CN113588767A - Biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry - Google Patents
Biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry Download PDFInfo
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Abstract
The invention provides a biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry, which adopts a thermal desorption sample injector to realize time-resolved sample injection of different biogenic amines based on different thermal desorption characteristics of various biogenic amines; meanwhile, the biogenic amine is detected by adopting a reagent molecule assisted positive ion photoionization ion mobility spectrometry, and the detection sensitivity and the identification accuracy of biogenic amine are improved through reagent molecule screening so as to realize the simultaneous and rapid detection of various biogenic amines. The method can realize the simultaneous accurate quantification of various mixtures, and has the advantages of high sensitivity, high detection speed and portable instrument.
Description
Technical Field
The invention relates to the technical field of food safety, in particular to a biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry, which realizes simultaneous and rapid detection of different biogenic amines.
Background
After the marine products are fished, if the marine products are not processed and preserved in time, the marine products are easy to decay under the action of endogenous and microorganisms, and the free amino acids in the marine products are decarboxylated to generate biogenic amine due to decay. Biogenic amines are nitrogen-containing low-molecular-weight alkaline compounds with biological activity, and common compounds include histamine, putrescine, cadaverine, tryptamine, phenylethylamine, spermine, spermidine and the like. Biogenic amine accumulated in human bodies to a certain degree can generate toxicity to human bodies, cause anaphylactic reactions such as headache, nausea, vomit, diarrhea, palpitation, respiratory disorder and the like, and seriously threaten life.
Currently, the techniques for detecting biogenic amines are mainly Gas Chromatography (GC), High Performance Liquid Chromatography (HPLC), Thin Layer Chromatography (TLC), Capillary Electrophoresis (CE), and the like. Although the method can accurately perform qualitative and quantitative analysis on the biogenic amine, the requirements on large instruments and the technical requirements on operators limit that the methods can only be performed indoors, and meanwhile, the requirements on sample pretreatment are high, the operation is complicated, time and labor are consumed, and the requirements on rapid field detection cannot be met.
Ion Mobility Spectrometry (IMS) is a gas-phase ion detection technology, and separation, analysis and detection are realized according to the difference in ion mobility of different gas-phase ions in a uniform electric field under atmospheric pressure. However, the existing ion mobility spectrometry technology mostly adopts radioactive ionization sources, and when a plurality of complex mixtures are detected simultaneously, due to the influence of competitive ionization, the simultaneous accurate quantification of the plurality of mixtures cannot be realized.
Disclosure of Invention
The invention provides a biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry, which aims to solve the problem of simultaneous and rapid detection of various biogenic amines.
In order to achieve the above object, the present invention provides a method for detecting biogenic amine based on time-resolved dynamic thermal desorption ion mobility spectrometry, comprising the following steps: carrying out time resolution by thermal desorption sample injection according to thermal desorption characteristics of different biogenic amines, and sequentially analyzing; and detecting biogenic amine by adopting a photo-ionization ion migration tube assisted by reagent molecules, and screening the reagent molecules to realize the detection of biogenic amine.
Further, the method comprises the following steps:
s1, putting the biogenic amine sample into a thermal desorption sample injection device for heating, and sequentially desorbing different biogenic amines according to different thermal desorption characteristics;
s2, the analyzed gas of the biogenic amine sample enters a reaction area of the ion migration tube along with carrier gas, the sample molecules and the reagent molecules are carried into the reaction area of the ion migration tube while the carrier gas is carried out, and after the biogenic amine sample molecules and the reagent molecules react in the reaction area, corresponding characteristic product ions are respectively formed;
and S3, enabling the characteristic product ions to enter a migration region under the pulse action of an ion gate, separating under the action of an electric field and a floating gas, and enabling different characteristic product ions to sequentially reach a Faraday disc to be detected to form different product ion peaks so as to realize the detection of biogenic amine.
Preferably, the preparation of the biological amine sample comprises: dissolving biogenic amine in a dissolving solvent, and dripping or wiping the biogenic amine on a sampling cloth until the dissolving solvent is completely volatilized.
Preferably, the dissolving solvent comprises methanol and ethanol.
Preferably, the flow rate of the carrier gas is 100-400 mL/min.
Preferably, the flow rate of the floating gas is 200-500 mL/min.
Preferably, the carrier gas and the float are purified and dehumidified air.
Preferably, the reagent molecules comprise one of a single aromatic compound or a volatile ketone with high photoionization efficiency.
Preferably, the monoaromatic compound includes toluene, xylene; the volatile ketone substances comprise acetone and 2-butanone.
Preferably, a plurality of metal rings are distributed at intervals in the migration tube and positive high voltage is applied, and the voltage difference between the two metal rings is 150-200V.
Preferably, the electric field intensity is 215-285V/cm, the resolving temperature is 80-130 ℃, and the temperature of the migration tube is 90-120 ℃.
The invention has the beneficial effects that:
the method is based on photoionization ion mobility spectrometry, and can be combined with a time-resolved dynamic thermal desorption sampling technology to realize simultaneous and rapid detection of various biogenic amines.
The invention utilizes the reagent molecule assisted photoionization positive ion mobility spectrometry technology and combines the time-resolved dynamic thermal desorption sampling technology, can realize the trace detection of ng-grade biogenic amine (biogenic amine can not be detected when no reagent molecule is added), improves the selectivity and sensitivity of detection, enables different sample molecules to enter a reaction area in sequence, and realizes the separation detection of biogenic amine. Compared with the traditional chromatographic technology, the technology has high analysis speed and can finish sample detection in less than 10 s.
Drawings
FIG. 1 is an ion mobility spectrum of example 1 of the present invention;
FIG. 2 is an ion mobility spectrum of example 2 of the present invention;
FIG. 3 is a schematic illustration of ion mobility in the apparatus of the present invention;
wherein, 1, a carrier gas inlet; 2. a reagent molecule generator; 3. a thermal desorption sample injector; 4. a vacuum ultraviolet light ionization source; 5. a reaction zone; 6. an ion gate; 7. a migration zone; 8. a Faraday disc; 9. a detector signal amplifier; 11. a carrier gas inlet; 12. a float gas inlet; 13. and a main air outlet.
Detailed Description
A biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry adopts thermal desorption sample injection, and realizes the time-resolved sample injection of different biogenic amines based on the thermal desorption characteristics of different biogenic amines; the method adopts a photo-ionization ion migration tube assisted by reagent molecules to detect the biogenic amine, improves the separation degree and sensitivity of detection by a reagent molecule screening and time-resolved dynamic thermal analysis sample introduction technology and utilizing specific reaction reagent ions formed by the ionization of the reagent molecules in an ionization region, and realizes the simultaneous and rapid detection of various biogenic amines.
The ion mobility spectrometry is a positive ion mode mobility spectrometry of reagent molecule assisted photoionization, a photoionization source is a non-radioactive vacuum ultraviolet lamp, positive high voltage is applied to an ion migration tube of the photoionization positive ion mobility spectrometry through stainless steel metal rings which are distributed at intervals, a plurality of metal rings are distributed at intervals in the migration tube of the photoionization positive ion mobility spectrometry and the positive high voltage is applied, the voltage difference between the two metal rings is 150-plus-200V, and the electric field intensity on the migration tube is 215-plus-285V/cm. A non-radioactive vacuum ultraviolet lamp is used as an ionization source, the vacuum ultraviolet lamp generates photons of 10.7eV, and the generated photons and added reagent molecule toluene generate single photon ionization to form toluene positive ions losing one electron; and acetone molecules firstly undergo single photon ionization to lose an electron to form acetone positive ions, and the formed acetone positive ions continue to react with the acetone molecules to form two acetone molecule protonated ions.
As shown in fig. 3, the ion mobility spectrometry tube is composed of a reagent molecule generator 2, a vacuum ultraviolet light ionization source 4, a reaction zone 5, an ion gate 6, a migration zone 7, a faraday disk 8, a signal amplifier 9, a carrier gas inlet 11, a floating gas inlet 12, a total gas outlet 13, and the like. The ion migration tube comprises a reaction area 5 at the front end and a migration area 7 at the tail end, an ion gate 6 is arranged between the reaction area 5 and the migration area 7, sample molecules and reaction reagent ions in the reaction area 5 react through specific molecular ions to form product ions, and the product ions enter the migration area 7 through the pulse-opened ion gate and sequentially reach a Faraday disc 8 through the migration area 7. The carrier gas inlet 11 is close to one end of the reaction area of the vacuum ultraviolet lamp and is responsible for carrying sample molecules and reagent molecules into the ion migration tube; a floating gas inlet 12 is arranged at one end of the migration area close to the Faraday disk 8, and a total gas outlet 13 is arranged at the other end of the reaction area between the floating gas inlet 12 and the carrier gas inlet 11 close to the migration area.
The specific method comprises the following steps:
(1) dissolving a certain amount of biogenic amine in a dissolving solvent to prepare a biogenic amine sample, dropwise adding the biogenic amine sample into a sampling cloth, and carrying out sample injection after the dissolving solvent is completely volatilized, or carrying out sampling by adopting a wiping method;
(2) the method comprises the steps of inserting a sampling cloth carrying a biological amine sample into a thermal analysis sample introduction device 3 for heating, sequentially analyzing different biological amines according to different thermal analysis characteristics, allowing analyzed biological amine gas to enter a reaction zone of an ion migration tube along with carrier gas (the carrier gas enters through a carrier gas inlet 1) and then through a carrier gas inlet 11 of the ion migration tube, allowing biological amine sample molecules and reagent molecules in the reaction zone to react to form corresponding characteristic product ions respectively, allowing the characteristic product ions to enter a migration zone 7 under the pulse action of an ion gate 6, separating under the action of an electric field and a floating gas in the migration zone 7, allowing different characteristic product ions to sequentially reach a Faraday disk 8 for detection, and forming different product ion peaks to realize detection of the biological amines.
The dissolving solvent comprises a solvent for dissolving biogenic amine such as methanol and ethanol.
Carrying the sample molecules and the reagent molecules into a reaction area of the ion migration tube while carrying out carrier gas, wherein the flow rate of the carrier gas is 100-400 mL/min. The direction of the gas flow of the sample molecules, the reagent molecules and the carrier gas in the reaction area is consistent, the direction of the gas flow in the migration area is opposite to that of the gas flow in the migration area, and all the gas in the ion migration pipe leaves the ion migration pipe through the total gas outlet 13.
The drift gas enters the ion migration tube from the end of the migration zone in the ion migration tube close to the Faraday disc, and the flow rate of the drift gas is set to be 200-500 mL/min.
The used carrier gas and the floating gas are the purified and dehumidified air, and the mass flow meter is connected with the carrier gas inlet 11 and the floating gas inlet 12 to control the flow rate of the carrier gas and the floating gas.
The adopted reagent molecules are one of monoaromatic compounds (toluene and xylene) or volatile ketone substances (acetone and 2-butanone) with high photoionization efficiency.
The sample molecules, the reagent molecules, the carrier gas and the gas for bleaching are all compressed air which is filtered by active carbon, silica gel and a molecular sieve in sequence.
The working conditions of the ion mobility spectrometry are as follows: the voltage difference between the two metal rings is 150-200V, the electric field intensity on the migration tube is 215-285V/cm, the analysis temperature is 80-130 ℃, and the temperature of the migration tube is 90-120 ℃.
Example 1
In positive ion mode, toluene is used asThe thermal desorption temperature of a reagent molecule is 120 ℃, the temperature of a migration tube is 100 ℃, the flow rate of carrier gas is 200mL/min, the flow rate of a bleaching gas is 300mL/min, the voltage difference between two metal rings is 178V, the electric field distribution on the migration tube is 250V/cm, and ion migration spectrograms of putrescine, cadaverine, spermine, spermidine, phenethylamine and histamine with different reduction mobility rates are respectively obtained, as shown in figure 1, toluene mainly forms the ion migration rate of 2.07cm2V-1s-1The reactant ions and 7 kinds of biogenic amines react to form corresponding characteristic product ions respectively. Wherein the reduction mobility of putrescine formation is 1.94cm2V-1s-1The product ion of (1); cadaverine formation approximate mobility of 1.82cm2V-1s-1Also small amounts of other product ions are formed; spermine mainly forms with an approximate mobility of 1.38cm2V-1s-1The product ion of (1); spermidine mainly forms with an reduced mobility of 1.63cm2V-1s-1Also forms two product ions having reduced mobility less than 1.63cm2V-1s-1The product ion of (1); phenylethylamine mainly forms and has an approximate mobility of 1.29cm2V-1s-1Simultaneously form two product ions with reduced mobility less than 1.29cm2V-1s-1The product ion of (1); tryptamine and histamine formed an approximate mobility of 1.29cm2V-1s-1And 1.84cm2V-1s-1The product ion of (1). The reaction ions formed by toluene can be well separated from the product ions of 7 kinds of biogenic amines, but the response sensitivity to cadaverine, tryptamine and histamine is not high.
Example 2
Under a positive ion mode, acetone is used as a reagent molecule, the thermal desorption temperature is 120 ℃, the temperature of a migration tube is 120 ℃, the flow rate of carrier gas is 200mL/min, the flow rate of a bleaching gas is 300mL/min, the voltage difference between two metal rings is 178V, and the electric field distribution on the migration tube is 250V/cm, so that the ionic migration of putrescine, cadaverine, spermine, spermidine, phenethylamine and histamine with different reduction mobility rates are respectively obtainedThe spectrum, as shown in FIG. 2, shows that acetone mainly forms an approximate mobility of 1.84cm2V-1s-1The reactant ions and 7 kinds of biogenic amines react to form corresponding characteristic product ions respectively. Wherein the reduction mobility of putrescine formation is 1.94cm2V-1s-1And 1.71cm2V-1s-1Two product ions; cadaverine formation approximate mobility of 1.82cm2V-1s-1And 1.64cm2V-1s-1Two product ions; spermine mainly forms with an approximate mobility of 1.38cm2V-1s-1The product ion of (1); spermidine mainly forms with an reduced mobility of 1.63cm2V-1s-1Also forms two reduced mobilities of less than 1.40cm2V-1s-1The product ion of (1); phenylethylamine mainly forms and has an approximate mobility of 1.29cm2V-1s-1While forming product ions of reduced mobility less than 1.29cm2V-1s-1The product ion of (1); tryptamine and histamine formed an approximate mobility of 1.14cm2V- 1s-1And 1.30cm2V-1s-1The product ion of (1). Compared with toluene, the detection sensitivity of histamine is obviously enhanced, but the detection sensitivity of phenylethylamine is weakened. In addition, putrescine, cadaverine, tryptamine and histamine all formed product ions completely different from those formed when toluene was used as the reagent molecule, the reactive ion formed by acetone and the cadaverine formed an approximate mobility of 1.82cm2V-1s-1The degree of separation is poor.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (10)
1. A biogenic amine detection method based on time-resolved dynamic thermal desorption ion mobility spectrometry is characterized by comprising the following steps: firstly, thermal desorption sample injection is carried out, time resolution is carried out according to thermal desorption characteristics of different biogenic amines, and sequential desorption is carried out; and then, detecting biogenic amine by adopting a photo-ionization ion migration tube assisted by reagent molecules, and screening the reagent molecules to realize the detection of biogenic amine.
2. The method for detecting biogenic amines according to claim 1, comprising the steps of:
s1, putting the biogenic amine sample into a thermal desorption sample injection device (3) for heating, and sequentially desorbing different biogenic amines according to different thermal desorption characteristics;
s2, the analyzed gas of the biogenic amine sample enters a reaction area (5) of the ion migration tube along with carrier gas, the sample molecules and the reagent molecules are carried into the reaction area (5) of the ion migration tube while the carrier gas is carried out, and after the biogenic amine sample molecules and the reagent molecules react in the reaction area, corresponding characteristic product ions are respectively formed;
s3, the characteristic product ions enter a migration region (7) under the pulse action of an ion gate (6), separation is carried out under the action of an electric field and a floating gas, different characteristic product ions sequentially reach a Faraday disc (8) to be detected, and different product ion peaks are formed to realize the detection of biogenic amine.
3. The method for detecting biogenic amines according to claim 2, wherein said preparation of a biogenic amine sample comprises: dissolving biogenic amine in a dissolving solvent, and dripping or wiping the biogenic amine on a sampling cloth until the dissolving solvent is completely volatilized.
4. The method according to claim 3, wherein the solvent comprises methanol or ethanol.
5. The method as claimed in claim 2, wherein the flow rate of the carrier gas is 100-400 mL/min; the flow rate of the floating gas is 200-500 mL/min.
6. The method of claim 2, wherein the carrier gas and the bleaching are purified, dehumidified air.
7. The method of claim 2, wherein the reagent molecule comprises one of a monoaromatic compound or a volatile ketone compound having high photoionization efficiency.
8. The method according to claim 7, wherein the monoaromatic compound comprises toluene, xylene; the volatile ketone substances comprise acetone and 2-butanone.
9. The method as claimed in claim 2, wherein a plurality of metal rings are distributed at intervals in the migration tube and a positive high voltage is applied, and the voltage difference between the two metal rings is 150-200V; the migration tube adopts a non-radioactive vacuum ultraviolet lamp as an ionization source.
10. The method as claimed in claim 2, wherein the electric field strength is 215-285V/cm, the temperature of the analysis is 80-130 ℃, and the temperature of the migration tube is 90-120 ℃.
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| US4551624A (en) * | 1983-09-23 | 1985-11-05 | Allied Corporation | Ion mobility spectrometer system with improved specificity |
| US20020120406A1 (en) * | 2001-02-01 | 2002-08-29 | Avraham Lorber | Diagnostic method and apparatus |
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| CN108088887A (en) * | 2016-11-21 | 2018-05-29 | 中国科学院大连化学物理研究所 | A kind of detection method of front three amine gas |
| CN111239234A (en) * | 2018-11-29 | 2020-06-05 | 中国科学院大连化学物理研究所 | Non-radioactive method for on-line monitoring of ammonia gas |
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- 2021-08-03 CN CN202110883912.8A patent/CN113588767A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4551624A (en) * | 1983-09-23 | 1985-11-05 | Allied Corporation | Ion mobility spectrometer system with improved specificity |
| US20020120406A1 (en) * | 2001-02-01 | 2002-08-29 | Avraham Lorber | Diagnostic method and apparatus |
| CN103884770A (en) * | 2012-12-19 | 2014-06-25 | 中国科学院大连化学物理研究所 | Method for detecting explosive |
| CN106226384A (en) * | 2016-09-22 | 2016-12-14 | 大连工业大学 | A kind of trimethylamine detection method based on Ion mobility spectrometry |
| CN108008001A (en) * | 2016-10-28 | 2018-05-08 | 中国科学院大连化学物理研究所 | Improve three peroxidating diamines of hexa-methylene（HMTD）The detection method of dosing accuracy |
| CN108088887A (en) * | 2016-11-21 | 2018-05-29 | 中国科学院大连化学物理研究所 | A kind of detection method of front three amine gas |
| CN111239234A (en) * | 2018-11-29 | 2020-06-05 | 中国科学院大连化学物理研究所 | Non-radioactive method for on-line monitoring of ammonia gas |
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