Patent US5310462A successfully refreshed from Google Patents.
US5310462A
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Quantitation of samples utilizing capillary electrophoresis
Assignee: Beckman Coulter Inc
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Filing: 1991-05-31
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Granted: 1994-05-10
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Expires: 2011-05-31
Abstract
Abstract Ionic species and method for the quantitation of sample constituents utilizing capillary zone electrophoresis analysis are disclosed. As disclosed, at least one ionic species having a charge density of betwen about 0.02 and about 0.001 is added to the sample. Benzoic acid is a preferred ionic species. Quantitation of a sample constituent can be derived by normalizing the constituent species with the ionic species.
Inventors
- 👤 Fu-Tai A. Chen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 14:48
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N27/00; G01N27/26; G01N27/416; G01N27/447; G01N27/44704; G01N27/44717; G01N27/44721; G01N27/44726
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
107 words
A method of quantifying a peptide-containing constituent species in a sample by capillary zone electrophoresis comprising the following steps: a) adding at least one ionic species to the sample to form a mixture, the ionic species having a charge density greater than substantially all of the peptide-containing constituent species in the sample, b) subjecting the mixture to capillary zone electrophoresis, c) detecting the ionic species and the peptide-containing constituent species in said mixture, and d) normalizing at least one peptide-containing constituent species relative to the detected ionic species, where the normalized value of step (d) is the quantified amount of said peptide-containing constituent species in the sample.
Claim 2
Independent Claim
107 words
A method of quantifying a peptide-containing constituent species in a sample by capillary zone electrophoresis comprising the following steps: a) adding at least one ionic species to the sample to form a mixture, the ionic species having a charge density greater than substantially all of the peptide-containing constituent species in the sample, b) subjecting the mixture to capillary zone electrophoresis, c) detecting the ionic species and the peptide-containing constituent species in said mixture, and d) normalizing at least one peptide-containing constituent species relative to the detected ionic species, where the normalized value of step (d) is the quantified amount of said peptide-containing constituent species in the sample.
Claim 3
Dependent Claim
20 words
The method of claim 1 wherein the charge density of the ionic species is between about 0.02 and about 0.001.
Claim 4
Dependent Claim
20 words
The method of claim 1 wherein the charge density of the ionic species is between about 0.01 and about 0.004.
Claim 5
Dependent Claim
20 words
The method of claim 1 wherein the charge density of the ionic species is between about 0.01 and about 0.006.
Claim 6
Dependent Claim
25 words
The method of claim 1 wherein the ionic species comprises at least one negative charge and has a molecular weight of at least about 50.
Claim 7
Dependent Claim
21 words
The method of claim 1 wherein said ionic species is soluble in an aqueous medium at pH greater than about 8.00.
Claim 8
Dependent Claim
18 words
The method of claim 1 wherein said ionic species has an absorbance at less then about 300 nm.
Claim 9
Dependent Claim
109 words
The method of claim 1 wherein the concentration of said ionic species added to the sample is determined by multiplying a dilution factor of the sample by the protein concentration for a normal control to obtain a first value; multiplying a desired integrated area percentage of the ionic species by the first value to obtain a second value; and multiplying the second value by a third value, the third value derived by dividing an absorbance value of a fixed amount of the ionic species measured at a fixed wavelength by an absorbance value of about the same fixed amount of the sample measured at about the same fixed wavelength.
Claim 10
Dependent Claim
18 words
The method of claim 8 wherein the desired integrated area percentage is a value less than about 0.50.
Claim 11
Dependent Claim
20 words
The method of claim 8 wherein the desired integrated area percentage is a value between about 0.05 and about 0.40.
Claim 12
Dependent Claim
14 words
The method of claim 8 wherein the desired integrated area percentage is about 0.30.
Claim 13
Dependent Claim
18 words
The method of claim 8 wherein the dilution factor is a value between about 0.05 and about 0.01.
Claim 14
Dependent Claim
12 words
The method of claim 8 wherein the dilution factor is about 0.02.
Claim 15
Dependent Claim
18 words
The method of claim 8 wherein the protein concentration is between about 10 μg/ml and about 60,000 μg/ml.
Claim 16
Dependent Claim
13 words
The method of claim 8 wherein the protein concentration is about 60 mg/ml.
Claim 17
Dependent Claim
65 words
The method of claim 1 wherein said ionic species is selected from the group consisting of formic acid, acetic acid, benzo-phosphoric acid, propionic acid, isopropionic acid, butyric acid, isobutyric acid, benzoic acid, benzo-sulfonic acid, ortho-chloro benzoic acid, meta-chloro benzoic acid, para-chloro benzoic acid, naphthyl sulfonic acid, benzo naphthalinic acid, chloro-benzo naphthalinic acid, chloro-naphthyl sulfonic acid, tetra-iodo benzo naphthyl sulfonic acid, and di-iodo anthracenyl sulfonic acid.
Claim 18
Dependent Claim
12 words
The method of claim 1 wherein said ionic species is benzoic acid.
Claim 19
Dependent Claim
19 words
The method of claim 17 wherein the concentration of said benzoic acid in said sample is about 0.10 mg/ml.
Claim 20
Dependent Claim
174 words
The method of claim 1 wherein said normalizing of at least one peptide-containing constituent species is determined by dividing an integrated area value of at least one peptide-containing constituent species by the total integrated area amounts from the analysis to obtain a first value; dividing the first value by a second value to obtain a third value, the second value derived by dividing the integrated area value of the ionic species by the total integrated area amounts from the analysis; multiplying the third value by a fourth value, the fourth value derived by dividing the dilution factor of the sample by the numeral 1.0 to obtain a fifth value; multiplying the fifth value by a sixth value, the sixth value derived by dividing an absorbance value of a fixed amount of the sample measured at a fixed wavelength by an absorbance value of about the same fixed amount of the ionic species measured at about the same fixed wavelength; and multiplying the sixth value by the concentration of the ionic species in the sample.
Claim 21
Independent Claim
255 words
A method of quantifying peptide-containing constituent species in a clinical sample by capillary zone electrophoresis comprising the following steps: a) adding at least one ionic species having a charge density of between about 0.02 and about 0.001 to the sample to form a clinical mixture, the charge density of the ionic species being greater than substantially all of the constituent species in the clinical sample; b) separating said clinical mixture into its constituent parts by capillary zone electrophoresis; c) detecting the ionic species and the peptide-containing constituent species in said mixture; and d) normalizing at least one peptide-containing constituent species by dividing the integrated area value of said peptide-containing species by the total integrated area values from all of the peptide-containing constituent species to obtain a first value; dividing the first value by a second value to obtain a third value, the second value being obtained by dividing the integrated area value of the ionic species by the total integrated area amounts from the constituent species; multiplying the third value by a fourth value, the fourth value being the dilution factor of the sample to obtain a fifth value; multiplying the fifth value by a sixth value, the sixth value being obtained by dividing the absorbance value of a fixed amount of the sample measured at a fixed wavelength by the absorbance value of about the same fixed amount of the ionic species measured at about the same fixed wavelength; and multiplying the sixth value by the concentration of the ionic species in the sample.
Claim 22
Independent Claim
255 words
A method of quantifying peptide-containing constituent species in a clinical sample by capillary zone electrophoresis comprising the following steps: a) adding at least one ionic species having a charge density of between about 0.02 and about 0.001 to the sample to form a clinical mixture, the charge density of the ionic species being greater than substantially all of the constituent species in the clinical sample; b) separating said clinical mixture into its constituent parts by capillary zone electrophoresis; c) detecting the ionic species and the peptide-containing constituent species in said mixture; and d) normalizing at least one peptide-containing constituent species by dividing the integrated area value of said peptide-containing species by the total integrated area values from all of the peptide-containing constituent species to obtain a first value; dividing the first value by a second value to obtain a third value, the second value being obtained by dividing the integrated area value of the ionic species by the total integrated area amounts from the constituent species; multiplying the third value by a fourth value, the fourth value being the dilution factor of the sample to obtain a fifth value; multiplying the fifth value by a sixth value, the sixth value being obtained by dividing the absorbance value of a fixed amount of the sample measured at a fixed wavelength by the absorbance value of about the same fixed amount of the ionic species measured at about the same fixed wavelength; and multiplying the sixth value by the concentration of the ionic species in the sample.
Claim 23
Dependent Claim
65 words
The method of claim 20 wherein said ionic species is selected from the group consisting of formic acid, acetic acid, benzo-phosphoric acid, propionic acid, isopropionic acid, butyric acid, isobutyric acid, benzoic acid, benzo-sulfonic acid, ortho-chloro benzoic acid, meta-chloro benzoic acid, para-chloro benzoic acid, naphthyl sulfonic acid, benzo naphthalinic acid, chloro-benzo naphthalinic acid, chloro-naphthyl sulfonic acid, tetra-iodo benzo naphthyl sulfonic acid, and di-iodo anthracenyl sulfonic acid.
Claim 24
Dependent Claim
12 words
The method of claim 20 wherein said ionic species is benzoic acid.
Claim 25
Independent Claim
105 words
A method of quantifying peptide-containing constituent species in a sample by capillary electrophoresis comprising the following steps: a) adding at least one ionic species to the sample to form a mixture, the ionic species having a charge density less than substantially all of the peptide-containing constituent species in the sample, b) subjecting the mixture to capillary zone electrophoresis, c) detecting the ionic species and the peptide-containing constituent species in said mixture, and d) normalizing at least one peptide-containing constituent species relative to the detected ionic species, where the normalized value of step (d) is the quantified amount of said peptide-containing constituent species in the sample.
Claim 26
Independent Claim
105 words
A method of quantifying peptide-containing constituent species in a sample by capillary electrophoresis comprising the following steps: a) adding at least one ionic species to the sample to form a mixture, the ionic species having a charge density less than substantially all of the peptide-containing constituent species in the sample, b) subjecting the mixture to capillary zone electrophoresis, c) detecting the ionic species and the peptide-containing constituent species in said mixture, and d) normalizing at least one peptide-containing constituent species relative to the detected ionic species, where the normalized value of step (d) is the quantified amount of said peptide-containing constituent species in the sample.
Claim 27
Independent Claim
261 words
A method of quantifying peptide-containing constituent species in a clinical sample by capillary zone electrophoresis comprising the following steps: a) adding at least one ionic species having a charge density of between about 0.02 and about 0.001 to the clinical sample to form a clinical mixture, the charge density of the ionic species being less than substantially all of the peptide-containing constituent species in the clinical sample; b) separating said clinical mixture into its constituent parts by capillary zone electrophoresis; c) detecting the ionic species and the constituent peptide-containing species in said mixture; and d) normalizing at least one peptide-containing constituent species by dividing the integrated area value of said peptide-containing species by the total integrated area values from all of the constituent peptide-containing species to obtain a first value; dividing the first value by a second value to obtain a third value, the second value being obtained by dividing the integrated area value of the ionic species by the total integrated area amounts from the constituent peptide-containing species; multiplying the third value by a fourth value, the fourth value being the dilution factor of the clinical sample to obtain a fifth value; multiplying the fifth value by a sixth value, the sixth value being obtained by dividing the absorbance value of a fixed amount of the clinical sample measured at a fixed wavelength by the absorbance value of about the same fixed amount of the ionic species measured at about the same fixed wavelength; and multiplying the sixth value by the concentration of the ionic species in the clinical sample.
Claim 28
Independent Claim
261 words
A method of quantifying peptide-containing constituent species in a clinical sample by capillary zone electrophoresis comprising the following steps: a) adding at least one ionic species having a charge density of between about 0.02 and about 0.001 to the clinical sample to form a clinical mixture, the charge density of the ionic species being less than substantially all of the peptide-containing constituent species in the clinical sample; b) separating said clinical mixture into its constituent parts by capillary zone electrophoresis; c) detecting the ionic species and the constituent peptide-containing species in said mixture; and d) normalizing at least one peptide-containing constituent species by dividing the integrated area value of said peptide-containing species by the total integrated area values from all of the constituent peptide-containing species to obtain a first value; dividing the first value by a second value to obtain a third value, the second value being obtained by dividing the integrated area value of the ionic species by the total integrated area amounts from the constituent peptide-containing species; multiplying the third value by a fourth value, the fourth value being the dilution factor of the clinical sample to obtain a fifth value; multiplying the fifth value by a sixth value, the sixth value being obtained by dividing the absorbance value of a fixed amount of the clinical sample measured at a fixed wavelength by the absorbance value of about the same fixed amount of the ionic species measured at about the same fixed wavelength; and multiplying the sixth value by the concentration of the ionic species in the clinical sample.
References Cited (Backward Citations)
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