Method for liberating and detecting nitric oxide from nitrosothiols and iron nitrosyls in blood

• US 9,063,160 B2
• Filed: 07/15/2008
• Issued: 06/23/2015
• Est. Priority Date: 07/26/2007
• Status: Active Grant

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Stamler, Jonathan S., Angelo, Michael
• Primary Examiner(s): Lankford, Blaine
• Assistant Examiner(s): CONSTANTINE, CHARLES Z
• Application Number:
  US12/668,211
• Publication Number:
  US 20110008815A1
• Time in Patent Office: 2,534 Days
• US Class Current: 1/1
• CPC Class Codes: A61B 5/0059 using light, e.g. diagnosis by transillumination, diascopy, fluorescence A61B5/0093, A61B5/1455, A61B5/02416 and A61B5/0261 take precedence
• CPC Class Codes: A61B 5/1405 Devices for taking blood samples devices for taking samples of other body liquids A61B10/0045
• CPC Class Codes: A61B 5/14546 for measuring analytes not otherwise provided for, e.g. ions, cytochromes
• CPC Class Codes: A61B 5/150022 for capillary blood or interstitial fluid
• CPC Class Codes: A61B 5/150251 Collection chamber divided into at least two compartments, e.g. for division of samples injection syringes having more than one chamber A61M5/19; syringes for injection of two or more media A61M5/2448, A61M5/284; sample preparation for further analysis A61B5/150755
• CPC Class Codes: A61B 5/150343 Collection vessels for collecting blood samples from the skin surface, e.g. test tubes, cuvettes sampling from within body cavities A61B5/150015, A61B5/150045
• CPC Class Codes: A61B 5/412 Detecting or monitoring sepsis
• CPC Class Codes: G01N 27/416 Systems G01N27/27 takes precedence
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH
• CPC Class Codes: Y10T 436/177692 Oxides of nitrogen

Abstract

Amount of combined nitric oxide or nitric oxide present as iron nitrosyls in a blood sample is determined by directing a low power electromagnetic radiation beam at a blood sample to liberate nitric oxide gas, dissolving the liberated nitric oxide gas and electrochemically detecting amount of dissolved nitric oxide gas.

Claims

1. A method for liberating nitric oxide gas from nitrosothiols and iron nitrosyls in a blood sample, comprising the steps of:

(a) introducing the blood sample into a sample containing zone having a front side which is electromagnetic radiation transparent and a rear side which is porous to the extent of permitting nitric oxide gas to pass therethrough while preventing protein from passing therethrough; and

(b) directing electromagnetic radiation of less than 100 milliwatts at said front side to cause liberation of nitric oxide gas from the nitrosothiols and iron nitrosyls and passage of the liberated nitric oxide gas from said rear side.

2. The method of claim 1 where the electromagnetic radiation is ultraviolet radiation.

3. The method of claim 2 where the ultraviolet radiation has a wavelength ranging from 300 to 400 nm.

4. The method of claim 3 where the ultraviolet radiation has a wavelength ranging from 325 to 355 nm.

5. The method of claim 2 where the ultraviolet radiation has a wavelength ranging from 210 to 220 nm.

6. The method of claim 1 where the electromagnetic radiation is visible electromagnetic radiation having a wavelength ranging from 500 to 600 nm.

7. The method of claim 1 where the electromagnetic radiation is near-infrared radiation.

8. The method of claim 1 where the electromagnetic radiation is from a laser.

9. The method of claim 1 where the electromagnetic radiation is from a light-emitting diode.

10. The method of claim 1 wherein the method further comprises the following steps:

(c) providing a solvent containing zone to dissolve the liberated nitric oxide gas that has passed through said rear side where the solvent is one that dissolves nitric oxide gas;

(d) electrochemically detecting the amount of dissolved nitric oxide gas in the solvent which corresponds to the total amount of nitric oxide present as nitrosothiols and iron nitrosyls in the sample.

11. The method of claim 1, where the electromagnetic radiation is in the range of from 30 to 60 milliwatts.

12. The method of claim 11, where the electromagnetic radiation is 50 milliwatts.