Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Antibiotic Assay Medium No. 3
Used as the broth medium in turbidimetric or serial dilution assay of a wide variety of antibiotics in accordance with US Pharmacopoeia.
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Composition**
| Ingredients | Gms / Litre |
|---|---|
| Peptone | 5.000 |
| Yeast extract | 1.500 |
| Beef extract | 1.500 |
| Dextrose | 1.000 |
| Sodium chloride | 3.500 |
| Dibasic potassium phosphate | 3.680 |
| Monobasic potassium phosphate | 1.320 |
pH after sterilization: 7.0±0.05
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 17.5 grams in 1000 ml purified/distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Advice: Recommended for the Microbiological assay of Amikacin, Capreomycin, Chloramphenicol, Chlortetracycline, Cycloserine, Demeclocycline, Dihydrostreptomycin, Doxycycline, Gramicidin, Kanamycin, Methacycline, Oxytetracycline Rolitetracycline, Streptomycin, Tetracycline, Tobramycin and Troleandomycin according to official methods.
Principle And Interpretation
Grove and Randall have elucidated the antibiotic assays and medias in their comprehensive treatise on antibiotic assays (1).
Antibiotic assay Medium No. 3 is used as the broth medium in turbidimetric or serial dilution assay of a wide variety of antibiotics. This medium is formulated in accordance with The United States Pharmacopoeia (2).
Turbidimetric antibiotic assay is based on the change or inhibition of growth of a test microorganims in a liquid medium containing a uniform concentration of an antibiotic. After incubation of the test organism in the working dilutions of the antibiotics, the amount of growth is determined by measuring the light transmittance using spectrophotometer. The concentration of antibiotic is determined by comparing amounts of growth obtained with that given by the reference standard solutions. Use of this method is appropriate only when test samples are clear.
Peptone, beef extract and yeast extract provide essential nutrients and growth factors for enhanced microbial growth. Sodium chloride maintains the osmotic equilibrium and retains the cell viability and cell intergrity. Phosphates in the medium provide good buffering action. Dextrose serves as the carbon and energy source for luxuriant growth.
Quality Control
Appearance
Cream to yellow coloured homogeneous free flowing powder
Colour and Clarity of prepared medium
Light yellow coloured clear solution without any precipitate.
Reaction
Reaction of 1.75% w/v aqueous solution at 25°C (after sterilization). pH: 7.0±0.05
pH
6.95-7.05
Growth Promotion Test
In Accordance with the harmonized method of USP.
Cultural Response
Cultural characteristics observed after incubation at specified temperature.
Cultural Response
| Organism | Inoculum (CFU) | Growth | Serial dilution with | Incubation temperature / period |
|---|---|---|---|---|
| Escherichia coli ATCC 10536 | 50-100 | luxuriant | Chloramphenicol | 32-35°C / 24 hours |
| Klebsiella pneumoniae ATCC 10031 | 50-100 | luxuriant | Capreomycin, Dihydrostreptomycin, Streptomycin, Troleandomycin | 36-37.5°C / 16-24 hours |
| Staphylococcus aureus ATCC 29737 | 50-100 | luxuriant | Amikacin, Chlortetracycline, Cycloserine, Demeclocycline, Doxycycline, Kanamycin, Lincomycin, Methacycline, Oxytetracycline, Rolitetracycline, Tetracyclin, Tobramycin, | 32-35°C / 24 hours |
| Enterococcus hirae ATCC 10541 | 50-100 | luxuriant | Gramicidin | 36-37.5°C / 16-18 hours |
| Staphylococcus aureus ATCC 9144 | 50-100 | luxuriant | Tylosin | 35-39°C / 16-18 hours |
Storage and Shelf Life
Store below 30°C in tightly closed container and use freshly prepared medium. Use before expiry date on the label.
| Product Name | Antibiotic Assay Medium No. 3 |
|---|---|
| SKU | MU042 |
| Customized Product Available | No |
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