CN114965773A - Method for determining volatile substance components in Liupao tea in ion source low-energy mode - Google Patents
Method for determining volatile substance components in Liupao tea in ion source low-energy mode Download PDFInfo
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- 241001122767 Theaceae Species 0.000 title claims abstract description 59
- 239000000126 substance Substances 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 35
- 238000004949 mass spectrometry Methods 0.000 claims abstract description 14
- 238000002470 solid-phase micro-extraction Methods 0.000 claims abstract description 11
- 238000004817 gas chromatography Methods 0.000 claims abstract description 8
- 239000002904 solvent Substances 0.000 claims abstract description 8
- 238000005259 measurement Methods 0.000 claims abstract description 7
- 150000002500 ions Chemical class 0.000 claims description 42
- 239000000463 material Substances 0.000 claims description 29
- 238000010438 heat treatment Methods 0.000 claims description 24
- 238000001819 mass spectrum Methods 0.000 claims description 18
- 238000001179 sorption measurement Methods 0.000 claims description 7
- 238000001228 spectrum Methods 0.000 claims description 7
- 238000010586 diagram Methods 0.000 claims description 6
- 150000001793 charged compounds Chemical class 0.000 claims description 5
- 230000005526 G1 to G0 transition Effects 0.000 claims description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 3
- 230000007547 defect Effects 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- -1 polysiloxane Polymers 0.000 claims description 3
- 229920001296 polysiloxane Polymers 0.000 claims description 3
- 238000010926 purge Methods 0.000 claims description 3
- 238000010791 quenching Methods 0.000 claims description 3
- 230000000171 quenching effect Effects 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 125000006850 spacer group Chemical group 0.000 claims description 2
- 239000012535 impurity Substances 0.000 description 4
- 239000011159 matrix material Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- IPMYMEWFZKHGAX-UHFFFAOYSA-N Isotheaflavin Natural products OC1CC2=C(O)C=C(O)C=C2OC1C(C1=C2)=CC(O)=C(O)C1=C(O)C(=O)C=C2C1C(O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-UHFFFAOYSA-N 0.000 description 1
- UXRMWRBWCAGDQB-UHFFFAOYSA-N Theaflavin Natural products C1=CC(C2C(CC3=C(O)C=C(O)C=C3O2)O)=C(O)C(=O)C2=C1C(C1OC3=CC(O)=CC(O)=C3CC1O)=CC(O)=C2O UXRMWRBWCAGDQB-UHFFFAOYSA-N 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 235000014620 theaflavin Nutrition 0.000 description 1
- IPMYMEWFZKHGAX-ZKSIBHASSA-N theaflavin Chemical compound C1=C2C([C@H]3OC4=CC(O)=CC(O)=C4C[C@H]3O)=CC(O)=C(O)C2=C(O)C(=O)C=C1[C@@H]1[C@H](O)CC2=C(O)C=C(O)C=C2O1 IPMYMEWFZKHGAX-ZKSIBHASSA-N 0.000 description 1
- 229940026509 theaflavin Drugs 0.000 description 1
- 235000008118 thearubigins Nutrition 0.000 description 1
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- 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
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- 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
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- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7206—Mass spectrometers interfaced to gas chromatograph
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- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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- 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
- G01N2030/062—Preparation extracting sample from raw material
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Abstract
The invention discloses a method for determining volatile substance components in Liupu tea in an ion source low-energy mode, which belongs to the technical field of tea component determination, wherein the method can be used for accurately determining whether a Liupu tea sample contains the volatile substance components in a volatile substance database; extracting a Liupao tea sample by using a solvent, adsorbing by using a solid phase microextraction needle, putting the Liupao tea sample into a gas chromatograph-mass spectrometer for measurement to obtain a map, and matching the map with data in a volatile substance database to obtain a measurement result, wherein the measurement in the gas chromatograph-mass spectrometer comprises gas chromatography measurement and mass spectrometry, and an ion source is in a low energy mode when the mass spectrometry is carried out.
Description
Technical Field
The invention relates to the technical field of tea component determination, in particular to a method for determining volatile substance components in Liupu tea in an ion source low-energy mode.
Background
The Liupao tea prepared by the modern process is dark brown in appearance and color, thick and bright in liquor color, mellow and smooth in mouthfeel, pure and old in aroma, and particularly has the characteristic of being more and more fragrant, the storage year has a great influence on the aroma, taste and mouthfeel of the Liupao tea, and the storage year becomes an important influence factor on the product price of the Liupao tea in the market. Researches show that different storage years have influences on the quality of Liupu tea, tea polyphenol, amino acid, theaflavin, thearubigin, theabrownin, volatile components and the like.
Volatile substances in the Liupu tea are characteristic components in the Liupu tea and play an important role in the taste and aroma of the Liupu tea, so how to effectively measure the volatile substance components contained in the Liupu tea becomes an important factor for evaluating the quality of the Liupu tea. The traditional determination method is that the Liupao tea is subjected to solid phase microextraction and then volatile components of the Liupao tea are adsorbed and enriched, the enriched components on the solid phase microextraction needle are desorbed through a gas phase sample inlet, and the volatile components are determined through a gas phase mass spectrometer and a standard ion source energy mode.
The energy of an ion source used in the traditional determination method is 70eV, but most substances in the nature cannot obtain a molecular ion peak of the substance under the energy, so that excessive fragments are generated, the molecular weight of the substance cannot be judged through the molecular ions, the error of determining the volatile substance component is large, and the determination time is long. The traditional determination method for determining the volatile substance needs to use a standard substance to determine the retention time of a chromatographic peak, and the difficulty in separating a target peak and an impurity peak in a complex matrix of tea is high. Therefore, it is highly desirable to invent a method for efficiently measuring the volatile substance components in Liupu tea so as to measure the volatile substance components contained in Liupu tea more quickly and accurately and to evaluate the quality of Liupu tea better.
Disclosure of Invention
The invention aims to provide a method for measuring volatile substance components in Liupu tea in an ion source low-energy mode, whether the Liupu tea sample contains the volatile substance components in a volatile substance database can be accurately measured by using the measuring method, and the measuring method is good in anti-interference performance, high in measuring speed and strong in specificity.
The technical scheme of the invention is as follows:
a method for determining volatile substance components in Liupu tea in an ion source low-energy mode includes the steps of extracting Liupu tea samples through a solvent, adsorbing the Liupu tea samples through a solid phase microextraction needle, putting the Liupu tea samples into a gas chromatograph-mass spectrometer for determination to obtain a graph, and matching the graph with data in a volatile substance database to obtain a determination result, wherein the determination in the gas chromatograph-mass spectrometer includes gas chromatography determination and mass spectrometry, and the ion source is in the low-energy mode when the mass spectrometry is carried out.
Further, putting 1-2 g of Liupao tea sample into a 20mL headspace bottle, adding 5-8 mL of deionized water, capping and sealing, placing under a heating device for heating, and inserting a solid phase microextraction needle for adsorption.
Further, the heating temperature range of the heating device is 70-90 ℃, and the adsorption time is 30-45 min.
Further, the chromatographic column in the gas chromatography determination is a nonpolar column, the stationary phase is 5% phenyl-methyl polysiloxane, and the specification is 30m multiplied by 0.25mm, 0.25 μm; sample inlet temperature: 250 to 280 ℃; purging the isolation pad by 3-5 mL/min; the split ratio is as follows: 7-10: 1; flow rate of column He: 1 mL/min; quenching gas He: 2.25 mL/min; collision gas N2: 1.5 mL/min; interface temperature: 280-300 ℃; directly heating to 40 ℃ by adopting a programmed heating method, keeping the temperature for 3min, heating to 300 ℃ at a heating rate of 5 ℃/min, keeping the temperature for 5min, keeping the temperature of a column incubator at 310 ℃, then operating for 2min, and carrying out chromatographic column flow He: 2 mL/min.
The mass spectrometry conditions are as follows:
the ion source is a high-sensitivity ion source equipped with a low-energy mode; the solvent delay time is 3 min; the scanning range is 30-500 m/z; the ion source energy is 5-15 eV, and the data acquisition mode is a centroid diagram and a profile diagram.
Further, the specific steps of matching the spectrum obtained by measurement in the gas mass spectrometer with the data in the volatile substance database are as follows:
(1) performing primary mass spectrum matching on the obtained mass center graph and outline graph and material component data in the volatile material database one by one, performing the next step if the matching degree of the molecular ion peaks of the material components in the matching result reaches more than 90 minutes, and performing the primary mass spectrum matching of the next material component if the material is not present;
(2) and (3) performing secondary ion matching on the obtained mass center graph and the profile graph and the material component data in the volatile material database, if the deviation generated by the secondary ions in the matching result is less than or equal to 10ppm, determining that the material component is contained, otherwise, returning to the step (1) to perform primary mass spectrum matching of the next material component.
Furthermore, the volatile substance database stores information of Chinese and English names, molecular formulas, CAS numbers, accurate molecular weights and mass defects of various volatile substances.
Compared with the prior art, the invention has the beneficial effects that:
the invention relates to a method for determining volatile substance components in Liupu tea in an ion source low-energy mode, which comprises the steps of extracting Liupu tea samples through a solvent, adsorbing the Liupu tea samples through a solid phase microextraction needle, putting the Liupu tea samples into a gas mass spectrometer for determination, matching the obtained spectrum with data in a volatile substance database to obtain a determination result, determining the Liupu tea samples in the gas mass spectrometer by gas chromatography determination and mass spectrometry, wherein the ion source is in a low-energy mode during mass spectrometry, and a high-resolution spectrum is obtained by using the low-energy mode in the method and is used for monitoring the accurate mass-to-charge ratio of a detected substance, so that the method is good in anti-interference performance, and can be used for rapidly determining the volatile substance components through the accurate mass-to-charge ratio under the condition that a target object in the sample is not separated from impurity peaks, and the resolution of the substance components is higher, and the specificity is strong.
Detailed Description
The technical solution of the present invention will be described in further detail with reference to the following embodiments, but the present invention is not limited thereto.
The method for determining volatile substance components in Liupu tea in an ion source low-energy mode comprises the steps of extracting Liupu tea samples through a solvent, adsorbing the Liupu tea samples through a solid phase microextraction needle, putting the Liupu tea samples into a gas chromatograph-mass spectrometer to determine a graph, and matching the graph with data in a volatile substance database to obtain a determination result, wherein the determination in the gas chromatograph-mass spectrometer comprises gas chromatography determination and mass spectrometry, and the ion source is in the low-energy mode during mass spectrometry.
The method screens and qualifies multiple volatile characteristic components in the Liupu tea by using a solid phase microextraction technology, combining a gas chromatography-tandem flight time mass spectrum and combining a self-built volatile substance database, and has the advantages that a low-energy mode is used in the method, a high-resolution spectrum can be obtained, the high-resolution mass spectrum is used for monitoring the accurate mass-to-charge ratio of a detected substance, and the high-resolution mass spectrum has a good anti-interference effect on the qualification of the volatile components in the Liupu tea matrix. The traditional low-resolution gas mass spectrometry method for determining the nature of the substances needs to use standard substances to determine the retention time of chromatographic peaks, and the difficulty in separating target peaks and impurity peaks in a complex matrix of tea leaves is high. The low-resolution mass spectrum is easily interfered by substances co-flowed with the low-resolution mass spectrum in the determination process, so that a long time is consumed for separating the co-flowed substances from the target substances, and when different Liupao tea matrixes are used for determining the low-resolution mass spectrum in a certain difference manner, a certain pre-experiment is firstly carried out on the to-be-determined sample, and then the detection can be formally carried out after the substances are completely separated by a temperature programmed gradient method. The method can also carry out rapid qualitative operation through accurate mass-to-charge ratio under the condition of not separating the target object and the impurity peak in the sample, and has higher resolution ratio of the substance components and strong specificity. The method can also be used for detecting volatile substance components in other tea samples except for Liupu tea.
The Liupu tea sample dissolving, extracting and adsorbing process comprises the steps of putting 1-2 g of Liupu tea sample into a 20mL headspace bottle, adding 5-8 mL of deionized water, capping and sealing, placing under a heating device for heating, and inserting a solid phase microextraction needle for adsorption. The heating temperature range of the heating device is 70-90 ℃, the adsorption time is 30-45 min, and after the Liupao tea sample is dissolved, extracted and adsorbed according to the requirement, more complete volatile substance components can be obtained through adsorption, so that the later detection can obtain more accurate results.
The chromatographic column in the gas chromatography determination is a nonpolar column, the stationary phase is 5% phenyl-methyl polysiloxane, and the specification is 30m multiplied by 0.25mm, 0.25 mu m; sample inlet temperature: 250 to 280 ℃; purging the spacer at a rate of 3-5 mL/min; the split ratio is as follows: 7-10: 1; flow rate of the column He: 1 mL/min; quenching gas He: 2.25 mL/min; collision gas N2: 1.5 mL/min; interface temperature: 280-300 ℃; directly heating to 40 ℃ by adopting a programmed heating method, keeping the temperature for 3min, heating to 300 ℃ at a heating rate of 5 ℃/min, keeping the temperature for 5min, keeping the temperature of a column incubator at 310 ℃, then operating for 2min, and carrying out chromatographic column flow He: 2 mL/min.
The mass spectrometry conditions are as follows:
the ion source is a high-sensitivity ion source equipped with a low-energy mode; the solvent delay time is 3 min; the scanning range is 30-500 m/z; the ion source energy is 5-15 eV, and the data acquisition mode is a centroid diagram and a profile diagram.
The specific steps of matching the spectrum obtained by measurement in the gas mass spectrometer with the data in the volatile substance database are as follows:
(1) performing primary mass spectrum matching on the obtained mass center graph and outline graph and material component data in a volatile material database one by one, performing the next step if the matching degree of the molecular ion peaks of the material components in the matching result reaches more than 90 minutes (primary mass spectrum matching, deviation is less than or equal to 10ppm), and performing primary mass spectrum matching on the next material component if the material is not present;
(2) performing secondary ion matching on the obtained mass center graph and the profile graph and the material component data in the volatile material database, if the deviation generated by the secondary ions in the matching result is less than or equal to 10ppm, determining that the material component is contained, and returning to the step (1) to perform primary mass spectrum matching on the next material component; otherwise, the substance is not considered to exist, and the step (1) is returned to carry out the first-stage mass spectrum matching of the next substance component.
The discrimination points of the secondary ions were calculated according to Table 5 in European Union non-mandatory Law 2002-657-EC, and when the number of discrimination points reached was 4 or more, it was confirmed that the sample contained the compound. The identification point number calculation mode is that the identification point number of the parent ion is 2, the identification point number of each product ion (daughter ion) obtained by parent ion conversion is 2.5, and the high-resolution mass spectrum at least comprises one parent ion and one product ion (daughter ion) converted by the parent ion according to the qualitative requirement.
The volatile substance database stores Chinese and English names, molecular formulas, CAS numbers, accurate molecular weights and mass defects of various volatile substances. The volatile material compositions stored in the volatile material database are shown in table 1.
TABLE 1 volatile substance ingredient Table
The above description is only exemplary of the invention, and any modification, equivalent replacement, and improvement made within the spirit and scope of the present invention should be considered within the scope of the present invention.
Claims (6)
1. A method for determining volatile substance components in Liupu tea in an ion source low-energy mode is characterized in that Liupu tea samples are extracted through a solvent, the Liupu tea samples are adsorbed by a solid phase microextraction needle, a spectrum obtained by determination is placed in a gas mass spectrometer and then matched with data in a volatile substance database to obtain a determination result, determination in the gas mass spectrometer comprises gas chromatography determination and mass spectrometry, and the ion source is in the low-energy mode when mass spectrometry is carried out.
2. The method for determining volatile substances in Liupu tea in an ion source low-energy mode is characterized in that 1-2 g of Liupu tea samples are placed in a 20mL headspace bottle, 5-8 mL of deionized water is added, the flask is covered and sealed, the flask is placed under a heating device for heating, and a solid phase microextraction needle is inserted for adsorption.
3. The method for measuring the volatile substances in the Liupu tea in the low energy mode of the ion source as claimed in claim 2, wherein the heating temperature of the heating device is 70-90 ℃, and the adsorption time is 30-45 min.
4. The method for measuring the volatile substance components in the Liupu tea in an ion source low energy mode is characterized in that a chromatographic column in gas chromatography measurement is a non-polar column, a stationary phase is 5% phenyl-methyl polysiloxane, and the specification is 30m x 0.25mm and 0.25 μm; sample inlet temperature: 250 to 280 ℃; purging the spacer at a rate of 3-5 mL/min; the split ratio is as follows: 7-10: 1; flow rate of column He: 1 mL/min; quenching gas He: 2.25 mL/min; collision gas N2: 1.5 mL/min; interface temperature: 280-300 ℃; directly heating to 40 ℃ by adopting a programmed heating method, keeping the temperature for 3min, heating to 300 ℃ at a heating rate of 5 ℃/min, keeping the temperature for 5min, keeping the temperature of a column incubator at 310 ℃, then operating for 2min, and carrying out chromatographic column flow He: 2 mL/min.
The mass spectrometry conditions are as follows:
the ion source is a high-sensitivity ion source equipped with a low-energy mode; the solvent delay time is 3 min; the scanning range is 30-500 m/z; the ion source energy is 5-15 eV, and the data acquisition mode is a centroid diagram and a profile diagram.
5. The method for determining the volatile substance components in the Liupu tea in the low energy mode through the ion source is characterized in that the specific steps of matching the spectrum obtained through determination in the gas chromatograph-mass spectrometer with the data in the volatile substance database are as follows:
(1) performing primary mass spectrum matching on the obtained mass center graph and outline graph and material component data in the volatile material database one by one, performing the next step if the matching degree of the molecular ion peaks of the material components in the matching result reaches more than 90 minutes, and performing the primary mass spectrum matching of the next material component if the material is not present;
(2) and (3) performing secondary ion matching on the obtained mass center graph and the profile graph and the material component data in the volatile material database, if the deviation generated by the secondary ions in the matching result is less than or equal to 10ppm, determining that the material component is contained, otherwise, returning to the step (1) to perform primary mass spectrum matching of the next material component.
6. The method for ion source low energy mode determination of volatile substance components in Liupu tea as claimed in claim 1, wherein said volatile substance database stores information of Chinese and English names, molecular formulas, CAS numbers, exact molecular weights and mass defects of each volatile substance.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115480017A (en) * | 2022-10-12 | 2022-12-16 | 梧州市食品药品检验所 | Method for determining amino acid compounds in Liupu tea through high-resolution mass spectrometry |
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