Method for non-invasively determining the relative levels of two biological substances

• US 7,555,337 B2
• Filed: 06/17/2002
• Issued: 06/30/2009
• Est. Priority Date: 06/22/2001
• Status: Active Grant

• Current Assignee: University of Bath
• Original Assignee: Maria Begona Delgado-Charro, Richard Henry Guy
• Inventors: Guy, Richard Henry, Delgado-Charro, Maria Begona
• Primary Examiner(s): Sirmons; Kevin C
• Assistant Examiner(s): Hall; Deanna K
• Application Number:
  US10/481,676
• Publication Number:
  US 20050074810A1
• Time in Patent Office: 2,570 Days
• Field of Search: 604/20, 600/345, 600/309
• US Class Current: 604/20
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• CPC Class Codes: A61B 5/1486 using enzyme electrodes, e.g. with immobilised oxidase
• CPC Class Codes: A61N 1/30 Apparatus for iontophoresis, i.e. transfer of media in ionic state by an electromotoric force into the body, or cataphoresis

Abstract

The method for non-invasively determining the relative levels of two substances present in a biological system includes the steps of extracting by reverse iontophoresis charged and uncharged substances from said biological system, and collecting said substances, analysing the collected amount of a first extracted substance and a second extracted substance and determining the extraction ratio of the first substance to the second substance to determine their relative levels in the biological system. When the first and second substances are both analytes susceptible to changes in their concentration in the biological system, this method provides information about the relative concentrations of the two substances in the biological system. When the first substance is an analyte susceptible to changes in its concentration in the biological system and the second substance has a substantially constant concentration in the biological system, the second subtance acts as an internal standard and the extraction ratio becomes a direct measurement of the physiological level of the first substance in the biological system. A iontophoretic sampling device for monitoring the relative levels of two substances present in a biological system is also disclosed.

Claims

1. A method for non-invasively determining the relative levels of two substances present in a biological system, said method comprising

contacting an anodal chamber and a cathodal chamber of an iontophoresis device comprising reversible electrodes with a biological system;

extracting by reverse iontophoresis charged and uncharged substances from said biological system, and collecting said charged and uncharged substances each independently into the anodal chamber or the cathodal chamber;

determining a first substance and a second substance from said extracted charged and uncharged substances

a) such that the first substance is different from the second substance and

b) such that i) a transport and/or a transference number of the first substance is independent of ii) a transport and/or a transference number of the second substance;

analysing the collected amount of the first substance and the second substance;

subsequently, determining the extraction ratio of the first substance to the second substance to determine their relative levels in the biological system;

determining at least one of a physiological and pathological condition based on the determined extraction ratio and without requiring blood sampling for calibration.

2. The method according to claim 1, characterised in that the electrodes of the iontophoretic device are silver/silver chloride electrodes.

3. The method according to claim 2, characterised in that the first substance analysed and the second substance analysed are contained in the same chamber.

4. The method according to claim 3, characterised in that the biological system is a human body.

5. The method according to claim 4, characterised in that the current density applied by the iontophoretic device during the iontophoretic extraction is not higher than 0.5 mA/cm².

6. The method according to claim 5, characterised in that the determination of the extraction ratio of the first substance to the second substance is made by calculating the ratio of the collected amount of the first extracted substance to the collected amount of the second extracted substance.

7. The method according to claim 5, characterised in that the determination of the extraction ratio of the first substance to the second substance is made by first calculating the flux of the first extracted substance and the flux of the second substance, based on the extracted amount of the first substance and the extracted amount of the second substance, respectively, and then by calculating the ratio of the flux of the first extracted substance to the flux of the second extracted substance.

8. The method according to claim 7, characterised in that the analysis of the two selected substances is made by techniques involving specific enzymes or biosensors, ion-selective chemistry, measurement of conductivity, and all other known analytical chemistry techniques.

9. The method according to claim 8, characterised in that the first and second analysed substances are susceptible to changes in their concentration in the biological system.

10. The method according to claim 8, characterised in that the first analysed substance is susceptible to changes in its concentration in the biological system and the second analysed substance has a substantially constant concentration in the biological system.

11. The method according to claim 10, characterised in that the first substance is glucose.

12. The method according to claim 10, characterised in that the first substance is lithium.

13. The method according to claim 12, characterised in that the second substance is sodium.

14. The method according to claim 13, characterised in that electrode polarity is reversed subsequent to each extraction/analysis cycle.

15. A method for non-invasively determining the relative levels of two substances present in a biological system, said method comprising

contacting an anodal chamber and a cathodal chamber of an iontophoresis device having reversible electrodes with a biological system;

extracting by reverse iontophoresis charged and uncharged substances from said biological system, and collecting said charged and uncharged substances each independently into the anodal chamber or the cathodal chamber;

determining first and second substances from said extracted charged and uncharged substances

a) such that i) a transport number and/or a transference number of the first substance is independent of ii) a transport number and/or a transference number of the second substance, and

b) such that the second substance has a substantially constant concentration in the biological system and acts as an internal standard;

analysing the collected amount of the first extracted substance and the second extracted substance; and

subsequently, determining an extraction ratio of the first substance to the second substance to determine their relative levels in the biological system

determining at least one of a physiological and pathological condition based on the extraction ratio.