Technical Data Sheet

Tryptic Soy Broth (TSB) non-animal origin,

irradiated
Ordering number: 1.00550.0500 / 1.00550.5000

Dehydrated culture medium with vegetable peptones as a non-animal origin alternative to Soybean
Casein Digest Medium (SCDM), also referred to as Tryptic Soy Broth. The Broth is cold filterable and
gamma-irradiated for the microbiological validation of aseptic filling processes (Media Fill Test).

The broth is suitable for media fill tests according to the recommendations of the “FDA Aseptic Guide”
in aseptic production lines. The exclusive use of raw materials of non-animal origin allows the elimination
of a potential contamination risk with TSE/BSE.

The formulation of the basic medium (Vegetable Peptone Broth) is prepared according to the
recommendations of the current European, Japanese and United States Pharmacopoeia (EP, 2.6.12.;
JP, 4.05 and USP, 61), but contains peptones of non-animal origin instead of the recommended
peptones.

Mode of Action

The microbiological performance of TSB non-animal origin is equivalent to the standard TSB.
The dehydrated culture medium is gamma-irradiated with 48-76 kGy. The intensity of irradiation
guarantees that Bacteria, spores, viruses and mycoplasma are destroyed. It provides a higher level of
security with regard to radiation-resistant microorganisms than other commercially available products.
Due to the rich nutrient base, this medium is also suitable for the cultivation of even fastidious
microorganisms.
Typical Composition

Peptone Non-Animal Origin 20 g/l

D(+)-Glucose Monohydrate 2.5 g/l
NaCl 5.0 g/l
K2HPO4 2.5 g/l

Preparation

Suspend 30 g in 1 l of demineralized water. Use according to the purpose required. The prepared
medium is clear and yellowish-brown. The pH value at 25 °C is in the range of 7.1-7.5.

The life science business of Merck KGaA, Darmstadt, Germany

operates as MilliporeSigma in the U.S. and Canada. 1 of 3
Experimental Procedure and Evaluation

The medium is very suitable for the simulation of aseptic filling processes. The product is gamma-
irradiated and triple-bagged allowing for safe introduction into controlled areas. It is granulated which
guarantees excellent solubility even in cold water. The previously aseptically prepared nutrient medium
can be used for the simulation of the aseptic filling of liquids. As sterile filtration is the key element to
achieve sterility autoclaving is not required. Cold filterability is achieved by extensive testing comprising
raw materials, pre-production samples, in-process and end product control. For the validation of the
aseptic filling at least 3.000 units per run are expected to be filled. The contamination rate should be
<1/1000 at a probability of 95 % (EU-GMP guidelines, Annex 1, ISO 13408-1). A growth control should
be carried out for each run. The strains listed in the table below serve as growth controls, whereby
bacteria should demonstrate good growth after 3 days of incubation at the latest and yeasts and moulds
after 5 days.

Storage

The product can be used for sampling until the expiry date if stored upright, protected from light and
properly sealed at +15 °C to +25 °C.

After first opening of the bottle the content can be used up to the expiry date when stored dry and tightly
closed at +15 to +25° C.

Disposal

Please mind the respective regulations for the disposal of used culture medium (e.g. autoclave for 20
min at 121 °C, disinfect, incinerate etc.).

Quality Control

Control Strains ATCC # Inoculum CFU Incubation Expected Results

Escherichia coli 8739 10-100 18-24 h at 30-35 °C Visible Growth
Staphylococcus 18-24 h at 30-35 °C
6538 10-100 Visible Growth
aureus 3 days at 20-25 °C
Staphylococcus
25923 10-100 3 days at 20-25 °C Visible Growth
aureus
Staphylococcus
12228 10-100 3 days at 20-25 °C Visible Growth
epidermidis
Streptococcus
6301 10-100 18-24 h at 30-35 °C Visible Growth
pneumoniae
18-24 h at 30-35 °C
Bacillus subtilis 6633 10-100 Visible Growth
3 days at 20-25 °C
Candida albicans 2091 10-100 5 days at 20-25 °C Visible Growth
Candida albicans 10231 10-100 5 days at 20-25 °C Visible Growth
Aspergillus
brasiliensis 16404 10-100 5 days at 20-25 °C Visible Growth
(formerly A. niger)
Pseudomonas
9027 10-100 18-24 h at 30-35 °C Visible Growth
aerugionas
Salmonella
14028 10-100 18-24 h at 30-35 °C Visible Growth
typhimurium

We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowledge and
ability, but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies 2 of 3
in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking
the suitability of our products for the envisaged purpose.
MilliporeSigma, GranuCult, Readytube and Readyplate are trademarks of Merck KGaA, Darmstadt, Germany. Anaeroclip, Anaerocult,
Bactident, ChromoCult, Duopath, FluoroCult, Readycult, Readybag and Singlepath are registered trademarks of Merck KGaA,
Darmstadt, Germany.
Please refer to the actual batch related Certificate of Analysis

Test Incubation / Method Results

Stability test 7 days at room temperature clear
Sterility test 2 weeks at 20-25 °C No growth
Sterility test 2 weeks at 30-35 °C No growth
Negative post
Mycoplasma qPCR
sterility testing

Literature

European Pharmacopoeia 8.0 (2014): 2.6.1. Sterility; 2.6.12. Microbial examination of non-sterile
products (total viable aerobic count).

Guidance for Industry (2004): Sterile Drug Products Produced by Aseptic Processing - Current Good
Manufacturing Practice.

Hedderich, A., Klees, A., Eiermann, K., Greulich, Y. and Müller, R. (2009): Growth promoting properties
of a vegetable peptone broth (VPB) in comparison to tryptic soy broth (TSB). Poster presentation at
PDA´s Annual Global Conference on Pharmaceutical Microbiology.

Japanese Pharmacopoeia 16th edition (2011): 4.05 Microbial Limit Test.

PDA Technical Report No. 13 (2014 Revised): Fundamentals of an Environmental Monitoring Program.

United States Pharmacopoeia 38 NF 33 (2015): <61> Microbiological examination of non-sterile

products: Microbial Enumeration tests; <71> Sterility Tests.

Ordering Information

Product Cat. No. Pack size

Tryptic Soy Broth (TSB) non-animal origin, irradiated 1.00550.0500 500 g
Tryptic Soy Broth (TSB) non-animal origin, irradiated 1.00550.5000 5 kg

MilliporeSigma Find contact information for your

290 Concord Road country at:
Billerica, MA 01821 www.EMDmillipore.com/offices

For Technical Service, please visit:

www.EMDmillipore.com/techservice

For more information, visit

www.EMDmillipore.com/biomonitoring

We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowledge and
ability, but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies 3 of 3
in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking
the suitability of our products for the envisaged purpose.
MilliporeSigma, GranuCult, Readytube and Readyplate are trademarks of Merck KGaA, Darmstadt, Germany. Anaeroclip, Anaerocult,
Bactident, ChromoCult, Duopath, FluoroCult, Readycult, Readybag and Singlepath are registered trademarks of Merck KGaA,
Darmstadt, Germany.