Method and device for carbonyl detection and quantitation

• US 9,404,836 B2
• Filed: 06/29/2015
• Issued: 08/02/2016
• Est. Priority Date: 06/27/2014
• Status: Expired due to Fees

• Current Assignee: Pulse Health LLC
• Original Assignee: Pulse Health LLC
• Inventors: Thomas, Gerald, Young, Brian, Lara, Juven, Noll, Charles, Ingle, James
• Primary Examiner(s): Mui, Christine T
• Application Number:
  US14/754,567
• Publication Number:
  US 20150377858A1
• Time in Patent Office: 400 Days
• Field of Search: 436/128, 436/127
• US Class Current: 1/1
• CPC Class Codes: A61B 2010/0087 Breath samples
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• CPC Class Codes: A61B 5/082 Evaluation by breath analysis, e.g. determination of the chemical composition of exhaled breath A61B5/083, A61B5/091 take precedence
• CPC Class Codes: A61B 5/087 Measuring breath flow
• CPC Class Codes: G01D 11/24 Housings ; Casings for instruments
• CPC Class Codes: G01N 1/22 in the gaseous state specially adapted for biological material G01N33/497; measuring breath flow A61B5/087
• CPC Class Codes: G01N 2001/2244 Exhaled gas, e.g. alcohol detecting
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 2033/4975 other than oxygen, carbon dioxide or alcohol, e.g. organic vapours
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 2201/022 Casings
• CPC Class Codes: G01N 2201/062 LED's
• CPC Class Codes: G01N 2201/068 Optics, miscellaneous
• CPC Class Codes: G01N 33/497 of gaseous biological material, e.g. breath
• CPC Class Codes: G01N 33/4972 Determining alcohol content for vehicle safety devices B60K28/06
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/64 involving ketones
• CPC Class Codes: Y10T 436/200833 Carbonyl, ether, aldehyde or ketone containing
• CPC Class Codes: Y10T 436/202499 Formaldehyde or acetone

Abstract

Fluorescence chromophores such as phenylene diamine derivatives can undergo oxidative coupling and polymerization to form optical, colorimetric and fluorogenic, multimers and polymers. The presence of carbonyl containing moieties such as aldehydes and ketones under favorable environmental conditions can initiate, catalyze, accelerate and modulate this reaction which in turn provides a mechanism for the detection and quantitation of such moieties. Selected phenylene diamine derivatives can be used for the detection and quantitation of aldehyde and ketones via measurement of the reaction and the associated spectroscopic transformation. In particular, the use of meta-phenylene diamine (mPDA) and related compounds for aldehyde detection and quantitation is described. The method provides a convenient means for monitoring aldehyde and ketone levels without use of separation steps. The method is applicable to kinetic and quasi-endpoint detection assay formats.

Claims

1. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein the detected fluorescence corresponds to the concentration of carbonyl containing moieties in the biological sample, and wherein the biological sample is human breath, the carbonyl containing moiety is an aldehyde and the phenylene diamine derivative is m-phenylene diamine.

2. The method of claim 1 wherein the carbonyl containing moiety is selected from the group consisting of aldehydes, ketones, carboxylic acids and mixtures thereof.

3. The method of claim 2, wherein the aldehydes are aliphatic.

4. The method of claim 1, wherein the biological sample is human breath.

6. The method of claim 1, wherein the fluorescence has a measurable emission band at 505 nm.

7. The method of claim 1, wherein the fluorescing solution is maintained at below about 35°

C.

16. The method of claim 1, wherein the carbonyl containing moiety is eluted from silica.

17. The method of claim 1, wherein the carbonyl containing moiety is a C1 to C12 carbonyl containing organic molecule.

21. The method of claim 1, wherein the phenylene diamine derivative comprises pyridinium,4-[2-[4-(diethylaminio)phenyl-ethenyl]-1-[1-(3,5-diminobenzamide)-pentylamino-5-oxyhextyl].

5. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein the phenylene diamine derivative is selected from the group of meta-phenylene diamine derivatives and mixtures thereof.

8. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein the aqueous salt solution comprises an alcohol, a salt, a surfactant and a buffer.

9. The method of claim 8 where the buffer is citrate.

10. The method of claim 8 wherein the alcohol is chosen from the group consisting of R—

OH, where R is a straight or branched chain aliphatic group, and mixtures thereof.

11. The method of claim 9, wherein the alcohol is ethanol.

12. The method of claim 8, wherein the salt is NaCl.

13. The method of claim 8, wherein the surfactant is sodium dodecyl sulfate.

15. The method of claim 8, wherein the molar ratio of the phenylene diamine derivative to the salt ranges from 0.03 to 0.5.

14. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein pH of the aqueous solution ranges from 2 to 4.5.

18. A method of detecting and measuring the concentration of a carbonyl containing moiety in a biological sample, the method comprising

a) isolating the carbonyl containing moiety from the biological sample;

b) adding the carbonyl containing moiety to an aqueous solution containing a phenylene diamine derivative to form a fluorescing solution; and

c) measuring the fluorescence emitted from the fluorescing solution at a pre-determined wave length.

19. The method of claim 18, wherein the biological sample is a breath sample.

20. The method of claim 18, wherein the concentration of the carbonyl containing moiety in the biological sample is determined using the fluorescence measurement.

28. The method of claim 18, wherein the fluorescing solution generates a broad optical absorbance band at 405 nm and an associated emission band at 505 nm.

29. The method of claim 18, wherein the aqueous solution contains a surfactant, and the level of fluorescence over time varies with the concentration of the surfactant.

22. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein the phenylene diamine derivative comprises any one or mixtures of the following;

23. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a fluorescence chromophore to an aqueous salt solution to thereby form a fluorescence chromophore solution;

adding a carbonyl containing moiety from the biological sample to the fluorescence chromophore solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution.

24. The method of claim 23, wherein the fluorescence chromophore is a phenylene diamine derivative.

25. The method of claim 24, wherein the fluorescence chromophore is m-phenylene diamine.

26. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein the phenylene diamine derivative is polymerized by the carbonyl containing moiety in the solution to form a fluorogenic species.

31. The method of claim 26, wherein there is no precipitate in the fluorescing solution which at the time of measurement, substantially interferes with the measurement of the fluorogenic species.

27. A method of detecting carbonyl containing moieties in a biological sample, the method comprising adding a phenylene diamine derivative to an aqueous salt solution to thereby form a phenylene diamine solution;

adding a carbonyl containing moiety from the biological sample to the phenylene diamine solution to thereby form a fluorescing solution; and

detecting fluorescence from the fluorescing solution, wherein the phenylene diamine derivative is housed in an ampule.

30. A method for detecting carbonyl containing moieties in a biological sample, the method comprising the steps of providing a substrate having an active reactive capture agent incorporated therein, capturing on said substrate carbonyl containing moieties from the biological sample, and forming a solution comprising painted carbonyl containing moieties.