Buffered Sodium Chloride Peptone Solution pH 7.00 w/ 0.1% Polysorbate 80 and 0.3% Soya lecithin

Intended use

Recommended as a diluent for carrying out microbial limit testing by harmonized methodology of pharmaceutical products in accordance with USP/EP/BP/JP/IP.

Composition**

Final pH (at 25°C): 7.00±0.2

**Formula adjusted, standardized to suit performance parameters

# Peptone (meat or casein)

Directions

Suspend 20.10 grams (the equivalent weight of dehydrated medium per liter) in 1000 ml purified /distilled water. Heat if necessary to dissolve the medium completely. Dispense in tubes or flasks or as desired and sterilize by autoclaving at 15 lbs pressure 121°C for 15 minutes or as per validated cycle.

Principle And Interpretation

The composition of this medium is in accordance with the harmonized methodology of USP/EP/BP/JP/IP (1-5). This medium is recommended for preparation of stable test strain suspension employed for validating the microbiological testing procedures of non-sterile products. The standardized stable suspensions are used so that the suitability of this test to detect microorganism in presence of product can be established. Non-fatty products insoluble in water and water-soluble products are diluted/dissolved using this solution.

HMC Peptone serves as nutrient source and maintains the cell viability. Phosphates in the medium act as good buffering agents. Sodium chloride maintains the osmotic balance and cell integrity. Polysorbates reduce surface tension and also inactivate phenolic compound, if present in the test sample. For preparation of non-fatty products insoluble in water, 0.1% w/v Polysorbate 80 is added to assist the suspension of poorly wettable substances. Lecithin is incorporated to neutralize any residual disinfectant activity.

Preparation of test strain is recommended in Buffered Sodium chloride-Peptone solution pH 7.0 (M2156) at 30-35°C wherein there is no multiplication of organisms or there is no decrease in count for upto 4 hours.

Type of specimen

Pharmaceutical samples

Specimen Collection and Handling

For pharmaceutical samples follow appropriate techniques for handling specimens as per established guidelines (1-5). 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

1. This medium contains less nutrients and is not recommended for the growth of microorganisms.

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: White to cream homogeneous free flowing powder

Colour and Clarity of prepared medium: Colourless to pale yellow slightly hazy solution

pH: 6.80 - 7.20

Growth Promotion Test: Growth Promotion is carried out in accordance with the harmonized method of ICH(USP/EP/BP/JP/IP).

Cultural response

Cultural characteristics observed after recovery on Soybean Casein Digest Agar after an incubation at 30-35°C for 18-24 hours for bacteria and Sabouraud Dextrose Agar at 30-35°C for 24-48 hours.

Key : (*) Corresponding WDCM Numbers

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle inorder 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 must be decontaminated and disposed of in accordance with current laboratory techniques (6,7).

Reference

1. The British Pharmacopoeia, 2022, Medicines and Healthcare products Regulatory Agency.
2. European Pharmacopoeia, 2022, 10 th volume, European Directorate for the quality of medicines & Healthcare.
3. Indian Pharmacopoeia, 2022, Indian Pharmacopoeia Commission, Ministry of Health and Family Welfare Government of India.
4. The United States Pharmacopoeia-National Formulatory (USP-NF), 2022.
5. The Japanese Pharmacopoeia, 17th edition, 2016, The Ministry of Health, Labour and welfare.
6. Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
7. Jorgensen,J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.