HiMedia Laboratories GmbH, Marie-Curie-Str. 3, 64683, Einhausen, Germany.
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Intended Use
Recommended for microbiological assay of antibiotics in accordance with USP.
Composition
| Ingredients | g/L |
|---|---|
| Peptone | 6.000 |
| Yeast extract | 3.000 |
| HM peptone B # | 1.500 |
| Agar | 15.000 |
| pH after sterilization (at 25°C) | 6.6±0.1 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract
Directions
Suspend 25.5 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 or dispense as desired.
Principle And Interpretation
This medium is commonly used as base agar for microbiological agar diffusion assays for wide variety of antibiotics. Agar diffusion assays can be performed by cylinders, punched-hole or paper disc tests. This medium is identical numerically with the name assigned by Grove and Randall (1). This medium is prepared according to the specifications detailed in the USP and CFR (2,3).
Peptone, yeast and HM peptone B nitrogenous, vitamins and mineral requirement for the growth of test organisms. This medium provides solidified substratum for growth of organisms and supports the over layering of soft agar.
Type of specimen
Antibiotics as per USP
Specimen Collection and Handling
To perform an antibiotic assay the Antibiotic assay medium No.2 is used as Base Agar. This medium should be prepared on the same day as the test. For the cylinder method, a base layer of 21 ml is required. Once the base medium has solidified, Antibiotic assay medium No.1 as seed agar, inoculated with the standardized culture can be overlaid. Even distribution of the layer is important. After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Freshly prepared plates must be used or it may result in erroneous results.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance: Cream to yellow coloured homogeneous free flowing powder
Gelling: Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium: Amber coloured slightly opalescent gel forms in Petri plates.
pH: 6.50-6.70
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Basal layer |
|---|---|---|---|---|
| Micrococcus luteus ATCC 10240 | 50-100 | luxuriant | >=70% | Bacitracin |
| Staphylococcus aureus ATCC 9144 (00035*) | 50-100 | luxuriant | >=70% | Tylosin |
| Staphylococcus aureus ATCC 29737 | 50-100 | luxuriant | >=70% | Amikacin, Cephalothin, Cephapirin, Cloxacillin, Cycloserine, Chlortetracycline, Demeclocycline, Doxycycline, Kanamycin, Methacycline, Nafcillin, Oxytetracycline, Rolitetracycline, Tetracycline |
| Staphylococcus epidermidis ATCC 12228 (00036*) | 50-100 | good-luxuriant | >=70% | Novobiocin |
| Klebsiella pneumoniae ATCC 10031 | 50-100 | luxuriant | >=70% | Capreomycin, Streptomycin, Troleandomycin |
| Enterococcus hirae ATCC 10541 (00011*) | 50-100 | luxuriant | >=70% | Gramicidin, Thiostrepton, Tobramycin |
| Escherichia coli ATCC 10536 | 50-100 | luxuriant | >=70% | Chloramphenicol, Spectinomycin |
Key: *- Corresponding WDCM numbers
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and use freshly prepared medium. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5).
Reference
- Grove and Randall, 1955, Assay Methods of Antibiotics Medical Encyclopedia, Inc. New York.1.
- 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).
- The United States Pharmacopoeia-National Formulatory (USP-NF), 2022.
- Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
- Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.
| Product Name | Antibiotic Assay Medium No. 2 |
|---|---|
| SKU | MU005 |
| Application/ Industry | Pharmaceutical |
| Product Type | Regular |
| Physical Form | Powder |
| Regulatory | FDA, CFR, USP |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Grove and Randall, 1955, Assay Methods of Antibiotics Medical Encyclopedia, Inc. New York.1. 2. Indian Pharmacopoeia 2017, Minstry of Health and Family Welfare, Govt. of India, Delhi. 3. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition. 4. Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1. |
| Customized Product Available | No |
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