Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Antibiotic Assay Medium No.10
As seed layer for antibiotic plate assay of carbenicillin, colistimethate sodium and polymyxin B in accordance with USP/IP.
Gms/Lit : 42.00 +
11.90 Lit/500G
10 ml of polysorbate 80 volume
Gms/Lit : 42.00 +
11.90 Lit/500G
10 ml of polysorbate 80 volume
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Intended use
Antibiotic Assay Medium No.10 (Antibiotic Assay Medium H) is used as seed layer for microbiological assay of Carbenicillin, Colistimethate sodium, Colistin sulphate and Polymyxin B in accordance with USP/IP.
Composition
| Ingredients | g/L |
|---|---|
| Tryptone | 17.000 |
| Soya peptone | 3.000 |
| Dextrose (Glucose) | 2.500 |
| Sodium chloride | 5.000 |
| Dipotassium hydrogen phosphate | 2.500 |
| Agar | 12.000 |
| Final pH (after sterilization) | 7.2±0.1 |
Formula adjusted, standardized to suit performance parameters
Pancreatic digest of casein
Papaic digest of soyabean
Directions
Suspend 42.0 grams in 1000 ml purified / distilled water containing 10 ml of Polysorbate 80. 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.
Principle And Interpretation
This medium is used as seed agar for assay of Carbenicillin, Colistimethate sodium, Colistin sulphate and Polymyxin B. The medium composition is in accordance to IP, USP and CFR (1,2,3) and numerically identical with the name assigned by Groove and Randall (4).
Combination of tryptone and soya peptone provides nitrogenous and carbonaceous compounds, long chain amino acids, vitamins and other essential nutrients for the growth of test organisms. Natural soya sugars enhance microbial growth. Dextrose (Glucose) provides the carbon source, enhances the growth of test organism. Phosphates in the medium enhance buffering action and sodium chloride maintains osmotic equilibrium. Polymyxin are reported to have slow diffusion in agar giving smaller zone of inhibition (5). Hence the reduced agar concentration (1.2%) in this medium improves the diffusion of polymyxin in the medium. Polysorbate 80 are reported to function synergistically with polymyxin on spheroplasts of Pseudomonas aeruginosa. Polysorbate 80 enhances the penetration of Polymyxin to the cytoplasmic membrane and hence is an appropriate ingredient in the medium used for assay of Polymyxin (6).
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. The use of standard culture media in the test is one of the important step for the good results.
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (1,2). 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 medium must be used for testing or it may result in erroneous results.
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
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 homogeneous free flowing powder
Gelling: Firm,comparable with 1.2% Agar gel.
Colour and Clarity of prepared medium: Medium amber coloured clear to slightly opalescent gel forms in Petri plates.
pH: 7.10-7.30
Cultural Response
Cultural characteristics observed after an incubation at
As per IP: different temperatures for 24 hours.
As per USP: 32-35°C for 24 hours
As per USP
| Organism | Inoculum (CFU) | Incubation Temperature | Growth | Recovery | Antibiotics assayed |
|---|---|---|---|---|---|
| Bordetella bronchiseptica ATCC 4617 | 50-100 | 32-35°C | luxuriant | >=70% | Polymyxin B, Colistimethate, Colistin |
As per IP
| Organism | Inoculum (CFU) | Incubation Temperature | Growth | Recovery | Antibiotics assayed |
|---|---|---|---|---|---|
| Bordetella bronchiseptica ATCC 4617 | 50-100 | 32-35°C | luxuriant | >=70% | Polymyxin B, Colistimethate sodium, |
| Pseudomonas aeruginosa ATCC 25619 | 50-100 | 36-37.5°C | luxuriant | >=70% | Colistin sulphate Carbenicillin |
| Escherichia coli ATCC 10536 | 50-100 | 35-39°C | luxuriant | >=70% | Colistimethate sodium, Colistin sulphate |
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 (7,8).
| Product Name | Antibiotic Assay Medium No.10 |
|---|---|
| SKU | MAP225 |
| Customized Product Available | No |
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