WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Priyanka et al. World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 7.632
Volume 9, Issue 12, 1584-1597 Research Article ISSN 2278 – 4357
A ANALYTICAL STUDY OF JWARA KESARIKA RASA AND ITS
EVALUATION OF ANTI-BACTERIAL ACTIVITY ON SALMONELLA
TYPHI
Dr. Priyanka S.*, Dr. Rajendra Prasad M. L. and Dr. V. Shreeshananda Sharma
1
P.G. Scholar, Department of Rasa Shastra and Bhaishajya Kalpana, JSS Ayurveda Medical
College, Mysuru, India.
2
Assistant Professor, Department of Rasa Shastra and Bhaishajya Kalpana, JSS Ayurveda
Medical College, Mysuru, India.
3
Head, Department of Rasa Shastra and Bhaishajya Kalpana, JSS Ayurveda Medical
College, Mysuru, India.
Article Received on
ABSTRACT
21 October 2020, Introduction: In the current scenario, demand for Ayurvedic drugs is
Revised on 10 Nov. 2020,
Accepted on 31 Nov. 2020 growing exponentially. But, there are innumerable formulations
DOI: 10.20959/wjpps202012-17820 mentioned in the classics of Rasashastra which are yet to be explored
and effectively utilized. Jwara kesarikarasa is one of the unique
*Corresponding Author
formulations mentioned in “Jwara roga- adhikara” of Rasendra sara
Dr. Priyanka S. sangraha which is claimed to treat the disease Vishama jwara
P.G. Scholar, Department of effectively. Jwara kesarikarasa is a Herbomineral mercurial Kajjali
Rasa Shastra and Bhaishajya
compound described in Rasashastra classics. Kajjali prepared with
Kalpana, JSS Ayurveda
Shoditha Parada and Shoditha Gandhaka and finely powdered herbs
Medical College, Mysuru,
India. viz., Vatsanabha, Jayapala, Triphala and Trikatu are triturated with
Bhringaraja swarasa and vati prepared. The reviews hints about two
of the visha dravyas have been advantageously used in this complex herbo mineral
compound which is worth notice. Objectives of the study: To carry out qualitative and
quantitative analysis of Jwara kesarika Rasa. Analytical study: The product was analysed for
physico-chemical, qualitative and quantitative parameters.
KEYWORDS: Vishama jwara, Typhoid fever, Jwara kesarika Rasa, Salmonella typhi, Anti-
microbial testing.
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INTRODUCTION
Rasoushadhis, due to mineral and metallic state, they do not get decayed under any
circumstances. The utility of Rasoushadhis is remarkable because these can be applicable
even in Oushadha prayoga nishidda and asadhya avastha and there is no need to follow
much specifications like prakriti, dosha, desha, kala etc, Rasoushadhis are considered
superior than herbal drugs because of their smaller doses, quick- effects, long shelf-life and
palatability.
Jwara is one of the common disease effects the population.[1] It is the most complex and most
fatal disease as many number of causes are responsible. Jwara is the first disease explained in
Ayurveda indicating it is a prime condition. Raised body temperature is a cardinal feature of
Jwara.[2] Mainly infection with microorganisms are one of the important cause of this raised
body temperature.
In Ayurveda fever treatment is very important and highlighted as this can be understood as in
Charaka Samhita, renowned text of chikitsa where treatment of disease begin with chapter
named Jwara chikitsa. Even after development of Rasashastra, there are quite number of
new formulations were added to improve the outcome of treatment. As it is quite appreciated
that Rasaushadis are quick in action, and results restoration of healthy condition in faster rate
compared with kashta aushadies. In the treatment of Jwara this has tremendous value as
Jwara can affect all systems of body equally from their respective functioning and often
becomes fatal. Even in modern medicine fever is a symptom which has highlighted and
thoroughly investigated in to very deep level so that severity and prognosis can be
determined. So for physicians it is important that fever should be managed aggressively. This
is also true in Ayurveda. Our Acharyas with supreme knowledge, understood the quite
difficulties in treating Jwara and formulated different formulation to overcome this adverse
situation, to help the suffering people and saving many lives.
In this view formulation named Jwara kesarika Rasa taken for study. The name Jwara
kesarika Rasa is repeated in many texts. It is mentioned in Jwara Chikitsa of Rasendra sara
sangraha text.
This preparation consists of Shuddha Parada, Shuddha Vatsanabha, Trikatu, Shuddha
Gandhaka, Trikatu, Shuddha Jayapala, Bhrungaraja Swarasa as a Bhavana dravya. This
formulation has less number of ingredients.
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This preparation is indicated in Sarva jwara, Pitta jwara, Sannipata jwara, Daha jwara,
Vishama jwara, Bhuta jwara, Pliha jwara, Agnimandhya, Ajeerna, Shotha, Shula, Gulma, 18
types of Kustha, Pittaja rogas and also in Taruna Jwara. By keeping in mind all related
factors, it has been decided to work on Pharmaceutico – Analytical study of Jwara kesarika
Rasa and as a first step of research, anti – bacterial activity against Salmonella typhi. Bacteria
was taken.
Hence through this study Jwara kesarika Rasa was prepared in the department of
Rasashastra and Antibacterial study was carried out in the department of Microbiology.
MATERIALS AND METHODS
Analytical Study
The Physico-chemical Analysis of the drug sample was carried out at Ganesh Consultancy
and Analytical Services, Mysuru. The Special Instrumental Analysis was conducted at the
Vignana Bhavan of Manasagangotri, Mysuru.
OBSERVATIONS AND RESULTS OF ANALYTICAL STUDY
1. Classical Parameters
Table no. 1: Showing the interpretations of Kajjali Pareekshas.
Sl.no Name of pareeksha Kajjali Interpretation
+ve after trituration
for 78 hours
1. Varna Jet-black- Physical mixing of HgS
Anjanasannibha
Varna
+ve after trituration Reduced particle size,
2. Rekhapurnatwa
for 78 hours fineness
Lightness and fineness
due to expanded surface
+ve after trituration
3. Varitara area. Specific gravity less
for 78 hours
than water, so does not
break the surface tension
+ve after trituration
4. Nischandrikatwa Absence of free metal
for 78 hours
+ve after trituration Fineness with uniform
5. Shlakshnatwa
for 78 hours distribution of particles
Organoleptic test
Sample - Jwara kesarika Rasa
Sparsha - Smooth, No perceptible coarse particles
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Roopa - Greyish black
Rasa - Katu
Gandha - Teekshna
2. Modern parameters
Table no. 2: Physico-chemical tests[3] of Jwara kesarika Rasa.
Sl.no Test Unit Results Test Method
pH Value 5% GCAS/FC/SOP-
1 -- 3.87
solution 21-M
2 Colour -- Black colour
Error! Filename not specified.
3 Odour -- Characteristics of odour
Error! Filename not specified.
Total Ash (on
4 % 4.55
dry basis)
Acid Error! Filename not specified. Error! Filename not specified. Lab SOP
5
insoluble Ash % 0.50
Water soluble Error! Filename not specified.
6 3.43
Ash %
Loss on Error! Filename not specified.
7 95.45
ignition[4] %
8 Particle size Microns 28-666
Loss on FSSAI Manual
9 % 8.14
drying of Methods
Table no. 3: Microbial contamination test of Jwarakesarika Rasa.
Sl.No Test Unit Result Test Method
IS 5402:2012
1 Total Bacterial count Cfu/g 150000
(RA 2018)
IS 5401 (part
2 Coliforms Cfu/g <10 1):2012
(RA 2018)
IS 5887 (part
3 E.coli /g Absent 1):1976
(RA 2018)
IS:5887 (part 8 sec
4 Staphylococcus aureus /g Absent
1):2002
IS 5887(part
5 Salmonella /25g Absent 3):1999
(RA 2018)
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Table no. 4: Tablet evaluation.[5]
Sl No. Tests Results
Circular, round, brownish black in
1 Appearance
color, rough in surface
2 Uniformity of weight 0.23gm
3 Disintegration test Tablet pass the test at 15 mins
Mechanical strength /
4 1.3 Kg/m2
Hardness
5 Friability 0.7%
3. Characterization
a) SEM[6]
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SEM yielded high resolution figures of the sample surfaces. It showed a characteristic three
dimensional appearance which was useful for determining the surface structure of the sample
in a six images.
b) EDAX7
Graph no 1: EDAX of Jwara kesarika Rasa.
Table no. 5: Quantitative Results for: Base(466) Jwara kesarika Rasa EDAX.
Element Weight % Weight % Atom %
Line Error
OK 80.89 ± 2.54 95.62
SK 5.19 ± 0.62 3.06
SL --- --- ---
Hg L --- --- ---
Hg M 13.92 ± 1.37 1.31
Total 100.00 100.00
It is an analytical technique used for the elemental analysis or chemical characterization of a
sample.
This analysis confirmed the presence of Oxygen, Sulphur and Mercury. It showed the
presence of Oxygen in 80.89% which was the highest component in the given sample.
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Presence of Oxygen element might be due to any herbal drug used either as the ingredient or
for Shodhana purpose.
C) FTIR[8]
Graph no.2: Spectrum Graph
Graph no.3: Peak Table Graph.
Table no. 6: Peak Table.
Y Y Y Y
Peak X (cm-1) Peak X (cm-1) Peak X (cm-1) Peak X (cm-1)
(%T) (%T) (%T) (%T)
1 3318.26 93.53 2 2920.28 92.62 3 1613.12 93.58 4 1017.17 85.71
5 658.91 85.13 6 625.11 84.42 7 604.16 86.14
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Table no. 7: FTIR correlation of Sample.
Standard Actual
Specific type
Bond Type of bond Absorption peak peak in cm- Appearance
of bond
cm-1 (nm) 1
(nm)
Alcohols, High
O─H 3200-3400 cm-1 3318.26 Broad
phenols concentration
Ammonium Multiple
N─H Any 2400-3200 cm-1 2920.28
ions broad peaks
Primary
N─H Any 1560-1640 cm-1 1613.12 Strong
amines
Aliphatic Often
C─N Any 1020- 1220 cm-1 1017.17
amines overlapped
Weak to
C─X Chloroalkanes Any 540-760 cm-1 658.91
medium
Weak to
C─X Chloroalkanes Any 540-760 cm-1 625.11
medium
Weak to
C─X Chloroalkanes Any 540-760 cm-1 604.16
medium
Interpretation
FTIR analysis of the sample showed 7 peaks which includes 3 peaks in the range of 540-760
cm-1 with the peak of 658.91, 625.11 & 604.16 cm-1 respectively with the single weak to
medium appearance of C─X Chloroalkanes bond. O─H bond having Alcohols, phenols with
high concentration in the range of 3200-3400 cm-1 of broad appearance at 3318.26 cm-1 level.
Nitrogen and Hydrogen single strong bond with Ammonium ions and Primary amines peak at
2920.28 & 1613.12 cm-1. C─N bond having Aliphatic amines with the range of 1020-1220
cm-1 often overlapping appearance at 1017.17 cm-1 level.
On the basis of obtained data from FTIR analysis, it can be interpreted that the prepared
sample has complex chemical structure molecules are bound strongly with Carbon, Oxygen,
-1
Nitrogen, Sulphur and weakly with C─X Chloroalkanes. Peak at 3318.26 cm (O─H bond)
with high concentrations of Alcohols, Phenols is reprehensive of presence of free radicals
and this may because of more herbal ingredients in the formulation. However the extent of
this activity can not be claimed based on FTIR analysis. The analytical finding are indicative
of organo-mineral nature of the formulation in which alkaline, Oxide and Hydrogen in nature
is in major phase. Detection of various functional groups is suggestive of the impregnation of
molecules from individual ingredients which may be the effect of Bhavana Samskara or
trituration.
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DISCUSSION ON ANALYTICAL STUDY
Analytical Study of Jwara kesarika Rasa was another objective of the present study and
hence, the formulation Jwara kesarika Rasa was subjected for Analysis using both Classical
and Modern Parameters.
Classical Parameters
Analysis using Classical Parameters play a vital role in assessing the completion of Mardana
& Bhavana procedures incorporated during the preparation of a particular formulation. It also
helps in assessing the authenticity of the preparation.
• Varna of Kajjali was jet black resembling Anjana
• Kajjali have passed through Rekhapurnatwa indicating the fineness and reduced particle
size.
• Kajjali have passed through Shlakshnatwa which helped to confirm the fineness with
uniform distribution of particles.
• Kajjali passed through Varitara indicating the lightness and fineness of particles due to
expanded surface area. Also, as the specific gravity of particle was less than water, it did
not break the surface tension of water.
• Kajjali passed through Nischandrikatwa which indicates the absence of free Mercury.
• It is the confirmatory test of Kajjali which indicates the proper completion of its
preparation.
Modern Parameters
By utilising the modern techniques and tests, the formulation Jwara kesarika Rasa was
analysed for its better understanding.
PHYSICO-CHEMICAL TESTS
Physico-chemical Analysis of the formulation Jwara kesarika Rasa was carried out as per
API. It helped to assess both physical and chemical profile of the formulation.
pH value (5 % solution)
pH Value (5% solution) of the drug Jwara kesarika Rasa was 3.87 %.
This indicates that the formulation Jwara kesarika Rasa is highly acidic in nature. Hence it
can be administered with ghrta.
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Ash value
The ash value of the preparation is 4.55 %, It indicates that the sample is having 4.55% of
inorganic matter in total.
• But as per this report, it can be inferred that rest of 95.45% would be the organic matter
present in the formulation which corresponds to the higher proportions of herbal drugs
present in the formulation.
Acid Insoluble ash
The acid insoluble ash value of the drug Jwara kesarika Rasa was 0.50%. This indicates the
presence of Silica and Silicaceous matter present in the formulation which may be imbibed
from the herbal drug during Bhavana process.
• The human metabolic system and pharmacokinetics of drug depends upon purity and human
acceptable form of drug. Silica and Silicaceous matter of the formulation indicates that part
which is not absorbable in human system. Here, this value corresponds only to the inorganic
part. So, 0.50% of inorganic part of the formulation might not get absorbed.
Water Soluble ash
The Water soluble ash value of the drug Jwara kesarika Rasa was 3.43%. This value
corresponds to the amount of inorganic part of a drug which can dissolve in water. It is very
less, which indicates that the drug is soluble in water.
Loss on ignition
Jwara kesarika Rasa was estimated for loss on ignition and it was 95.45%, it indicate that
there is maximum loss on heating the drug. But in body not so much temperature is there that
which will reduce the efficacy of the drug.
Loss on drying-
The moisture content of any pharmaceutical agent spoils not only the drug activity but
everything. Loss on drying at 1100C is physical test to detect the percentage of least moisture
content and hence the shelf life of the sample, the least on loss of drying at 1100C the better
will be the drug. In the present study on Jwarakesarika rasa was found to posses 8.14% loss
on drying atv1100C hence it can be stated that it posses least moisture and hence very rare
chance of bacterial and fungal growth and also the drug is having least or null hydroscopic
activity and the drug detoriation chance or contamination chances etc. are very less. Also, it
enhances the shelf life of the formulation.
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Microbial contamination
The total bacterial count is 150000 Cfu/g in a sample. Coliforms count is <10 Cfu/g as it is
least count. E.coli, Staphylococcus aureus, Salomonella organisms are absent. As the
organisms are absent in a sample there is no contamination of growth of bacteria.
Disintegration
Disintegration refers to the mechanical break up of a compressed tablet into small granules
upon ingestion and therefore it is characterised by the breakdown of the interparticulate
bonds, which were forged during the compaction of the tablet. The disintegration time taken
in this drug Jwarakesarika rasa was 15 minutes, it indicates all the tablets pass the test at
desired time.
Mechanical strength / Hardness
Hardness of a tablet indicates the breaking point and structural integrity of a tablet and find
out how it changes "under conditions of storage, transportation, packaging and handling
before usage", as 1.3 Kg/m2 was the hardness of the drug Jwarakesarika rasa it implies that
the hardness was less because no binding agent was used while preparing tablets and as it was
manually prepared.
Friability
Friability indicates to test the durability of tablets during transit. 0.7% was the Friability of
the tablet Jwarakesarika rasa, it implies the tablet can withstand wear and tear during
transportation and handling.
Particle Size
Analysis of Particle size is an objective parameter for assessing the subjective property called
„Rekhapurnatwa‟ being mentioned in Rasa Shastra classics. Smaller the particle size, larger is
the surface area and hence, greater are the chances of absorption.
The particle size of the final product is 28-666 microns; this fineness may be due to mardana
and bhavana process done.
It increases the surface area of the compound for better action.
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SEM
Interpretation
SEM images of the drug Jwarakesarika Rasa showed particles which were not uniform in size
and having irregular shape and rough particle surface. It might be due to the presence of
herbal drugs like Trikatu Choorna and Triphala Choorna in higher proportions. Also,
presence of Mercury & Sulphur components and manual handling might have contributed for
irregular shape and rough surface of particles.
SEM study suggested the fineness of the drug with particle size ranging between 28-666
microns. For the effective role in therapeutic bio-availability of metallic and mineral
preparation, particle size is one of the important factors, as it decides the permeability of drug
through cells, tissues and blood capillaries.
EDAX
Interpretation
On scanning, Jwara kesarika Rasa showed peaks of Oxygen, Sulphur and Mercury. All
these elements corresponded to the ingredients of the formulation.
It showed the presence of Oxygen in 80.89% which was the highest component in the
given sample.
Presence of Oxygen might be from the herbal drugs used in the formulation either as an
ingredient or for Shodhanadi procedure.
K, L and M are the vacancy of element, sometime element may lose electron and vacancy
changes. K is inner vacancy, L is Middle vacancy and M is outer vacancy.
Element proportion can be identifies along with its vacancy and vacancy is related with
reactivity.
FTIR
Interpretation
FTIR is an effective analytical instrument for detecting functional groups and characterizing
covalent bond. FTIR identifies chemical bonds in a molecule by producing an infrared
absorption spectrum. The functional group region runs from 4000 cm-1 to 500 cm-1 and the
functional group region contains relatively few peaks.
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FTIR analysis of the sample showed 7 peaks which includes O─H bond having Alcohols,
phenols with high concentration in the range of 3200-3400 cm-1 of broad appearance at
3318.26 cm-1 level.
Nitrogen and Hydrogen single strong bond with Ammonium ions and Primary amines
peak at 2920.28 & 1613.12 cm-1.
C─N bond having Aliphatic amines with the range of 1020-1220 cm-1 often overlapping
appearance at 1017.17 cm-1 level.
3 peaks in the range of 540-760 cm-1 with the peak of 658.91, 625.11 & 604.16 cm-1
respectively with the single weak to medium appearance of C─X Chloroalkanes bond.
On the basis of obtained data from FTIR analysis, it can be interpreted that the prepared
sample has complex chemical structure molecules are bound strongly with Carbon,
Oxygen, Nitrogen, Sulphur and weakly with C─X Chloroalkanes.
Peak at 3318.26 cm-1 (O─H bond) with high concentrations of Alcohols, Phenols is
reprehensive of presence of free radicals and this may because of more herbal ingredients
in the formulation.
The analytical finding are indicative of organo-mineral nature of the formulation in which
alkaline, Oxide and Hydrogen in nature is in major phase. Detection of various functional
groups is suggestive of the impregnation of molecules from individual ingredients which may
be the effect of Bhavana Samskara or trituration.
CONCLUSION
Jwara kesarika Rasa is a Herbo-mineral classical Khalvi Rasayana preparation.
Khalvi rasayana is a unique scientific pharmaceutical method to bind herbal ingredients
to the metallo mineral ingredients without direct Agnipaka. Hence it has less
instrumentation, but based on Bhavana principle.
Thereby, the ingredients will form a mono molecular compact form.
The particle size of the formulation ranged between 28-666 microns
pH Value (5% solution) of the drug „Jwara kesarika Rasa’ was 3.87 % indicating the
highly acidic in nature of the formulation.
SEM images of the drug ‘Jwara kesarika Rasa’ showed particles with irregular shape and
rough particle surface which were not uniform in size.
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EDAX analysis confirmed the presence of Oxygen, Sulphur and Mercury. Oxygen was
the highest component of the formulation (80.89%).
FTIR identifies chemical bonds in a molecule by producing an infrared absorption
spectrum. As organic compounds are more in number in this formulation Oxide and
Hydrogen in nature is in major phase. The above physico-chemical tests were within the
standard limit, hence proving the safety and genuinity.
REFERENCES
1. Vaidhya Trikamji Yadavji Acharya, Caraka samhita of Agnivesha, Chaukhamba
Surbharathi Prakashan, Varanasi, Reprint-2013, Nidana sthana, Jwara Nidana, Chapter 1,
Shloka-35, 202-203, 738.
2. aidya davji Trikamji charya and r yan R m ch rya yatirtha, Su ruta
Samhit of Su ruta, houkamba Surbharati Prakashan, aranasi, Reprint- , hikitsa
sth nam, Athuropadrava chikitsitam, hapter -39, Shloka-, 824.
3. https://en.m.wikipedia.org/wiki/Analysis
4. https://en.wikipedia.org/wiki/Loss_on_ignition accessed on 16/9/2019
5. https://pharmawiki.in/pdf-tablet-evaluation-pharmaceutics-pharmaceutical- apparatus-
material-ppt/amp/ accessed on 6/3/2020
6. https://en.m.wikipedia.org/wiki/Scanning-Electron-Microscope accessed on 18/10/2019
7. https://en.m.wikipedia.org/wiki/Energy-dispersive-Xray- spectroscopy/accessed on
18/10/2019
8. https://www.intertek.com/analysis/ftir/ accessed on 18/10/2019
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