Methods, Systems, and Compositions for Detection of Aldehydes
Methods, systems and reagents are provided for detecting and quantifying carbonyl containing moieties in a variety of sample types. The amount of time elapsed from capturing of the carbonyl containing moieties from a sample to the detection of the carbonyl containing moieties is less than about 2 hours. Compounds are provided to facilitate labeling and detection of the carbonyl containing moieties.
1 . A method for detecting the presence of at least one aldehyde in a sample, the method comprising the steps of:
exposing the sample to a substrate to capture the aldehyde;
eluting the aldehyde off the substrate;
mixing the aldehyde with a reactive labeling agent;
injecting the labeled aldehyde onto a column;
eluting the labeled aldehyde from the column in an organic solvent; and
detecting the labeled aldehyde;
wherein the method of detecting is complete in less than about 2 hours.
2 . The method of claim 1 , further comprising measuring the concentration of the at least one aldehyde.
3 . The method of claim 1 , wherein the at least one aldehyde is selected from the group consisting of a C1 aldehyde, a C2 aldehyde, C3 aldehyde, a C4 aldehyde, a C5 aldehyde, a C6 aldehyde, a C7 aldehyde, a C8 aldehyde, a C9 aldehyde, a C10 aldehyde, and mixtures thereof.
4 . The method of claim 1 , wherein the at least one aldehyde is aliphatic, a di-aldehyde, or an aromatic aldehyde, or mixtures thereof.
5 . The method of claim 1 , wherein the sample comprises two or more aldehydes of different carbon chain lengths, and wherein the step of detecting the labeled aldehyde resolves each aldehyde.
6 . The method of claim 1 , wherein the sample is a biological sample.
7 . The method of claim 1 , wherein the sample is an environmental sample.
8 . The method of claim 1 , wherein the sample is selected from the group consisting of a breath sample, a urine sample, a blood sample, a plasma sample, and a sample of the headspace in a culture.
9 . The method of claim 1 , wherein the sample is a breath sample.
10 . The method of claim 1 , wherein the substrate is selected from the group consisting of silica, polysaccharides, cellulose acetate, nitrocellulose, polyether sulfone, polyethylene, nylon, polyvinylidene fluoride (PVDF), polyester, polypropylene, silica, deactivated alumina, diatomaceous earth, MgSO 4 , porous matrix, vinyl chloride, vinyl chloride-propylene copolymer, vinyl chloride-vinyl acetate copolymer, cloth, cotton, nylon, rayon, porous gels, silica gel, agarose, dextran, gelatin, polymeric film, and polyacrylamide.
11 . The method of claim 1 , wherein the step of capturing the aldehyde comprises collecting at least one aldehyde on a filter assembly.
12 . The method of claim 1 , wherein the reactive labeling agent comprises a fluorophore, a linker, and a reactive group.
13 . The method of claim 12 , wherein the fluorophore is selected from the group consisting of tetramethyl rhodamine (TAMRA), aminooxy 5(6) TAMRA, aminooxy 5 TAMRA, aminooxy 6 TAMRA, rhodamine X (ROX), rhodamine 6G (R6G), rhodamine 110 (R110), and a coumarin.
14 . The method of claim 12 , wherein the linker is selected from the group consisting of substituted alkyl-diamines (C2-C10), substituted amino-carboxylic acids (C2-C10), and substituted polyethylene glycols (N=1-10).
15 . The method of claim 12 , wherein the linker is selected from the group consisting of hexanoic acid, aminohexanoic acid, cadavarine, polyethylene glycol, and polyglycol.
16 . The method of claim 12 , wherein the reactive group is selected from the group consisting of a hydrazine moiety, a carbohydrazide moiety, a hydroxylamine moiety, a semi-carbazide moiety, an aminooxy moiety, and a hydrazide moiety.
17 . The method of claim 1 , wherein the reactive labeling agent is selected from the group consisting of
and mixtures thereof.
18 . The method of claim 1 , wherein the step of detecting the aldehyde comprises measuring fluorescence emission produced by excitation of the fluorophore.
19 . The method of claim 1 , wherein the step of detecting the aldehyde comprises measuring fluorescence absorbance produced by excitation of the fluorophore.
20 . The method of claim 1 , wherein the step of detecting the aldehyde comprises directing light within a predetermined wavelength range to the labeled aldehyde, thereby producing a fluorescence, and detecting the fluorescence.
21 . The method of claim 2 , wherein the concentration of the aldehyde is determined by calculating the fluorescence absorption or emission relative to a standard curve, wherein the fluorescence signal is proportional to the concentration of the aldehyde.
22 . The method of claim 1 , wherein the column is a reverse phase column.
23 . The method of claim 1 , wherein the organic solvent is selected from the group consisting of methanol, isopropanol, acetonitrile, and ethanol.
24 . A method for detecting the presence of at least one carbonyl containing moiety in a sample, the method comprising the steps of:
exposing the sample to a substrate to capture the carbonyl containing moiety,
eluting the carbonyl containing moiety off the substrate,
mixing the carbonyl containing moiety with a reactive labeling agent,
injecting the labeled carbonyl containing moiety onto a column,
eluting the labeled carbonyl containing moiety from the column in an organic solvent, and
detecting the labeled carbonyl containing moiety,
wherein the method of detecting is complete in less than about 2 hours.
25 . The method of claim 24 , wherein the carbonyl containing moiety is selected from the group consisting of aldehydes, ketones, carboxylic acids and mixtures thereof.
26 . A method of detecting carbonyl containing moieties in a gas sample, the method comprising:
isolating carbonyl containing moieties from a sample;
mixing the carbonyl containing moieties with a reactive labeling agent, wherein the carbonyl containing moieties associate with the reactive labeling agent;
passing the labeled carbonyl containing moieties through a column;
exciting the labeled carbonyl containing moieties exiting the column; and
detecting the carbonyl containing moieties by measuring the fluorescence emitted from or absorbed by the reactive labeling agent associated with the carbonyl containing moieties,
wherein the step of detecting resolves the carbonyl containing moieties based on the carbon chain length, and
wherein the time elapsed from isolating the carbonyl containing moieties from the sample to detecting the carbonyl containing moieties is less than about 2 hours.
27 . A compound comprising a fluorophore, a linker, and a reactive group.
28 . The compound of claim 27 , wherein the fluorophore is selected from the group consisting of ao-5-TAMRA, ao-6-TAMRA, and mixtures thereof.
29 . The compound of claim 27 , wherein the linker is selected from the group consisting of hexanoic acid, aminohexanoic acid, cadavarine, polyethylene glycol, and polyglycol.
30 . The compound of claim 27 , wherein the reactive group is selected from the group consisting of a hydrazine moiety, a carbohydrazide moiety, a hydroxylamine moiety, a semi-carbazide moiety, an aminooxy moiety, and a hydrazide moiety.
31 . The compound of claim 27 , comprising:
32 . The compound of claim 27 , comprising:
33 . A system for detecting the presence of at least one carbonyl containing moiety in a sample, the system comprising:
a substrate to capture the carbonyl containing moiety;
one or more reagents for eluting the carbonyl containing moiety off the substrate;
one or more reagents for associating the carbonyl containing moiety with a reactive labeling agent;
a column for resolving the labeled carbonyl containing moiety;
one or more solvents for eluting the labeled carbonyl containing moiety from the column; and
a light and detector for generating fluorescence excitation, absorbance, and/or emission to detect the labeled carbonyl containing moiety.
34 . The system of claim 33 , wherein the system completes one cycle in less than about 2 hours.
35 . The system of claim 33 , further comprising one or more standards for measuring the concentration of the at least one carbonyl containing moiety.