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Method to determine a diagnostic indicator of blood sugar condition, and, a liquid chromatographic microcolumn therefor

Assignee: Revvity Health Sciences Inc
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Filing: 1977-12-02
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Granted: 1979-03-06
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Expires: 1997-12-02
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Abstract

Abstract A method to determine a numerical percentage value as a diagnostic indicator of the blood sugar condition of a specific person. A whole blood sample is lysed and introduced into an improved ion exchange cellulose (Whatman CM-52)liquid column for microchromatographic separation of hemolysates. Amounts of hemoglobin species, particularly Hb-A 1a-c , are detected and measured by spectometric (color) analysis. A mathematical computation using integer factors corresponding to amounts of various hemoglobin species produces the numerical percentage value.

Inventors

  • 👤 Kenneth J. Acuff

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-23 12:39
CPC Classifications
G; G01; G01N; G01N30/00; G01N30/96; G01N30/02; G01N30/60; G01N30/6095

Claims Analysis

Total Claims: 9
Claim 1
Independent Claim 302 words
A method to determine a numerical percentage value as a diagnostic indicator of the blood sugar condition of a specific person, wherein a whole blood sample is taken from said person and thereafter prepared as a test sample containing a red blood cell hemolysate solution, and thereafter, a quantity of said test sample is introduced into an end of a column bed of ion exchange cellulose particles, said column bed comprising a suspension of particles of a microgranular ion exchange cellulose, modified by removal of the amorphous regions and stabilized by cross-linking, containing negatively charged carboxymethyl groups and having a size less than 400 mesh and an equilibrated pH of substantially 6.8 at 22.5° C. which will adsorb the blood components of said test sample, and then, a quantity of a phosphate-cyanide solution is introduced into an end of said column bed to eluate therefrom a fraction of said test sample and thereafter, an aliquot quantity of a first eluate fraction is collected from the other end of said column bed, and then, a quantity of wash solution is introduced into an end of said column bed to desorb substantially all the remaining blood components of said test sample from said cellulose particles, and thereafter, an aliquot quantity of a second eluate fraction is collected from the other end of said column bed, and then, the hemoglobin species present in each said eluate fraction are separately detected and measured by spectrometric analysis with the respective amounts thereof being expressed as an integer, and thereafter, the amount of hemoglobin species in said first eluate fraction is compared to the sum of the amounts of hemoglobin species in each fraction, according to the mathematical formula -- ##EQU5## said numerical percentage value being a diagnostic indicator of the blood sugar characteristics of said specific person.
Claim 2
Independent Claim 302 words
A method to determine a numerical percentage value as a diagnostic indicator of the blood sugar condition of a specific person, wherein a whole blood sample is taken from said person and thereafter prepared as a test sample containing a red blood cell hemolysate solution, and thereafter, a quantity of said test sample is introduced into an end of a column bed of ion exchange cellulose particles, said column bed comprising a suspension of particles of a microgranular ion exchange cellulose, modified by removal of the amorphous regions and stabilized by cross-linking, containing negatively charged carboxymethyl groups and having a size less than 400 mesh and an equilibrated pH of substantially 6.8 at 22.5° C. which will adsorb the blood components of said test sample, and then, a quantity of a phosphate-cyanide solution is introduced into an end of said column bed to eluate therefrom a fraction of said test sample and thereafter, an aliquot quantity of a first eluate fraction is collected from the other end of said column bed, and then, a quantity of wash solution is introduced into an end of said column bed to desorb substantially all the remaining blood components of said test sample from said cellulose particles, and thereafter, an aliquot quantity of a second eluate fraction is collected from the other end of said column bed, and then, the hemoglobin species present in each said eluate fraction are separately detected and measured by spectrometric analysis with the respective amounts thereof being expressed as an integer, and thereafter, the amount of hemoglobin species in said first eluate fraction is compared to the sum of the amounts of hemoglobin species in each fraction, according to the mathematical formula -- ##EQU5## said numerical percentage value being a diagnostic indicator of the blood sugar characteristics of said specific person.
Claim 3
Dependent Claim 51 words
A method according to claim 1 wherein said suspension of particles is prepared by treating said cellulose particles with a phosphate-cyanide solution, said treated particles being further treated with an acid solution, said treatments with said phosphate-cyanide solution and said acid solution being repeated until said suspension has said equilibrated pH.
Claim 4
Independent Claim 277 words
A method to determine a numerical percentage value as a diagnostic indicator of the blood sugar condition of a specific person, wherein a whole blood sample is taken from said person and thereafter prepared as a test sample containing red blood cell hemolysate solution, and thereafter, a quantity of said test sample is introduced into an end of a column bed of ion exchange cellulose particles, said column bed comprising a suspension of particles of a microgranular ion exchange cellulose, modified by removal of the amorphous regions and stabilized by cross-linking, containing negatively charged carboxymethyl groups and having a size less than 400 mesh and an equilibrated pH of substantially 6.8 at 22.5° C. which will adsorb the blood components of said test sample, and then, a quantity of a phosphate-cyanide solution is introduced into an end of said column bed to elute therefrom a fraction of said test sample and thereafter, an aliquot quantity of a first eluate fraction is collected from the other end of said column bed, also, a quantity of said test sample is significantly diluted to provide a red blood cell hemolysate fraction which may be conveniently detected by spectrometric analysis; and then, the hemoglobin species present in said first eluate fraction and said hemolysate fraction are separately detected and measured by spectrometric analysis with the respective amounts thereof being expressed as an integer, and thereafter, the amount of hemoglobin species in said first eluate fraction is compared to the amount of hemoglobin species in said hemolysate fraction according to the mathematical formula -- ##EQU6## said numerical percentage value being a diagnostic indicator of the blood sugar characteristics of said specific person.
Claim 5
Independent Claim 277 words
A method to determine a numerical percentage value as a diagnostic indicator of the blood sugar condition of a specific person, wherein a whole blood sample is taken from said person and thereafter prepared as a test sample containing red blood cell hemolysate solution, and thereafter, a quantity of said test sample is introduced into an end of a column bed of ion exchange cellulose particles, said column bed comprising a suspension of particles of a microgranular ion exchange cellulose, modified by removal of the amorphous regions and stabilized by cross-linking, containing negatively charged carboxymethyl groups and having a size less than 400 mesh and an equilibrated pH of substantially 6.8 at 22.5° C. which will adsorb the blood components of said test sample, and then, a quantity of a phosphate-cyanide solution is introduced into an end of said column bed to elute therefrom a fraction of said test sample and thereafter, an aliquot quantity of a first eluate fraction is collected from the other end of said column bed, also, a quantity of said test sample is significantly diluted to provide a red blood cell hemolysate fraction which may be conveniently detected by spectrometric analysis; and then, the hemoglobin species present in said first eluate fraction and said hemolysate fraction are separately detected and measured by spectrometric analysis with the respective amounts thereof being expressed as an integer, and thereafter, the amount of hemoglobin species in said first eluate fraction is compared to the amount of hemoglobin species in said hemolysate fraction according to the mathematical formula -- ##EQU6## said numerical percentage value being a diagnostic indicator of the blood sugar characteristics of said specific person.
Claim 6
Dependent Claim 51 words
A method according to claim 3 wherein said suspension of particles is prepared by treating said cellulose particles with a phosphate-cyanide solution, said treated particles being further treated with an acid solution, said treatments with said phosphate-cyanide solution and said acid solution being repeated until said suspension has said equilibrated pH.
Claim 7
Dependent Claim 148 words
A microcolumn for use in the determination of a numerical percentage value as a diagnostic indicator of the blood sugar condition of a person providing a whole blood sample: having a reservoir discharging into a barrel terminating in a discharge tip, the juncture between said reservoir and said barrel and the juncture between said barrel and said tip each being closed by a transverse disc, and ion exchange cellulose particles positioned in said barrel between said transverse discs to provide a column bed, said discs being permeable to a red blood cell hemolysate solution prepared from said whole blood sample; said column bed comprising a suspension of particles of a microgranular ion exchange cellulose, modified by removal of the amorphous regions and stabilized by cross-linking, containing negatively charged carboxymethyl groups and having a size less than 400 mesh and an equilibrated pH of substantially 6.8 at 22.5° C.
Claim 8
Dependent Claim 148 words
A microcolumn for use in the determination of a numerical percentage value as a diagnostic indicator of the blood sugar condition of a person providing a whole blood sample: having a reservoir discharging into a barrel terminating in a discharge tip, the juncture between said reservoir and said barrel and the juncture between said barrel and said tip each being closed by a transverse disc, and ion exchange cellulose particles positioned in said barrel between said transverse discs to provide a column bed, said discs being permeable to a red blood cell hemolysate solution prepared from said whole blood sample; said column bed comprising a suspension of particles of a microgranular ion exchange cellulose, modified by removal of the amorphous regions and stabilized by cross-linking, containing negatively charged carboxymethyl groups and having a size less than 400 mesh and an equilibrated pH of substantially 6.8 at 22.5° C.
Claim 9
Dependent Claim 50 words
A microcolumn according to claim 5 wherein said suspension of particles is prepared by treating said cellulose with a phosphate-cyanide solution, said treated particles being further treated with an acid solution, said treatments with said phosphate-cyanide solution and said acid solution being repeated until said suspension has said equilibrated pH.

References Cited (Backward Citations)

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USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1982-03-05 1982-09-04 1983-03-06 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1986-03-05 1986-09-04 1987-03-06 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1990-03-05 1990-09-04 1991-03-06 $7700.00 Window Closed / Lapsed

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