Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Antibiotic Assay Medium No.1
For microbiological assay of ß-lactam and other antibiotics in pharmaceutical and food related preparations in accordance with US Pharmacopoeia.
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Intended Use
Recommended for microbiological assay of B-lactam and other antibiotics in pharmaceutical and food related preparations in accordance with USP.
Composition**
| Ingredients | g/L |
|---|---|
| Peptone | 6.000 |
| Tryptone | 4.000 |
| Yeast extract | 3.000 |
| HM peptone B # | 1.500 |
| Dextrose (Glucose) | 1.000 |
| Agar | 15.000 |
| pH after sterilization | 6.6±0.1 |
**Formula adjusted, standardized to suit performance parameters
# 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 or as per validated cycle. Cool to 45-50°C. Mix well and pour into sterile Petri plates or dispense as desired.
Note: Recommended for the microbiological assay of Bacitracin, Carbenicillin, Cloxacillin, Colistimethate, Colistin, Erythromycin, Gentamicin, Neomycin, Nafcillin, Novobiocin, Paromomycin, Penicillin-G, Polymycin B, Sisomicin, Capreomycin, Chloramphenicol and Chlortetracycline.
Principle And Interpretation
This medium is also used as inoculum and maintenance medium for different test organisms for antibiotic assays. Composition of this medium is in accordance with US Pharmocopoeia (1) and is recommended by FDA (2) and identified numerically with the name assigned by Grove and Randall (3).
Essential nutrients, vitamins, mineral, trace elements and growth factors are supplied by peptone, pancreatic digest of casein, yeast and HM peptone 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.
Type of specimen
Antibiotics as per USP
Specimen Collection and Handling
Freshly prepared plates should be preferably used for assaying antibiotics. Test organisms is 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. One of the critical and important step for obtaining good results is use of appropriate standard culture media. 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
Yellow coloured slightly opalescent gel forms in Petri plates.
pH
6.50-6.70
Cultural Response
Cultural characteristics observed after an incubation at specified temperature and period.
| Organism | Inoculum Growth (CFU) | Recovery | Inoculum medium | Incubation temperature / period | Antibiotics assayed | |
|---|---|---|---|---|---|---|
| Bordetella bronchiseptica ATCC 4617 | 50-100 | good-luxuriant | >=50% | Colistimethate sodium, Colistin, Polymyxin B | 32-35°C/24 hours | |
| Escherichia coli ATCC 10536 | 50-100 | luxuriant | >=70% | Chloramphenicol | 32-35°C/24 hours | |
| Klebsiella pneumoniae ATCC 10031 | 50-100 | good-luxuriant | >=50% | Capreomycin, Dihydrostreptomycin, Neomycin, Streptomycin, Troleandomycin | 36-37.5°C/ 16-24 hours | |
| $ Kokuria rhizophila ATCC 9341 | 50-100 | luxuriant | >=70% | Erythromycin | 32-35°C/24 hours | |
| Micrococcus luteus ATCC 10240 | 50-100 | good-luxuriant | >=70% | Bacitracin | 32-35°C/24 hours | Bacitracin |
| Pseudomonas aeruginosa ATCC 25619 | 50-100 | luxuriant | >=70% | Carbenicillin | 36-37.5°C/ 16-24 hours | |
| Staphylococcus epidermidis ATCC 12228 (00036*) | 50-100 | good-luxuriant | >=70% | Gentamicin, Netilmicin, Neomycin, Novobiocin, Paromomycin, Sisomycin | 32-35°C/24 hours | Novobiocin |
| Staphylococcus aureus ATCC 29737 | 50-100 | luxuriant | >=70% | Amikacin, Cephalothin, Cephaperin, Chlortetracycline, Cloxacillin, Cycloserine, Demeclocycline, Kanamycin, Methacycline, Nafcillin, Penicillin-G, Rolitetracycline, Tetracycline, Tobramycin, Tylosin | 32-35°C/24 hours | Cephalothin, Cephaperin, Cloxacillin, Nafcillin, Penicillin-G, |
*- Corresponding WDCM numbers
$ Formerly known as Micrococcus luteus
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.
| Product Name | Antibiotic Assay Medium No.1 |
|---|---|
| SKU | MU003 |
| Application/ Industry | Pharmaceutical |
| Product Type | Regular |
| Physical Form | Powder |
| Regulatory | FDA, CFR, USP |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Pelczar M. J. Jr., Reid R. D., Chan E. C. S., 1977, Microbiology, 4th Edi, Tata McGraw-Hill Publishing Company Ltd,New Delhi 2. The United States Pharmacopoeia, 2018, The United States Pharmacopoeial Convention,Rockville, MD. 3. Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Eds.), 2003, Manual of Clinical Microbiology,8th Ed., American Society for Microbiology, Washington, D.C. 4. Grove and Randall, 1955, Assay Methods of Antibiotics Medical Encyclopedia, Inc.New York.5.European Pharmacopoeia, 2017, European Department, for the Quality of Medicines6.British Pharmacopoeia, 2016,The Stationery office British Pharmacopoeia7.Tests and Methods of Assay of Antibiotics and Antibiotic containing Drugs, FDA, CFR, 1983 Title 21, Part 436, SubpartD, Washington, D.C.: U.S. Government Printing Office, paragraphs 436, 100-436, 106, p. 242- 259 (April 1).8.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.9.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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