Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Antibiotic Assay Medium D is used for microbiological assay of antibiotics in accordance with Indian Pharmacopoeia.
Composition**
| Ingredients | Gms / Litre |
|---|---|
| Peptone | 6.000 |
| Pancreatic digest of casein | 4.000 |
| Yeast extract | 3.000 |
| Beef extract | 1.500 |
| Dextrose | 1.000 |
| Agar | 15.000 |
| pH after sterilization | 7.9±0.1 |
**Formula adjusted, standardized to suit performance parameters
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.
Advice: Recommended for the Microbiological assay of Erythromycin, Chlortetracycline, Framycetin, Gentamicin, Kanamycin sulphate, Neomycin .
Advice: Cup plate method is carried out using
B. pumilis / Kanamycin and
M. luteus / Erythromycin
1) Dilution : 16 mg Kanamycin in 10 ml distilled water Stock: 1:10 dilution of above solution
| Concentration | Stock (ml) | D/W(ml) | Zone of Inhibition |
|---|---|---|---|
| 5 | 0.25 | 4.75 | 15 mm |
| 20 | 1.00 | 4.00 | 20 mm |
| 10 | 0 | 5.00 | 25 mm |
2) Dilution : 9 mg Erythromycin in 10 ml distilled water Stock: 1:10 dilution of above solution.
| Concentration | Stock (ml) | D/W(ml) | Zone of Inhibition |
|---|---|---|---|
| 5 | 0.25 | 4.75 | 22 mm |
| 10 | 0.50 | 4.50 | 32 mm |
| 10 | 0 | 5.00 | 41 mm |
Principle And Interpretation
This medium is forumlated in accordance to IP and CFR; and is employed to analyze the Neomycin, Erythromycin content as per FDA and the IP (1,2). This medium provides a pH range of 8 while Antibiotic assay medium A provides pH range of 6.5-6.7.
Peptone, pancreatic digest of casein, yeast and beef extract supplies essential nutrients, vitamins, mineral, trace elements and growth factors. 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. Higher pH provides the optimal conditions for activity of antibiotic and also supports the growth of test organisms.
Freshly prepared plates should be used for antibiotic assays. Test organisms are inoculated in sterile seed agar pre-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.
Quality Control
Appearance: Cream to yellow homogeneous free flowing powder
Gelling: Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium: Light yellow coloured clear to slightly opalescent gel forms in Petri plates.
Reaction: Reaction of 3.05% w/v aqueous solution. pH: 7.9±0.1
pH: 7.80-8.00
Growth Promotion Test: As per Indian Pharmacopoeia.
Cultural Response: Cultural characteristics observed after an incubation at .....
| Organism | Inoculum (CFU) | Growth | Recovery | Antibiotics assayed | Incubation temperature/ Period |
|---|---|---|---|---|---|
| Micrococcus luteus ATCC 9341 | 50-100 | luxuriant | >=70% | Erythromycin | 32-35°C/24 hrs |
| Staphylococcus epidermidis ATCC 12228 | 50-100 | luxuriant | >=70% | Gentamycin, Neomycin | 32-35°C/24 hrs |
| Bacillus pumilis ATCC 14884 | 50-100 | luxuriant | >=70% | Chlortetracycline, Kanamycin sulphate, Framycetin | 32-35°C/5 days |
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 D |
|---|---|
| SKU | MM004 |
| Application/ Industry | Pharmaceutical |
| Product Type | Regular |
| Physical Form | Powder |
| Regulatory | IP, FDA, CFR |
| 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, (April 1). |
| Customized Product Available | No |
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