Method of quantitatively analyzing analyte contained in whole blood sample
First Claim
1. In the method of quantitatively analyzing an analyte in a whole blood sample by using a multi-layered analysis element including at least a coloring reagent layer in which said analyte reacts with a reagent composition to exhibit an optically detectable color change or color generation and a blood cell separating layer for separating the blood cells in the whole blood to feed blood plasma to said reagent layer, an improved method comprising the steps of:
- (1) spotting a whole blood sample onto said blood cell separating layer;
(2) allowing said multi-layered analysis element to stand until the optical density of the reflected light due to the coloring matters contained in the blood cells detected from the coloring reagent layer side reaches a substantially constant level;
(3) measuring the rate of change in optical density within a unit time in said coloring reagent layer from the coloring reagent layer side; and
(4) colorimetrically determining the quantity of said analyte depending on the rate of change in optical density measured by the preceding step (3).
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Abstract
A method of quantitatively analyzing an analyte contained in a whole blood sample, wherein a dry multi-layered analysis element is used. The method provides particular merits when the used multi-layered analysis element has no light-shielding layer which is interposed, in the conventional analysis element, between a coloring reagent layer and a blood cell separating layer, so that red coloring matters of blood cells are detected from the support side during the step of measuring the optical density of the reflected light. After the optical density due to coloring matters of blood cells trapped by the blood cell separating layer has reached a constant level or background density, the changing rate in optical density is measured and then the measured changing rate is converted to the corresponding content or activity of the analyte through a colorimetric operation, or the total change in optical density is measured and then the substantially constant background density is subtracted therefrom to know the change in optical density caused by a coloring dye or like material formed in the coloring reagent layer in the presence of the analyte, followed by a similar conversion operation performed on the basis of the principle of colorimetry, to determine the content or activity of the analyte.
34 Citations
18 Claims
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1. In the method of quantitatively analyzing an analyte in a whole blood sample by using a multi-layered analysis element including at least a coloring reagent layer in which said analyte reacts with a reagent composition to exhibit an optically detectable color change or color generation and a blood cell separating layer for separating the blood cells in the whole blood to feed blood plasma to said reagent layer, an improved method comprising the steps of:
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(1) spotting a whole blood sample onto said blood cell separating layer; (2) allowing said multi-layered analysis element to stand until the optical density of the reflected light due to the coloring matters contained in the blood cells detected from the coloring reagent layer side reaches a substantially constant level; (3) measuring the rate of change in optical density within a unit time in said coloring reagent layer from the coloring reagent layer side; and (4) colorimetrically determining the quantity of said analyte depending on the rate of change in optical density measured by the preceding step (3). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. In the method of quantitatively analyzing an analyte in a whole blood sample by using a multi-layered analysis element including at least a coloring reagent layer in which said analyte reacts with a reagent composition to exhibit an optically detectable color change or color generation and a blood cell separating layer for separating the blood cells in the whole blood to feed blood plasma to said reagent layer, an improved method comprising the steps of:
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(1) spotting a whole blood sample onto said blood cell separating layer; (2) allowing said multi-layered analysis element to stand until a measurement of optical density of the reflected light due to the coloring matters contained in the blood cells measured from the coloring reagent layer side reaches a substantially constant level; (3) measuring the total optical density of the reflected light from the coloring reagent layer side, said total optical density being the sum of the optical density of the reflected light due to the color change or color generation of said analyte and the optical density of the reflected light due to the coloring matters contained in the blood cells; (4) determining the difference in optical density between said total optical density measured in said step (3) and the optical density measured in said step (2); and (5) colorimetrically determining the quantity of said analyte depending on the difference in optical density determined by the preceding step (4). - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18)
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Specification