture to stand for 15 min. Eg, common tea in a
teapot or cup.
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INTRODUCTION CONTINUED
 For
a long time, efforts towards utilisation of
HMPs were geared towards –
 Isolation
of finished products containing pure
chemical entities
 As
source of lead compounds for the synthesis
of commercial drugs.
 Even
in the said era, large population of
communities in Africa, Europe and other
continents, still depend on fresh juice, powder,
decoction, hot and cold infusion obtained
directly from traditionally medicinally active
plants for treatment of common ailment
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INTRODUCTION CONTINUED
It is no gainsaying the fact that HMPs are
gradually gaining a substantial part of the
global market as a result of the increasing
demand for them.
Hence the issues that pertain to the quality
of HMPs have become critical
Have also become the focus of many
international and national regulatory and
health institutions

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INTRODUCTION CONTINUED
In
this discussion therefore, we will look at
the analytical procedures of HMPs as it
pertains to
Sampling,
Sample preparation
Analysis
20
SAMPLING OF HMPs
•
•
Sampling approach for HMPs analysis
depends on the form and the source.
The source may be
-shipped imported consignment
awaiting entry
-fresh collection for extemporaneous
HMPs
-wholesale samples in the market or
from suppliers
-Retail samples from vendors
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SAMPLING OF HMPs
o
Manner of selection of HMP samples affects quality of
assessment exercise
o
It is known that precision and reliability of analytical
procedures is directly related to sampling.
o
The extent to which the sample taken are true
representative of the material is the bottom line.
o
Medicinal plant materials have peculiars problem in
relation to homogeneity.
o
Hence,USP(1994)
procedures.
prescribed
definite
sampling
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SAMPLING OF HMPs
o
o
o
o
o
For small consignments, all parts should be
examined
Where not possible, portions should be taken
from different parts of the consignment.
For powders or pieces size less than 1cm, cores
are extracted from top to bottom in each
direction
For samples less than 100kg, at least 250g of the
material is taken.
For consignments with material size greater than
100kg,samples are taken, repeatedly mixed and
quartered as shown below
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SAMPLING OF HMPs
No of pkgs in shipment
1-10
10-25
25-50
50-75 `
75-100
No of pkgs to be sampled
1-3
2-4
3-6
6-8
8-10
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SAMPLING OF HMPs
•
Quartering consist of placing the sample adequately
mixed and formed into an even and squared shaped heap
and dividing diagonally into four equal parts.
•
Two diagonal quarters are rejected, the remaining two
mixed and re-quartered until size near to but not less
than 250 mg is gotten
•
For consignment size greater than 100kg, repeated
samples are taken and mixed and quartered as in the
procedure above until two of the quarters weigh as near
as possible to but less than 500g
•
25
SAMPLING OF MATERIAL IN BULK
Inspect each container/packing unit for
conformity with pharmacopoeia monograph or
other requirement regarding packaging and
labeling.
• -Check the integrity of the outer package to note
any defects which may influence the quality or
stability of the contents (e.g. physical damage,
moisture others).
• - damaged containers are sampled separately and
individually.
• -When the above initial inspection process
indicates uniformity of the batch, the sampling
proceeds further as follows
•
26
SAMPLING •
For a batch comprising 5 containers/units, sample is
taken from all of them.
•
From a batch made up of 6-50 units, a sample from
5 packages is taken. For batch of over 50, 10% of the
units is sampled, rounding up the no of units to the
next highest figure of ten e.g. 51 units would be
sampled as for 60.
•
After opening, containers are inspected for the
following parameters before sampling.
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SAMPLING Organoleptic characteristics (Color ,texture and Odour)
Presentation of the material( raw, cut, crushed, compressed).
Possible presence of admixture and/or foreign matter.
(Sand, glass particles, dirt, mould or signs of decay)
Presence of insects
Presence of packaging materials originating from poor or degraded
containers
 Three
original samples are taken from each container,
while taking care to avoid fragmentation.
 Samples are taking from the upper, middle and lower
parts.
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SAMPLING For sacks and packages, 3 separate samples are taken by hand from a depth of not
less than 10cm from the top, and after cutting into the side of package from the
middle and lower parts.
When the material in question is seed, the samples are withdrawn
with a grain probe.
When the material is contained in a box, it is first sampled from
the upper layer then first the content is removed and new samples are
taken again. After further removal of the material then another
sample is taken from the bottom.
Sample should be as uniform as possible in mass. Finally the
separate samples are combined into a pooled sample and are mixed
thoroughly.
Quartering technique is used to obtain the average sample.
.The process is repeated until the required quantity is obtained.
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SAMPLING  The
pooled samples are quartered until the required amount
remains which should be within 10 % ( 100-200g) for
flowers and up to 10kg for certain roots.
 The remaining materials are returned to the bulk batch.
 The
average sample quartered again and final samples are
assembled and tested for the following features:
• Degree of fragmentation(Sieve test)
• Identification of level of impurities
• Determination of moisture and ash content
• Assay of active ingredients where possible
A
portion of the final sample is retained to serve as reference
material which may also be used for purposes of quality
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control tests.
SAMPLING OF MATERIAL IN RETAIL
 Two
consumer packages are usually taken at
random from each wholesale container such as
boxes, cartons etc.
 Ten
consumer packages are taken from small
boxes.
 The
contents of the selected consumer packages
are mixed and the procedure followed as for the
final sample above
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SAMPLING PREPARATION DURING
ANALYTICAL PROCEDURES FOR HMP
 Sample
preparation is a critical step during analysis
and for the accuracy of results of analysis.
 It is as crucial as correct sampling procedure in
guaranteeing the reliability and authenticity of
analytical result.
 Approach to sample preparation for analytical
procedures depend on the nature of the HMP and
can involve one or more of the following procedures:
 Harvesting /collection, Drying, Extraction, Distillation,
Sublimation
Fractionation,
Concentration,
Fermentation, Expression, Percolation, Filtration and
Purification
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SAMPLING PREPARATION
Hence, HMP may take any of the following forms
– Crude Herbal drug (crude Herbal substance)
– Dry powdered extract
– Liquid extracts (Galenicals) (tinctures, decoction
infusion, cold infusion and maceration )
– Dry solvent fractions
– Formulated Herbal products (tablets, capsules,etc)
• The preparatory operations to which the herbal drug will
be subjected depend on its form
• Samples should be sourced from approved suppliers
•
33
SAMPLING PREPARATION
•
•
•
•
In order to contain active ingredient with known concentration limits,
herbal drug products must be prepared by following strictly the
standardized procedures such as maceration percolation decoction etc.
The detail of such procedures are laid down in the pharmacopoeia
which specify weight of the drug to be used,
– the solvent volume during the extraction procedure.
– Precautionary measures necessary during extraction are
specified and should be followed to arrive at desired
result and avoid deterioration.
At times, it is needed that the initial extract be concentrated or diluted
to arrive at the desired concentration
e.g ‘prepared digitalis’ for therapeutic use is actually digitalis leaf
powder diluted with an inert leaf powder to give a standard
concentration of one unit of actively per 100 mg.
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SAMPLING PREPARATION
 Prototype
:2g of the powdered sample (crude drug)
was weighed and taken in a 250ml beaker. 50ml 0f the
specified solvent was added and refluxed on a water
bath for 15 minutes.
•
It was cooled and filtered through whatman filter
paper.
•
The marc was further extracted two times with the
same solvent and the combined filtrate from three
refluxes was concentrated to dryness under vacuum in
case of successive extraction each solvent extraction is
carried out in the above said manner
35
ANALYTICAL PROCEDURE FOR
HMPs
 Basically, there are three common types of
analytical procedures . These are
o identification tests
o Test for impurities (Quantitative tests or Limit
tests)
o Quantitative tests (Assay for a substance or
element in a given sample or bioassay for activity
 Type of analysis
to be conducted depends on
Form or type of the HMP
Type of method prescribed
36
ANALYTICAL PROCEDURES
1. Physical methods of analysis
•These includes



•




Refraction index for oils, foam- forming index for
saponins,
Determination of bitter activity for bitter principles,
Determination of total solids.
Spectrophotometric methods as applied to quality
control of pure synthetic substances.
Determination of swelling index for gums and those
containing an appreciable amount of mucilage, pectin
or hemicelluloses,
Determination of Hemolytic index of saponins.
Determination of Tannins
Determination of Arsenic/Heavy metals.
37
ANALYTICAL PROCEDURES
2. Physico- chemical methods;
 These include the determination of volatile oils in
essential oil – containing plants


All types of chromatographic procedures◦ Gas chromatography
◦ Column chromatography,
◦ High performance liquid chromatography,
◦ Electrophoresis and
◦ Spectrophotometry
38
ANALYTICAL PROCEDURES CONTINUED
•
•
•
•
•
•
•
•
•
3. Pharmacognostic methodsDetermination of foreign matter
Specification of orgoleptic properties of the plan or extract.
-small, taste, colour, texture, fracture.
-microscopic and macroscopic characters including
-the diagnostic features
Macromorphological and cytomorphological evaluation of Herbal drugs.
-Determination of ash value
–sulphated ash,
acid insoluble ash
-Determination of solvent extractive value
eg water extractive values,
alcohol extractive values.
In determination of water extractive, chloroform water is used to determine
deterioration that occur when water is used to extract plant that contain a lot of
sugars or other carbohydrate which can ferment during prolonged extraction
procedures.
Determination of crude fibre.
-Determination of moisture content
39
ANALYTICAL PROCEDURES
4 Chemical methods.
• These include the various identification reactions
• They include the various phyto-chemical tests that are
employed in qualitative analysis to ensure that the
crude drug, the plant extract or fraction or the herbal
drug preparation still contains the expected secondary
metabolites
• e.g. identification tests for alkaloids, glycosides,
tannins, flavonoids etc , the Borntrager’s test for
anthraquinone containing plants or the Keller killiani test for deoxy sugar in cardiac glycosides,
saponification value for oils, acid-base titration of total
extract to determine the total alkaloids
40
ANALYTICAL PROCEDURES
5. Biological methods
 These are the microbiological assays for
antimicrobial activity in plant or the determination
of cardio activity in cardio active plant.
6. Quality control/ standardization of the
finished dosage form.
 Here, the quality control tests usually prescribed for
the formulated pure synthetic drugs are usually
required for Herbal drug preparation in form of
tablet capsules or others.

41
ANALYTICAL PROCEDURES
Certain
adjuvants and excipients added to act as
binders, disintegrants, lubricants etc affect the
release of the active constituents of the Herbal
preparation.
These tests are:
Weight uniformity
Tablet Hardness,
Friability test,
Disintegration test ,
Dissolution rate test and
Absolute drug content test
42
ANALYTICAL PROCEDURES
7 Stability Studies
 This is done to provide evidence on how the
quality of an HMP will vary with time under the
influence of several factors including
temperature,
◦ humidity and
◦ light.

Such studies enable establishment of expiry dates
and recommendation of ideal storage conditions
43
ANALYTICAL PROCEDURES
•
•
8. Toxicological analysis
Certain drug regulatory bodies in Europe allow
the anecdotal and traditional proof that a herbal
drug has been used as a medicine without reports
of toxicity as satisfactory for its continued use,
provided the mode of use is not altered (except
plant powder)
Nonetheless, proof of non -toxicity is desirable
before a plant is incorporated into herbal
medicinal use .
44
Toxicological analysis Continues
• Both acute and chronic toxicity test are required of
herbal drugs and their preparation .
• These tests help to determine the upper dose limit of
administration.
• Knowledge of the therapeutic index of the drug is
needful and also usually required in other to specify the
highest dose that can be administered without
encountering toxicity.
• Methods of acute toxicity testing
-Miller and Tainter probit method (1944)
-Lorke method (1983)
45
ANALYTICAL PROCEDURES CONTD
Biopharmaceutical methods
• These are methods that relate to bioavailability and
pharmacokinetics of medicinal agents.
• Attention is beginning to be given to formulated
medicinal plant product because excipients and
additives used in such formulation will affect the
solubility of the active ingredient and indirectly affect
the amount of active principles which the body can
utilize.
• In –vivo concentration against –time curve will
generate the necessary pharmacokinetics data namely
AUC, Cmax,Tmax, Half- life etc.
•
46
ANALYTICAL PROCEDURES
For Africa and other developing countries,
emphasis should be on simplicity of method while
maintaining tolerable accuracy
 Among the chromatographic procedure for the
analysis of marker compounds, TLC has gained
prominence because of its

•
•
•
•
simplicity,
cost effectiveness,
reproducibility and
visualizability .
47
ANALYTICAL PROCEDURES


Fingerprinting of the TLC profile and quantitative
analysis of marker has been made possible by using
densitometry scanners
A set of guideline for the assessment of herbal
remedies has been published by WHO. These
guidelines specify criteria to be used in evaluating
commercial crude drugs intended for use in making
MPP. In –house procedures designed for convenience
and simplicity are expected to be evaluated against
the stipulated official methods (where possible)
48
USE OF MARKERS IN QUALITY CONTROL
OF HMPS
 The
use of markers in the quality control is gaining
increasing relevance and utility.
 Because HMPs are usually multi- component preparations,
unlike conventional medicines,
 analysis of them in the drug matrix presents a peculiar
problem not encountered in orthodox formulations.
 At times, the phytochemical constituents of HMP cover a
wide range of secondary metabolites like Alkaloids,
triterpenoids, tannins, sapanin.
 For this, a chromatographic or spectroscopic method is
evolved that depend on the response of one or two of
chemical constituents for the analysis.
49
EMA SELECTION CRITERIA FOR MARKERS









Markers
Selection to be justified
should be appropriate for the intended purpose
should connect steps of production process and the quality control
measure
used for quantitative and qualitative purpose and are important to link
the active substance with HMP
Acceptance limit for the content of a proposed marker to be specified
and justified
The mere determination of the stability of the marker will not suffice
for the proving the stability of the HMP
since the Herbal substance or preparation in its entirety is regarded as
active substance.
The stability of other substances present in the herbal substance should
also be demonstrated,
for example by use of chromatographic appropriate finger printing, and
that their proportional content remains comparable to the initial
fingerprint
50
The two prototype analytical monographs
given below were taken from “ Quality
control of herbal drugs” and represents
in-house standards of NATURAL
REMEDIES PVT LTD, Bangalore,
India(www.Indianherbs.com)
51
ACORUS CALAMUS LINN
(a) Classification;
Kingdom : Plantae
Division
:Angiospermae
Class
Order
Family
Genus
Species
:Monocotyledoneae
:Spathiflorae
:Araceae
:Acorus
:calamus linn
(b) Part used :
Rhizomes(dried)
52
(c) Phyochemistry: Dry rhizomes 1.5-3.5% of yellow aromatic
volatile oil of which asarone is the major component. Other
constituents include –asarone, calamenol, calamene,
calamenone, methyl eugenol etc.; sesquiterpene ketones viz,
acrone, calarene,calacone etc, sesquiterpene alcohols viz.,
isocalamendiol, preisocalamendiol.
(d) Marker constituent:
β-Asarone*
OCH3
H3CO
H
CH3
C 12H16O3
H3CO
H
Mol.Wt.208.26
β-Asarone is only a marker compound. Not an active and is carcinogenic.
53
ANALYTICAL SPECIFICATIONS
OF THE CRUDE DRUG
Macroscopic characters:
Colour and appearance
Taste
Odour
:Rhizome is brown in colour. The rhizomes
are cylindrical and branched. They are
somewhat shrunken and with deep
longitudinal wrinkles. The leaf `scars` are
more prominent on upper surface and
encircle the rhizome. The under surface of
rhizome bear very small, but raised circular
root scars. 5 to 15 cm in length and 1 to 2 in
thickness.
: Bitter, pungent and disagreeable
: Characteristic
54
Tests
Limits
Tests for extraneous material
Foreign matter
Sand &silica
Insect infestation
Rodent contamination
Physico-chemical analysis
Moisture content
Ash content
Acid insoluble ash
Volatile oil
Alcohol soluble extractive value
Water soluble extractive value
Protocols
<1.0%
Absent
Nil
Nil
<5.0%w/w
<8.0%w/w
<1.0%w/w
2.0-5.0%w/w
>15.0%w/w
>25.0%w/w
WHO1
Successive extractive value
Petroleum ether extractive value 1.0-4.5%w/w
Chloroform extractive value
0.5-1.5%w/w
Methanol extractive value
14.0-20.0%w/w
Phytochemical analysis
Total Glycosides
>35.0% By Gravimetry
-
55
IDENTIFICATION OF CRUDE DRUG BY
TLC/HPTLC
Sample
:
Acorus calamus (Rhizomes)
Solvent system
:
Toluene : Ethylacetate
93
:
7
The powdered sample successively
extracted
with
petroleum
ether,
chloroform
and
methanol.
The
solutions separately concentrated to 10ml.
Anisaldehyde sulfuric acid reagent (Fig 1)
Sample preparation :
Detection
:
Deensitometer scan :
(Fig – 1)
223nm (Fig 2, 3, 4)
(Fig – 2)
56
T2(Fig – 1)
T3 (Fig – 2)
HPLTC chromatogram of successive extracts
Rf value of B-asarone = 0.56
T1 – Petroleum ether extract
T2 – Successive chloroform extract
T3 – Successive methanolic extract
57
ADHATODA VASICA NEES.
(a) Classification :
Kingdom
:
Plantae
Division
:
Angiospermae
Class
:
Dicotyledoneae
Order
:
Tubiflorae
Family
:
Acanthaceae
Genus
:
Adhatoda
Species
:
vasica Nees
(b) Part used : Leaves
(c) Phytochemistry: Leaves contain vasicine as the major bioactive
pyrralazoquinazoline alkaloid. Also contain other alkaloids viz., vasicol,
adhatonine, vasicinone, vasicinol, vasicinolone; aliphatic hydroketones
viz., 37- hydroxyhexatracont-1-en-15 one and 37 – hydroxyl
hentetracontan – 19- one.
OH
N
N
C11H12N2O
Mol.Wt.188.23
58
ANALYTICAL SPECIFICATIONS OF
THE CRUDE DRUG
Macroscopic characters:
Colour & appearance
:Leaves are simple, petiolate, broad,
glabrous and mostly elliptic lanceolate to
ovate
lanceolate. They have slightly
crenate or entire margin. Dorsal surface is
green in colour but the
ventral one is
slightly pale. 8-12 pairs of bilateral veins
which are reticulate.
Taste
Odour
: Bitter
: Characteristic
59
Tests
Limits
Tests for extraneous material
Foreign matter
Sand &silica
Insect infestation
Rodent contamination
Protocols
<1.0%
Absent
Nil
Nil
WHO
Physico-chemical analysis
Moisture content
Ash content
Acid insoluble ash
Alcohol soluble extractive value
Successive extractive value
Petroleum ether extractive value
Chloroform extractive value
Methanol extractive value
Phytochemical analysis
Vasicine content
<8.0% w/w
<15.0% w/w
<2.0% w/w
<8-12% w/w
0.7-2.0% w/w
0.7-2.0% w/w
6-11% w/w
0.4% w/w
BY HPLC
60
IDENTIFICATION OF CRUDE DRUG
BY TLC/HPTLC
Sample
:
Adhatoda vasica (Leaves)
Solvent System
:
1,4- Dioxane
: Ammonia solution
9
:
7
Powdered sample extracted with methanol. The
extract Concentrated and the residue dissolved in
methanol.
Sample preparation :
Detection
:
Deensitometer scan :
S
T
Fig -1
Drangendorff reagent (Fig. 1).
254 nm (Fig.)
Fig -2
61
THANK YOU FOR THE
APT ATTENTION
62