Antibiotic HiVeg™ Assay Medium No. 35

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SKU:
MV798
Used for the microbiological assay of Bleomycin using Mycobacterium smegmatis.


Intended Use

Recommended for microbiological assay of Bleomycin using Mycobacterium smegmatis.

Composition

Ingredients g/L
HiVeg® peptone 10.000
HiVeg® extract 10.000
Sodium chloride 3.000
Agar 17.000
Final pH (at 25°C) 7.0±0.2

Formula adjusted, standardized to suit performance parameters

Directions

Suspend 40.0 grams in 1000 ml purified/distilled water containing 10 ml of glycerol. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

This medium is formulated in accordance to CFR (1). This medium is employed widely as base agar for agar diffusion assay of Bleomycin using Mycobacterium smegmatis. Antibiotic HiVeg® Assay Medium No. 35 is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. The nutrients essential for growth of test organism is provided by HiVeg® extract and HiVeg® peptone in this medium. Agar provides excellent solid substratum for support and overlayering of seed agar, for the assay of Bleomycin. Addition of glycerol is important for provision of carbon to the test organism. To perform the antibiotic assay the Base Agar should be prepared on the same day as the test. For the cylinder method, a base layer of 10 ml is required. Once the base medium has solidified, seed layer inoculated with the standardized culture can be overlaid. Even distribution of the layer is important.

Type of specimen

Pharmaceutical sample

Specimen Collection and Handling

For pharmaceutical sample samples follow appropriate techniques for handling specimens as per established guidelines (1). 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. Freshly prepared medium plates must be used or it may result in erroneous results.

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 1.7% agar gel.

Colour and Clarity of prepared medium
Medium amber coloured clear to slightly opalescent gel forms in Petri plates

Reaction

Reaction of 4.0% w/v aqueous solution containing 1% glycerol at 25°C. pH: 7.0±0.2

pH

6.80-7.20

Cultural Response

Cultural characteristics observed after an incubation at 36-37.5 for 18-48 hours.

Organism Inoculum
(CFU)
Growth Recovery Antibiotics
assayed
Mycobacterium smegmatis
ATCC 607
50-100 luxuriant >=50% Bleomycin

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and use freshly prepared medium. 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 (2,3).

More Information
Product Name Antibiotic HiVeg™ Assay Medium No. 35
SKU MV798
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Tests and Methods of Assay of Antibiotics and Antibiotic containing Drugs, FDA, CFR, 1983Title 21, Part 436, Subpart D,Washington, D.C.: U.S. Government Printing Office, paragraphs 436, 100-436, 106, p. 242-259, (April 1).
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