CN111398463A - Method for detecting content of aldehydes and ketones in exhaled air - Google Patents
Method for detecting content of aldehydes and ketones in exhaled air Download PDFInfo
- Publication number
- CN111398463A CN111398463A CN202010252958.5A CN202010252958A CN111398463A CN 111398463 A CN111398463 A CN 111398463A CN 202010252958 A CN202010252958 A CN 202010252958A CN 111398463 A CN111398463 A CN 111398463A
- Authority
- CN
- China
- Prior art keywords
- detecting
- exhaled breath
- content
- human body
- adsorption
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 29
- 150000002576 ketones Chemical class 0.000 title claims description 21
- 150000001299 aldehydes Chemical class 0.000 title claims description 20
- 239000000126 substance Substances 0.000 claims abstract description 31
- -1 aldehyde ketone Chemical class 0.000 claims abstract description 29
- 238000004817 gas chromatography Methods 0.000 claims abstract description 14
- 238000004885 tandem mass spectrometry Methods 0.000 claims abstract description 10
- 238000001179 sorption measurement Methods 0.000 claims description 58
- 238000005070 sampling Methods 0.000 claims description 44
- 239000007788 liquid Substances 0.000 claims description 35
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 18
- 238000000576 coating method Methods 0.000 claims description 18
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 16
- 239000011248 coating agent Substances 0.000 claims description 16
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 15
- 230000005526 G1 to G0 transition Effects 0.000 claims description 14
- 238000002470 solid-phase micro-extraction Methods 0.000 claims description 13
- 239000003153 chemical reaction reagent Substances 0.000 claims description 12
- 239000000945 filler Substances 0.000 claims description 12
- 238000011049 filling Methods 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 11
- 238000010438 heat treatment Methods 0.000 claims description 9
- 238000001212 derivatisation Methods 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 238000012360 testing method Methods 0.000 claims description 7
- 239000003480 eluent Substances 0.000 claims description 6
- BYCUWCJUPSUFBX-UHFFFAOYSA-N (2,3,4,5,6-pentafluorophenyl)hydrazine Chemical compound NNC1=C(F)C(F)=C(F)C(F)=C1F BYCUWCJUPSUFBX-UHFFFAOYSA-N 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- HORQAOAYAYGIBM-UHFFFAOYSA-N 2,4-dinitrophenylhydrazine Chemical group NNC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O HORQAOAYAYGIBM-UHFFFAOYSA-N 0.000 claims description 4
- HVMVKNXIMUCYJA-UHFFFAOYSA-N hydron;o-[(2,3,4,5,6-pentafluorophenyl)methyl]hydroxylamine;chloride Chemical compound Cl.NOCC1=C(F)C(F)=C(F)C(F)=C1F HVMVKNXIMUCYJA-UHFFFAOYSA-N 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 238000007664 blowing Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 34
- 239000012855 volatile organic compound Substances 0.000 abstract description 11
- 239000007789 gas Substances 0.000 description 40
- 239000012086 standard solution Substances 0.000 description 34
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 18
- 229910052757 nitrogen Inorganic materials 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 239000004205 dimethyl polysiloxane Substances 0.000 description 8
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 8
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 8
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 8
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000012417 linear regression Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 238000003795 desorption Methods 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000004445 quantitative analysis Methods 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 230000005264 electron capture Effects 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 2
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 229920000058 polyacrylate Polymers 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- XYHKNCXZYYTLRG-UHFFFAOYSA-N 1h-imidazole-2-carbaldehyde Chemical compound O=CC1=NC=CN1 XYHKNCXZYYTLRG-UHFFFAOYSA-N 0.000 description 1
- GWYFCOCPABKNJV-UHFFFAOYSA-M 3-Methylbutanoic acid Natural products CC(C)CC([O-])=O GWYFCOCPABKNJV-UHFFFAOYSA-M 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- GWYFCOCPABKNJV-UHFFFAOYSA-N beta-methyl-butyric acid Natural products CC(C)CC(O)=O GWYFCOCPABKNJV-UHFFFAOYSA-N 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 238000001871 ion mobility spectroscopy Methods 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 238000001184 proton transfer reaction mass spectrometry Methods 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/067—Preparation by reaction, e.g. derivatising the sample
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention provides a method for detecting the content of aldehyde ketone substances in exhaled breath, and belongs to the field of content detection. The method solves the problems that the existing exhaled breath cannot be quantitatively detected basically when the concentration range of some VOCs in the exhaled breath is small, and the like, and comprises the following steps: s01: collecting exhaled breath of a human body; s02: and detecting by gas chromatography or gas chromatography-tandem mass spectrometry. The method has the advantages of being capable of quantitatively detecting the aldehyde ketone substances in the exhaled breath of the human body and the like.
Description
Technical Field
The invention belongs to the field of content detection, and particularly relates to a method for detecting the content of aldehyde ketone substances in exhaled breath.
Background
The exhaled air of human body is directly exhaled by lung, and the exhaled air of normal person is collected in 1971 by pauling et al, and 250 volatile organic compounds in the exhaled air are detected. More and more researchers have subsequently conducted systematic studies on these organic substances, which were found to be mainly the products of lipid peroxidation processes.
In recent years, the analysis of exhaled breath has become the frontier of international research, and technologies such as sensors, proton-transfer reaction mass spectrometry, gas chromatography-mass spectrometry, ion mobility spectrometry and the like are reported to be used for detecting related potential tumor markers in exhaled breath VOCs (volatile organic compounds). Studies have shown that the volatile organic compounds contained in exhaled breath are closely related to the metabolic state of human diseases (i.e. by detecting volatile organic compounds, representing the risk of having a certain disease, further diagnosis is required). But the content of the VOCs in the human body is low and even lower than the lowest quantitative error requirement of instrument detection. For example: some VOCs in exhaled breath range only between ppt-ppb, which is already below the quantification requirements of many instruments. Some components in the exhaled air such as water vapor and CO2The content of the isovaleric acid is extremely high, and the quantitative determination of an instrument is interfered. Finally, exhaled breath also contains a mixture of chemical diversity features such as alcohols, ketones, aldehydes, etc., which complicates the identification of disease biomarkers.
Therefore, most of the existing methods for detecting the volatility of the exhaled breath of the human body are qualitative detection methods, quantitative detection is difficult, and especially when the concentration ranges of some VOCs in the exhaled breath are small, quantitative detection of the VOCs cannot be basically carried out.
Disclosure of Invention
The invention aims to provide a method for detecting the content of aldehyde ketone substances in exhaled air of a human body, aiming at the problems in the prior art.
The purpose of the invention can be realized by the following technical scheme: a method for detecting the content of aldehyde ketone substances in exhaled air is characterized by comprising the following steps:
s01: collecting exhaled breath of a human body;
s02: and detecting by gas chromatography or gas chromatography-tandem mass spectrometry.
Preferably, the method comprises the following steps:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s001: introducing the human body exhaled air into an adsorption device at a constant flow rate under the condition of constant-temperature heating by the air bag, eluting the adsorption device by using an eluant, and filtering to obtain a liquid to be detected;
s02: and detecting the liquid to be detected by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
Preferably, in step S001, the adsorption device is an adsorption sampling tube, and the preparation method of the adsorption sampling tube is as follows:
(1) preparing a coating liquid containing 1% of derivatization reagent by using an organic solvent;
(2) filling a stationary phase filler on a blank adsorption column;
(3) coating the coating liquid on an adsorption column filled with a stationary phase, finally drying by blowing, sealing, refrigerating and storing;
the chemical formula of the derivatization reagent is R-NH2Or R-O-NH2The preparation process is carried out in a low-aldehyde ketone environment, and the content of formaldehyde is required to be less than or equal to 0.1ug, and the content of acetone is required to be less than or equal to 0.25 ug.
Preferably, the derivative reagent is selected from one of 2, 4-dinitrophenylhydrazine, pentafluorophenylhydrazine, and O- (2,3,4,5, 6-pentafluorobenzyl) hydroxylamine hydrochloride.
Preferably, in step S001, the adsorption device is a physical adsorption sampling tube, the physical adsorption sampling tube is a self-made adsorption sampling tube or a commercial physical adsorption sampling tube, and the self-made adsorption sampling tube is prepared by the following steps: filling the stationary phase filler on a blank adsorption column for later use.
Preferably, the eluent is one or a mixture of n-hexane and ethyl acetate.
Preferably, in step S001, the temperature of the constant temperature treatment of the air bag is selected from 40-70 deg.C, and the constant flow rate of the exhaled air of the human body is selected from 5-1000 ml/min.
Preferably, the method comprises the following steps:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s002: inserting the solid phase micro-extraction column into an air bag collecting exhaled breath of a human body;
s02: and taking out the solid phase microextraction column, and detecting by using a thermal desorption-gas chromatography or gas chromatography-mass spectrometer.
Preferably, the method comprises the following steps:
s01: a micro-injector is adopted to collect the exhaled air of the human body in real time;
s02: directly feeding sample, and detecting volatile aldehyde ketone substances in human exhaled breath by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
Preferably, the background test in the pretreated airbag meets the requirements that the aldehyde substance is less than or equal to 0.1ug calculated by formaldehyde and the ketone substance is less than or equal to 0.25ug calculated by acetone.
The working principle of the invention is as follows: the present invention includes four parallel schemes.
The first scheme is as follows: the method comprises the following steps:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s001: introducing the human body exhaled breath into a chemical derivative adsorption sampling tube at a constant flow rate (the constant flow rate of the human body exhaled breath is selected from 5-1000ml/min) under the condition of constant-temperature heating of the air bag (the constant temperature is selected from 40-70 ℃), eluting the adsorption sampling tube by using an eluent (the eluent is one or a mixture of n-hexane and ethyl acetate), and filtering to obtain a liquid to be detected;
s02: and detecting the liquid to be detected by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
The preparation method of the chemical derivative adsorption sampling tube comprises the following steps: (1) preparing a coating liquid containing 1% of derivatization reagent by using an organic solvent; (2) filling a stationary phase filler on a blank adsorption column; (3) coating the coating liquid on a filling stationary phase (stationary phase filler)Selected from silica gel, C18tenax-GR, etc.), finally drying, sealing and refrigerating for storage; the chemical formula of the derivatization reagent is R-NH2Or R-O-NH2The preparation process is carried out in a low-aldehyde ketone environment, and the content of formaldehyde is required to be less than or equal to 0.1ug, and the content of acetone is required to be less than or equal to 0.25 ug. Further, the derivative reagent is selected from one of 2, 4-dinitrophenylhydrazine, pentafluorophenylhydrazine, and O- (2,3,4,5, 6-pentafluorobenzyl) hydroxylamine hydrochloride.
Scheme II: the method comprises the following steps:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s001: under the condition of constant temperature heating (constant temperature selected from 40-70 deg.C), eluting the physical adsorption sampling tube with eluent (one or two of n-hexane and ethyl acetate) at constant flow rate (constant flow rate of human body exhalation selected from 5-1000ml/min), and filtering to obtain solution to be detected;
s02: and detecting the liquid to be detected by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
Wherein, the physical sampling pipe is self-made absorption sampling pipe or commercial physical absorption sampling pipe, and the preparation method of self-made absorption sampling pipe is as follows: filling the stationary phase filler on a blank adsorption column for later use. Commercially available physical sampling tubes are available directly from the market.
The third scheme is as follows: the method comprises the following steps:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s002: inserting the solid phase micro-extraction column into an air bag collecting exhaled breath of a human body;
s02: taking out the solid phase micro-extraction column (the filler of the solid phase micro-extraction column has the filler with equivalent models and performances such as PDMS, Carboxen/PDMS, PDMS/DVB, Polyacrylate, DVB/CAR/PDMS, Stableflex and the like), and detecting by a thermal desorption-gas chromatography or gas chromatography-mass spectrometer.
And the scheme is as follows: the method comprises the following steps:
s01: a micro-injector is adopted to collect the exhaled air of the human body in real time;
s02: directly feeding sample, and detecting volatile aldehyde ketone substances in human exhaled breath by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
The air bag background tests in the first scheme, the second scheme and the third scheme all meet the condition that the aldehyde substance is less than or equal to 0.1ug calculated by formaldehyde, and the ketone substance is less than or equal to 0.25ug calculated by acetone.
The solution to be detected in the first scheme contains aldehyde ketone derivatives, namely, the first scheme is to convert aldehyde ketone substances into aldehyde ketone derivatives for detection; correspondingly, the standard solution used for detection is a standard solution of a plurality of aldehyde ketone derivative products.
The solution to be tested in the second scheme contains aldehyde ketone substances. The second scheme is that the aldehyde ketone substance under the eluent is directly detected; correspondingly, the standard solution used for detection is a standard solution of a plurality of aldehydes and ketones.
In the third scheme, a solid phase microextraction column is adopted to enrich exhaled air of a human body, and then a thermal desorption sampling mode is adopted; correspondingly, the standard solution used for detection is a standard solution of a plurality of aldehydes and ketones.
The fourth scheme is that human body exhaled breath collected by a micro-injector is directly injected for detection; correspondingly, the standard solution used for detection is a standard solution of a plurality of aldehydes and ketones.
Compared with the prior art, the invention has the following advantages:
1. the invention aims at the quantitative detection of the aldehyde ketone content in the exhaled air of human body, and the aldehyde ketone content in the exhaled air of human body can be detected even if the aldehyde ketone content is very small. The invention can quantitatively detect the content of at least 25 aldehydes and ketones simultaneously.
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the present invention is not limited to these examples.
Example 1
Pretreatment of gas bags
The air bag can be a Taidela air bag or a nalophen or PFV air bag or an air bag made of other materials. Pretreating the air bag, putting the air bag into a constant temperature control box, setting the temperature at 60 ℃, filling nitrogen into the air bag, keeping the nitrogen in the air bag for 3-5 minutes, extracting the nitrogen, introducing the nitrogen again, repeating the steps for 4 times, and then carrying out cleaning verification on the air bag. The temperature, number of purges, gas introduced can be varied, and the other steps are as above.
Cleaning and verifying the air bag, namely filling 3L nitrogen into the air bag after cleaning the air bag, heating the air bag at the constant temperature of 50 ℃, absorbing the air by using an absorption sampling tube, completely absorbing the air in the bag, eluting the absorption sampling tube by using a solvent, fixing the volume to 2ml, filtering a test background, and enabling the bag to be used after the background test meets the requirement (the quantitative result of a marker measured in the background of the sampling bag is that aldehyde substances are counted by formaldehyde, the converted result is less than or equal to 0.1ug, the ketone substances are counted by acetone, and the converted result is less than or equal to 0.25 ug).
Experiments prove that: when the temperature is 60-80 ℃, introducing nitrogen or helium into the air bag, staying in the air bag for 30 minutes, extracting and introducing, and repeating for 4-8 times, wherein the background test can meet the requirements.
Preparation of selective adsorption sampling tube
Preparing a coating solution containing a derivatization reagent with the content of 1% by using an organic solvent such as n-hexane or ethyl acetate; then filling 200mg of stationary phase filler (silica gel, C18, tenax-GR, etc.) on the blank adsorption column; and finally, coating the coating liquid on an adsorption column filled with a stationary phase, wherein the coating method is carried out by adopting a direct coating method.
As the derivatizing reagent, 2, 4-Dinitrophenylhydrazine (DNPH), pentafluorophenylhydrazine (PFPH), O- (2,3,4,5, 6-pentafluorobenzyl) hydroxylamine hydrochloride (PFBHA) can be used.
Before coating, a blank adsorption column is firstly rinsed by solvents such as normal hexane or ethyl acetate, a negative pressure solid phase extraction system is used for removing rinsed organic reagents, then 1% of coating liquid is directly coated on the adsorption column, after the coating is saturated, the negative pressure solid phase extraction system is used for removing redundant coating liquid, a selective adsorption sampling tube capable of selectively adsorbing the exhaled breath ketones is prepared, the adsorption column is dried by nitrogen, and the selective adsorption sampling tube is sealed and stored at 4 ℃ for standby. The preparation process needs to be carried out in a low-aldehyde ketone environment, and the content of formaldehyde is required to be less than or equal to 0.1ug, and the content of acetone is required to be less than or equal to 0.25 ug.
Collection and treatment of exhaled air from human body
Collecting human body exhaled air by using a pretreated air bag, wherein the human body exhaled air is 0.1L-10L, heating the collected human body exhaled air at a constant temperature, wherein the treatment temperature is 40-70 ℃, enabling the treated human body exhaled air to flow through the adsorption sampling tube for selectively absorbing the aldehyde ketone substances, which is prepared by the method, at a constant flow rate (the flow rate is selected to be 5-1000ml/min), eluting the adsorption sampling tube by using an eluant (the eluant is one or both of n-hexane and ethyl acetate), filtering, and fixing the volume to 2ml by using the eluant to be detected.
Detection of a liquid to be detected
Before detection, preparing a mixed standard solution of a liquid to be detected, wherein the mixed standard solution is a standard solution of various aldehyde ketone derivative products, measuring the mixed standard solution by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, drawing a linear regression working curve by using the concentration content of each corresponding aldehyde ketone derivative marker in the mixed standard solution as an abscissa and the corresponding measured peak area as an ordinate, and taking the amount at a signal-to-noise ratio of 3 times as a detection limit and the amount at a signal-to-noise ratio of 9 times as a quantification limit; and detecting the liquid to be detected by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, and bringing the liquid to be detected into a working curve to obtain the liquid to be detected.
The selection of 25 markers in the mixed standard solution is shown in table 1:
table 125 markers and corresponding aldehyde ketone derivatives
In the table, the name butanone-DNPH of the standard solution, butanone as the corresponding substance in the respiratory gas, and the other similar reasons.
The selected quantitative analysis instruments were: the gas chromatograph is provided with a sample introduction system (manual and automatic sample introduction), a column temperature system, a detection system (provided with a detector comprising a hydrogen flame ionization detector FID, a flame brightness detector FPD, a nitrogen and phosphorus detector NPD and an electron capture detector ECD), a data processing system, a separation system and the like. And the gas chromatography-mass spectrometer (comprising a tandem mass spectrometer) is provided with a sample introduction system (manual and automatic sample introduction), a column temperature system, a detection system (an electron impact ion source EI and an electron ionization source of a configured detector), a data processing system, a separation system and the like.
In conclusion, the detection method of the embodiment 1 has high sensitivity, the lowest quantifiable exhaled breath aldehyde ketone is as low as 0.05 ng/L (calculated by aldehyde ketone, 3L exhaled breath), the stability is good, the RSD% of all aldehyde ketone markers is controlled within 5% (all the markers are controlled by standard quality), the precision is high, and the relative standard deviation of all the markers is controlled within 15% (all the markers are controlled by standard quality) in the quantitative result of different time tests.
Example 2
Pretreatment of gas bags
The airbag was treated as in example 1
Preparation of adsorption sampling tube
Self-made adsorption sampling pipe or commercial adsorption sampling pipe
The preparation method of the self-made adsorption sampling tube comprises the following steps: and filling 50mg of stationary phase filler into a blank adsorption column to prepare an adsorption sampling tube for later use. The optional stationary phase packing is silica gel, C18, tenax-GR, tenax-TA, etc.
Commercially available adsorption sampling tubes are available directly from the market.
Collection and treatment of exhaled air from human body
Collecting 0.1L-10L of human body exhaled breath by using a pretreated air bag, heating the collected human body exhaled breath at a constant temperature of 40-70 ℃, enabling the treated human body exhaled breath to flow through the adsorption sampling tube at a constant flow rate (the flow rate is selected to be 5-1000ml/min) (physical adsorption can be carried out on volatile organic compounds), eluting the adsorption sampling tube by using an eluant (the eluant is one or a mixture of n-hexane and ethyl acetate), and filtering to obtain a liquid to be detected;
detection of a liquid to be detected
Before detection, preparing a mixed standard solution of a liquid to be detected, wherein the mixed standard solution is a standard solution of various aldehydes and ketones, measuring the mixed standard solution by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, drawing a linear regression working curve by using the concentration content of each corresponding aldehyde and ketone in the mixed standard solution as an abscissa and the corresponding measured peak area as an ordinate, and taking the amount at a signal-to-noise ratio of 3 times as a detection limit and the amount at a signal-to-noise ratio of 9 times as a quantification limit; and detecting the liquid to be detected by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, and bringing the liquid to be detected into a working curve to obtain the liquid to be detected.
The selection of 25 markers in the mixed standard solution is shown in table 2:
table 225 markers and corresponding aldehyde ketone substances
The selected quantitative analyzer was the same as in example 1.
Example 3
Pretreatment of gas bags
The airbag was treated as in example 1
Commercial adsorption sampling tube
And (4) purchasing an adsorption sampling pipe from the market for later use.
Collection and treatment of exhaled air from human body
Collecting human body exhaled air 0.1L-10L with pretreated air bag, heating the collected human body exhaled air at 40-70 deg.C, and flowing the treated human body exhaled air at constant flow rate (selected 5-1000 ml/min).
Detection of a liquid to be detected
Firstly, dripping the aldehyde-ketone mixed standard solution (namely the various aldehyde-ketone standard solutions in the embodiment 2) on a commercial adsorption sampling tube according to a static mass solution mode, and then carrying out thermal desorption and direct sample injection analysis. Measuring the mixed standard solution by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, drawing a linear regression working curve by using the concentration content of each corresponding aldehyde ketone in the mixed standard solution as an abscissa and the corresponding measured peak area as an ordinate, and taking the amount of the signal-to-noise ratio of 3 times as a detection limit and the amount of the signal-to-noise ratio of 9 times as a quantitative limit; and detecting the liquid to be detected by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, and bringing the liquid to be detected into a working curve to obtain the liquid to be detected.
The selected quantitative analysis instruments were: the gas chromatograph is provided with a sample introduction system (manual and automatic sample introduction) or a thermal desorption sample introduction system, a column temperature system, a detection system (a detector is provided with a hydrogen flame ionization detector FID, a flame brightness detector FPD, a nitrogen and phosphorus detector NPD, an electron capture detector ECD), a data processing system, a separation system and the like. And the gas chromatography-mass spectrometer (comprising a tandem mass spectrometer) is provided with a sample introduction system (manual and automatic sample introduction), a column temperature system, a detection system (an electron impact ion source EI and an electron ionization source of a configured detector), a data processing system, a separation system and the like.
Example 4
Pretreatment of gas bags
The airbag was treated as in example 1
Commercial solid phase micro-extraction column
The solid phase micro-extraction column is purchased from the market for standby. The solid phase micro-extraction column is filled with fillers with equivalent models and performances such as PDMS, Carboxen/PDMS, PDMS/DVB, Polyacrylate, DVB/CAR/PDMS, Stableflex and the like.
Collection and treatment of exhaled air from human body
Collecting 0.1L-10L of human body exhaled air by the pretreated air bag, heating the collected human body exhaled air at a constant temperature of 40-70 ℃, placing the treated human body exhaled air bag at a constant temperature, inserting the column head of the solid phase micro-extraction column into the air bag placed at the constant temperature, standing for adsorption for a period of time, and taking out.
Detection of a liquid to be detected
Firstly, dripping aldehyde-ketone mixed standard solution (namely the various aldehyde-ketone standard solutions in the embodiment 2) on a commercial solid phase microextraction column according to a static mass solution mode, and then carrying out thermal desorption and direct sample injection analysis. Measuring the mixed standard solution by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, drawing a linear regression working curve by using the concentration content of each corresponding aldehyde ketone in the mixed standard solution as an abscissa and the corresponding measured peak area as an ordinate, and taking the amount of the signal-to-noise ratio of 3 times as a detection limit and the amount of the signal-to-noise ratio of 9 times as a quantitative limit; and detecting the liquid to be detected by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, and bringing the liquid to be detected into a working curve to obtain the liquid to be detected.
Selected quantitative analysis apparatus as in example 3
Example 5
The micro-injector is directly adopted to collect the exhaled air of the human body in real time. And directly injecting a sample for analysis after collection.
Detection of a gas to be detected
Before detection, preparing a mixed standard solution of a solution to be detected, wherein the mixed standard solution is a standard solution of various aldehydes and ketones (same as the example 2), measuring the mixed standard solution by using a corresponding gas chromatograph or gas chromatograph-mass spectrometer, drawing a linear regression working curve by using the concentration content of each corresponding aldehyde and ketone in the mixed standard solution as an abscissa and the corresponding measured peak area as an ordinate, and taking the amount at a signal-to-noise ratio of 3 times as a detection limit and the amount at a signal-to-noise ratio of 9 times as a quantification limit; and detecting the gas to be detected by using a corresponding gas chromatography or gas chromatography-mass spectrometry combination instrument, and bringing the gas to be detected into a working curve to obtain the gas detection device.
The selected quantitative analyzer was the same as in example 1.
In conclusion, the methods of examples 2 to 5 can also be used for quantitative detection of aldoketones in exhaled breath. But the sensitivity of detection was lower than in example 1. Example 1 is the best solution.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (10)
1. A method for detecting the content of aldehyde ketone substances in exhaled air is characterized by comprising the following steps:
s01: collecting exhaled breath of a human body;
s02: and detecting by gas chromatography or gas chromatography-tandem mass spectrometry.
2. The method for detecting the content of aldehydes and ketones in exhaled breath according to claim 1, comprising the steps of:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s001: introducing the exhaled air of the human body into an adsorption sampling device at a constant flow rate under the condition of constant-temperature heating by the air bag, eluting the adsorption device by using an eluant, and filtering to obtain a liquid to be detected;
s02: and detecting the liquid to be detected by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
3. The method for detecting the content of the aldehydes and ketones in the exhaled breath according to claim 2, wherein in step S001, the adsorption device is an adsorption sampling tube, and the preparation method of the adsorption sampling tube is as follows:
(1) preparing a coating liquid containing 1% of derivatization reagent by using an organic solvent;
(2) filling a stationary phase filler on a blank adsorption column;
(3) coating the coating liquid on an adsorption column filled with a stationary phase, finally drying by blowing, sealing, refrigerating and storing;
the chemical formula of the derivatization reagent is R-NH2Or R-O-NH2The preparation process is carried out in a low-aldehyde ketone environment, and the content of formaldehyde is required to be less than or equal to 0.1ug, and the content of acetone is required to be less than or equal to 0.25 ug.
4. The method according to claim 3, wherein the derivatizing reagent is selected from 2, 4-dinitrophenylhydrazine, pentafluorophenylhydrazine, and O- (2,3,4,5, 6-pentafluorobenzyl) hydroxylamine hydrochloride.
5. The method for detecting the content of the aldehyde ketone substances in the exhaled breath according to claim 2, wherein in step S001, the adsorption device is a physical adsorption sampling tube, the physical adsorption sampling tube is a self-made adsorption sampling tube or a commercial physical adsorption sampling tube, and the self-made adsorption sampling tube is prepared by the following steps: filling the stationary phase filler on a blank adsorption column for later use.
6. The method for detecting the content of the aldehydes and ketones in the exhaled breath according to claim 2, wherein the eluent is one or a mixture of n-hexane and ethyl acetate.
7. The method according to claim 2, wherein in step S001, the temperature of the constant temperature treatment of the air bag is selected from 40 to 70 ℃, and the constant flow rate of the exhaled breath is selected from 5 to 1000 ml/min.
8. The method for detecting the content of aldehydes and ketones in exhaled breath according to claim 1, comprising the steps of:
s01: collecting the exhaled air of the human body by using the pretreated air bag;
s002: inserting the solid phase micro-extraction column into an air bag collecting exhaled breath of a human body;
s02: and taking out the solid phase microextraction column, and detecting by using a thermal desorption-gas chromatography or gas chromatography-mass spectrometer.
9. The method for detecting the content of aldehydes and ketones in exhaled breath according to claim 1, comprising the steps of:
s01: a micro-injector is adopted to collect the exhaled air of the human body in real time;
s02: directly feeding sample, and detecting volatile aldehyde ketone substances in human exhaled breath by adopting gas chromatography or gas chromatography-tandem mass spectrometry.
10. The method according to any one of claims 2 or 8, wherein the background test in the pretreated air bag satisfies that the aldehyde substance is less than or equal to 0.1ug calculated as formaldehyde and the ketone substance is less than or equal to 0.25ug calculated as acetone.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010252958.5A CN111398463A (en) | 2020-04-02 | 2020-04-02 | Method for detecting content of aldehydes and ketones in exhaled air |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010252958.5A CN111398463A (en) | 2020-04-02 | 2020-04-02 | Method for detecting content of aldehydes and ketones in exhaled air |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111398463A true CN111398463A (en) | 2020-07-10 |
Family
ID=71433338
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010252958.5A Pending CN111398463A (en) | 2020-04-02 | 2020-04-02 | Method for detecting content of aldehydes and ketones in exhaled air |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111398463A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112098534A (en) * | 2020-11-18 | 2020-12-18 | 必睿思(杭州)科技有限公司 | Method for detecting metabolite and kit thereof |
| CN112255354A (en) * | 2020-09-11 | 2021-01-22 | 广州医科大学附属第一医院 | Characteristic substance for diagnosing Marneffei panuliasis and screening method and application thereof |
| CN112857925A (en) * | 2021-01-27 | 2021-05-28 | 暨南大学 | Method and device for collecting and analyzing aldehyde and ketone substances in biogas sample |
| CN114720590A (en) * | 2022-03-18 | 2022-07-08 | 北京安胜瑞力科技有限公司 | Derivatization-gas chromatography detection method of 1, 3 dicarbonyl compound |
| WO2024110879A1 (en) * | 2022-11-22 | 2024-05-30 | Anika Sterilis Private Limited | Method and apparatus for breath screening for tuberculosis |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104020246A (en) * | 2014-06-24 | 2014-09-03 | 上海大学 | Method for simultaneously detecting plurality of volatile trace carbonyl compounds in atmosphere |
| CN205643257U (en) * | 2016-03-10 | 2016-10-12 | 北京爱康远创医疗科技有限公司 | Expired gas analysis and detection appearance |
| WO2018052689A1 (en) * | 2016-09-13 | 2018-03-22 | Michael Phillips | A method and apparatus for identification of biomarkers in breath and methods of using same for prediction of lung cancer |
-
2020
- 2020-04-02 CN CN202010252958.5A patent/CN111398463A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104020246A (en) * | 2014-06-24 | 2014-09-03 | 上海大学 | Method for simultaneously detecting plurality of volatile trace carbonyl compounds in atmosphere |
| CN205643257U (en) * | 2016-03-10 | 2016-10-12 | 北京爱康远创医疗科技有限公司 | Expired gas analysis and detection appearance |
| WO2018052689A1 (en) * | 2016-09-13 | 2018-03-22 | Michael Phillips | A method and apparatus for identification of biomarkers in breath and methods of using same for prediction of lung cancer |
Non-Patent Citations (5)
| Title |
|---|
| MAGDALENA LIGOR等: "Determination of volatile organic compounds in exhaled breath of patients with lung cancer using solid phase microextraction and gas chromatography mass spectrometry", 《CLIN CHEM LAB MED》 * |
| T LOMONACO1等: "Determination of carbonyl compounds in exhaled breath by onsorbent derivatization coupled with thermal desorption and gas chromatography-tandem mass spectrometry", 《JOURNAL OF BREATH RESEARCH》 * |
| YUMEI LIN等: "Protocol for Collection and HPLC Analysis of Volatile Carbonyl Compounds in Breath", 《CLINICAL CHEMISTRY》 * |
| 杨静: "呼出气丙酮在慢性射血分数保留的心力衰竭中的临床价值研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 胡燕婕: "肺癌呼气特征性VOCs的筛选及诊断价值研究", 《中国优秀博硕士学位论文全文数据库(博士) 医药卫生科技辑》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112255354A (en) * | 2020-09-11 | 2021-01-22 | 广州医科大学附属第一医院 | Characteristic substance for diagnosing Marneffei panuliasis and screening method and application thereof |
| CN112255354B (en) * | 2020-09-11 | 2021-09-10 | 广州医科大学附属第一医院 | Characteristic substance for diagnosing Marneffei panuliasis and screening method and application thereof |
| CN112098534A (en) * | 2020-11-18 | 2020-12-18 | 必睿思(杭州)科技有限公司 | Method for detecting metabolite and kit thereof |
| CN112857925A (en) * | 2021-01-27 | 2021-05-28 | 暨南大学 | Method and device for collecting and analyzing aldehyde and ketone substances in biogas sample |
| CN112857925B (en) * | 2021-01-27 | 2023-09-12 | 暨南大学 | A collection method, analysis method and device for aldehydes and ketones in biogas samples |
| CN114720590A (en) * | 2022-03-18 | 2022-07-08 | 北京安胜瑞力科技有限公司 | Derivatization-gas chromatography detection method of 1, 3 dicarbonyl compound |
| WO2024110879A1 (en) * | 2022-11-22 | 2024-05-30 | Anika Sterilis Private Limited | Method and apparatus for breath screening for tuberculosis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111398463A (en) | Method for detecting content of aldehydes and ketones in exhaled air | |
| Schieweck et al. | Analytical procedure for the determination of very volatile organic compounds (C3–C6) in indoor air | |
| Ochiai et al. | Analysis of volatile sulphur compounds in breath by gas chromatography–mass spectrometry using a three-stage cryogenic trapping preconcentration system | |
| CA2526439C (en) | Method for verifying the integrity of thermal desorption sampling tubes | |
| CN111398460A (en) | Method for detecting content of aldehydes and ketones in human exhaled air | |
| Savareear et al. | Comprehensive comparative compositional study of the vapour phase of cigarette mainstream tobacco smoke and tobacco heating product aerosol | |
| CN103675134B (en) | Method for detecting benzene series pollutants in gas by using internal standard method | |
| Biagini et al. | Using labelled internal standards to improve needle trap micro-extraction technique prior to gas chromatography/mass spectrometry | |
| Gorlo et al. | Calibration procedure for solid phase microextraction—gas chromatographic analysis of organic vapours in air | |
| Turner et al. | Gas Chromatography–Mass Spectrometry: How Do I Get the Best Results? | |
| CN101539549A (en) | Photoionization gas chromatography detection method of organic volatile compounds in air | |
| Pawliszyn | Solid phase microextraction | |
| Lim et al. | A simple sampling method for quantification of hazardous volatile organic compounds in mainstream cigarette smoke: method development and prestudy validation | |
| Liedtke et al. | Hyphenation of a MEMS based pre-concentrator and GC-IMS | |
| Sanchez et al. | On‐line multi‐bed sorption trap for VOC analysis of large‐volume vapor samples: injection plug width, effects of water vapor and sample decomposition | |
| CN102539556A (en) | Method for detecting pollutants in air by using ionic liquid headspace sampling | |
| Rodinkov et al. | Potassium fluoride as a selective moisture trapping agent for SPE-TD-GC-FID determination of volatile organic compounds in air | |
| US20130000485A1 (en) | Flow Control System, Device and Method for Thermal Desorption | |
| Peng et al. | Optimization of two-stage thermal desorption combined with pentafluorophenyl hydrazine derivatization-gas chromatography/mass spectrometry analytical method of atmospheric carbonyl compounds | |
| Zou et al. | Automated method for short-chain aldehydes emission measurement by dynamic solid-phase microextraction on-fiber derivatization GC-MSD coupled with a flow-cell | |
| US6174732B1 (en) | Analytical methodology for qualitative and quantitative determination of chemical agent vapor | |
| Iannone et al. | A technique for atmospheric measurements of stable carbon isotope ratios of isoprene, methacrolein, and methyl vinyl ketone | |
| Warren | Needle trap device and solid phase microextraction combined with portable GC-MS for on-site applications | |
| Dudek et al. | The application of an open tubular trap in analysis of organic air pollutants | |
| CN115166111B (en) | Determination method for deodorizing performance of paper diaper |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200710 |
|
| RJ01 | Rejection of invention patent application after publication |