By-

Dr. Manojkumar Patil

P.G.Scholar Dept Of Rasashastra

UNDER THE GUIDANCE OF -

Dr M.S.GavimathM.D
Dr (Smt)P .P. Dindore. MD
Dr R.S. Hiremath M.D

K.L.E’s Shri B M K Ayurveda Mahavidyalaya. Belgaum,

Karnataka.
 CONTENTS
 INTRODUCTION

 OBJECTIVES

 METHODOLOGY

 RESULTS

 DISCUSSION

 CONCLUSION
 INTRODUCTION

 Anandbhairava rasa is one among,mentioned for

the cure of Jwaratisara.

 Adopting the preparation by good manufacturing

practice,optimum quality of drug can be prepared

 Numerous diseases arise by various micro-

organisms,among infectious diarrhea is one
where,E.Coli and S.Aureus are the main causative
organisms.
 OBJECTIVES

 Preparation of Anandabhairava rasa as per

classical reference.

 Physico-Chemical analysis of the Anandabhairava

rasa.

 Anti-microbial study of prepared standard

formulation in comparison of two different market
samples of Anandabhairava rasa to evaluate it’s
sensitivity on micro-organisms.
 METHODOLOGY
It is in two groups -

A) Material and methods of Anandabhairava rasa.

B) Material and methods of Antimicrobial study

PREPARATION OF ANANDA BHAIRAVA RASA
 Equipment :Khalwayantra,Patra

 Ingredients :
1. Shuddha Hingula - 100 gms
2. Shuddha Vatsanabha - 100 gms
3. Trikatu - 100 gms
4. Shuddha Tankana - 100 gms
5. Shuddha Gandhaka - 100 gms
6. Jambeera Swarasa - 600 ml

 Procedure :
- All the ingredients are taken in khalwayantra in above
mentioned proportion.
- Jambeera swarasa is added to it.
- Trituration is done carefully.
- Bhavana is given for 6 Hrs.
- Then gunja pramana vati are prepared.
- Vatis are placed on a patra and allowed to shadow dry for
one day.
- Then collected and preserved in a plastic container.
 Organoleptic features of Anandabhairava Rasa :

Features
Sl. No.
1. Colour Reddish orange

2. Smell Pleasant
3. Touch Fine powder
Quantitative Analysis of Raw drugs :

Sl. Samples
N Tests
o I II III IV V VI VII

1 Hg% 86.5% 66.6% - - - - -

w/w w/w

2. S% 12.64% 8.0% - - - 11.17% 10.48%

w/w w/w w/w

3. Borax % - - 16.6% 17.55% 16.7% - -

w/w w/w w/w

4. Na % - - 3.8% 4.2% 3.8% - -

w/w w/w w/w

I  Ashudda Hingula V  Nirmaleekruta Tankana

II Shudha Hingula VI  Ashudha Gandhak
III  Ashudda Tankana VII  Shuddha Gandhak
IV  Shudha Tankana
 Analysis of prepared Anandabhairava rasa

Sl. Sampl Tests

N e
o.

Hg % S% Free Borax Fe as Ca % Phosphate Silica K+

S Fe2 as PO4
% O3

01. VIII 12.3% 18.73% Nil 1.01% 0.17% 0.977% Nil 0.95% 0.48%
w/w w/w w/w w/w w/w w/w w/w

Ash value Acid Insoluble Ash Volatile matter LOD at 110OC P

H

15.0% w/w 1.47% w/w 15.61% w/w 12.38% w/w 7.24

VIII  Anadabharava Rasa

 Method:
The Antimicrobial study was designed in two different methods as :
 Macro dilution broth susceptibility test.(M.B.S.T) for more accuracy
 Cup- plate diffusion method.

1) Cup-plate diffusion method

 Standard drug
 Test organism
 Preparation of test solution
 Preparation of inoculum
 Culture medium
 Sterilization
 Preparation of agar plate
 Experimental procedure

2) Macro dilution Broth Susceptibility Test (M.B.S.T)

 Preparation of stock solution
 Experimental procedure
 ANTIBACTERIAL AND ANTIFUNGAL SCREENING RESULTS

Results of cup-plate diffusion method

Zone of inhibition in mm :
Compound code S. Aureus (+ ve) E-coli (-ve) Candida Albicans
Sl No. (Fungal )
0.1 mg/ml 0.5 mg/ml 0.1 mg/ml 0.5 mg/ml 0.1 mg/ml 0.5 mg/ml
01. Standard (prepared) 12 (0.66) 11 (0.61) 14 (0.64) 14 (0.68) 1 (0.05) 1
(0.05)
02. Market – I 8 (0.44) 10 (0.55) 11 13 (0.59) 0 2
(0.50) (–) (0.12)
03. Market – II 6 11 (0.61) 12 (0.55) 13 (0.59) 1 1
(0.33) (0.05) (0.05)

Results of Standard Allopathic Drug on organisms :

Standard drugs S. Aureus E.Coli Candida

Sl. No. Albicans
01. Oflaxacine (2 mcg/ml) 20 22 -
02. Ampicillin (10 mcg/ml) 18 19 -
03. Gresiofulvin - - 17
 Macrodilution Broth Susceptibility Method

Result of STAPHYLOCOCCUS AUREUS :

10 5 mg/ml 2.5 1.25 0.612 0.3 0.15 MIC

Sl. No. mg/ mg/ mg/ mg/ mg/ mg/ Valu
ml ml ml ml ml ml e
Mkt I S R R R R R R 10
Mkt II S R R R R R R 10
III S S S S R R R 1.25

Result of ESHERICHIA COLI :

10 5 mg/ml 2.5 1.25 0.612 0.3 0.15 MIC

Sl. No. mg/ mg/ mg/ mg/ mg/ mg/ Valu
ml ml ml ml ml ml e
Mkt I S S R R R R R 5
Mkt II S S S R R R R 2.5
III S S S R R R R 2.5

Mkt. I  Market Sample I III  Standard drug

Mkt II Market sample II
Zone of inhibition
Macro dilution Broth Susceptibility Test Method
 DISCUSSION

1st Method

 All the three show activity on organisms.

 Two concentration of 0.1mg/ml & 0.5 mg/ml were

studied

 Zone of inhibition of prepared drug is little more,

especially on E.coli at 14mm

 Anti fungal activity on C.Albicans was very minimal

 Cont…
2nd Method

 Studied in 7 different concentration

 Prepared drug shown activity for S.Aureus at 1.25 mg/ml

conc.where as both market drug shown activity at 10 mg/ml

 Prepared drug shown activity on E.Coli at 2.5 mg/ml conc.

Market –II also shown at the same, where as market –I
shown activity at 5 mg/ml conc.

 Still drug concentration to be improved as compared

standard allopathic drugs
 CONCLUSION

 Anandabhairava rasa is economic, easy for preparation (raw

drugs are easily available)

 Qualitative & Quantitative values are within the range of

standard values

 Overall effect of Anandabhairava rasa on micro-organisms is

sensitive

 More significant activity on S.Aureus.

 Prepared drug is more potent on micro-organisms as

compared to market samples
THANK YOU

THANK YOU