Technical Specification Sheet

Tryptic Soy Broth (Soybean-Casein Digest Broth) (NCM0004)

Intended Use
Tryptic Soy Broth (Soybean-Casein Digest Broth) is used for the cultivation of a wide variety of
microorganisms. Tryptic Soy Broth conforms to Harmonized USP/EP/JP requirements and is not intended
for use in the diagnosis of disease or other conditions in humans.

Description
Tryptic Soy Broth, a general-purpose medium, is commonly referred to as Casein Soya Bean or Soybean-
Casein Digest broth, and abbreviated as TSB. This medium was originally developed for use without
blood in determining the effectiveness of sulfonamides against pneumococci and other organisms.
Clostridia and non-sporulating anaerobes grow luxuriantly in this broth when incubated under anaerobic
conditions. TSB is recommended for testing bacterial contaminants in cosmetics 5 and complies with
established standards in the food industry. TSB was chosen by the USDA Animal and Plant Health
Inspection Service for detecting viable bacteria in live vaccines .

TSB conforms to Harmonized United States Pharmacopoeia (USP), European Pharmacopoeia (EU), and
Japanese Pharmacopoeia (JP). The rich nutritional base of TSB, supplemented with SPS and CO2, is an
excellent broth for blood cultures. With the addition of 6.5% NaCl, TSB can be used for the selective
growth of group D streptococci.

Enzymatic Digest of Casein and Enzymatic Digest of Soybean Meal are nitrogen sources in TSB. Dextrose
is the carbon energy source that facilitates organism growth. Sodium Chloride maintains osmotic balance;
Effective Date: 2/7/2020

Dipotassium Phosphate is a buffering agent.

Formula / Liter
Enzymatic Digest of Casein 17.0 g/L
Enzymatic Digest of Soybean 3.0 g/L
Sodium Chloride 5.0 g/L
Dipotassium Hydrogen Phosphate 2.5 g/L
Glucose Monohydrate 2.5 g/L
Final pH: 7.3 ± 0.2 at 25°C
Formula may be adjusted and/or supplemented as required to meet performance specifications.

Precaution
Refer to SDS

Preparation
1. Dissolve 30 g of the medium in one liter of purified water.
2. Mix thoroughly.
3. Autoclave at 121°C for 15 minutes.
Revision: 2

Test Procedure
Refer to appropriate references for specific procedures using Tryptic Soy Broth.

Quality Control Specifications

Dehydrated Appearance: Powder is homogeneous, free flowing, and light beige.

Prepared Appearance: Prepared medium is brilliant to clear, yellow to amber, with none to light
precipitate.
 Technical Specification Sheet

Expected Cultural Response and USP/EP/JP Growth Promotion Testing: Cultural response in Tryptic
Soy Broth tested at Harmonized USP/EP/JP specified temperatures and incubation times. precipitate.
Microorganism Approx. Incubation Expected Growth
Inoculum Period
(CFU)
Aspergillus brasiliensis ATCC® 16404 10 – 100 Within 5 days Growth
Bacillus subtilis ATCC® 6633 10 – 100 18 – 72 hours Growth
Candida albicans ATCC® 10231 10 – 100 18 – 72 hours Growth
Clostridium sporogenes ATCC® 19404 10 – 100 18 – 72 hours Growth
Escherichia coli ATCC® 8739 10 – 100 18 – 24 hours Growth
Pseudomonas aeruginosa ATCC® 9027 10 – 100 18 – 24 hours Growth
Salmonella typhimurium ATCC® 14028 10 – 100 18 – 24 hours Growth
Staphylococcus aureus ATCC® 6538 10 – 100 18 – 24 hours Growth
Streptococcus pneumoniae ATCC® 6305 10 – 100 18 – 72 hours Growth
The organisms listed are the minimum that should be used for Growth Promotion testing.

Results
Refer to appropriate references for test results. Growth is indicated by turbidity.

Expiration
Refer to expiration date stamped on the container. The dehydrated medium should be discarded if not
free flowing, or if the appearance has changed from the original pale to light beige. Expiry applies to
Effective Date: 2/7/2020

medium in its intact container when stored as directed.

Limitation of the Procedure

Due to nutritional variation, some strains may grow poorly or fail to grow on this medium.

References
1. European Pharmacopoeia 10th Edition (2020)
2. United States Pharmacopeia National Formulary Reference 2018: USP 41 NF 36
3. Curry, A. S., G. G. Joyce, and G. N. McEwen, Jr. 1993. CTFA Microbiology guidelines. The Cosmetic,
Toiletry, and Fragrance Association, Inc. Washington, D.C.
4. www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalytical
manualBAM/default.htm.
5. Cunnif, P. 2016. Official methods of analysis AOAC International, 20th ed. AOAC International,
Arlington, VA.
6. Federal Register. 1992. Detection of viable bacteria and fungi except in live vaccine. Fed. Regist.
21:113.26.
Revision: 2