CONTENTS

Microbiological Assay Of Antibiotics.

Types of Microbiological Assay
Preparation Of The Test
Methods Of Microbiological Assay
Microbiological assay of Antibiotics
The microbiological assay of an antibiotics is based
upon a comparison of the inhibition of growth of
microorganism by measered concentrationsof the
antibiotics under examination with that produced by
known concentration of a standard preparation of the
antibiotic having a known activity.
Types Of Microbiological Assay
The Cylinder-Plate(or cup plate)Method:- The
cylinder plate method depends upon diffusion of the
antibiotics from a vertical cylinder through a
solidified agar layer nin the petridish or plate to and
extend such that growth of the added micro-organism
is presented entirely is a zone around the cylinder
containg a solution of antibiotics.
The Turbidimetric Method or Diffusion
Method:
The method depends upon the inhibition of growth
of a microbial culture in a uniform solution of the
antibiotic in a fluid medium that is favourable to its
rapid growth in the observe of the antibiotics.
Preparation Of The Test
Culture Media:-
Media has to be prepared for the specified test
organism for the ingredients listed in IP according to
the individual requirement of the test organism.
Preparation of the test
*preparation of the standard solution

The stock solution is preparation by dissolving a

quantity of the Standard preparation of a given
antibiotic as per IP
Preparation of the test
Preparation of the sample solution

 From the information available for the substance

under examination, a stock solution is prepared as
specified in IP
Preparation of the test
Test Organism

 The test organism for each antibiotic in listed in

Table , Together with its identification number in the
American Type Culture Collection (ATCC)
Antibiotic Test organism Assay method Medium Incubation
temp.(°C)
Ampicillin Microccus A D 32-35
luteus

ol
.
Chloramphenic Escherichia coli B C 35-37

Doxycycline Staphylococcus B C 35-37

areus
Erythromycin Micrococcus A D 35-37
luteus
Oxytetracycline Bacillus A F 32-35
mycoides
Rifampicin Bacillus subtilis A B 32-35

Streptomycin Bacillus subtilis A E 32-35

Sulp[hate
Tetracycline Bacillus subtilis A F 32-35

Tetracycline Staphylococcus B C 35-37

aureus
.METHOD A-CYLINDER PLATE METHOD, METHOD B-TURBIDIMETRIC METHOD
METHODS OF MICROBIOLOGICAL ASSAY
Serial Dilution Method:-
The dilution of the antibiotic under test that will
inhibit the growth of a suspectable organism is
compared with the