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Most-Probable-Number (MPN) Method

The Most Probable Number (MPN) method is used to determine the viable bacterial count in samples where other methods are difficult. It relies on statistical probabilities to estimate concentrations from the number of positive and negative growth results in serial dilutions. The procedure involves making serial dilutions of a sample and inoculating aliquots into broth tubes. The tubes are incubated and observed for growth. The number of positive tubes at each dilution is used to estimate the MPN from statistical tables. The method is less precise than membrane filtration or plate counting but is useful for samples with low bacterial concentrations like pharmaceutical products and foods. An example is given of using the MPN method and tables to determine if a cream passes criteria for aerobic microbes.

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Ahmad Azmi
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86 views

Most-Probable-Number (MPN) Method

The Most Probable Number (MPN) method is used to determine the viable bacterial count in samples where other methods are difficult. It relies on statistical probabilities to estimate concentrations from the number of positive and negative growth results in serial dilutions. The procedure involves making serial dilutions of a sample and inoculating aliquots into broth tubes. The tubes are incubated and observed for growth. The number of positive tubes at each dilution is used to estimate the MPN from statistical tables. The method is less precise than membrane filtration or plate counting but is useful for samples with low bacterial concentrations like pharmaceutical products and foods. An example is given of using the MPN method and tables to determine if a cream passes criteria for aerobic microbes.

Uploaded by

Ahmad Azmi
Copyright
© © All Rights Reserved
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Most-Probable-Number

(MPN) Method
❖ Principle:
● The most probable number is a method used to determine the viable count of bacteria in a sample,
where other viable count methods are difficult to perform.
● The precision and accuracy of the MPN Method is less than that of the Membrane Filtration method or
the Plate-Count Method.
● This method is particularly useful for samples which contain low concentrations of bacterial cells; for
example, non sterile pharmaceutical products (Like viable count methods), in addition to raw materials
in the pharmaceutical industry, milk, water, etc.
● MPN relies on a statistical method based on probability theory. According to the USP, there are
statistical tables designed to find MPN values for a sample. These tables show the results at 95%
confidence limits (‫ بكون متاكد من صحة النتائج‬% 95 ‫)بنسبة‬.
● Unreliable results are obtained particularly for the enumeration of molds. For these reasons, the MPN
Method is reserved for the enumeration of TAMC (Total Aerobic Microbial Count).
❖ Procedures (According to USP):
1- Prepare a series of three serial 10-fold dilutions from the sample solution provided using the provided
sterile pipettes, saline, and empty sterile tubes.
2- From each level of dilution, three aliquots of 1 mL are used to inoculate three tubes
with 9 mL of the provided Soybean–Casein Digest Broth.

3- Incubate all tubes at 30° to 35° for not more than 3 days.
•If reading of the results is difficult or uncertain owing to the nature of the product to
be examined, subculture in the same broth or in Soybean–Casein Digest Agar for 1 to
2 days at the same temperature, and use these results.
From USP Table, determine the most probable number of microorganisms per g or
mL of the product to be examined.
Example: (Please explain this example in the section)
You are a pharmacist working in the quality control of industrial
pharmaceutical company. You performed the MPN for a cream
using 1 g from it after dissolving in the suitable solvent. Later,
you obtained the following result:

According to the USP, cutaneous non-sterile pharmaceutical


products should contain no more than 100 cfu/g. Please
determine whether this cream is Passed for use or not according
to the USP criteria.
Answer: from the table MPN is 150/g and the cream is not passed

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