Evaluation of anti bacterial activity of arka taila 5 1- 16
1. EVALUATION OF ANTI BACTERIAL
ACTIVITY OF ARKA TAILA PREPARED
WITH AND WITHOUT MURCHANA
PRESENTER
Dr.D. SEETARAMA KISHORE
Final YEAR PG SCHOLAR
GUIDED BY
Dr. GAZALA HUSSAIN
ASSOCIATE PROFESSOR
Department of RS &
BK
S.D.M. COLLEGE OF AYURVEDA & HOSPITAL
HASSAN
2. CONTENTS
• Introduction
• Research question
• Hypothesis
• Aim & Objectives
• Pharmaceutical study
• Analytical study
• Experimental study
• Discussion
• Conclusion
2
3. Introduction
• Prevalence of Skin disorders is more in present
era
• Indication of external medication is more for skin
ailments
• Vicharchika (Eczema) is one of the common
disorders and one of the main cause of it is micro
organisms
• Arka taila is a classical external medication with
ingredients having Krimihara action
3
5. Hypothesis
5
• Arka taila shows
Antibacterial activity with
and without murchana
Research
hypothesis
• Arka taila does not show
Antibacterial activity with
and without murchana
Null
hypothesis
6. 6
Arka taila
Murchita arka taila
Amurchita arka
taila
1/8th
Kalka
1/4th
Kalka
1/8th
Kalka
1/4th
Kalka
Murchana of
Sarshapa taila
Samples A B
C D
1st
Sample
2nd
Sample
7. Aim & objectives
AIM :
• To evaluate Antibacterial activity of Arka taila prepared with & without Murchita
Sarshapa taila
OBJECTIVES :
• To prepare Arka taila with Murchita Sarshapa taila (2 Samples )
• To prepare Arka taila with Amurchita Sarshapa taila (2 Samples )
• Analytical study of four samples of Arka taila
• In vitro study to assess Anti bacterial activity of Four samples of Arka taila
7
8. Plan of study
8
Review of
literature
Pharmaceutical
study
Analytical
study
Experimental
study
Discussion Conclusion
9. Pharmaceutical study
9
Collection of
the drug
Authentication
of the raw
drugs
Murchana of
Sarshapa taila
Preparation of
Murchita arka
taila
Preparation of
Amurchita
arka taila
10. INGREDIENTS
Sl.no Drug Latin name Part used
1 Arka Calotropis
procera
Leaf -
swarasa
2 Haridra Curcuma longa Rhizome
3 Sarshapa taila Brassica
campestris
Seeds
ARKA TAILA PREPARATION
10
11. MURCHANA OF SARSHAPA TAILA
Sl.no Drug Latin name Part used
1 Hareetaki Terminalia chebula Fruit
2 Haridra Curcuma longa Rhizome
3 Mustha Cyperus rotundus Rhizome
4. Bilva Aegele marmelous Fruit pulp
5. Dadima Punica granatum Seeds
6. Naga kesara Musea ferra Flower
7. Krishna jeera Carum carvi Fruits
8. Hribera Pavonia odorata Roots
9. Nalika Cinnamomum tamala Leaf
10. Vibheetaki Terminalia bellerica Fruit
11. Manjishta Rubia cardifolia Root
11
12. PRE PROCEDURE OF MURCHANA
OF SARSHAPA TAILA
• All drugs were pounded in Khalva yantra
measuring
Height external – 15 cm
Height internal – 10.5 cm
Thickness – 2.5 cm
Length – 37 cm
External breadth – 22 cm
Internal breadth – 16 cm
KHALVA YANTRA PESHANI
Length – 28 cm
Circumference of base – 25 cm
12
13. MEASUREMENT OF VESSEL
• Circumference – 101 cm
• Depth at centre - 18 cm
• Thickness – 2 cm
13
14. PROCEDURE OF MURCHANA OF
SARSHAPA TAILA
• Equipments - Vessel, Stirrer, Stove, Measuring jar, Thermometer,
Plate
• Mandagni
• Drava dravya (Jala) – 3 litres 75 ml
• Sneha dravya (Sarshapa taila ) – 768 ml (approx. 770ml)
• Kalka dravya (all pounded drugs) – 216 g
• Quantity obtained - 750 ml
2 Samples Date of commencement Date of completion
1st Sample 14 – 05 – 15 15 – 05- 15
2nd Sample 25 – 06 -15 27– 06 15
14
18. 18
Samples Date of
commenceme
nt
Date of
completion
Murchita
Sarshapa
taila
Arka patra
swarasa
Haridra kalka
Sample – A 19 – 5 – 2015 21 – 5- 2015 500 ml 2 litres 63 g
Sample – B 26- 5- 2015 28 - 5- 2015 500 ml 2 litres 63 g
Sample - C 9- 7- 2015 11 - 7- 2015 300 ml 1200 ml 75 g
Sample- D 16 - 7 - 2015 18 - 7- 2015 300 ml 1200 ml 75 g
19. Contd..
19
Samples Quantity Taken Quantity Obtained
Sample A 500 ml 300 ml (60 %)
Sample B 500 ml 400 ml (80 %)
Sample C 300 ml 200 ml (66%)
Sample D 300 ml 100 ml (33%)
Samples Total Time taken for process
Sample A 5 hrs
Sample B 5 hrs 50 mins
Sample C 3 hrs 58 mins
Sample D 4 hrs
20. Analytical study
• In present study Analytical evaluation of Arka
taila was carried out to develop preliminary
standards
• Four Samples of prepared medicine were
analysed following parameters as per the
references available in protocol for testing
published by CCRAS.
20
21. Organoleptic characters of the Four samples
Parameter SA SB SC SD
Colour Brown Yellow Brown Yellow
Consistency Viscous Liquid Viscous
Liquid
Viscous
Liquid
Viscous
Liquid
Odour Slightly Aromatic Aromatic Characteristic Characteristic
21
22. Results of standardization parameters
Parameter Results n = 3 %w/w
SA SB SC SD
Refractive
Index
1.47156 1.47206 1.47206 1.47006
Saponification
Value
159.06 149.469 180.741 184.594
Acid Value 3.287 3.217 2.275 3.249
Iodine Value 92.486 101.52 100.014 107.611
Specific
gravity
0.916 0.911 0.917 0.919
Viscosity 54.718 64.136 55.132 65.322
22
23. TLC photo documentation of Chloroform extract of S-A with Moorchita Taila
1:8 (SAWM ), S-B without Moorchita Taila 1:8 (SBWM), S-C with Moorchita
Taila 1:4 (SCWM), S-D without Moorchita Taila 1:8 (SDWM
At 254 nm At 366 nm
At 540 nm Post Derivatisation
23
24. Densitometric scan of all the Sample At 254 nm
SAMPLE A SAMPLE B
SAMPLE C
SAMPLE D
24
25. Densitometric scan of all the Sample At 366 nm
SAMPLE A SAMPLE B
SAMPLE C SAMPLE D
25
26. Densitometric scan of all the Sample At 540 nm
SAMPLE A
SAMPLE C
SAMPLE B
SAMPLE D
26
27. 3-D Display of all the samples
At 540 nm
At 366 nm
At 254 nm
Post derivatisation 27
28. Experimental study
• Objective :
• In the present study in-vitro evaluation of Anti
bacterial activity of 4 Samples of Arka taila was
conducted.
• Place of study :
Research Center for Ayurveda and Allied sciences,
SDMCA Udupi, Karnataka.
28
29. Culture media for the organism
• Preparation of nutrient broth:
• Dissolve beef extract (1 g), yeast extract (2 g), peptone
(5 g), Sodium Chloride (5 g) in 990 ml of distilled water.
The pH was adjusted to 7.2 and make up the volume to
1000 ml and autoclaved at 121˚C for 20 minutes.
• Preparation of nutrient agar media:
• Dissolve beef extract (1 g), yeast extract (2 g), peptone
(5 g), Sodium chloride (5 g) in 990 ml of distilled water.
The pH was adjusted to 7.2 and make up the volume to
1000 ml. Finally add (15 g) agar to the media and
autoclaved at 121˚C for 20 minutes.
29
30. PROCEDURE
• The work place in laminar air flow was cleaned using
70 % ethyl alcohol and switch on the UV for 20
minutes
• Inoculation of one loop of Staphylococcus aureus
from the culture into 10 ml of nutrient broth was
done & mixed well
• 15 ml of the agar medium was poured uniformly
over the sterile petri dish
• 1 ml of nutrient broth containing the organism was
added uniformly over petri dish, mixed well and
allowed the media to solidify
30
31. Contd ..
• Five equidistant wells on the plate were made.
• Different volumes of test sample 4 Samples (25
µl, 50 µl, 75 µl and 100 µl) to different wells
were added
• 100 µl of standard (Ampicillin) separately into
different wells was taken
• All the petridish at 37˚C for 24 hrs were
incubated. After the incubation period, the
zone of inhibition was measured
31
36. RESULTS
Volumes
(µl)
Zone of inhibition
(mm) Murchita
Arka taila with
1/8th kalka
(Sample – A)
Zone of
inhibition (mm)
Amurchita Arka
taila with 1/8th
kalka
(Sample – B)
Zone of
inhibition (mm)
Murchita Arka
taila with 1/4th
kalka
(Sample – C)
Zone of
inhibition (mm)
Amurchita Arka
taila with 1/4th
kalka
(Sample – D)
25 µl 05 0 05 0
50 µl 05 0 05 0
75 µl 05 0 05 0
100 µl 06 0 05 0
36
37. Experimental study at S.D.M.Hassan
• A trail was done to check the Antibacterial effect
of Arka taila of all four samples on the culture
taken from patients
• Diagnosed Vicharchika O.P.D. cases in
S.D.M.C.A.H HASSAN were taken for the present
study
• The swab was collected in a sterile bottle and
subjected to Culture and sensitivity test to check
the growth of bacteria
• 5 patients were taken for the study
37
38. • Only one sample showed the growth of Bacteria
& Gram -ve Bacteria was found
• The procedure followed for the study was
similar to method adopted at SDMCRAAS,
Udupi
38
39. 39
Volumes
(µl)
Zone of inhibition
(mm) Murchita
arka taila with
1/8th kalka
(Sample – A)
Zone of
inhibition (mm)
Amurchita arka
taila with 1/8th
kalka
(Sample – B)
Zone of
inhibition (mm)
Murchita arka
taila with 1/4th
kalka
(Sample – C)
Zone of
inhibition (mm)
Amurchita arka
taila with 1/4th
kalka
(Sample – D)
25 µl - - - -
50 µl - - - -
75 µl - - - -
100 µl - - - -
RESULTS
• It was observed that there was no Zone of inhibition of test
standard drug in Gram negative Bacteria at 25 µl, 50 µl , 75 µl ,
100 µl.
40. • Sneha kalpana is one such dosage form in which both the lipid &
water soluble active principles will be extracted.
• In Gudartha deepika commentery of Sharangadhara Samhita it is
opined to prepare Arka taila with 1/4th quantity of kalka dravya
• While other literature review on Sneha kalpana opine the use of
1/8th quantity of kalka dravya when Swarasa is used as the drava
dravya
• Nalika is a controversial drug, it is mentioned as Nelumbo
nucifera in Indian Medicinal Plants & in A.P.I as Cinnamomum
tamala
40
Discussion
41. LITERARY REVIEW
Literary review suggested that the common cause of
eczema is staphylococcus aureus hence t is taken
in present study for anti bacterial study.
PHARMACEUTICAL STUDY
• In the present study Pharmaceutical study has 3
steps
• Murchana of Sarshapa taila
• Extraction of Arka patra swarasa
• Preparation of Arka taila by 4 methods
41
42. Extraction of Arka patra swarasa
• Pilot study of Arka patra swarasa carried out
suggested that Swedana method is better to extract
Arka patra swarasa compared to Puta method &
hence Swedana method was adopted to extract
arka patra swarasa
• Swedana method yielded more Swarasa & time
taken for extraction is Less
• T.S.S of swarasa is 6
42
43. Preparation of Arka taila by 4 methods
• Preparation of taila paka was done for three
days as swarasa is the drava dravya
• Yield of Sample B is more (80%) compared to
Sample A (60%)
• Sample C (66%) has more yield compared to
Sample D (33 %)
43
44. contd...
• The time taken for the preparation of Amurchita arka
taila is more than the Muchita arka taila
• As swarasa is drava dravya which has suspened solid
particles will combine with kalka dravya which leads to
less yield
• The range of Agni for preparation of taila was
maintained at 90 – 94 0 C by using Mandagni for all the
4 samples to incorporate the active principles of the
drugs used into the media
44
45. ANALYTICAL
• Refractive index of 4 samples are almost similar with
minute difference
• Sample A is 1.47156, Sample D is having least as 1.47006
and Sample B, C are having as 1.47207 i.e. A variation of
0.002 was observed. This minute difference is because of
the density of the taila as Sample D is Amurchita arka
Taila with more quantity of Kalka
• Saponification value of Sample A (159.06) is more than
Sample B (149.469), Sample D (184.594) is more than
Sample C (180.741)
• Saponification value is less in Sample A & Sample B with
1/8th kalka which facilitates more shelf life
45
46. Contd..
• Acid value of Sample A (3.287) is more than
Sample B (3.217), Sample D (3.249) is more than
Sample C (2.275)
• The more the acid value indicates more free fatty
acids in taila responsible for rancidity of the
compound
• Iodine value of Sample B (101.52) is more than
Sample A ( 92.486), Sample D ( 107. 611) is more
than Sample C (100.014)
• It is less in Sample A indicates less chance of
Rancidity
46
47. Contd..
• Specific gravity of sample A is 0.916. sample B is 0.911,
sample C is 0.917 and sample D is 0.919. In these 4
samples there is no much difference in specific gravity
values.
• Minimal increase in sample D is observed may be due to
more solute particles in that sample.
• Viscosity is an index of a liquid to flow it is found that the
viscosity of sample A is 54. 718, sample B is 64.136,
sample C is 55.132 and sample D is 65.322.
• With this it can be mentioned that murchita arka taila i.e.
Sample A and sample C shows better absorption than
Amurchita arka taila i.e. Sample B and D.
47
48. • Murchita Arka taila has showed more number of
Bands in T.LC. Report comparing to Amurchita Arka
taila
• Post derivatisation of H.P.T.L.C. Report showed
more number of peaks (Drugs) in Murchita arka
taila comparing to Amurchita arka taila
• The difference in bands are because of the
ingredients added during the process of murchana
of sarshapa taila.
• The Height of the peaks resembles the quantity of
the drug respectively
48
49. EXPERIMENTAL
• In the present study the Anti Bacterial activity of Arka
taila prepared with and without murchana was assayed
against microorganism Staphylococcus aureus
• Amurchita arka taila prepared with both 1/8th and 1/4th
kalka did not show any zone of inhibition at 25, 50, 75,
100 μl concentrations on staphylococcus aureus
• Murchita arka taila prepared with both 1/8th and 1/4th
kalka showed 05 mm zone of inhibition at 25, 50,75 μl and
06 mm at 100 μl concentrations on Staphylococcus aureus
It is observed that activity of standard drug Ampicillin was
15mm at 1 mg/ml concentration
49
50. Contd...
• i.e. Murchita arka taila showed mild anti bacterial
effect as the drugs used for murchana are also
having anti bacterial activity
• Due to the process of murchana Sneha dravya
incorporates the active Principles from the Kalka
dravyas that yield to better action of Murchita Arka
taila
• Patient sample did not show any Zone of inhibition in
all concentrations. Though the culture showed as a
Gram – ve bacteria the exact Bacteria was unknown
may be Arka taila is efficacious in Staphylococcus
aureus & not in other Bacteria
50
51. Conclusion
• Most of the drugs employed for Murchana & Arka Taila
preparation are Tikta kashaya rasa pradhana dravyas
• The Drugs used for murchana have Krimighna property
• Drugs used for both murchana process and for Arka taila
are easily available
• Pharmaceutical preparation is also simple and easy
• The Physico chemical parameters of Murchita & Amurchita
Arka taila and H.P.T.L.C findings generated have provided
preliminary standards
51
52. • In T.LC. Murchita Arka taila Shown Better results
comparing to Amurchita Arka taila at 254nm ,366nm
,540 nm respectively
• In H.P.T.L.C At Post derivatisation Murchita Arka taila
Shown better results comparing to Amurchita Arka taila
• Murchita Arka taila with both 1/8th & 1/4th kalka have
shown remarkable anti bacterial activity
• Zone of Inhibition was more in Sample A at 100 μl
indicates that Murchita arka taila with 1/8th Kalka
shown better result against Staphylococcus aureus
52
53. Limitations of the study
• Different peaks were observed in HPTLC , the
peaks or constituents can be identified by
adopting GLC technique
• The specific bacteria of the patient sample was
not identified
53
54. Scope for further study
• To check the efficacy of same formulations on
different bacteria
• To prepare Arka patra kashaya in place of
swarasa and check the efficacy in vicharchika
54
