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Standardization Of Homoeopathic
Preparations
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Standardization
Of Drugs
Quality
Control
Quality
Assurance
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• Standardization is a process of having a product in
uniform standard as mentioned in the pharmacopoeia.
•
• It includes a Quality Control System with various
tests to perform at different stages of manufacture, from
raw materials to finished products.
• It also involves an Assurance & assessment of quality
of products with ref. to a pharmacopoeial monograph.
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• Supply of essential drugs of good quality which is the
prerequisite for the delivery of any Health Care
• Functioning of national drug regulation and control
systems which is the only means to assure safety and
quality of medicines
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Standardization of Drugs
Depends on
• To assure pure & quality end products in a proper
container bearing the correct label
• In order to find its relevance to homoeopathic
pharmaceutics
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Aims
• Confirmation of Drug identity
• Determination of Drug quality & purity ( impurity, if any)
• Determination of Vehicle quality & purity ( impurity, if any)
• Detection of nature of adulteration
• Estimating active constituents present in crude drug
All these steps constitute the Evaluation of Drugs
Steps
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• Biochemical variation in the drug
• Deterioration due to treatment & storage
• Substitution & adulteration, as a result of mishandling,
ignorance or fraud
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Evaluation of Drugs is necessary for three reasons:
Drug Adulteration
Original crude drugs are partially/wholly adulterated by
substitution with substandard similar looking substance
i.e. adulterant
Type of Adulterants:
• Substitution with substandard commercial varieties e.g.
nux blanda for nux vomica; medicinal ginger for Cochin
ginger
Cochin ginger
Medicinal ginger
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Substitution with similar inferior drugs e.g. ailanthus
leaves for belladonna leaves; Japan wax for beeswax
bell
ailanthus
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Type of Adulterants
• Substitution with artificially manufactured substances
e.g. yellow paraffin for beeswax; compressed chicory for
coffee
• Substitution with exhausted drugs e.g. clove, coriander
• Substitution with synthetic substances e.g. benzyl
benzoate to balsam of Peru
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Type of Adulterants:
• Substitution with vegetable matter from the same plant
e.g. moss, liverworts and epiphytes growing in bark
mixed with cinchona / cascara; stem mixed with leaves
of stramonium, lobelia
• Substitution with harmful adulterants e.g. limestones in
asafoetida, leadshot in opium
• Adulteration of Powders e.g. dextrin in ipecac, ginger
powder in colocynth
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Guidelines for Pharmaceutical Operations & Record
Keeping
• Proper sanitary measures
• Cleansing & calibration of equipments on regular basis
• Maintaining record of receipt, identification, processing,
yield of manufacture, storage, handling, sampling, testing
of labelling & packaging
• Keeping record of any deviation from these written
procedures & its justification & approval
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Stability of Drugs
a. Environmental factors such as heat, moisture, light,
oxygen, vibration, radiation, & other physical stress &
changes
b. Product-related factors such as:
i. Chemical & physical properties of crude & active drug
substances & of pharmaceutical aids (excipients) used
ii. Dosage form & its composition
iii. Manufacturing process
iv. Nature of container & other packaging
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Standardization of Homoeopathic Products
In India, Drugs and Cosmetics Act 1940 controls quality of
homoeopathic medicines.
The Homoeopathic Pharmacopoeia Laboratory (HPL) was
established in 1975 under the Ministry of Health and Family
Welfare, Government of India as a quality monitoring apex
body.
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Worked out standards are released in form of
Homoeopathic Pharmacopoeia of India (HPI).
The problems of quality control with homoeopathic
potencies serve one of the areas of greatest
challenge.
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The extremely high dilutions of homoeopathic potencies
make it almost impossible to apply analytical tests by
conventional methods in the laboratory.
Only mother tinctures, apart form the raw material are
subjected to a comprehensive analysis, both qualitative
and quantitative.
Good Manufacturing Practices (GMP) must be in force to
assure & assess the quality of homoeopathic preparations
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Quality Control of Raw Material
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The pharmacopoeia indicates in the monograph of each
drug, details for identification and collection, the part to
be used, the method of preparation, assessment of
purity and limits of impurity.
It is essential that these instructions be strictly followed
to obtain mother tinctures of desired standards.
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Evaluation involves determination of identity,
purity and quality or activity of drug ...
• Identity is determined by morphological characters,
sensory characters, microscopic characters, chemical and
micro-chemical tests and certain physical constants.
• Purity is determined by estimating foreign inorganic
matter, foreign organic matter and moisture content.
• Quality is determined by physical, chemical & biological
methods.
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• Organoleptic evaluation
• Microscopic evaluation
• Physical evaluation
• Chemical evaluation
• Biological evaluation
Methods
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O r g a n o l e p t i c E v a l u a t i o n
It refers to qualitative evaluation of drugs by means of organs of
senses, e.g. eyes (to see); nose (for odour); tongue (to taste); skin
(by touch) and includes the macroscopic appearance, its odor,
taste and the feel of drug to touch.
Occasionally, the fracture of plant material is also considered.
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• External morphology (Shape and Size)
• External colour
• External markings
• Fracture
• Odour
• Taste
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M i c r o s c o p i c E v a l u a t i o n
It helps in identifying organized drugs & searching
adulterants in powdered drugs.
• Histology
• Micro-chemistry:
- Isolation of drug constituents by micro-sublimation,
micro-extraction, micro-filtration, micro-crystallization
- Identification of constituents of drugs by
crystallography, melting point determination, physical test,
chemical test, colour reaction test
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C h e m i c a l E v a l u a t i o n
It covers isolation, identification, purification and characteristic
determination of drugs having active principles.
It helps in detection of alkaloids, carbohydrates & glycosides,
fixed oil & fats, volatile oil, saponins, proteins & free amino
acids, gums & mucilages, phenolic compounds & tannins,
phytosterols
Analytic chemistry may be qualitative or quantitative.
Quantitative analysis may be gravimetric,
volumetric, titrimetric, colorimetric analysis
& etc
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The chemical assays include :
• Color reaction test
• Acid value
• Iodine value
• Saponification value
• Ester value
• Ash value
– Sulphated ash value
– Acid-insoluble ash value
– Water-soluble ash value
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P h y s i c a l E v a l u a t i o n
•Chromatographic study of drugs
•Spectroscopy
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•Fluorescence test :
The reaction of certain drugs either in powdered
form or on sectioned surfaces with filtered UV light help
in Identification and purity.
Aconitine - light blue
Berberine - yellow
Emetine - orange
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The application of physical constants is largely employed to
the active principle of drugs.
• Solubility
• Specific gravity
• Density
• Viscosity
• Melting point
• Congealing point
• Refractive index
• Optical rotation
• Water content or loss
on drying
• Foreign organic matter
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B i o l o g i c a l E v a l u a t i o n
Pharmacological activity of certain drugs is applied to
evaluate and standardize them.
The assays on living animals and on their intact or cut-off
organs can indicate the strength of the drug or their
preparation.
As living organisms are used, these assays are known as
biological assays or bioassays.
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Biological Evaluation
Toxic Method Symptomatic Method Tissue Method
Live animals Live animals Isolated organs/
used used tissues used
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Analysis
Chemical information – exists between drug & tissues of
organism depending upon the chemical affinity
Mechanical information – includes signs of efforts by
organism to reject the offending substances from its inside
Dynamic information – includes changes depending upon the
vitality & susceptibilities of living, healthy/diseased organism
in relation to the peculiar properties of drug in small qty
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Few examples of Bioassays
• Cardiac glycosides containing drugs like digitalis are bio-
assayed on cat, frog or pigeons.
• Evaluation of ergot is done on rabbit intestine or uterus.
• Saponin is assayed by haemolytic index on fish.
• Curare causes relaxation of muscles and is carried out by rabbit
head-drop method.
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Quality Control of Finished Products
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An analysis of homoeopathic potencies neither reveals
the identity of the medicine, nor the potency.
Only mother tinctures can be subjected to a
comprehensive analysis, both qualitative and
quantitative.
“In-process” quality control at every step of
preparation, from raw material to finished products,
is critical in ensuring the purity.
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Mother Tincture Analysis
• Alcohol content determination
• Weight per ml
• pH value
• Total solids
• λ Max
• Fluorescence analysis
• Chromatography
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Specific Gravity
The specific gravity of a substance is the weight of a
given volume of that substance at a stated temperature
as compared with the weight of an equal volume of
water at the same temperature, all weighings being
taken in air.
A suitable pycnometer is used
for the determination.
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Alcohol content
Estimation of alcohol content of mother tincture is a test
for the potency of the mother tincture and an evaluation
of the extraction process.
Specific gravity of the distillate is determined
pycnometrically and the corresponding alcohol contents
in percent by volume read off in the
alcoholometric tables.
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Calculation of Alcohol Content
• Sample taken at 25oC = 25 ml
• Wt of empty specific gravity bottle = A
• Wt of specific gravity bottle + distillate at 25oC = B
• Wt of specific gravity bottle + water at 25oC = C
• Specific gravity = B – A / C – A
Alcohol Content, expressed as % v/v is obtained from
reference table in pharmacopoeia.
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Weight per ml
Weight per milliliter of a liquid is determined by dividing the
weight in air, expressed in grams, of the quantity of the liquid
which fills a pycnometer at 20o or 25o by the capacity of the
pycnometer at 20o or 25o respectively, expressed in milliliters.
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pH Determination
The pH is defined
as the common
logarithm of the
reciprocal of the
hydrogen ion
concentration
expressed in
gram per liter.
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• Storage, exposure to temperature and pressure
tend to alter the pH, thus making the liquid
unstable.
• It also helps to detect adulteration.
• The two most important methods of measuring
pH are by the use of colored indicators or of
special electrodes.
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Total Solids
• Weigh accurately or measure an
accurate quantity of the
preparation. Place it in a tarred
dish, evaporate at as low a
temperature as possible until
the alcohol is removed. Heat on
a water-bath until the residue is
apparently dry. Transfer to an
oven and dry to constant weight
at 105o. Cool in an efficient
desiccator.
The term 'total solids' is applied to the residue obtained
when the prescribed amount of the preparation is dried to
constant weight under specified conditions.
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Calculation of Total Solids
Weight of empty glass dish = A
Sample taken = 10 ml
Weight of dish + dry residue = B
Total solids = (B – A) X 100 / 10, expressed as percentage
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λ Max
Molecules have translational, rotational, or vibrational
modes. Molecules as result of these movements have
energy.
The energy from ground level gets excited if light or
electromagnetic waves are passed through it. It is
possible to effect a change in a particular type of
molecular energy using appropriate frequency and
wavelength of the incident radiation and measure them.
This is the basis of Spectroscopy..
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Result of these leads to change in absorption.
Intensity of emitted light will decrease as the thickness
or concentration of absorbing media (drug solution)
increase.
The wavelength corresponding to maximum absorptivity
is denoted by λ max.
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Fluorescence Analysis
Many substances when suitably illuminated, emit light of a
different color from that which falls on them. This fluorescence
ceases when the exciting light is removed.
Aconitine - light blue
Emetine - orange
Inner surface of cinchona bark + dilute sulphuric acid - blue
Location of separated compounds on paper and TLC by use of
UV light is extensively employed.
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Chromatography
Chromatography is a separation process based upon the
differential distribution of a mixture between two phases, one
of which is percolated through other.
There are various methods of chromatography study –
paper, thin layer, columnar, gas, etc.
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The fixed phase is called stationary phase and the
other is termed as mobile phase.
If the stationary phase is a solid, the process is called
adsorption chromatography.
If the stationary phase is a liquid, the process is called
partition chromatography.
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Rf value :
The ratio of distance travelled on the medium by a given
compound to the distance travelled by the front of the
mobile phase, from the point of the application of the
test substance, is the Rf value of the compound.
Rr value :
The ratio between the distances travelled by a given
compound and a reference substance is the Rr value.
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Location of the Spots :
• Direct inspection or ultraviolet light
• Inspection after treatment with reagents that will make
the spots visible – for example, Dragendorff's agent,
iodine vapour, antimony trichloride in chloroform, etc.
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Paper Chromatography :
In paper chromatography the adsorbant
is a sheet of paper of suitable texture and
thickness.
The paper chromatography is of following types:
Descending Chromatography
Separation of chemical compounds by descending
chromatography is accomplished by a procedure of allowing the
mobile phase to flow downward on the chromatographic sheet.
Ascending Chromatography
In ascending chromatography the lower edge of the sheet is
dipped into the mobile phase, to permit the mobile phase to rise on
the chromatographic sheet.
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Thin Layer Chromatography (TLC)
It consists of preparing, on a suitable glass plate, a thin
layer of material.
The mother tincture is dissolved in a suitable solvent and
placed as a series of spots on the film towards one end
of the plate; this end is dipped in a suitable solvent
mixture and the whole enclosed in an airtight container.
Solvent front travels up the film and after a suitable time
plate is removed, solvent front marked, solvent allowed
to evaporate and positions of separated compounds
determined.
www.experthomeo.com
Dr.A.B.Ram Jyothis MD (Hom)
Reader
Department of Homoeopathic Pharmacy
Athurasramam NSS Homoeopathic Medical College
Phone No: 9497053705
www.experthomeo.com

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Standardization Of Homeopathic Preprations

  • 3. • Standardization is a process of having a product in uniform standard as mentioned in the pharmacopoeia. • • It includes a Quality Control System with various tests to perform at different stages of manufacture, from raw materials to finished products. • It also involves an Assurance & assessment of quality of products with ref. to a pharmacopoeial monograph. www.experthomeo.com
  • 4. • Supply of essential drugs of good quality which is the prerequisite for the delivery of any Health Care • Functioning of national drug regulation and control systems which is the only means to assure safety and quality of medicines www.experthomeo.com Standardization of Drugs Depends on
  • 5. • To assure pure & quality end products in a proper container bearing the correct label • In order to find its relevance to homoeopathic pharmaceutics www.experthomeo.com Aims
  • 6. • Confirmation of Drug identity • Determination of Drug quality & purity ( impurity, if any) • Determination of Vehicle quality & purity ( impurity, if any) • Detection of nature of adulteration • Estimating active constituents present in crude drug All these steps constitute the Evaluation of Drugs Steps www.experthomeo.com
  • 7. • Biochemical variation in the drug • Deterioration due to treatment & storage • Substitution & adulteration, as a result of mishandling, ignorance or fraud www.experthomeo.com Evaluation of Drugs is necessary for three reasons:
  • 8. Drug Adulteration Original crude drugs are partially/wholly adulterated by substitution with substandard similar looking substance i.e. adulterant Type of Adulterants: • Substitution with substandard commercial varieties e.g. nux blanda for nux vomica; medicinal ginger for Cochin ginger Cochin ginger Medicinal ginger www.experthomeo.com
  • 9. Substitution with similar inferior drugs e.g. ailanthus leaves for belladonna leaves; Japan wax for beeswax bell ailanthus www.experthomeo.com
  • 10. Type of Adulterants • Substitution with artificially manufactured substances e.g. yellow paraffin for beeswax; compressed chicory for coffee • Substitution with exhausted drugs e.g. clove, coriander • Substitution with synthetic substances e.g. benzyl benzoate to balsam of Peru www.experthomeo.com
  • 11. Type of Adulterants: • Substitution with vegetable matter from the same plant e.g. moss, liverworts and epiphytes growing in bark mixed with cinchona / cascara; stem mixed with leaves of stramonium, lobelia • Substitution with harmful adulterants e.g. limestones in asafoetida, leadshot in opium • Adulteration of Powders e.g. dextrin in ipecac, ginger powder in colocynth www.experthomeo.com
  • 12. Guidelines for Pharmaceutical Operations & Record Keeping • Proper sanitary measures • Cleansing & calibration of equipments on regular basis • Maintaining record of receipt, identification, processing, yield of manufacture, storage, handling, sampling, testing of labelling & packaging • Keeping record of any deviation from these written procedures & its justification & approval www.experthomeo.com
  • 13. Stability of Drugs a. Environmental factors such as heat, moisture, light, oxygen, vibration, radiation, & other physical stress & changes b. Product-related factors such as: i. Chemical & physical properties of crude & active drug substances & of pharmaceutical aids (excipients) used ii. Dosage form & its composition iii. Manufacturing process iv. Nature of container & other packaging www.experthomeo.com
  • 14. Standardization of Homoeopathic Products In India, Drugs and Cosmetics Act 1940 controls quality of homoeopathic medicines. The Homoeopathic Pharmacopoeia Laboratory (HPL) was established in 1975 under the Ministry of Health and Family Welfare, Government of India as a quality monitoring apex body. www.experthomeo.com
  • 15. Worked out standards are released in form of Homoeopathic Pharmacopoeia of India (HPI). The problems of quality control with homoeopathic potencies serve one of the areas of greatest challenge. www.experthomeo.com
  • 16. The extremely high dilutions of homoeopathic potencies make it almost impossible to apply analytical tests by conventional methods in the laboratory. Only mother tinctures, apart form the raw material are subjected to a comprehensive analysis, both qualitative and quantitative. Good Manufacturing Practices (GMP) must be in force to assure & assess the quality of homoeopathic preparations www.experthomeo.com
  • 17. Quality Control of Raw Material www.experthomeo.com
  • 18. The pharmacopoeia indicates in the monograph of each drug, details for identification and collection, the part to be used, the method of preparation, assessment of purity and limits of impurity. It is essential that these instructions be strictly followed to obtain mother tinctures of desired standards. www.experthomeo.com
  • 19. Evaluation involves determination of identity, purity and quality or activity of drug ... • Identity is determined by morphological characters, sensory characters, microscopic characters, chemical and micro-chemical tests and certain physical constants. • Purity is determined by estimating foreign inorganic matter, foreign organic matter and moisture content. • Quality is determined by physical, chemical & biological methods. www.experthomeo.com
  • 20. • Organoleptic evaluation • Microscopic evaluation • Physical evaluation • Chemical evaluation • Biological evaluation Methods www.experthomeo.com
  • 21. O r g a n o l e p t i c E v a l u a t i o n It refers to qualitative evaluation of drugs by means of organs of senses, e.g. eyes (to see); nose (for odour); tongue (to taste); skin (by touch) and includes the macroscopic appearance, its odor, taste and the feel of drug to touch. Occasionally, the fracture of plant material is also considered. www.experthomeo.com
  • 22. • External morphology (Shape and Size) • External colour • External markings • Fracture • Odour • Taste www.experthomeo.com
  • 23. M i c r o s c o p i c E v a l u a t i o n It helps in identifying organized drugs & searching adulterants in powdered drugs. • Histology • Micro-chemistry: - Isolation of drug constituents by micro-sublimation, micro-extraction, micro-filtration, micro-crystallization - Identification of constituents of drugs by crystallography, melting point determination, physical test, chemical test, colour reaction test www.experthomeo.com
  • 24. C h e m i c a l E v a l u a t i o n It covers isolation, identification, purification and characteristic determination of drugs having active principles. It helps in detection of alkaloids, carbohydrates & glycosides, fixed oil & fats, volatile oil, saponins, proteins & free amino acids, gums & mucilages, phenolic compounds & tannins, phytosterols Analytic chemistry may be qualitative or quantitative. Quantitative analysis may be gravimetric, volumetric, titrimetric, colorimetric analysis & etc www.experthomeo.com
  • 25. The chemical assays include : • Color reaction test • Acid value • Iodine value • Saponification value • Ester value • Ash value – Sulphated ash value – Acid-insoluble ash value – Water-soluble ash value www.experthomeo.com
  • 26. P h y s i c a l E v a l u a t i o n •Chromatographic study of drugs •Spectroscopy www.experthomeo.com
  • 27. •Fluorescence test : The reaction of certain drugs either in powdered form or on sectioned surfaces with filtered UV light help in Identification and purity. Aconitine - light blue Berberine - yellow Emetine - orange www.experthomeo.com
  • 28. The application of physical constants is largely employed to the active principle of drugs. • Solubility • Specific gravity • Density • Viscosity • Melting point • Congealing point • Refractive index • Optical rotation • Water content or loss on drying • Foreign organic matter www.experthomeo.com
  • 29. B i o l o g i c a l E v a l u a t i o n Pharmacological activity of certain drugs is applied to evaluate and standardize them. The assays on living animals and on their intact or cut-off organs can indicate the strength of the drug or their preparation. As living organisms are used, these assays are known as biological assays or bioassays. www.experthomeo.com
  • 30. Biological Evaluation Toxic Method Symptomatic Method Tissue Method Live animals Live animals Isolated organs/ used used tissues used www.experthomeo.com
  • 31. Analysis Chemical information – exists between drug & tissues of organism depending upon the chemical affinity Mechanical information – includes signs of efforts by organism to reject the offending substances from its inside Dynamic information – includes changes depending upon the vitality & susceptibilities of living, healthy/diseased organism in relation to the peculiar properties of drug in small qty www.experthomeo.com
  • 32. Few examples of Bioassays • Cardiac glycosides containing drugs like digitalis are bio- assayed on cat, frog or pigeons. • Evaluation of ergot is done on rabbit intestine or uterus. • Saponin is assayed by haemolytic index on fish. • Curare causes relaxation of muscles and is carried out by rabbit head-drop method. www.experthomeo.com
  • 33. Quality Control of Finished Products www.experthomeo.com
  • 34. An analysis of homoeopathic potencies neither reveals the identity of the medicine, nor the potency. Only mother tinctures can be subjected to a comprehensive analysis, both qualitative and quantitative. “In-process” quality control at every step of preparation, from raw material to finished products, is critical in ensuring the purity. www.experthomeo.com
  • 35. Mother Tincture Analysis • Alcohol content determination • Weight per ml • pH value • Total solids • λ Max • Fluorescence analysis • Chromatography www.experthomeo.com
  • 36. Specific Gravity The specific gravity of a substance is the weight of a given volume of that substance at a stated temperature as compared with the weight of an equal volume of water at the same temperature, all weighings being taken in air. A suitable pycnometer is used for the determination. www.experthomeo.com
  • 37. Alcohol content Estimation of alcohol content of mother tincture is a test for the potency of the mother tincture and an evaluation of the extraction process. Specific gravity of the distillate is determined pycnometrically and the corresponding alcohol contents in percent by volume read off in the alcoholometric tables. www.experthomeo.com
  • 38. Calculation of Alcohol Content • Sample taken at 25oC = 25 ml • Wt of empty specific gravity bottle = A • Wt of specific gravity bottle + distillate at 25oC = B • Wt of specific gravity bottle + water at 25oC = C • Specific gravity = B – A / C – A Alcohol Content, expressed as % v/v is obtained from reference table in pharmacopoeia. www.experthomeo.com
  • 39. Weight per ml Weight per milliliter of a liquid is determined by dividing the weight in air, expressed in grams, of the quantity of the liquid which fills a pycnometer at 20o or 25o by the capacity of the pycnometer at 20o or 25o respectively, expressed in milliliters. www.experthomeo.com
  • 40. pH Determination The pH is defined as the common logarithm of the reciprocal of the hydrogen ion concentration expressed in gram per liter. www.experthomeo.com
  • 41. • Storage, exposure to temperature and pressure tend to alter the pH, thus making the liquid unstable. • It also helps to detect adulteration. • The two most important methods of measuring pH are by the use of colored indicators or of special electrodes. www.experthomeo.com
  • 42. Total Solids • Weigh accurately or measure an accurate quantity of the preparation. Place it in a tarred dish, evaporate at as low a temperature as possible until the alcohol is removed. Heat on a water-bath until the residue is apparently dry. Transfer to an oven and dry to constant weight at 105o. Cool in an efficient desiccator. The term 'total solids' is applied to the residue obtained when the prescribed amount of the preparation is dried to constant weight under specified conditions. www.experthomeo.com
  • 43. Calculation of Total Solids Weight of empty glass dish = A Sample taken = 10 ml Weight of dish + dry residue = B Total solids = (B – A) X 100 / 10, expressed as percentage www.experthomeo.com
  • 44. λ Max Molecules have translational, rotational, or vibrational modes. Molecules as result of these movements have energy. The energy from ground level gets excited if light or electromagnetic waves are passed through it. It is possible to effect a change in a particular type of molecular energy using appropriate frequency and wavelength of the incident radiation and measure them. This is the basis of Spectroscopy.. www.experthomeo.com
  • 45. Result of these leads to change in absorption. Intensity of emitted light will decrease as the thickness or concentration of absorbing media (drug solution) increase. The wavelength corresponding to maximum absorptivity is denoted by λ max. www.experthomeo.com
  • 46. Fluorescence Analysis Many substances when suitably illuminated, emit light of a different color from that which falls on them. This fluorescence ceases when the exciting light is removed. Aconitine - light blue Emetine - orange Inner surface of cinchona bark + dilute sulphuric acid - blue Location of separated compounds on paper and TLC by use of UV light is extensively employed. www.experthomeo.com
  • 47. Chromatography Chromatography is a separation process based upon the differential distribution of a mixture between two phases, one of which is percolated through other. There are various methods of chromatography study – paper, thin layer, columnar, gas, etc. www.experthomeo.com
  • 48. The fixed phase is called stationary phase and the other is termed as mobile phase. If the stationary phase is a solid, the process is called adsorption chromatography. If the stationary phase is a liquid, the process is called partition chromatography. www.experthomeo.com
  • 49. Rf value : The ratio of distance travelled on the medium by a given compound to the distance travelled by the front of the mobile phase, from the point of the application of the test substance, is the Rf value of the compound. Rr value : The ratio between the distances travelled by a given compound and a reference substance is the Rr value. www.experthomeo.com
  • 50. Location of the Spots : • Direct inspection or ultraviolet light • Inspection after treatment with reagents that will make the spots visible – for example, Dragendorff's agent, iodine vapour, antimony trichloride in chloroform, etc. www.experthomeo.com
  • 51. Paper Chromatography : In paper chromatography the adsorbant is a sheet of paper of suitable texture and thickness. The paper chromatography is of following types: Descending Chromatography Separation of chemical compounds by descending chromatography is accomplished by a procedure of allowing the mobile phase to flow downward on the chromatographic sheet. Ascending Chromatography In ascending chromatography the lower edge of the sheet is dipped into the mobile phase, to permit the mobile phase to rise on the chromatographic sheet. www.experthomeo.com
  • 52. Thin Layer Chromatography (TLC) It consists of preparing, on a suitable glass plate, a thin layer of material. The mother tincture is dissolved in a suitable solvent and placed as a series of spots on the film towards one end of the plate; this end is dipped in a suitable solvent mixture and the whole enclosed in an airtight container. Solvent front travels up the film and after a suitable time plate is removed, solvent front marked, solvent allowed to evaporate and positions of separated compounds determined. www.experthomeo.com
  • 53. Dr.A.B.Ram Jyothis MD (Hom) Reader Department of Homoeopathic Pharmacy Athurasramam NSS Homoeopathic Medical College Phone No: 9497053705 www.experthomeo.com