Method and apparatus for quantitatively determining the level of hemoglobin in a biological sample
AssigneeRegents of the University of Minnesota
InventorSamuel Schwartz
Life of this patent
Estimating the term…
Abstract
A method of quantitatively determining the level of hemoglobin in a biological material which includes the steps of preparing a test sample of the biological material, converting the heme portion of the hemoglobin in the test sample to porphyrin, assaying the fluorescence of the converted porphyrin and comparing the flourescence of the converted porphyrin to a standard. The invention also relates to an apparatus for performing the above method and a sampler device for collecting and preparing a test sample of the biological material.
Claims42
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with an effective quantity of oxalic acid and a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate;
- assaying the fluorescence of the converted porphyrin;
- and
- comparing the fluorescence of the converted porphyrin to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice including weighing said test sample and homogenizing said test sample in a known volume of a salt solution wherein the quantity of test sample homogenized in the known volume of salt solution results in a test sample concentration of approximately 2.5% to 5.0%;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with an effective quantity of oxalic acid and a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate;
- assaying the fluorescence of the converted porphyrin;
- and
- comparing the fluorescence of the converted porphyrin to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with a two molar oxalic acid and a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate wherein the test sample is combined with a quantity of the reducing salt sufficient to form approximately a 0.3% to 3% solution with the two molar oxalic acid;
- assaying the fluorescence of the converted porphyrin;
- and
- comparing the fluorescence of the converted porphyrin to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining the test sample with an effective quantity of oxalic acid and ferrous oxalate or ferrous sulfate in the presence of heat;
- assaying the fluorescence of the converted porphyrin;
- and
- comparing the fluorescence of the converted porphyrin to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with an effective quantity of oxalic acid and a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate;
- combining the mixture containing the converted porphyrin with a solution of ethyl acetate and glacial acetic acid and centrifuging such combination;
- assaying the supernatent of such centrifuged combination for fluorescence;
- and
- comparing the fluorescence to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with an effective quantity of oxalic acid and a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate;
- assaying the fluorescence of the converted porphyrin by assaying the fluorescence of the test sample in which the heme portion of the hemoglobin has been converted to porphyrin, assaying the fluorescence of a duplicate blank test sample combined with a quantity of citric acid in which the heme portion of the hemoglobin has not been converted significantly to porphyrin, and subtracting the fluorescence of said blank test sample from the fluorescence of said test sample;
- and
- comparing the fluorescence of the converted porphyrin to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with an effective quantity of oxalic acid and reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate in a carrier resulting in a mixture having a gel-like consistency at room temperature;
- assaying the fluorescence of the converted porphyrin;
- and
- comparing the fluorescence of the converted porphyrin to the fluorescence of a standard.
A method of quantitatively determining the level of hemoglobin in a fecal, urine or gastric juice specimen comprising the following steps:
- preparing a test sample of a fecal, urine or gastric juice specimen;
- converting the heme portion of the hemoglobin in said test sample to porphyrin by combining said test sample with a reaction mixture comprising a combination of oxalic acid, a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate and a vehicle for maintaining said reaction mixture in a gel-like consistency at room temperature and heating the combined test sample and reaction mixture to a temperature sufficient to liquify said reaction mixture and convert substantially all of the heme to porphyrin;
- and
- comparing the fluorescence of the converted porphyrin to a standard.
A method of quantitatively determining the level of hemoglobin in feces, urine or gastric juice comprising the following steps:
- preparing a first test sample of feces, urine or gastric juice;
- converting the heme portion of the hemoglobin in said first test sample to porphyrin by combining said first test sample with an effective quantity of oxalic acid and a reducing salt selected from the group consisting of ferrous oxalate and ferrous sulfate;
- assaying the fluorescence of said first test sample;
- assaying the fluorescence of a second test sample of feces, urine or gastric juice combined with a quantity of a non-reducing acid which does not significantly convert the heme portion of the hemoglobin in said second test sample to porphyrin;
- and
- comparing the fluorescence of said first and second test samples.
Dependencies are inferred from the claim text.