Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for the selective isolation of Pseudomonas aeruginosa from pharmaceutical products in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP/IP.
Composition
| Ingredients | g/L |
|---|---|
| Gelatin peptone # | 20.000 |
| Magnesium chloride | 1.400 |
| Dipotassium sulphate | 10.000 |
| Cetrimide | 0.300 |
| Glycerol | 10.000 |
| Agar | 13.600 |
| pH after sterilization (at 25°C) | 7.2±0.2 |
**Formula adjusted, standardized to suit performance parameters
# Pancreatic digest of gelatin
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
Cetrimide Agar was described by King et al (1). This media formulation is in accordance with the harmonized method of USP/EP/BP/JP/IP (2,3,4,5,6). It is used as a selective medium for the isolation of Pseudomonas aeruginosa from pharmaceutical products. This medium is also used for microbial limit testing for non- sterile products. Lowburry first reported the use of cetrimide as an agent for selective isolation of Pseudomonas (7). This medium is also used for determining the ability of an organism to produce fluorescein and pyocyanin. Cetrimide (N-acetyl-N,N,N-trimethylammonium bromide) is incorporated in the medium to inhibit bacteria other than Pseudomonas aeruginosa. This compound a cationic detergent acts as a quaternary ammonium compound, which causes nitrogen and phosphorus to be released from bacterial cells other than Pseudomonas aeruginosa. Magnesium chloride and potassium sulphate incorporated in the medium enhances the production of pigment pyocyanin, which is a blue-green pigment, diffusing into the medium. This improves detection of Pseudomonas on this medium. Presence of magnesium ions can also neutralizes EDTA, if present in the sample. Gelatin peptone provides the essential nutrients for growth of Pseudomonas, while glycerin serves as slow and continuous carbon source for the growing cell.
For the isolation of Pseudomonas aeruginosa, plates of Cetrimide Agar should be inoculated from non-selective medium such as Soybean Casein Digest Medium (MH011). If the count is high the test sample can be directly inoculated onto this medium. Pseudomonas aeruginosa colonies may appear pigmented greenish (under uv light also).
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (2,3,4,5,6). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions:
Read the label before opening the pack. 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
- This medium is a selective medium, some strains may show poor growth as cetrimide is highly toxic.
- 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.
- It is recommended to store the plates at 24-30°C to avoid minimum condensation.
- Further biochemical tests must be carried out for complete identification.
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: Sterile Cetrimide Agar in 90 mm disposable plates with smooth surface and absence of black particles/ cracks/ bubbles.
Colour of medium: Light amber coloured medium
Quantity of medium: 25 ml of medium in 90 mm disposable plates.
pH: 7.00-7.40
Sterility Check: Passes release criteria
Growth Promotion Test
Growth Promotion is carried out in accordance with the harmonized method of USP/EP/BP/JP/IP. Cultural response was observed after an incubation at 30-35°C for specified time. Recovery rate is considered as 100% for bacteria growth on Soybean Casein Digest Agar.
Growth promoting properties
Growth of microorganism comparable to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not more than the shortest period of time specified inoculating <=100 cfu (at 30-35°C for <=18 hours).
Inhibitory properties
No growth of the test microorganism occurs for the specified temp for not less than longest period of time specified inoculating >=100 cfu (at least 100 cfu) (at 30-35°C for >= 72 hours).
Cultural Response
Cultural characteristics observed after incubation at 30-35 °C for 18-72 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.
| Organism | Inoculum Growth (CFU) | Recovery | Incubation temperature | Incubation period | Pigment production |
|---|---|---|---|---|---|
| Growth promoting | |||||
| ^Pseudomonas paraeruginosa ATCC 9027 (00026*) | 50-100 luxuriant | >=50% | 30-35 °C | <=18 hrs | Yellow green |
| Inhibitory | |||||
| Escherichia coli ATCC 8739 (00012*) | >=10³ inhibited | 0% | 30-35 °C | >=72 hrs | |
Key: (*) Corresponding WDCM numbers.
^Formerly known as Pseudomonas aeruginosa.
Storage and Shelf Life
On receipt store between 20-30°C. Use before expiry date on the label. 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 (8,9).
| Product Name | Cetrimide Agar Plate |
|---|---|
| SKU | MPH024 |
| Customized Product Available | No |
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