Measuring device with electrodes fabricated on porous membrane substrate in whole

• US 6,210,907 B1
• Filed: 09/22/1999
• Issued: 04/03/2001
• Est. Priority Date: 03/31/1997
• Status: Expired due to Fees

• Current Assignee: Samduck International Corporation
• Original Assignee: Samduck International Corporation
• Inventors: Cha, Geun-Sig
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): PHAM, MINH QUAN K
• Application Number:
  US09/381,788
• Time in Patent Office: 559 Days
• Field of Search: 422/68.1, 422/69, 422/70, 422/82.01, 422/82.02, 422/255, 422/261, 435/4, 435/7.1, 435/7.72, 435/11, 435/962, 436/501, 436/518, 436/524, 436/528, 436/529, 436/530, 436/71, 436/806, 204/400, 204/403
• US Class Current: 435/7.1
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: C12Q 1/60 involving cholesterol
• CPC Class Codes: G01N 2030/285 electrically driven carrier
• CPC Class Codes: G01N 2030/8822 involving blood
• CPC Class Codes: G01N 27/3274 Corrective measures, e.g. error detection, compensation for temperature or hematocrit, calibration coding of calibration information G01N33/48771
• CPC Class Codes: G01N 30/64 Electrical detectors
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property

Abstract

The present invention relates to a measuring device which comprises electrodes fabricated on porous membrane substrate in which the sample migrates chromatographically; and the method of quantifying material in the sample by using the device. The sample material for measuring can be quantified by the measuring device of this invention, which is consists pretreatment bands in the lower part of porous membrane substrate and electrodes in the upper prt of pretreatment bands, by the procedure as foolows: The sample material for measuring is chromatographically migrated in the porous membrane substrate by applying the sample on the lower part of the porous membrane substrate; and then the changes of electric signal by the material at the electrode are measured to quantify the material. The analyzing method using the measuring device of this invention has merits that no additional preparation of the sample is needed, simple analyzing process, quantitative analysis of the material in a short time, and the economical efficiency because of the dispensability of skilled personnel due to easy manipulation.

Claims

1. A measuring device which comprises a porous membrane substrate having a lower part, center part, and upper part in which a sample can migrate chromatographically from the lower part in the direction of the upper part;

pretreatment bands constructed in the lower part of the porous membrane substrate; and

electrodes constructed in the upper part of the porous membrane substrate, the electrodes comprise a working electrode, a counter electrode and a reference electrode, and a connection pad for connecting the electrodes with a circuit electrometer and having an insulating layer which prevents short circuit of the electrodes, wherein the porous membrane substrate is selected from the group consisting of nitrocellulose paper, filter paper, organic polymers comprising a hydrophilic polymer, and inorganic polymers comprising a hygroscopic ceramic and has first and second insulating layers provided in the center part and the lower part of the porous membrane substrate, respectively;

the pretreatment bands comprise (1) a layer for removing interferents, (2) a layer for converting a sample material to be detected by the electrodes, and (3) a layer for detecting the sample material by the electrodes by at least one of controlling the pH and dispersing the sample material to be detected.

2. The measuring device of claim 1, wherein the electrodes are selected from the group consisting of voltammetric, amperometric, potentiometric and conductometric electrode system.

3. The measuring device of claim 1, wherein the layer for removing the interferents comprises at least one of an antibody layer and an ethylenediaminetetraacetic acid (EDTA) layer.

4. The measuring device of claim 1, wherein the layer for converting a sample to be detected by the electrodes comprises an enzyme layer.

5. The measuring device of claim 1, wherein the layer providing the optimum condition for the electrode comprises a buffer solution layer or a surfactant layer.

6. The measuring device of claim 1, wherein the working electrode is made of a material selected from the group consisting of conducting carbon pastes and modified conducting carbon pastes comprising at least one of heavy metal oxides, organometallic compounds, and noble metal pastes.

7. The measuring device of claim 6, wherein the heavy metal oxides is ruthenium oxide (Ru₂O), the organometallic compounds is cobalt (II) phthalocyanine and the noble metal pastes are pastes of gold and platinum.

8. The measuring device of claim 1, wherein the counter electrode is made of a material selected from the group consisting of conducting carbon pastes and noble metal pastes comprising at least one of gold and platinum.

9. The measuring device of claim 1, wherein the reference electrode is made of a material selected from the group consisting of silver, silver epoxy, silver/silver chloride, and silver/silver ion.

10. The measuring device of claim 1, wherein the pretreatment bands comprise (1) high density lipoprotein (HDL) and very low density lipoprotein (VLDL) antibody layers (2) a surfactant layer, and (3) cholesterol esterase (CED) and cholesterol oxidase (COD) enzyme layers.

11. Process for preparation of the measuring device of claim 1, which comprises forming the measuring device by screen printing in each step of the following steps:

(1) fabricating the first and second insulating layers on the porous membrane substrate to prevent the oxidation of the connection pad due to the penetration of a sample;

(2) constructing the working electrode and the counter electrode on the porous membrane substrate by using conducting materials, and constructing the reference electrode on the porous membrane substrate;

(3) constructing the connection pad by using conducting materials for connecting the electrodes to the electrometer;

(4) wrapping the connection pad with the third insulating layer to prevent the short circuit; and

(5) constructing pretreatment bands on the porous membrane substrate.

12. The process for preparation of claim 1, wherein the process comprises a heat treatment step at 50 to 150°

C. for 15 to 30 minutes after each of steps (1)-(4) for the densification of the materials of each component of the measuring device.

13. A method of quantifying by using the measuring device of claim 1 as a sensor, which comprises pretreating the sample in the pretreatment bands constructed in the lower part of the porous membrane substrate, and quantifying the pretreated material in the electrode part constructed in an upper part of the porous membrane substrate, by measuring the changes of electric signal.

14. The method of quantifying of claim 13, wherein the sample is selected from the group consisting of organic materials, inorganic materials, biosamples comprising blood and urine;

environmental samples;

industrial samples; and

foodstuffs.

15. The method of quantifying of claim 14, wherein the pretreatment bands are selected from the group consisting of antibody layers, detergent layers and enzyme layers (CED and COD), and a method for determining LDL concentration is characterized by:

fixing HDL and VLDL in the blood in antibody layers,

converting LDL to cholesterol ester during migration of detergent layers chromatographically,

converting cholesterol ester to cholesterol by hydrolysis in CED,

generating H₂O₂ and cholesterol-4-en-3-one by the reaction of the cholesterol and COD, and

generating an electric signal by the redox reaction of H₂O₂ with amperometric electrometer.