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Letheen Broth, Modified, Granulated (Modified Letheen Broth, Granulated)
Sterility Testing#CC293D
Intended Use
This medium is recommended for screening cosmetic products for microbial contamination.
Composition
| Ingredients | Gms / Litre |
|---|---|
| Peptic digest of animal tissue | 20.000 |
| Casein enzymic hydrolysate | 5.000 |
| Beef extract | 5.000 |
| Yeast extract | 2.000 |
| Sodium chloride | 5.000 |
| Sodium bisulphite | 0.100 |
| Lecithin | 0.700 |
| Polysorbate 80 | 5.000 |
| Final pH (at 25°C) | 7.0±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 42.8 grams in 1000 ml distilled water. Heat if necessary to ensure complete dissolution. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Mix well and dispense in tubes or flasks or as desired.
Principle And Interpretation
In the early 40s, Weber and Black recommended the use of lecithin and polysorbates to neutralize the antimicrobial action of the quaternary ammonium compounds (5). In 1965, the methodology was accepted by AOAC for the antimicrobial assays and extended their use to all the cationic detergents. In 1978, the FDA incorporated it as pre-enrichment medium for every microbial examination of cosmetics.
Letheen Broth, Modified is prepared as per FDA (1) for screening cosmetic products for microbial contamination. There are great chances of altering the chemical composition of cosmetics by the metabolism of organism s thereby spoiling and causing harm to the users (2, 3, 4). Direct colony counts and enrichment culturing are the methods of choice for isolating microorganisms from cosmetic products. The word Letheen represents a combination of lecithin and polysorbate (Tween) 80.
Peptic digest of animal tissue, casein enzymic hydrolysate, beef extract and yeast extract provide nitrogenous nutrients, carbon compounds and trace elements to the microorganisms. Incorporation of lecithin and polysorbate 80 to the medium enables the recovery of bacteria from materials containing residues of disinfectant compounds or preservatives used in cosmetics. Polysorbate 80 is added to nullify phenolic compounds, hexachlorophene, formalin and along with lecithin neutralizes ethyl alcohol (6). Lecithin also neutralizes quaternary ammonium compounds present in the cosmetics. Sodium chloride maintains the osmotic balance of the medium.
Enrichment in this medium should be done for 7 days at 30-32°C and then subcultured on Letheen Agar, Modified (GM946/ M946) and / or MacConkey Agar (GM081/M081).
Quality Control
Appearance
Cream to yellow coloured granular medium
Colour and Clarity of prepared medium
Yellow coloured, clear solution in tubes
Reaction
Reaction of 4.28% w/v aqueous solution at 25°C. pH: 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth |
|---|---|---|
| Escherichia coli ATCC 25922 | 50-100 | luxuriant |
| Staphylococcus aureus ATCC 25923 | 50-100 | luxuriant |
| Staphylococcus aureus ATCC 6538 | 50-100 | good-luxuriant |
Storage and Shelf Life
Store below 30°C in tightly closed container and prepared medium at 2-8°C. Use before expiry period on the label.
| Product Name | Letheen Broth, Modified, Granulated (Modified Letheen Broth, Granulated) |
|---|---|
| SKU | GM976 |
| Product Type | Granulated |
| Physical Form | Granular |
| Origin | Animal |
| Packaging type | HDPE |
| References | 1. Bacteriological Analytical Manual, 1995, Food and Drug Administration, 8th Ed., AOAC International, Gaithersburg, MD,U.S.A. |
| Customized Product Available | No |











