Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Antibiotic Assay Medium A
For microbiological assay of ß-lactam and other antibiotics in pharmaceutical and food related preparations in accordance with Indian Pharmacopoeia.
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Composition
| Ingredients | Gms / Litre |
|---|---|
| Peptone | 6.000 |
| Tryptone # # | 4.000 |
| Yeast extract | 3.000 |
| Meat extract B # | 1.500 |
| Dextrose | 1.000 |
| Agar | 15.000 |
| pH after sterilization | 6.55±0.05 |
**Formula adjusted, standardized to suit performance parameters
# # Pancreatic digest of casein
# Equivalent to Beef extract
Directions
Suspend 30.5 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. Cool to 45-50°C. Mix well and pour into sterile Petri plates.
Recommended Applications
Recommended for the microbiological assay of Bacitracin, Novobiocin, Streptomycin, Vancomycin and Netilmicin sulphate and as a maintenance medium for the assay of Amikacin, Bacitracin, Carbenicillin, Capreomycin, Chlortetracycline, Colistimethate sodium, Colistin sulphate, Doxycycline, Erythromycin, Framycetin, Gentamicin, Gramicidin, Kanamycin B sulphate, Neomycin, Netilmicin sulphate, Novobiocin, Oxytetracycline, Polymyxin-B, Sisomicin, Spiramycin, Streptomycin, Teicoplanin, Tetracycline, Tobramycin, Tylosin, Tyrothricin, Vancomycin.
Principle And Interpretation
This medium is widely employed as seed agar in the preparation of plates for microbiological agar diffusion assays by FDA cylinder plate method for wide variety of antibiotics. This medium is also used as inoculum and maintenance medium for several test organisms. Composition of this medium is in accordance to Indian Pharmocopeia (1) and CFR (2)
Essential nutrients, vitamins, mineral, trace elements and growth factors are supplied by peptone, tryptone yeast extract and meat extract B. Dextrose in the medium serves as the carbon source for stimulating the growth of the test microorganism. Agar provides excellent medium for antibiotic diffusion and gives well defined zones of inhibition.
Freshly prepared plates should be used for antibiotic assays. Test organisms are inoculated in sterile seed agar cooled to 40-45°C and spread evenly over the surface of solidified base agar. All conditions in the microbiological assay must be controlled carefully. The use of standard culture media in the test is one of the important step for the good results.
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Yellow coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
After sterilization, reaction of 3.05% w/v aqueous solution. pH: 6.55±0.05
pH
6.50-6.60
Growth Promotion Test
As per Indian Pharmacopoeia
Cultural Response
Cultural characteristics observed after an incubation at specified temperature for specified period.
| Organism | Inoculum Growth (CFU) | Inoculum medium | Antibiotics assayed | Incubation temperature / period |
|---|---|---|---|---|
| Bordetella bronchiseptica ATCC 4617 | 50-100 | good-luxuriant | Polymyxin B | 32-35°C / 24 hours |
| Bacillus cereus var mycoides ATCC 11778 | 50-100 | good-luxuriant | Oxytetracycline, Tetracycline | 32-35°C / 5 days |
| Bacillus pumilis ATCC 14884 | 50-100 | good-luxuriant | Chlortetracycline, Framycetin, Kanamycin sulphate | 32-35°C / 5 days |
| Bacillus subtilis ATCC 6633 | 50-100 | good-luxuriant | Streptomycin | Streptomycin 32-35°C / 5 days |
| Klebsiella pneumoniae ATCC 10031 | 50-100 | good-luxuriant | Streptomycin, Capreomycin | 36-37°C / 24 hours |
| Micrococcus luteus ATCC 9341 | 50-100 | good-luxuriant | Erythromycin | 32-35°C / 24 hours |
| Micrococcus luteus ATCC 10240 | 50-100 | good-luxuriant | Bacitracin | Bacitracin 32-35°C / 24 hours |
| Pseudomonas aeruginosa ATCC 25619 | 50-100 | good-luxuriant | Carbenicillin | 36-37.5°C / 24 hours |
| Staphylococcus aureus ATCC 29737 | 50-100 | good-luxuriant | Amikacin, Doxycycline, Kanamycin sulphate, Oxytetracycline, Tetracycline, Tobramycin, Tylosin | 32-35°C / 24 hours |
| Staphylococcus aureus ATCC 6538 | 50-100 | good-luxuriant | Gramicidin, Netilmicin sulphate | 32-35°C / 24 hours |
| Staphylococcus epidermidis ATCC 12228 | 50-100 | good-luxuriant | Novobiocin, Gentamicin, Neomycin, Sisomicin | Novobiocin 32-35°C / 24 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 A |
|---|---|
| SKU | MM003 |
| Application/ Industry | Pharmaceutical |
| Product Type | Regular |
| Physical Form | Powder |
| Regulatory | IP, FDA, CFR |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Indian Pharmacopeia 2010, Ministry of Health and Family welfare, Govt 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, (April 1). |
| Customized Product Available | No |
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