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

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

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 gms3. Trikatu - 100 gms4. Shuddha Tankana - 100 gms5. Shuddha Gandhaka - 100 gms6. 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 :

Sl. No.Features

1. Colour Reddish orange

2. Smell Pleasant

3. Touch Fine powder

Quantitative Analysis of Raw drugs :

Sl. No

Tests

Samples

I II III IV V VI VII

1 Hg% 86.5% w/w

66.6% w/w

- - - - -

2. S% 12.64% w/w

8.0% w/w

- - - 11.17% 10.48% w/w

3. Borax % - - 16.6% w/w

17.55% w/w

16.7% w/w

- -

4. Na % - - 3.8% w/w

4.2% w/w

3.8% w/w

- -

I Ashudda Hingula V Nirmaleekruta TankanaII Shudha Hingula VI Ashudha GandhakIII Ashudda Tankana VII Shuddha Gandhak

IV Shudha Tankana

Analysis of prepared Anandabhairava rasa

Sl. No.

Sample

Tests

Hg % S % Free S%

Borax Fe as Fe2

O3

Ca % Phosphate as PO4

Silica K+

01. VIII 12.3% w/w

18.73% w/w

Nil 1.01% w/w

0.17% w/w

0.977% w/w

Nil 0.95% w/w

0.48% w/w

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

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 accuracyCup- 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 :

Sl No.Compound code S. Aureus (+ ve) E-coli (-ve) Candida Albicans

(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 (0.50)

13 (0.59) 0 (–)

2 (0.12)

03. Market – II 6 (0.33)

11 (0.61) 12 (0.55) 13 (0.59) 1 (0.05)

1 (0.05)

Results of Standard Allopathic Drug on organisms :

Sl. No.Standard drugs S. Aureus E.Coli Candida

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 :

Sl. No.10

mg/ml

5 mg/ml 2.5 mg/ml

1.25 mg/ml

0.612 mg/ml

0.3 mg/ml

0.15 mg/ml

MIC Valu

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 :

Sl. No.10

mg/ml

5 mg/ml 2.5 mg/ml

1.25 mg/ml

0.612 mg/ml

0.3 mg/ml

0.15 mg/ml

MIC Valu

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 drugMkt 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

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