Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for microbiological assay of Amphotericin B and Nystatin using Saccharomyces cerevisiae & Candida tropicalis respectively.
Composition
| Ingredients | Gms/Litre |
|---|---|
| Peptone | 9.400 |
| Yeast extract | 4.700 |
| HM peptone B # | 2.400 |
| Sodium chloride | 10.000 |
| Dextrose (Glucose) | 10.000 |
| Agar | 23.500 |
| Final pH (at 25°C) | 6.0±0.2 |
**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 or dispense as desired.
Advice: Recommended for the microbiological assay of Amphotericin B and Nystatin.
Principle And Interpretation
Grove and Randall have elucidated the antibiotic assays and medias in their comprehensive treatise on antibiotic assays (1). Antibiotic assay Medium F is recommended for the microbiological assay of Nystatin and Amphotericin B using Saccharomyces cerevisiae and Candida tropicalis. This medium is formulated in accordance with the European Pharmacopoeia and British Pharmacopoeia (2,3). Freshly prepared plates should be used for antibiotic assays. Test organisms are inoculated in sterile seed agar cooled to 40-45°C and spread evenly over the surface of solidified base agar. After incubation the concentration of the antibiotic being assayed is determined by measuring the zone of inhibition obtained, with that of reference standard antibiotic. All conditions in the microbiological assay must be carefully controlled. The use of standard culture media in the test is one of the important steps for good results. Peptone, yeast extract and HM Peptone B supply essential nutrients, minerals and growth factors for the growth of the test organisms. Glucose monohydrate in the medium provides enhanced source of carbon and energy. Osmotic equilibrium in the medium is provided by sodium chloride thereby maintaining the cell viability and integrity. Higher agar concentration provides solid substratum for growth of colonies and controls the diffusion of antibiotics.
Type of specimen
Pharmaceutical sample
Specimen Collection and Handling
For pharmaceutical sample samples follow appropriate techniques for handling specimens as per established guidelines (1,2). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precaution
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 plates must be used or it may result in erroneous results.
- Use of this method is appropriate only when test samples are clear.
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 2.35% Agar gel.
Colour and Clarity of prepared medium: Light yellow coloured clear to slightly opalescent gel forms in Petri plates.
pH: 5.80-6.20
Growth promotion test: In accordance with EP/BP
Cultural Response: Cultural characteristics observed after an incubation at specified temperature for 18 - 48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Antibiotics assayed | Incubation Temperature |
|---|---|---|---|---|---|
| Saccharomyces cerevisiae ATCC 9763 | 50-100 | luxuriant | >=70% | Amphotericin B, Nystatin | 35-37°C 30-32°C |
| Candida tropicalis CIP 1433-83 | 50-100 | luxuriant | >=70% | Nystatin | 30-37°C |
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).
| Product Name | Antibiotic Assay Medium F |
|---|---|
| SKU | M923 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Grove and Randall, 1955, Assay Methods of Antibiotics Medical Encyclopedia, Inc, New York. |
| Customized Product Available | No |
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