Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Antibiotic Assay Medium No. 3
Used as the broth medium in turbidimetric or serial dilution assay of a wide variety of antibiotics in accordance with USP/IP.
Gms/Lit : 17.50
28.57 Lit/500G
Gms/Lit : 17.50
28.57 Lit/500G
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Intended Use
Used as the broth medium in turbidimetric or serial dilution assay of a wide variety of antibiotics in accordance with USP/IP.
Composition
| Ingredients | g/L |
|---|---|
| Peptone | 5.000 |
| Yeast extract | 1.500 |
| HM peptone B # | 1.500 |
| Dextrose (Glucose) | 1.000 |
| Sodium chloride | 3.500 |
| Dipotassium hydrogen phosphate | 3.680 |
| Potassium dihydrogen phosphate | 1.320 |
| pH after sterilization | 7.0±0.05 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract
Directions
Suspend 17.5 grams in 1000 ml purified/distilled water. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes.
Advice: Recommended for the Microbiological assay of Amikacin, Capreomycin, Chloramphenicol, Chlortetracycline, Cycloserine, Demeclocycline, Dihydrostreptomycin, Doxycycline, Gramicidin, Kanamycin, Methacycline, Oxytetracycline Rolitetracycline, Streptomycin, Tetracycline, Tobramycin and Troleandomycin according to official methods.
Principle And Interpretation
Grove and Randall have elucidated the antibiotic assays and medias in their comprehensive treatise on antibiotic assays (1).
Antibiotic assay Medium No. 3 is used as the broth medium in turbidimetric or serial dilution assay of a wide variety of antibiotics. This medium is formulated in accordance with USP and IP (2,3).
Turbidimetric antibiotic assay is based on the change or inhibition of growth of a test microorganims in a liquid medium containing a uniform concentration of an antibiotic. After incubation of the test organism in the working dilutions of the antibiotics, the amount of growth is determined by measuring the light transmittance using spectrophotometer. The concentration of antibiotic is determined by comparing amounts of growth obtained with that given by the reference standard solutions. Use of this method is appropriate only when test samples are clear.
Peptone, HM peptone B and yeast extract provide essential nutrients and growth factors for enhanced microbial growth. Sodium chloride maintains the osmotic equilibrium and retains the cell viability and cell intergrity. Phosphates in the medium provide good buffering action. Dextrose serves as the carbon and energy source for luxuriant growth.
Type of specimen
Antibiotics as per USP & IP
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (2,3).
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
- 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 coloured homogeneous free flowing powder
Colour and Clarity of prepared medium: Light yellow coloured clear solution without any precipitate.
pH: 6.95-7.05
Growth Promotion Test: In accordance with USP/IP.
Cultural Response
Cultural characteristics observed after incubation at specified temperature.
| Organism | Inoculum (CFU) | Growth | Inoculum medium for maintenance | Antibiotics assayed | Incubation temperature / period |
|---|---|---|---|---|---|
| As per USP | |||||
| Enterococcus hirae ATCC 10541 (00011*) | 50-100 | luxuriant | Gramicidin | Gramicidin | 36-37.5°C/16-18h |
| Klebsiella pneumoniae ATCC 10031 | 50-100 | luxuriant | Capreomycin, Dihydrostreptomycin, | 36-37.5°C/16-24h | |
| Escherichia coli ATCC 10536 | 50-100 | luxuriant | Chloramphenicol | 32-35°C/24h | |
| Staphylococcus aureus ATCC 29737 | 50-100 | luxuriant | Chlortetracycline, Oxytetracycline, Tetracycline | 32-35°C/24h | |
| Staphylococcus aureus ATCC 9144 (00035*) | 50-100 | luxuriant | Tylosin | 35-39°C/16-18 h | |
| As per IP | |||||
| Escherichia coli ATCC 9637 | 50-100 | luxuriant | Colistimethate sodium | 35-37°C/24h | |
| Entrococcus hirae ATCC 10541 (00011*) | 50-100 | luxuriant | Gramicidin Tyrothricin | 36-38°C/16-18h | |
| Klebsiella pneumoniae ATCC 10031 | 50-100 | luxuriant | Capreomycin Streptomycin | 36-37°C/24h | |
| Staphylococcus aureus ATCC 6538 (00032*) | 50-100 | luxuriant | Gramicidin Vancomycin |
32-35°C/16-18h 37-39°C/24h |
|
| Staphylococcus aureus ATCC 29737 | 50-100 | luxuriant | Amikacin, Doxycycline, Kanamycin sulphate, Oxytetracycline, Tetracycline, Tobramycin, Tylosin | 32-35°C/24h | |
Key:-*- Corresponding WDCM numbers
Storage and Shelf Life
Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°C. 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 No. 3 |
|---|---|
| SKU | MAP042 |
| Customized Product Available | No |
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