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Antibiotic Assay Medium G
For the microbiological assay of Amphotericin B and Nystatin using Saccharomyces cerevisiae as the test organisms in accordance with Indian Pharmacopoeia.
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Composition**
| Ingredients | Gms / Litre |
|---|---|
| Peptone | 9.400 |
| Yeast extract | 4.700 |
| Beef extract | 2.400 |
| Dextrose | 10.000 |
| Sodium chloride | 10.000 |
| Agar | 23.500 |
| pH after sterilization | 6.1±0.1 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 60.0 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Principle And Interpretation
The medium composition is in accordance to IP and CFR (1,2). This medium is used as seed agar for assay of antifungal agents like Amphotericin B and Nystatin. This medium is used for maintenance and inoculum development of Saccharomyces cerevisiae, indicator organism. This medium is also used for assaying mycostatic activity in pharmaceutical formulations. This medium is forumulated as reported by Kirshbam and Arret (3)
Ingredients like peptone, yeast extract and beef extract supplements essential nutrients, minerals and growth factors for the growth of test organism. Dextrose in the medium provides enhanced source of carbon and energy. Osmotic equilibrium in the medium is maintained by sodium chloride which retains the cell intergrity and viability.
Freshly prepared plates should be used for antibiotic assays. Test organisms are inoculated in sterile seed agar precooled to 40-45°C and spread evenly over the surface of solidified base agar. Prediffusion of antibiotics for 20 minutes in the agar by incubating at temperature below the optimal growth temperature for microorganism would facilitate better diffusion of antibiotic, followed by incubation of the plates for microbial growth.
Quality Control
Appearance: Cream to yellow coloured homogeneous free flowing powder
Gelling: Firm, comparable with 2.35% Agar gel.
Colour and Clarity of prepared medium: Yellow coloured clear to slightly opalescent gel forms in Petri plates.
pH: 6.00-6.20
Cultural Response
MM101: Cultural characteristics observed after an incubation at 29-31°C for 48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Antibiotics assayed |
|---|---|---|---|---|
| Saccharomyces cerevisiae ATCC 2601 | 50-100 | luxuriant | >=70% | Nystatin |
| Saccharomyces cerevisiae ATCC 9763 | 50-100 | luxuriant | >=70% | Amphotericin B |
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 G |
|---|---|
| SKU | MM101 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Indian Pharmacopoeia 2010, Ministry of Health and Family welfare, Government of India, New Delhi. 2.Tests and Methods of Assay of Antibiotics and Antibiotic containing Drugs, FDA, CFR, 1983 Title 21, Part 436,Subpart D,Washington, D.C : U.S. Government Printing Office, paragraphs 436, 100-436, 106, p. 242-259, (April1). 3.Krishbam A and ArretB, 1967, J.Pharma. Sci. 56:512 |
| Customized Product Available | No |
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