Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Antibiotic Assay Medium No.19
For microbiological assay of Amphotericin B, Natamycin and Nystatin using Saccharomyces cerevisiae as the test organisms in accordance with USP/IP.
Gms/Lit : 60.00
8.33 Lit/500G
Gms/Lit : 60.00
8.33 Lit/500G
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Intended Use:
Recommended for microbiological assay of Amphotericin B and Nystatin using Saccharomyces cerevisiae as the test organisms in accordance with USP/IP.
Composition**
| Ingredients | g/L |
|---|---|
| Peptone | 9.400 |
| Yeast extract | 4.700 |
| HM peptone B # | 2.400 |
| Dextrose | 10.000 |
| Sodium chloride | 10.000 |
| Agar | 23.500 |
| pH after sterilization | 6.1±0.1 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract
Directions
Suspend 60.0 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.
Principle And Interpretation
The medium composition is in accordance to USP, IP and CFR (1,2,3). This medium is used as seed agar for assay of antifungal agents like Amphotericin B and Nystatin. This medium is used for maintenance and inoculum development of Saccharomyces cerevisiae. This medium is also used for assaying mycostatic activity in pharmaceutical formulations. This medium is formulated as reported by Kirshbam and Arret (4).
Ingredients like peptone, yeast extract and HM peptone B supplement essential nutrients, minerals and growth factors for the growth of test organism. Dextrose in the medium provides enhanced source of carbon and energy. Osmotic equilibrium in the medium is maintained by sodium chloride which retains the cell intergrity and viability. This medium is used as both base and seed medium for agar diffusion assay for antibiotics like Amphotericin B and Nystatin.
Type of specimen
Antibiotics as per USP & IP (1,2)
Specimen Collection and Handling
Freshly prepared plates should be used for antibiotic assays. Test organisms are inoculated in sterile seed agar precooled to 40-45°C and spread evenly over the surface of solidified base agar. Prediffusion of antibiotics for 20 minutes in the agar by incubating at temperature below the optimal growth temperature for microorganism would facilitate better diffusion of antibiotic, followed by incubation of the plates for microbial growth. 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 2.35% Agar gel.
Colour and Clarity of prepared medium
Yellow coloured clear to slightly opalescent gel forms in Petri plates
pH
6.00-6.20
Growth Promotion Test
As per USP/IP
Cultural Response
Cultural characteristics observed after an incubation at 29-31°C for 48 hours.
| Organism | Inoculum (CFU) |
Growth | Recovery | Antibiotics assayed |
|---|---|---|---|---|
| #Saccharomyces kudriavzevii ATCC 2601 | 50-100 | luxuriant | >=70% | Nystatin |
| Saccharomyces cerevisiae ATCC 9763 (00058*) | 50-100 | luxuriant | >=70% | Amphotericin B |
Key: *- Corresponding WDCM numbers,
# Formerly known as Saccharomyces cerevisiae
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 (5,6).
| Product Name | Antibiotic Assay Medium No.19 |
|---|---|
| SKU | MAP101 |
| Customized Product Available | No |
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