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

• US 9,562,913 B2
• Filed: 05/22/2014
• Issued: 02/07/2017
• Est. Priority Date: 07/26/2007
• Status: Expired due to Fees

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Stamler, Jonathan S., Angelo, Michael
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US14/285,106
• Publication Number:
  US 20140332416A1
• Time in Patent Office: 992 Days
• Field of Search: 435/287.1, 436/68, 436/116, 436/120, 436149-150
• 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 presented 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 determining amount of combined nitric oxide in a blood sample, comprising the steps of:

(a) introducing a sample of the blood to be analyzed for amount of combined nitric oxide therein, 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;

(b) directing electromagnetic radiation of less than 100 milliwatts from at least one of a laser and a light emitting diode at said front side to cause liberation of nitric oxide gas from combined nitric oxide and passage of the liberated nitric oxide gas from said rear side;

(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 amount of dissolved nitric oxide gas in the solvent which corresponds to the total amount of nitric oxide present as combined nitric oxide, present as nitrosothiols and iron nitrosyls, in the sample.

2. The method of claim 1 where the front side of the sample containing zone is ultraviolet radiation transparent and the electromagnetic radiation of step (b) is ultraviolet radiation.

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

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

5. The method of claim 2 where the ultraviolet radiation of step (b) has a wavelength ranging from 210-220 nm.

6. The method of claim 1 where the front side of the sample containing zone is visible radiation transparent and the electromagnetic radiation of step (b) is visible electromagnetic radiation having a wavelength ranging from 500 to 600 nm.

7. The method of claim 1 where the front side of the sample containing zone is near infrared radiation transparent and the electromagnetic radiation of step (b) is near infrared radiation.

8. A method for determining amount of nitric oxide present as combined nitric oxide and also amount of nitric oxide present as iron nitrosyls in blood comprising:

(a) obtaining two samples of blood from the same source, each comprising combined nitric oxide present as nitrosothiols and iron nitrosyls and where one of the samples is denoted as the first sample and the other of the samples is denoted as the second sample;

(b) treating the second sample with a nitrosothiols degrading agent to cause decomposition of nitrosothiols to nitrous acid and leaving iron nitrosyls in the sample;

(c) analyzing for amount of nitric oxide present as combined nitric oxide in the first sample by steps comprising (i) introducing the first sample into a first sample containing zone which has 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, (ii) directing electromagnetic radiation at said front side of the first sample containing zone to cause liberation of nitric oxide gas from combined nitric oxide and passage of the liberated nitric oxide gas through said rear side, (iii) providing a first 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, (iv) electrochemically detecting amount of dissolved nitric oxide in the first solvent containing zone which corresponds to the total amount of nitric oxide present as combined nitric oxide in said first sample, (d) analyzing for amount of nitric oxide present in the iron nitrosyls left in the step (b) treated second sample by steps comprising (i) introducing the treated second sample into a second sample containing zone which has 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, (ii) directing electromagnetic radiation of less than 100 milliwatts at said front side of the second sample containing zone to cause liberation of nitric oxide gas from said iron nitrosyls and passing of the liberated nitric oxide gas through said rear side, (iii) providing a second 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, (iv) electrochemically detecting amount of dissolved nitric oxide in the second solvent containing zone which corresponds to the amount of nitric oxide present as iron nitrosyls in said second sample.

9. The method of claim 8 where in (c)(i) and (d)(i), the front side of the sample containing zone is ultraviolet radiation transparent and where in (c)(ii) and (d)(ii), the electromagnetic radiation is ultraviolet radiation.

10. The method of claim 9 where the ultraviolet radiation has a wavelength ranging from 300 to 400 nm.

11. The method of claim 10 where the ultraviolet radiation has a wavelength ranging from 325 to 355 nm.

12. The method of claim 9 where the electromagnetic radiation has a wavelength ranging from 210-220 nm.

13. The method of claim 8 where in (c)(i) and (d)(i), the front side of the sample containing zone is visible radiation transparent and where in (c)(ii) and (d)(ii), the electromagnetic radiation is visible electromagnetic radiation having a wavelength ranging from 500 to 600 nm.

14. The method of claim 8 where in (c)(i) and (d)(i), the front side of the sample containing zone is near infrared radiation transparent and where in (c)(ii) and (d)(ii), the electromagnetic radiation is near infrared radiation.

15. The method of claim 8 where the electromagnetic radiation of (c)(ii) and (d)(ii) is from a laser.

16. The method of claim 8 where the electromagnetic radiation of (c)(ii) and (d)(ii) is from a light-emitting diode.