This document discusses various analytical parameters that can be used to analyze bhasma formulations in Ayurveda. It begins by explaining the importance of analyzing bhasma quality given its increased demand. It then discusses several analytical techniques that can be used, including conventional chemical analyses, physico-chemical analyses using modern instrumentation, X-ray diffraction for characterization, SEM-EDX for elemental analysis and mapping, FTIR for identifying functional groups, and particle size analysis. The document provides details on the principles, procedures, advantages, and applications of these various analytical techniques for studying bhasma quality in a scientific manner.

1. BY-Dr.Ranjith.B.M.2nd year P.G scholar
Dept of RS&BK
T.G.A.M.C
Bellary
GUIDE
Dr.SHANKAR
GOWDA
HOD(UG)
Dept of RS & BK,
TGAMC,
BELLARY
HOD
Dr.M.S.DODDAMANI
Dept of RS & BK,
TGAMC,
BELLARY
SEMINAR ON ANALYTICAL PARAMATERS
OF BHASMA

2. Contents
Analytical paramaters
Bhasma
Analytical paramaters in bhasma
conclusion

3. IMPORTANCE OF TOPIC
In the present scenario, where Rasashastra
preparations are having an increased demand in
market, the need to judge the quality of the Raw
material or finished product is the need of the hour,
Analytical study provides us the objective parameters
for standardization.

4. INTRODUCTION

5. INTRODUCTION
ANALYTICAL STUDY
The analytical study reveals the chemical
composition of the formulations as well as their
concentration. By this it helps to ensure safety
limits and accuracy of the drug. Physico-
chemical analyses of the drugs are carried out
by using current analytical methodologies for
understanding and interpretation of physico-
chemical changes occurring during and after
pharmaceutical processing.

6. Analytical Study in a research helps to explore the topic
in-depth. It involves evaluation of the concepts &
information relative to the research being conducted.
In a study it can be used to prove a hypothesis or
support an idea by giving supportive evidence to the
current research being done, which makes the work
more reliable.
The important aim of Analytical study is to know the
particular chemical configuration and to know the
Physico-chemical changes and effect of different
Samskaras (Shodhana, Marana etc.) and the probable
role of a media during these pharmaceutical processes.

7. Concept of Bhasma:
The word ‘Bhasma is formed from the words
‘bha’ which means shine or luster & suffix ‘Sma’
indicates past tense. So the entire term ‘bhasma’
means ‘Shining in the past’ or “one which has lost the
luster”.
The prime aim of converting minerals or metals
into suitable form for their effective absorption is due
to the fact that the pre-requisite for eliciting any
pharmacodynamic response by any substance on the
human system, is its absorption in the system. After
the marana process metals or minerals elements get
transformed into organo-metalic compounds which
are easily assimilable in the biological system.

8. Purpose behind this preparation of Bhasma
To make metals and other mineral substances useful
for mankind therapeutically.
To reduce or destroy their toxic effect if any.
To convert them into organo – metallic compounds
which are acceptable by the body.
The fineness of final product ensures easy absorption.
To plot or enhance required virtues in the substance.
To obtain stable form of the material.

9. Properties of Bhasma:
-Very fine therefore easy for absorption &
assimilation
-Quick action
-Minimal dosage
-Properties of curing many diseases are
developed when this prakriya is done in
combination with mercury.

10. Parameters for assessment of Bhasma:
Qualitative Analysis: Information
regarding the presence or absence of one
or more components of the sample.
Quantitative Analysis: Information which
is finally obtained by measuring the
physical property that is characteristically
related to the component.

11.  Methods of Analysis:
Conventional Chemical analysis by
volumetric & Gravimetric method
analysis.
Physical analysis.
Instrumental methods of analysis by
using analytical instrumentation.

12. The Bhasmas are mainly examined in terms of
physical test and chemical test. The tests
mentioned below are applicable totally or
partially to the Bhasmas of different drugs. All
the tests are not applicable to all Bhasmas;
selected tests are applicable for particular
Bhasmas.

13. Sl.No. 3. Tests
1 Description/Colour/Odour
2 Identification –chemical composition
3 Particle size mesh size — 200 - 300
4 Loss on drying at 105 C
5 Total – ash
6 Acid – insoluble ash
7 Water soluble ash
8 Assay of element (s)
9 Ayurvedic specifications
10 Lustreless (Nishchandrica)
11 Fine enough to enter the crevices of finger (Rekha purnatva)
12 Floats on water (Varitara)
13 Smokeless (Nirdhoom)
14 Tasteless (Niswadu)
15 Irreversible (Apunar bhav)
16.NPST

14. Bhasma Varna: If the colour of prepared
Bhasma coincides with that of textual
description it can be considered as one of
the signs of properly prepared Bhasma.
The color of the Bhasmas of the same drug
may vary according to the media used, so
this test can be taken as supportive one.

15. Identification –chemical composition
It has been carried out to estimate the
concentration of elements present in
it. Chemical tests are carried out
basically by Volumetric, Gravimetric
analysis and analytical instruments

16. TITRIMETRIC / VOLUMETRIC ANALYSIS
The term titrimetric analysis refers to quantitative chemical analysis carried out by
determining the volume of a solution of accurately known concentration which is
required to react quantitatively with a measured volume of a solution of the
substance to be determined.
Procedure:
The standard solution is added from a long graduated tube called burette.
The process of adding the standard solution until the reaction is just complete is
called a titration.
The completion of the titration is detected by some physical change, produced by
the standard solution itself.
The reaction must fulfill the following conditions for use in titrimetric analysis:
The substance to be determined should react completely with the reagent.
The reaction should be accurately fast. (most of the ionic reaction satisfy this
condition)
There must be an alteration in some physical or chemical property of the solution at
the equivalence point.

17. Advantages:
Equipment does not require constant
re-calibration.
Methods are relatively inexpensive.
High accuracy.
Applied aspect: It is a convenient means of qualitative and
quantitative estimation of the elements.

18. GRAVIMETRIC ANALYSIS
Fundamentals:
Gravimetric analysis deals with the weighing of a substance
that has been either precipitated from solution or volatilized
and absorbed.
The factors which determine successful analysis by
precipitation :
The precipitate must be so insoluble that no appreciable loss
occurs when it is collected by filtration.
The physical nature of the precipitate must be such that it can
be readily separated from the solution by filtration and can be
washed free of soluble impurities.
The precipitate must be convertible into a pure substance of
definite chemical composition either by ignition, evaporation,
etc.

19.  Precipitation of the desired
constituent
 Filtration
 Drying
 Weight of the precipitate.
Applications:
•Analysis of the standards which has to be used for the testing
or calibration of instrumental techniques.
Analysis requiring high occurrence, although the time
consuming nature of gravimetric limits this application to
small numbers of determinations
Steps involved in Gravimetric analysis:

20. X-RAY DIFFRACTION METHOD
Introduction: About 95% of all solid materials
can be described as crystalline. When X-rays
interact with a crystalline substance, one gets a
diffraction pattern. The X-ray diffraction pattern
of a pure substance is, therefore, like a
fingerprint of the substance.
Definition: X-ray diffraction is a method of
determining the arrangement of atoms within a
crystal with the help of x-rays.

21. Principle: Solid matter can be described as:
Amorphous: The atoms are arranged in a random way.
Crystalline: The atoms are arranged in a regular pattern.
When a beam of X-ray is incident upon a crystalline substance, the
electrons constituting the atoms of that substance start oscillating at the
same frequency as that of incident X-ray and emits electromagnetic
radiations in all directions. Every crystalline substance scatters the X-rays
in its own unique diffraction pattern producing a fingerprint of its
atomic and molecular structure.
The distance between each set of atomic planes (i.e. inter atomic
space‘d’) is determined with the help of wave length (λ) of x-ray beam
and angle of diffraction (θ)by applying Bragg’s Law (n λ= 2d sin θ). This
law relates the wavelength of electromagnetic radiation to the
diffraction angle and the lattice spacing in a crystalline sample.

22. Instrumentation: X-ray Tube: X-rays are generated in a cathode ray tube
by heating a filament to produce electrons.
Collimator: In order to get a narrow beam of x-rays, the x-rays
generated by the target material are allowed to pass through a
collimator which consists of two sets of closely packed metal plates
separated by a small gap.
Monochromators: Filters or crystal monochromators, is required to
produce monochromatic X-rays.
X-ray detector: The x-ray intensities can be measured and recorded
either by photographic or counter methods. The photographic method
is mainly used in diffraction studies since it reveals the entire diffraction
pattern on a single film.

23. Procedure: The X-ray generated by a cathode ray tube, is filtered to
produce monochromatic radiation, collimated to concentrate, and
directed toward the sample. The interaction of the incident rays with
the sample produces constructive interference (and a diffracted ray)
when conditions satisfy Bragg's Law (nλ=2d sin θ). These diffracted X-
rays are then detected, processed and counted. By scanning the sample
through a range of 2θangles, all possible diffraction directions of the
lattice should be attained due to the random orientation of the
powdered material. Conversion of the diffraction peaks to d-spacing
allows identification of the mineral because each mineral has a set of
unique d-spacing. Typically, this is achieved by comparison of d-spacing
with standard reference patterns.

24. Applied aspect:
Characterization of crystalline materials.
Measurement of sample purity.
Useful to make distinction between the allotropic
modifications of the same substance.
Inorganic qualitative analysis, where both the compounds
themselves and their chemical derivatives can be identified.
Differentiation among various oxides. For e.g. difference
between FeO, Fe2O3 & Fe3O4 can be identified.
Determining relative abundance and actual state of chemical
combination.

25. Advantages:
Powerful and rapid (<20 min) technique for
identification of an unknown mineral
It is a non-destructive method.
Minimal sample preparation is required.
Data interpretation is relatively straight
forward.
Limitations:
XRD cannot help in the case of amorphous
solids.
Trace element detection is often difficult.

26. XRD
MACHINE

27. SEM-EDX: ENERGY-DISPERSIVE X-RAY
SPECTROSCOPY ANALYSIS
CONDUCTED BY MEANS OF SCANNING
ELECTRON MICROSCOPY

28. Introduction: A Scanning Electron Microscope (SEM) is essentially a high magnification
microscope, which uses a focused scanned electron beam to produce images of the
sample. The electrons interact with the atoms that make up the sample producing
signals that contain information about the sample's surface topography, composition,
and other properties such as electrical conductivity.
Principle: Accelerated primary electrons in an SEM carry significant amounts of kinetic
energy, and this energy is dissipated as a variety of signals produced by electron-
sample interactions. The primary electron beam interacts with the sample in a number
of key ways:-
Primary electrons generate low energy secondary electrons, which tend to emphasis
the topographic nature of the specimen.
Primary electrons can be backscattered which produces images with a high degree of
atomic number (Z) contrast.
Ionized atoms can relax by electron shell-to-shell transitions, which lead to either X-ray
emission or Auger electron ejection. The X-rays emitted are characteristic of the
elements and are measured by the EDX detector.

29. Instrumentation: Essential
components of all SEMs include the
following:
Electron Source ("Gun")
Electron Lenses
Sample Stage
Detectors for all signals of interest
Display / Data output devices

30. Sample Preparation :For conventional imaging in the SEM, specimens
must be electrically conductive, at least at the surface, and electrically
grounded to prevent the accumulation of electrostatic charge at the
surface. Metal objects require little special preparation for SEM except
for cleaning and mounting on a specimen stub.
Nonconductive specimens tend to charge when scanned by the electron
beam, this causes scanning faults and other image artefacts. They are
therefore usually coated with an ultrathin coating of electrically
conducting material, deposited on the sample either by low-vaccum
sputter coating or by high-vaccum evaporation. Conductive materials
in current use for specimen coating include gold, palladium alloy,
platinum, osmium, iridium, tungsten, chromium, and graphite.
Additionally, coating may increase signal/noise ratio for samples of
low atomic number (Z). The improvement arises because secondary
electron emission for high-Z materials is enhanced.

31. Limitations:
EDS detectors on SEM's cannot detect very light elements (H,
He, and Li), and many instruments cannot detect elements
with atomic numbers less than 11 (Na).
Most SEMs use a solid state x-ray detector (EDS), while these
detectors are very fast and easy to utilize, they have relatively
poor energy resolution and sensitivity to elements present in
low abundances when compared to wavelength dispersive x-
ray detectors (WDS).
An electrically conductive coating must be applied to
electrically insulating samples for study in conventional
SEM's, unless the instrument is capable of operation in a low
vacuum mode.

32. Application:
The SEM is routinely used to generate high-
resolution images of shapes of objects and to
show spatial variations in chemical composition.
Acquiring elemental maps or spot chemical
analysis using EDS.
Precise measurement of very small features and
objects down to 50 nm in size is also
accomplished using the SEM.
Back scattered electron images (BSE) can be
used for rapid discrimination of phases in
multiphase samples.

33. Energy Dispersive X-Ray Spectroscopy
(EDS or EDX): is a technique used in
conjunction with chemical microanalysis by
scanning electron microscopy (SEM). EDS
technique detects X-rays emitted from the
sample during bombardment by an
electron beam to characterize the
elemental composition of the volume
analyzed. Functions or steps as small as 1
micron or less can be analyzed.

34. Information Analysis:
Qualitative analysis – The sample values from x-ray energy
spectrum are compared with known characteristic x-ray
energy values to determine the presence of an element in the
sample. Elements with atomic numbers ranging from
beryllium to uranium can be detected.
Quantitative analysis – can be obtained from the count on the
x-ray energy levels characteristic for components of the
sample. Semi-quantitative results are readily available,
without using the standard mathematical corrections based
on the analysis parameters and the composition of the
sample.

35. Applications:
Analysis of foreign materials
Evaluation of corrosion
Analysis of composition of coating
Phase identification and distribution

36. FOURIER TRANSFORM INFRARED
SPECTROSCOPY [F.T.I.R]
It is a powerful tool for identifying
types of chemical bonds in a molecule
by producing an infrared absorption
spectrum that is like a molecular
"fingerprint".

37. Instrumentation
1. The Source
2. The Interferometer
3. Beam splitter
4. Sample
5. The Detector
6. The Computer

38. Advantages of FT-IR:
Speed: Because all of the frequencies are measured
simultaneously, most measurements by FT-IR are
made in a matter of seconds rather than several
minutes.
Sensitivity: The detectors employed are much more
sensitive, the optical throughput is much higher
(referred to as the Jacquinot Advantage) which results
in much lower noise levels, and the fast scans enable
the co-addition of several scans in order to reduce the
random measurement noise to any desired
level(referred to as signal averaging).

39. Mechanical Simplicity: The moving mirror in
the interferometer is the only continuously
moving part in the instrument. Thus, there is
very little possibility of mechanical breakdown.
Internally Calibrated: These instruments
employ a HeNe laser as an internal wavelength
calibration standard (referred to as the Connes
Advantage). These instruments are self-
calibrating and never need to be calibrated by
the user.

40. Application:
Provides special information about chemical
bonding and molecular structures
Capable of identifying organic functional groups
and often specific organic compounds.
Extensive spectral libraries for compound
identification.
It can identify unknown materials
It can determine the quality or consistency of a
sample
It can determine the amount of components in
a mixture

41. 3. PARTICLE SIZE ANALYSIS
Introduction: The Zeta Pals is an automatic
instrument designed for use with suspensions of
particles or solutions of macromolecules. Particles
with diameters from 10nm to 30μm (depending on
particle density) can be measured. The software for
instrument control and data analysis is written for use
in the Microsoft Windows environment though a DOS
version is also available. The technique employed -
electrophoretic light scattering (ELS) - is based on
reference beam (modulated) optics and a dip-in
(Uzgiris type) electrode system. It is also known as
Laser Doppler Velocimetry (LDV).

42. Specifications:-
Zeta Potential Range : -150 to + 150 mV
Size Range : 10nm to 30μm (depending on particle density)
Accuracy : ± 2%
Repeatability : ± 2% with dust free samples
Laser: 35 mW solid state laser, red (660 nm wavelength).
Optional 50mW green (532nm) laser, ~10x sensitivity
Temperature Control : 6 °C to 74 °C in steps of 0.1 °C
Particle Size: Size range (radius)< 0.3 nm to > 3 microns
(*) Sample volume10 µL, 40 µL, 1 - 3 mL
Concentration range0.1 mg/mL to 10%

43. Zeta
pals

44. Loss on drying at 105 degree
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 about 2 mm in diameter
Spread the sample on the crucible so that the layer is not thicker than 5
mm and weigh it accurately.
Dry in an air over at 1100 till a constant weight is obtained.
If the sample melts at a temperature lower than the specified drying
temperature, dry it at a temperature 5 －100 lower than the melting
temperature for 1 to 2 hours, and dry it under the specified conditions.
The difference in the two weighing gives loss on drying. Calculate the %
of loss on drying.
Applied aspect: This method is used to measure the amount of water
content and other volatile material in a sample upon drying or heat
treatment.

45. Total ash
The residue remaining after incineration of the crude drug is designated
as ash. The residue obtained usually represents the inorganic salts
naturally occurring in the drug and adhering to it. It may also include
inorganic matter deliberately added for the purpose of adulteration. The
total ash usually consists of carbonates, phosphates, silicates and silica.
Method:
Accurately weighed about 3 grams of air dried powdered drug is taken
in a tarred silica crucible.It is incinerated by gradually increasing the
temperature to make it dull red hot until free from carbon.
Cooled and weighed, repeated for constant value. If a carbon free ash
cannot be obtained, exhaust the charred mass with hot water.
Residue should be collected on ashless filter paper.Incinerate the
residue collected on the filter paper until ash is white or nearly so.
Then the percentage of total ash is calculated with reference to the air-
dried drug.

46. Acid insoluble ash
Acid insoluble ash is a part of total ash insoluble in dilute
hydrochloric acid.
Method:
The ash obtained as directed under total ash is boiled with 25
ml of 2N hydrochloric acid for 5 minutes.
The insoluble matter is collected on an ash less filter paper,
washed with hot water, dried and ignited to constant weight.
Then calculate the percentage of acid insoluble ash with
reference to the air-dried drug.
Application: A higher limit of acid-insoluble ash is imposed,
especially in cases where silica may be present or when the
calcium oxalate content of the drug is higher

48. Water soluble ash
Determination of Water soluble ash
Method:
Boil the ash for 5 minutes with 25ml of water.
Collect the insoluble matter in a Gooch crucible or on ash less
filter paper;
Wash with hot water and ignite for 15 minutes at a
temperature not exceeding 4500C.
Subtract the weight of the insoluble matter from the weight
of the ash; the difference in the weight represents the water
soluble ash.
Calculate the percentage of water soluble ash with reference
to the air dried drug.
Applied aspect: This water insoluble ash particularly indicates
absorbable percentage of any drug.

49. Ayurvedic specifications

50. Nischandrata:The Bhasma should be
Nishchandra (lustreless) before therapeutic
application. Chandratva (lustre) is the
character of a metal. After proper
incineration, the lustre of the metal should
not remain. This test indicates the change
of specific metallic lustre to a lustreless
compound after incineration. This test is
applicable to Lohas and Abhraka Bhasma

51. Rekhapurnatva: When the Bhasma is
rubbed between index finger and
thumb, particles of Bhasma enters into
the grooves in between the ridges on
the fingertips. This signifies the
microfineness

52. Varitara: When a Mritadravya is
capable of floating on water it is called
varitara

53. Nirdhooma: The bhasma is sprinkled
on red hot coal, if no smoke is emitted
then it can be considered as genuine.
The emission of smoke indicates
presence of some organic substances
in Bhasma & should be given further
Putas.

55. Tasteless (Niswadu)
alpamaatropa yogitvat arucher
aprasangataam|
kshipramaroogya daayitvaata
oushadebhyo adhiko rasah||

56. Irreversible (Apunar bhav)
gudagunjasukasparshamadvajyai saha
yojitam|
nayaati prakrutim
dhmanaadapunarbhavamucchate|

57. Slakshnatvam and Mrudutva: The hard
materials convert to soft and smooth
ash form. These qualities can be felt by
simple touch.

58. Uttama: Mrita substance is considered
Uttama when the bhasma which floats
on water can also carry a relatively
heavier material like dhanya. The
dhanya is said to stay afloat over the
layer of bhasmalika a swan(Hamsa)

59. Niruttha: The Bhasma mixed with
silver, Kept in a crucible & subjected to
intense heat. If there is no weight
changes in silver, it proves that the
Bhasma has been properly prepared &
does not contain any free metal

60. Niruttha pareeksha of vanga

61. NPST
NAMBURI PHASED SPOT TEST
Introduction: Namburi phased spot test was introduced in
1970 by Dr.Namburi Hanumantha Rao for the analysis of
coded Sindoora and Bhasma. The main aim behind
commencing this innovative method is the identification of
Bhasmas and Sinuduras by their specific names as known in
Ayurveda by virtue of their quality difference and not by their
chemical names alone.
Principle: This technique is based on the principles of liquid
chromatography, which helps for the differential identification
of each Bhasma from the other Bhasmas having same
element as the main constituent.

62. Vanga bhasma Haratala bhasma
Taleshwara rasa

63. Procedure: Whatman paper No.1 should be impregnated with 2.5%
Potassium Ferrocyanide and dried carefully on a clean glass sheet.
About 2 gm of the sample (to be tested) should be taken in a test tube
and heated till the bottom appears red, it is cooled & 4ml of 5N HNO3 is
added. The solution is mixed well and heated again for a while. Shake
the test tube now and then for 2hrs. Allow it to react with the reagent
for 40hrs. A drop of this supernatant solution should be carefully put
with the help of dropper on the impregnated Whatman paper. As the
drop comes in contact with the paper an instantaneous characteristic
spot begins to develop and changes with time. The change of colours
and the pattern of the spot at 3 different phases at 3 different time
intervals i.e.,5 minutes, 20 minutes & 4 hours are to be recorded.

64. Application: In Namburi Phased Spot test
sensitivity of reactions at different time
intervals is measured unlike the
chromatography of chemistry.
The continuous chemical reactions taking place
gradually between 2 chemical substances on a
static media at a fraction of second are easily
detected by their distinct colour changes and
the pattern of spot which is specific to each rasa
formulation.

65. CONCLUSION
Its quite important for an Rasa tantragna to
have knowledge regarding analytical
paramaters of bhasma as in this era is of
globalization everything should be explained in
universal language. Also these kinds of
previously recorded data play the role for new
research scholars. Knowledge of these adopted
technologies helps in better understanding and
interpretation of the drug.

66. Thank you
Dr. RANJITH. B. M.