Ayurvedic ointment, lepa, malahara, gandhaka malahara, suppurative fomentation, ointment for skin diseases, cracked heels.

1. Seminar on Analytical
Parameters of Lepa and
Malahara Kalpana
By
Dr.Gajendra.D.R
1st Yr Pg Scholar
RS&BK
Guide
Dr.Laxmi.B.Kurle
Asso Professor
Dept RS & BK
HOD
Dr.M.S.Doddamani
Professor & HOD
Dept RS & BK

2. Contents
• Introduction
• Concept of Lepa
• Concept of Malahara
• Analytical parameters of Lepa and Malahara
• Conclusion

3. Introduction
Topical applications are the main route of drug
administration on dermatological conditions.
Importance of these applications were
highlightened by ancient scholars, Lepa and
Malahara plays a major role in Twakgata
vikara's( skin disorders).
Lepa and Malahara treatment modality comes
under Bhahir parimarjana chikitsa.

4. CHIKITSA
ANTAR
PARIMARJANA
BAHIR
PARIMARJANA
LEPA
MALAHARA

5. Lepa Kalpana
Definition:
The Medicines in the form of a Paste used for external
application are called as lepas.
Synonyms:
Alepa, Lipta, Lepana,Lepa.

6. Malahara Kalpana
Malahara = Malham or Marham from Unani
Introduced in Ayurveda by Yogaratnakara
Usually with a base like Sarjarasa, Taila,
Ghrita, siktha.
Malahara are Considered as ointment, creams.

7. Analytical Parameters
As the demand of various Ayurvedic preparations is going on increasing in
market, manufactures are tempted to or actually compromising with the quality
of the product in urge of fulfilling all demands. Here to judge the quality of the
raw material or finished product purchased from market we need some
standards for each product and for these standards we need on analytical study.
Analytical study provides us the objective parameters for standardization. It
helps in understanding and interpretation of a drug and its structure. It assists
us to conduct the comparative study among various samples during drug
manufacturing, by offering objective parameter to judge the exact status of the
drug and to decide the future work plan. By advanced analytical techniques,
now one can also deduce the volatile content or organic content of a Herbo-
mineral drug.

8. Analytical Specifications of Lepa
and Malahara… acc to PLIM
• Organoleptic characters
• Uniformity of Content
• Microscopic
• Rancidity Test
• Identifications. TLC / HPLC/GLC
• Assay
• Viscosity
• pH
• Particle size (mesh size 125-150)
• Total fatty matter
• Loss on drying at 1050c
• Spreadability.
• Test for heavy metals:
Lead, Cadmium , Mercury, Arsenic.
• Microbial contamination:-
Total Bacterial Count
Total Fungal Count
• Test for specific pathogen:-
E. coli
Salmonella spp
S.aureus
Pseudomonas aeruginosa

9. Organoleptic characters
 Colour:- This represents the Colour of the lepa. It is mainly
depends on ingredients which are used in that. Change in
colour indicates improper preparation of lepa or detoriation of
lepa.
ex:- Rasadi lepa -Black colour
 Smell:- This shows smell of the product.
ex:- Rasadi lepa- Sarshapa Gandha.
 Texture :- This percives surface quality of the product .
It is Soft for lepa's and Malahara .

10. Content Uniformity
Most important parameter governing product
stability and process control of the disperse
system.
In ointment/cream formulation are more
dependent on particle size, shear rate, and mixing
efficiency in order to attain and maintain
uniformity of the active drug component(usually
the internal phase).

11. Method :
Drug content -1gm of lepa was accurately weighed, in
a 50ml of volumetric flask to which 20ml purified water
was added with continuous shaking. Volume was
adjusted with a mixture of 10% methanol in water.
Absorbance of the solution with the blank was
measured at 360nm using UV-spectrophotometer.
Homogeneity of drug content -For homogeneity of
drug contents, six tubes were taken randomly and
assayed for the drug content as stated above.
The usual sample size for testing ranges between 0.5
and 1.5 g per sample assay.

12. Microscopic
Microscopic identification of the botonical ingradients is a
standard for statutory purposes in several solid and semisolid
composed formulations.
Microscopic identification tests are confined to those
formulations where the botonical ingradients are not more then
ten and where they are added in situ in power form as prakshepa
dravys. such comminuted ingradients land themselves for
microscopic identification as they are not drastically changed in
cell structure or content while processing and appear intact in
microscopic slide preparation after proper treatement.
Method of preparing specimens of crude materials of vegetable
drugs for microscopical studies vary, depending on the
morphological groups of drugs to be examined and also on the
natures of the materials i.e entire cut or powdered.

13. Rancidity Test
Rancidity testing determines the level of oxidation in a sample.
when lipids (fats and oils ) go rancid, its nutritional values is
compromised, and the lipids will taken on a rancid taste and
odour. Proper rancidity testing is an essential component in
determining the shelf life of the product.
The test depends upon the formation of a red colour when
oxidized fat is treated with con. HCl and a solution of
phloroglucinol in ether. The compound of Rancid fats responsible
for the colour reaction is epihydrin aldehyde. All oxidised fats
responds to the kreis test and the intensity of the colour produced
is roughly proportional to the degree of oxidative rancidity.

14. TLC
Thin-layer chromatography (TLC) is
a chromatography technique used to
separate non-volatile mixtures.
Thin layer chromatography is done exactly
as it says –
 using a thin uniform layer of silica gel
or
 Alumina coated onto a piece of glass,
metal or rigid plastic.

15. TLC (Thin Layer Chromatography)
TLC in which a solute undergoes distribution between two
phases, a stationary phase acting through adsorption and a
mobile phase in the form of a liquid. The adsorbent is a
relatively thin, uniform layer of dry finely powdered material
applied to a glass, plastic or metal sheet or plate. Glass
plates are most commonly used. Separation may also be
achieved on the basis of partition or a combination of
partition and adsorption, depending on the particular type
of support , its preparation and its use with different solvent.
Identification can be effected by observation of spots of
identical Rf value and about equal magnitude obtained,
respectively with an unknown and a reference sample
chromatographed on the same plate. A visual comparison
of the spot usually serves for semi-quantitative estimation.

16. TLC Applications
 Thin-layer chromatography can be used to monitor the progress of a reaction,
identify compounds present in a given mixture.
 To determine the purity of a substance.
 TLC assays are done as general screening for group of drugs like steroids,
amphetamines & other stimulants , NSAIDS, antihistamines, anti depressants,
barbiturates.
 For quantitative analysis of drugs
 For separation of active ingredients present in the drugs
 For fingerprinting of drugs.
 For quantification of compound drug.
 For standardization of compound drugs.
 To research adulterants present in the drug.
 For identification of drug.

17. Assays
• Drugs which cannot be assayed by chemical, or physical
means are evaluated by biological methods.
• A crude drug may be assayed for a particular group of
constituents(e.g. The total alkaloids in belladonna or the
total glycosides of digitalis) or specific components (
e.g. reserpine content of Rauwolfia spp.)

18. Bio - Assays
When the estimation of crude drug or its preparation
is done by means of its effect on living organism like bacteria,
fungi, or animal tissue or entire animal it is known as
BIOASSAY.
Indication For Bio Assay Evaluation
• This is true for the substances having an Interfering obstacles
• When quantity is too small.
• No specific chemical test is available
• When the action of drug is due to a mixture of substance
• Purification of drug is not possible

19. VISCOSITY
Viscosity is the property of a fluid that resists
deformation. It is a property of a resistance offered by the fluid
material to a movement.
 Viscosity of liquid may be determined by any method that will
measure the resistance to shear offered by the liquid.
 Absolute Vicosity can be measured directly if accurate
dimensions of the measuring instruments are known but it is
more common practice to calibrate the instrument with a liquid
of known viscosity and to determine the viscosity of the
unknown fluid by comparison with that of the known.
 Choice of Viscometer: ostwald viscometer
 Eg. Castor Oil 1000 cp

20. Viscometer

21. Method :
The Viscometer- Calibrated to measure the apparent
viscosity of the disperse system at equilibrium at a given
temperature to establish system reproducibility.
Consistency type Approximate
viscosity in cps at
25°C
Pharmaceutical
example
Soft, spreadable 100,000-300,000 W/O, O/W CREAM
Plastic flow, spreadable 300,000-1,000,000 Ointment
Application :
It is a resistance of a liquid to flow.
Higher the value indicates more the solute or more
concentration of substance. Thus viscosity of lepa and
malahara should be more .

22. pH determination
Topically applied drug products should be tested of pH
at the time of batch release and at designated stability time
points for batch to batch monitoring. Because some topically
applied drug products contain very limited quantities of water
or aqueous phase, pH measurements may not always be
warranted.
This test is generally firnulation dependent. Therefore it
is not included in compendial drug product monograph but is
part of the manufacturer's specification for the drug product.

23. Conti……
The pH value of an aqueous liquid may be defined as the
common logarithm of the reciprocal of the hydrogen ion
concentration expressed in g/l .
Although this definition provides a useful practical means
for the quantitative indication of the acidity or alkalinity of
the solution, it is less satisfactory from a strictly theoretical
point of view.
No definition of pH as a measurable quantity can have a
simple meaning. which is also fundamental and exact.
The pH value of a liquid is determined potentially by means
of glass, electrode and a suitable pH meter.
Eg:- pH of Darvyadi Malahara is :- 3.7 to 4.2

24. Importance of pH
 To determine the Physical, chemical and biological stability of crude drugs.
 Qualitative and quantitative analysis of the samples.
 Solubility of the drug:- pH of the solution affects its solubility. solubility of weak
acids increase (to a limit) as pH increase and solubility of weak bases increase (to a
limit) as pH decrease.
 Stability of the drug :-The pH of the solution can affect the degradation rate of the
drug. pH of stability varies from drug to drug. Example, aspirin is most stable (has
lowest degradation rate) at pH = 2.5.
 Permeability of the drug through biological membranes:-This depends on the
extent of ionization of the drug. The pH of the solution can affect the extent
of ionization of weak acidic or weak basic drugs.
 Tissue irritation -pH of a solution can’t be too acidic or too basic. Greater the pH
than the physiological pH greater the irritation.

25. pH Meter
Potentiometric ph meter
Digital ph meter

26. Particle size Consideration
Control of particle morphology and particle
size are important parameters to attain high
quality drug product manufacture and control
procedure.
Particle size distribution for most
disperse system should be in the range of
0.2-20 microns.

27. Conti….
Applications
- Size and surface area of a particle can be related
in a significant way to the physical, chemical, and
pharmacological properties of a drug.
-Clinically, the particle size of a drug can affect its
release from dosage forms that are administered orally,
parenterally, rectally and topically.

28. FAT CONTENT
To estimate fatty content present in the product
Method :-
• Extract accurately weighed air dried sample ,with
petroleum ether (40-600C) in Soxhlet apparatus.
• Dry the extract over anhydrous sodium sulphate and
remove solvent under vacuum at 400C.
• Weigh residue and calculate percentage with
reference to the weight of sample material used.

29. Loss on drying
Method is used to measure the amount of water content and other
volatile material in a sample upon drying or heat treatement.
method :-
Weigh accurately about 2gm of drug in a nickel or silica crucible . if
the sample is large crystal or lump, promptly crush it into particles
not larger than 2mm in diameter.
Spread the sample on the crucible so that the layer is not thicker
than 5mm, and weigh it accurately.
Dry in an air over at 1050 till a constant weight is obtained.
If the sample melts at a temperature lower than the specified drying
temperature, dry it at a tempereture 5-100 lower than the melting
temperature for 1to2 hours and dry it under the specified conditions.
The difference in the two weighing gives loss on drying . Calculate
the % of loss on drying

30. Spreadability
The efficasy of a topical therapy depends on the patient
spreading the drug formulation in an even layer to
administer a standard dose.
Spreadability is therefore an important characteristic of
these formulations and is responsible for currect dosage
transfer to the currect target site, ease of application on
the substrate, excrudability from the package and most
important, consumer prefarence.

31. Spreadability method
Determined by an apparatus suggested by Muttimer
(1999) Consist of a wooden block having a pulley at
one end with fixed glass slide on block.
Spreadability was expressed in terms of time in seconds
taken by two slides to slip of from the cream/ ointment,
place in b/w the slides under certain load. Lesser the
time taken for separation of the two slides, better the
spreadability.

32. A shorter interval indicates better spreadability. Spreadability is
measured as S = m × l /t.
S:- Spreadability, m:- weight placed over the upper glass plate.
l:- length of the glass, t:-Time for spreading in seconds.

33. Test for Heavy Metals
The test for Heavy Meatals is designed to determine the
content of metallic impurities that are coloured by
sulphide ion, under specified conditions. The limit for
Heavy metal is indicated in the individual monographs
in terms of the parts of lead per million parts of the
substance ( by weight), as determined by visual
comparision of the colour prodused by the substance
with that of a control prepared from a standard lead
solution.

34. Microbial limits
The following tests are designed for the estimation of
the number of visible aerobic micro-organisms present
and for detecting the presence of designated microbial
species in phamaceutical substances. The term 'growth'
is used to designate the presence and presumed
proliferation of viable micro-organisms.

35. Limits for ASU products
Parameter Specifications
Total bacterial count 1 X 105 CFU/gm
Yeast & mould 1 X 103 CFU/gm
E.Coli Absent
Salmonella Absent
P. Aeruginosa Absent
S. Aureus Absent
2. Pesticide residue – organo-chloro group Less than 1 ppm
3.Heavy metals
Lead 10 ppm
Mercury 01 ppm
Arsenic 03 ppm
Cadmium 0.3 ppm
4.Aflatoxin B1 – 0.5 ppm
G1 – 0.5 ppm
B2 – 0.1 ppm
G2 – 0.1 ppm

36. Conclusion
• No doubt that Analytical Parameters play vital
role in standardization of lepa and malahara
kalpana.
• Ayurvedic drugs are comes under the drug
and cosmetic act, the need for the quality
control methods for the Ayurvedic drugs has
become very necessary.
• Many classical preparations of Lepa and
Malahara are yet to be standardized which are
used in day to day practice.