EP0122148A1
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Method of measuring the number of cells of microorganisms
Assignee: Ajinomoto Co Inc
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Filing: 1984-04-10
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Granted: —
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Expires: 2004-04-10
Abstract
Abstract A method of measuring the number of microorganism cells in a sample which comprises preparing a solution or suspension containing a sample of a medicine, food, drink, cosmetic or water, adding to said solution or suspension a 7-amino-4-methyl-coumarin derivative represented by formula (1): wherein R is an alkyl group, an allyl group, an aralkyl group, or a heterocyclic group, or R-CO- is an amino acid or peptide residue, said derivative not inhibiting the hydrolysis of the amine bond of formula (1) by microorganism hydrolases contained in the samples; and measuring the fluorescence of 7-amino-4--methyl-coumarin released by the microorganism hydrolases.
Inventors
- 👤 Hideo Kanou
- 👤 Masahiro Kamata
- 👤 Shigeru Yamanaka
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-30 21:40
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/34; C12Q1/02; C12Q1/04; C12Q1/06; C12Q2334/00; C12Q2334/20; C12Q2334/22; C12Q2337/00; C12Q2337/20; C12Q2337/22; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/911
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
78 words
A method of measuring the number of microorganisms cells in a sample which comprises; if necessary, preparing the sample by making a solution or suspension of it; adding to said sample a 7-amino-4-methyl- coumarin derivative represented by formula (1) wherein R is an alkyl group, an allyl group, an aralkyl group, or a heterocyclic group, and measuring the fluorescence of 7-amino-4-methyl- coumarin released by hydrolysis of the amide bond of the derivative by the hydrolases of the microorganism.
Claim 2
Independent Claim
78 words
A method of measuring the number of microorganisms cells in a sample which comprises; if necessary, preparing the sample by making a solution or suspension of it; adding to said sample a 7-amino-4-methyl- coumarin derivative represented by formula (1) wherein R is an alkyl group, an allyl group, an aralkyl group, or a heterocyclic group, and measuring the fluorescence of 7-amino-4-methyl- coumarin released by hydrolysis of the amide bond of the derivative by the hydrolases of the microorganism.
Claim 3
Dependent Claim
30 words
The method of claim 1 wherein R-CO- in formula (1) represents a naturally occurring amino acid residue or a peptide residue, said peptide residue formed from naturally-occurring amino acid residues.
Claim 4
Dependent Claim
22 words
The method of claim 1 wherein R-CO- in formula (1) represents D or L-leucyl-, or D or L-arginyl- or D or L-pyroglutamyl.
Claim 5
Dependent Claim
12 words
The method of claim 1 wherein R-CO- in formula (1) represents Boc-Phe-Ser-Arg-.
Claim 6
Dependent Claim
31 words
The method of Claim 1 wherein desoxychlic acid is added to the sample prior to the addition of the 7-amino-4-methyl-coumarin derivative in an amount sufficient to enhance the growth of Coliforms.
Claim 7
Dependent Claim
38 words
The method of Claim 1 wherein an antibiotic which prevents the growth of microorganisms other than Eumycetes but does not prevent the growth of Eumycetes is added to the sample prior to the addition of the 7-amino-4-methyl-coumarin derivative.
Claim 8
Dependent Claim
12 words
The method of Claim 6 wherein said antibiotic is pencillin or chloramphenicol.
Claim 9
Dependent Claim
40 words
The method of Claim 1 wherein the solution is maintained at a temperature of more than 40°C for a time sufficient to kill most or all of the organisms which are not thermotolerant before the addition of the 7-amino-4-methyl-coumarin derivative.
Claim 10
Dependent Claim
57 words
The method of Claim 1 wherein one or more derivatives of 4-methyl-umbelliferone, with the general formula (2): wherein X-O- is an alcohol radical, a sugar radical or a phosphate radical which does not inhibit the hydrolysis of the X-O bond in formula (2) by microorganism hydrolases contained in said solution or suspension, is added to the sample.
Claim 11
Independent Claim
37 words
The method of any one of the preceding Claims wherein before adding the 7-amino-4-methyl-coumarin derivative a solution or suspension containing the sample is-centrifuged to produce a precipitated material and said precipitated material is used for subsequent steps.
References Cited (Backward Citations)
US4146604A
Cited
DE3117241A1
Cited
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