TRYPTIC SOY AGAR (TSA), USP

Cat. no. G60 TSA, USP, 15x100mm Plate, 18ml 10 plates/bag
Cat. no. G60BX TSA, USP, 15x100mm Plate, 18ml  100 plates/box
Cat. no. J330 TSA / CET (Cetrimide Selective Agar) / MSA (Mannitol Salt Agar), USP, 15x100mm Triplate, 7ml/section 10 plates/bag
Cat. no. H19 TSA, USP, 15x150mm Plate, 69ml 10 plates/bag
Cat. no. W64 TSA, USP, 15x100mm Plate, 26ml 10 plates/bag
Cat. no. Q58 TSA, USP, 20x125mm Tube, 18ml Deep 20 tubes/box
Cat. no. Q80 TSA 1.5x, USP, 20x150mm Tube, 20ml Deep 100 tubes/box
Cat. no. U49 TSA, USP, 8oz. Glass Bottle, 150ml 20 bottles/box
Cat. no. U60 Tryptic Soy Agar (TSA), USP, 1L Polypropylene Bottle, 700ml 10 bottles/box
Cat. no. U158 Tryptic Soy Agar (TSA), USP, 500ml Glass Bottle, 500ml 1 each
Cat. no. U260 TSA, USP, 8oz. Glass Bottle, 200ml 12 bottles/box
Cat. no. U360 TSA, USP, 16oz. Glass Bottle, 400ml 12 bottles/box
Cat. no. U361 Tryptic Soy Agar (TSA), USP, 500ml Polycarbonate Bottle, 500ml 10 bottles/box
Cat. no. U373 Tryptic Soy Agar (TSA), USP, 1L Polycarbonate Bottle, 1000ml 10 bottles/box

INTENDED USE

Hardy Diagnostics Tryptic Soy Agar (TSA), USP is recommended for use as a general growth medium for the detection and enumeration of microorganisms from non-clinical samples as specified by the United States Pharmacopoeia (USP).(1) In addition, the medium complies with the harmonized European, U.S. and Japanese Pharmacopoeias for determining the microbial quality of non-sterile products.(1)

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

Tryptic Soy Agar (TSA), USP is formulated in accordance with the U.S. Pharmacopoeia standard formula for Soybean-Casein Digest Agar and contains digests of soybean meal and casein, which provide amino acids and other nitrogenous compounds to promote microbial growth. Sodium chloride is added to help cells maintain osmotic equilibrium. Dextrose is added as an energy source. Agar is the solidifying agent.

FORMULA

Ingredients per liter of deionized water:*

Pancreatic Digest of Casein 15.0gm
Peptic Digest of Soybean Meal 5.0gm
Sodium Chloride 5.0gm
Agar 15.0gm

Final pH 7.3 +/- 0.2 at 25ºC.

In addition:

TSA 1.5X, USP (Cat. no. Q80) contains 1.5X the above ingredients.

Prepared in accordance with USP <62>.(1)

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store Cat. nos. G60, G60BX, H19, and W64 at 2-8ºC away from direct light. Store Cat. nos. Q58, Q80, U49, U60, U158, U260, U360, U361, and U373 at 2-30ºC . Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Before Use: The medium should be warmed to room temperature and the surface dry prior to inoculating. To reduce the potential for cross-contamination, it is strongly suggested that appropriate gowning and glove procedures, designated aseptic processing areas, appropriate sporicidal disinfectants, and environmental monitoring procedures be strictly enforced to reduce the likelihood of accidental contamination.(1) Use of stringent aseptic techniques, appropriate sporicidal agents, and a laminar clean bench are recommended in accordance with USP Microbiological Best Laboratory Practices <1117> and Sterility Testing - Validation of Isolator Systems <1208>.

Environmental Monitoring: Consult USP Microbiological Control and Monitoring of Aseptic Processing Environments <1116>.(1)

Sedimentation (Settling) Plate Method: Place the plate on a clean piece of paper and expose the agar by removing the lid. Do not invert the lid while removed to avoid exposure to falling sediment. Expose the agar for 15 minutes or longer, depending upon established procedures, and replace the lid. Incubate according to laboratory protocol.

Impact Air Sampling Method: Use the plate size specified for the impact air sampling unit. Remove the sampler head and place the plate, lid up, into the slot. Aseptically remove the lid and expose the agar; do not invert the lid while removed to avoid exposure to falling sediment. Place the sampler head back on the unit and turn the unit on; sample a specific volume of air according to laboratory procedure. After sampling, remove the sampler head, aseptically return the lid of the plate, and remove the plate from the sampling unit; incubate per laboratory protocol.

For re-melting solid tube and bottle media: Autoclave containers with slightly loose caps at 121ºC for 1-3 minutes or until melted. Do not heat media longer than 3 hours at 45-50ºC. Alternatively, solid agar in capped containers can be racked and placed in a covered, boiling water bath (100ºC) before use. There should be enough water in the water bath to reach the top of the media line. A covered water bath will maintain consistent temperature of the media until melted. Cool media to 45-50ºC and aseptically dispense into sterile containers. Note: Sterile solidified media can be re-melted only once. In addition, the use of microwaves to melt media is not advised.

Performance Testing and Preparation of Test Strains: Use stable standardized suspensions of test strains per reference method. Use appropriate diluent for making test suspensions and use suspensions within the specified time period or maintain under appropriate storage practices.(1)

Testing Growth Promotion or Inhibitory Properties of Media:
Growth Promotion, Liquid Media Inoculate a portion of the appropriate medium with a small number (not more than 100cfu) of the appropriate test microorganism.
Growth Promotion, Solid Media Perform surface spread or plate-count methods, inoculating each plate with a small number (not more than 100cfu) of the appropriate microorganism.
Inhibitory Properties, Liquid/Solid Media Inoculate the appropriate medium with at least 100cfu of the appropriate test microorganism.
Indicative Properties Perform surface spread or plate-count methods, inoculating each plate with a small number (not more than 100cfu) of the appropriate microorganism.

Perform membrane filtration or the plate count method, as required.(1)

Incubate media using appropriate atmospheric, temperature, and duration conditions as outlined by the test or reference method.(1) Place plates in an inverted position until growth is evident. Incubate bacterial cultures at 30-35ºC for up to 3 days; for fungal cultures, incubate at 20-25ºC for up to 5 days.

INTERPRETATION OF RESULTS

Clearly visible growth in the form of colonies constitutes a positive result. Note the inoculum dilution with the smallest and largest quantity of growth and determine the probable number of bacterial cells per gram or milliliter of sample. Growth of the microorganism should not differ by a factor greater than two from the calculated value for a standard inoculum and should be comparable to that obtained from a previously tested and approved batch of the same medium.(1)

For environmental monitoring procedures, consult USP <1116> and count the number of colonies: report as the number of colony forming units (CFU).

For growth promotion, enumeration, or sterility testing procedures, consult USP <61>, <62>, or <71>.

Because of the inherent variability of environmental sampling methods, it is more useful to trend contamination recovery results rather than focus on the number of colonies recovered from a single sample. Action should be required when the contamination recovery rate trends above the recommended action levels for a significant time.

If action levels have been identified, a thorough investigation into the adequacy of personnel work practices, operational procedures, cleaning procedures and solutions, and air filtration efficiency within the processing area must be made. Once changes have been made, monitoring procedures must be repeated to determine if the changes made were effective. Documentation of all monitoring results, remedial action, and follow-up monitoring must be maintained. Consult listed reference for more detailed information concerning plate count methods.(1)

LIMITATIONS

Accurate counting may be difficult with molds or spreading colonies.

Sampling challenges may occur with irregular, porous, rough, or textured media surfaces.

Rare, fastidious microorganisms may not grow on general non-selective media formulations.

Refer to the document "Limitations of Procedures and Warranty" on the Hardy Diagnostics Technical Document website for more information.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, applicator sticks, other culture media, incinerators, incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Staphylococcus aureus
ATCC® 6538
J 1-3 days 30-35°C Aerobic Growth
Pseudomonas aeruginosa
ATCC® 9027
J 1-3 days 30-35°C Aerobic Growth
Bacillus subtilis
ATCC® 6633
J 1-3 days 30-35°C Aerobic Growth
Candida albicans
ATCC® 10231
J 1-5 days 30-35°C Aerobic Growth
Aspergillus brasiliensis
ATCC® 16404
J 1-5 days 30-35°C Aerobic Growth
In addition to the above, Cat. no. G60 and G60BX also include the following:
Staphylococcus aureus
ATCC® 25923
A 18-24 hrs 35°C Aerobic Growth
Escherichia coli
ATCC® 25922
A 18-24 hrs 35°C Aerobic Growth
Escherichia coli
ATCC® 8739
J 1-3 days 30-35°C Aerobic Growth

* Refer to the document "Inoculation Procedures for Media QC" on the Hardy Diagnostics Technical Document website for more information.

Tested in accordance with USP <61>.(1)

USER QUALITY CONTROL

PHYSICAL APPEARANCE

Tryptic Soy Agar (TSA), USP should appear translucent, and light amber in color.

Tryptic Soy Agar

Uninoculated plate of Tryptic Soy Agar, USP (Cat. no. G60).

REFERENCES

1. United States Pharmacopoeia and National Formulary (USP-NF). Rockville, MD: United States Pharmacopeial Convention.

Microbiological Tests
<61> Microbial Examination of Nonsterile Products: Microbial Enumeration Tests
<62> Microbial Examination of Nonsterile Products: Tests for Specified Microorganisms
<71> Sterility Tests

General Information
<1116> Microbiological Control and Monitoring of Aseptic Processing Environments
<1117> Microbiological Best Laboratory Practices
<1208> Sterility Testing - Validation of Isolator Systems

ATCC is a registered trademark of the American Type Culture Collection.

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