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Nutrient Agar Medium
A general culture medium which may be used as enriched medium by incorporating blood or other biological fluids in accordance with Indian Pharmacopoeia.
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Intended Use
A general culture medium which may be used as enriched medium by incorporating blood or other biological fluids in accordance with as per I.P.1996.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Peptone | 10.000 |
| HM peptone B # | 10.000 |
| Sodium chloride | 5.000 |
| Agar | 12.000 |
| pH after sterilization | 7.3±0.1 |
**Formula adjusted, standardized to suit performance parameters
# Equivalent to Beef extract
Directions
Suspend 37.0 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at A115°C for 30 minutes or alternatively at 15 lbs pressure (121°C) for 15 minutes or as per validated cycle. A Corresponds to 10 lbs pressure.
Note: Directions specified are as per the concurrent edition of pharmacopoeia in force. Specified expiry period corresponds to this. User must ensure its compatibility with the latest edition.
Principle And Interpretation
Nutrient Agar is a basic culture medium used for maintaining microorganisms (1), for purity checking prior to biochemical or serological testing. It is used for the cultivation and enumeration of bacteria, which are not particularly fastidious. In semisolid form it is used for maintenance of control or standard organisms. Indian Pharmacopoeia has recommended it for microbial limit tests of viable aerobic microorganism present in pharmaceutical substances (2).
Peptone and HM peptone B provide the necessary nitrogen compounds, carbon, vitamins and also some trace ingredients. Sodium chloride maintains osmotic equilibrium. Nutrient media may be used as enriched media by the addition of 10% v/v blood or other biological fluids like ascitic fluid, serum etc.
Type of specimen
Pharmaceutical samples
Specimen Collection and Handling
For pharmaceutical products, follow appropriate techniques for sample processing in case of viscous materials as mentioned under sterility. (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
- 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.
- Other biochemical tests must be carried out for confirmation.
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: Light yellow coloured clear to slightly opalescent gel forms in Petri plates
pH: 7.20-7.40
Growth Promotion Test
Growth Promotion is carried out as per Indian Pharmacopoeia
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
| Organism | Inoculum (CFU) |
Observed Lot value (CFU) |
Recovery | Incubation temperature |
Incubation period |
|---|---|---|---|---|---|
| Escherichia coli ATCC 8739 (00012*) |
50-100 | 35-100 | >=70% | 35-37 °C | 18-24 hrs |
| Staphylococcus aureus subsp aureus ATCC 6538 (00032*) |
50-100 | 35-100 | >=70% | 35-37 °C | 18-24 hrs |
| Salmonella Typhimurium ATCC 14028 (00031*) |
50-100 | 35-100 | >=70% | 35-37 °C | 18 -24 hrs |
| Salmonella Abony NCTC 6017 (00029*) |
50-100 | 35-100 | >=70% | 35-37 °C | 18-24 hrs |
| Peudomonas aeruginosa ATCC 9027 (00026*) |
50-100 | 35-100 | >=70% | 35-37 °C | 18-24 hrs |
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 (3,4).
| Product Name | Nutrient Agar Medium |
|---|---|
| SKU | MM012 |
| Product Type | Regular |
| Physical Form | Powder |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Lapage S., Shelton J. and Mitchell T., 1970, 'Methods in Microbiology, Norris J. and Ribbons D. (ed.), Vol. 3A., AcademicPress, London. 2.Indian Pharmacopoeia, 1996, Govt. of India, The Controller of Publications, Delhi. |
| Customized Product Available | No |
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