Device and methods of using device for detection of hyperammonemia

• US 9,952,199 B2
• Filed: 09/02/2014
• Issued: 04/24/2018
• Est. Priority Date: 08/30/2013
• Status: Active Grant

• Current Assignee: Children's National Medical Center, The United States of America as Represented by the Government of the Department of Health and Human Services, University of Maryland, College Park
• Original Assignee: Children's National Medical Center, United States Department Of Health And Human Services, University of Maryland, College Park
• Inventors: Ayyub, Omar Bilal, Behrens, Adam Michael, Kofinas, Peter, Summar, Marshall Lynn, Cabrera-Luque, Juan Manuel, Cunningham, Gary, Simeonov, Anton, Marugan, Juan
• Primary Examiner(s): Kosson, Rosanne
• Application Number:
  US14/915,602
• Publication Number:
  US 20160231310A1
• Time in Patent Office: 1,330 Days
• CPC Class Codes: G01N 21/78 producing a change of colour
• CPC Class Codes: G01N 27/327 Biochemical electrodes , e.g. electrical or mechanical details for in vitro measurements
• CPC Class Codes: G01N 27/3275 Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
• CPC Class Codes: G01N 2800/04 Endocrine or metabolic disorders
• CPC Class Codes: G01N 33/4925 measuring blood gas content, e.g. O2, CO2 , HCO3
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH

Abstract

The present disclosure relates to a bio-sensor capable of measuring the total concentration of one or a plurality of ammonia or ammonium ions with the use of indophenol reagents in the presence of an ionomer. In some embodiments, the biosensor comprises a perflurinated membrane that comprises an ionomer in contact with an alkaline buffer in a vessel configured to receive a sample, such as whole blood. The disclosure also relates to a method of detecting or quantifying the ammonia or ammonium ion concentration in whole blood in a point of care biosensor without reliance on gas chromatography or any measurement that takes more than about twenty minutes.

Claims

1. A biosensor comprising:

(i) at least a first and second vessel;

(ii) a fluid exchange opening positioned between the first and the second vessel;

(iii) at least one conduit in fluid communication with the at least first vessel, the at least one conduit configured to receive a fluid from a point external to the biosensor; and

(iv) a membrane positioned at the fluid exchange opening;

wherein the membrane comprises an ionomer;

wherein the at least one conduit is configured to hold a sample volume from about 5 μ

l to about 100 μ

l; and

wherein the first vessel or the second vessel comprises, individually or in combination;

a hypohalite, an alkali buffer, and at least one;

phenolic reagent or indophenol related compound.

2. The biosensor of claim 1, wherein either the first or the second vessels individually comprises hypohalite, an alkali buffer, a catalyst, and at least one phenolic reagent or indophenol related compound.

3. The biosensor of claim 1 further comprising at least a first electrically conductive support, the electrically conductive support in fluid communication with the at least second vessel, the electrically conductive support operably connected by at least one wire to an amperometer, voltmeter, spectrophotometer, or combination thereof.

4. The biosensor of claim 3 wherein the electrically conductive support further comprises a hydrogel and at least one metabolic enzyme or functional fragment thereof.

5. The biosensor of claim 1, wherein the biosensor does not comprise one or more of the following:

(i) uricase or a functional fragment thereof;

(ii) a hydrogel comprising dextran or a derivative thereof;

(iii) a bacterial cell;

(iv) an electronic dipole configured for electrophoresis;

(v) 3,4-Dihydroxybenzoic acid (3,4-DHB); and

(vi) a vaporizer, gas chromotograph, or a heating element configured for converting liquid ammonia into a gaseous state.

6. The biosensor of claim 1, wherein the biosensor is not functionally dependent upon exposure to any stimulus external to the biosensor.

7. The biosensor of claim 3, further comprising a circuit comprising the at least one wire, and a digital display operably connected to a processor configured to receive digital information from the spectrophotometer and to send digital information to the digital display.

8. The biosensor of claim 1, wherein the biosensor further comprises a test strip comprising the first conduit configured for receiving a volume of bodily fluid.

9. The biosensor of claim 1, wherein the alkali buffer comprises:

(i) from about 0.1 M to about 5 M sodium acetate;

or (ii) from about 0.1 M to about 5 M sodium chloride.

10. The biosensor of claim 1, wherein the hypohalite is selected from:

hypochlorite or bleach.

11. The biosensor of claim 1, wherein the at least one indophenol reagent or indophenol related compound is selected from:

phenol, 2-phenylphenol, or napthol.

12. The biosensor of claim 1, wherein the alkali buffer comprises about 1.0 M of any buffer selected from:

sodium chloride, calcium chloride, zinc chloride, sodium acetate, calcium acetate, and zinc acetate.

13. The biosensor of claim 1, wherein the membrane comprises Nafion.

14. A system comprising the biosensor of claim 1 further comprising a computer processor in operable connection to at least one computer storage memory.

15. The system of claim 14 further comprising a digital display in operable connection to at least one electrically conductive support by an electrical circuit capable of carrying an a electrical signal corresponding to a measurement of a wavelength, current, and/or voltage differential from a diode, spectrophotometer, voltmeter and/or amperometer to the digital display, wherein the digital display is configured to display a concentration value of ammonia, ammonium ion and/or an amino acid in a sample when the at least one electrically conductive support is in contact with the sample for a time period sufficient for at least one catalyst to catalyze an indophenol reaction.

16. A kit comprising a solid support that comprises:

(i) at least a first and second vessel;

(ii) a fluid exchange opening positioned between the at least first and second vessel;

(iii) at least one conduit in fluid communication with the at least first vessel, the at least one conduit configured to receive a fluid from a point external to the solid support; and

(iv) a membrane positioned at the fluid exchange opening;

wherein the membrane comprises an ionomer;

wherein the at least one conduit is configured to hold a sample volume from about 5 μ

l to about 100 μ

l; and

wherein the first vessel or the second vessel comprise, individually or in combination;

hypohalite, an alkali buffer, and at least one indophenol reagent or indophenol related compound.

17. A test strip comprising a solid support comprising:

(i) at least a first and second vessel;

(ii) a fluid exchange opening positioned between the at least first and second vessel;

(iii) at least one conduit in fluid communication with the at least first vessel, the at least one conduit configured to receive a fluid from a point external to the biosensor; and

(iv) a membrane positioned at the fluid exchange opening;

wherein the membrane comprises an ionomer, and

wherein the first vessel or the second vessel comprise, individually or in combination;

hypohalite, an alkali buffer, and at least one;

phenolic reagent, indophenol reagent or indophenol related compound.

18. The test strip of claim 17, wherein the test strip is adapted for a portable device comprising a diode, spectrophotometer, voltmeter and/or amperometer and a digital display such that, when the test strip is contacted to the device, the first and second electrodes become operably connected to a closed electrical circuit comprising the voltmeter and/or amperometer and the digital display, and, upon contact with a sample, hypohalite, an alkali buffer, and at least one indophenol reagent or indophenol related compound catalyze an indophenol reaction resulting in a current on the first electrode corresponding to a concentration value of ammonia in the sample, such concentration value readable on the display of the portable device.