TECHNICAL DATA SHEET

BUFFERED PEPTONE WATER

DILUENT
ENRICHMENT BROTH

1 INTENDED USE

Buffered Peptone Water is a general use diluent destined for numerous operations and standards including sample
preparation, the preparation of stock suspensions and serial dilutions of samples (standard ISO 6887).
This media is also used for the pre-enrichment of Salmonella and for Cronobacter sakazakii, by allowing the
resuscitation of microorganisms having undergone sublethal treatments as in the case of spray drying, pasteurization,
action of conservators, elevated osmotic pressure and high acidity (NF EN ISO 6579-1, ISO 22964...).

2 PRINCIPLES

Sodium chloride maintains the osmotic balance.

The medium is buffered with sodium and potassium phosphates.

3 TYPICAL COMPOSITION

The composition can be adjusted in order to obtain optimal performance.

For 1 liter of media:

- Peptone ................................................ 10.00 g
- Sodium chloride ...................................... 5.00 g
- Disodium phosphate, anhydrous............. 3.57 g
- Monopotassium phosphate, anhydrous . 1.50 g
pH of the ready-to-use media at 25 °C: 7.0 ± 0.2.

For 25.5 g of media BK018 For 20 g of media BK131

Peptone ............................................................. 10.0 g Peptone ............................................................. 10.0 g
Sodium chloride ................................................... 5.0 g Sodium chloride ................................................... 5.0 g
Disodium phosphate, dodecahydrated .............. 9.0 g Disodium phosphate, anhydrous...................... 3.57 g
Monopotassium phosphate, anhydrous ............. 1.5 g Monopotassium phosphate, anhydrous ............. 1.5 g

4 PREPARATION

• Dissolve 25.5 g of dehydrated medium BK018 or 20.0 g BK131 in 1 liter of Reconstitution:

distilled or demineralized water. - BK018: 25.5 g/L
• Stir slowly until complete dissolution. - BK131: 20.0 g/L
• Dispense in vials or tubes.
• Sterilize in an autoclave at 121°C for 15 minutes. Sterilization:
• Cool to room temperature. 15 min at 121 °C

NOTE: For the double strength buffered peptone water dissolve 51.0 g of dehydrated medium BK018 or 40.0 g of
dehydrated medium BK131 in 1 liter of distilled or demineralized water.

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5 INSTRUCTIONS FOR USE

Preparation of stock solutions:

• Aseptically add 10 or 25 g of the product to analyze to a tared flask containing 90 or 225 mL of media prepared as
above (or to the ready-to-use media BM057 or BM010).
• Homogenize thoroughly with an appropriate mixer in order to obtain a stock suspension or a pre-enrichment
broth.
• For Salmonella or enterobacteria enrichment notably, incubate by carefully respecting the appropriate analytical
protocol being followed.

Preparation of serial dilutions:

• Introduce 1 mL of the stock suspension into a tube containing 9 mL of media prepared as described or using
ready-to-use media (BM056).
• Mix well.
• Redo the operation as many times as needed to achieve the proper dilution.

NOTE: For the analyze of acidic products (pH between 4.5 and 3.5), the double-strength buffered peptone water may
be used (ISO 6887).

6 QUALITY CONTROL

Dehydrated media: cream-white powder, free-flowing and homogeneous.

Prepared media: amber solution, limpid, can have a slight precipitate after prolonged storage.
Typical culture response (NF EN ISO 11133):

Microorganisms Growth
(1)
Salmonella Typhimurium WDCM 00031 Positive, score 2
(1)
Salmonella Enteritidis WDCM 00030 Positive, score 2
(1) Escherichia coli WDCM 00012 Positive, score 2
(2)
Listeria monocytogenes 4b WDCM 00021 ± 30 % colonies / T0
(3)
Escherichia coli WDCM 00012 ± 30 % colonies / T0
(3)
Staphylococcus aureus WDCM 00034 ± 30 % colonies / T0
(1)
After 18 hours of incubation at 37 °C (inoculum ≤ 102 microorganisms)
(2)
After 60 minutes of incubation at 20 °C
(3)
After 45-60 minutes of incubation at 20-25 °C

7 STORAGE / SHELF LIFE

Dehydrated media: 2-30 °C.

Ready-to-use media, in tubes, vials or flexible bags: 2-25 °C.
The expiration dates are indicated on the labels.

Prepared media in vials or tubes (*): 180 days at 2-25 °C.

(*) Benchmark value determined under standard preparation conditions, following manufacturer’s instructions.

8 PACKAGING

Dehydrated media (25.5 g/L):

500 g bottle........................................................................................................................................................... BK018HA
5 kg drum ............................................................................................................................................................. BK018GC

Dehydrated media (20 g/L):

500 g bottle........................................................................................................................................................... BK131HA
5 kg drum ............................................................................................................................................................. BK131GC

Ready-to-use media:
50 x 9 mL tubes .................................................................................................................................................... BM05608
10 x 225 mL vials ................................................................................................................................................. BM01008

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10 x 90 mL vials ................................................................................................................................................... BM05708
3 x 3 liter flexible bags .......................................................................................................................................... BM13108
2 x 5 liter flexible bags .......................................................................................................................................... BM13208
40 x 5 liter flexible bags ........................................................................................................................................ BM21408

9 BIBLIOGRAPHY

NF U 47-100. July 2007. Animal health analysis methods - Isolation and identification of Salmonella or search for
particular serovar's in the animal production sector.
NF U 47-101. November 2007. Animal health analysis methods - Isolation and identification of any salmonella
serotypes or of specified salmonella serotypes among birds.
NF U 47-102. January 2008. Animal health analysis methods - Isolation and identification of any salmonella serotype
or of specified salmonella serotypes among mammals.
NF V 08-407. December 2011. Microbiology of food products - Raw materials coming into the composition of canned
foods - Enumeration of the heat-resisting spores of Bacillus and Clostridium thermophilus - Most probable number
technique.
NF EN ISO 19250. June 2013. Water quality - Detection of Salmonella spp.
NF EN ISO 11133. July 2014. Microbiology of food, animal feed and water - Preparation, production, storage and
performance testing of culture media.
NF EN ISO 11133/A1. March 2018. Microbiology of food, animal feed and water - Preparation, production, storage and
performance testing of culture media – Amendment 1.
NF EN ISO 11133/A2. Mai 2020. Microbiology of food, animal feed and water - Preparation, production, storage and
performance testing of culture media – Amendment 2.
NF EN ISO 6579-1. April 2017. Microbiology of the food chain - Horizontal method for the detection, enumeration and
serotyping of Salmonella - Part 1: detection of Salmonella spp..
NF EN ISO 6579-1/A1. March 2020. Microbiology of the food chain - Horizontal method for the detection, enumeration
and serotyping of Salmonella - Part 1: detection of Salmonella spp..- Amendment 1.
NF EN ISO 22964. June 2017. Microbiology of the food chain - Horizontal method for the detection of Cronobacter
spp.
NF EN ISO 6887-1. June 2017. Microbiology of the food chain - Preparation of test samples, initial suspension and
decimal dilutions for microbiological examination - Part 1: general rules for the preparation of the initial suspension and
decimal dilutions
NF EN ISO 6887-2. June 2017. Microbiology of the food chain - Preparation of test samples, initial suspension and
decimal dilutions for microbiological examination - Part 2: specific rules for the preparation of meat and meat products.
NF EN ISO 6887-3. June 2017. Microbiology of the food chain - Preparation of test samples, initial suspension and
decimal dilutions for microbiological examination - Part 3: specific rules for the preparation of fish and fishery products.
NF EN ISO 6887-4. June 2017. Microbiology of the food chain - Preparation of test samples, initial suspension and
decimal dilutions for microbiological examination - Part 4: specific rules for the preparation of miscellaneous products.
NF EN ISO 6887-5. May 2020. Microbiology of the food chain - Preparation of test samples, initial suspension and
decimal dilutions for microbiological examination - Part 5: specific rules for the preparation of milk and milk products.
NF EN ISO 11290-2. July 2017. Microbiology of the food chain - Horizontal method for the detection and enumeration
of Listeria monocytogenes and of Listeria spp. - Part 2: enumeration method.
NF EN ISO 21528-1. July 2017. Microbiology of the food chain - Horizontal method for the detection and enumeration
of Enterobacteriaceae - Part 1: Detection of Enterobacteriaceae.

10 ADDITIONAL INFORMATION

The information provided on the labels take precedence over the formulations or instructions described in this
document and are susceptible to modification at any time, without warning.

Document code : BUFFERED PEPTONE WATER_v20 (en)

Creation date : 06-2003
Updated : 10-2023
Origin of revision : Review of quality control and bibliography

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