CN111175420B - Separation and analysis method for aroma components in tobacco leaves - Google Patents
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Abstract
The application provides a separation and analysis method for aroma components in tobacco leaves, which specifically comprises the following steps: (1) Extracting the tobacco leaves by using water, an organic solvent or a mixed solvent of the water and the organic solvent to obtain a tobacco leaf extracting solution, and concentrating the tobacco leaf extracting solution to obtain a tobacco leaf extract; (2) Separating the tobacco leaf extract in the step (1) by using a chromatography method, grouping according to the aroma characteristics of each fraction, and concentrating respectively to obtain grouped concentrates; (3) And taking deuterated naphthalene and deuterated anthracene as internal standards, qualitatively analyzing the grouped concentrate by adopting a full-two-dimensional gas chromatography-mass spectrometry combined method, and screening to obtain the aroma components in the tobacco leaves. The method can effectively carry out qualitative and semi-quantitative analysis on the aroma components in the tobacco leaves, provides basis for further understanding the association between the aroma components of the tobacco leaves and the aroma style characteristics, and has a relatively high application prospect.
Description
Technical Field
The invention belongs to the technical field of separation and analysis, and particularly relates to a separation and analysis method for aroma components in tobacco leaves.
Background
The tobacco leaf aroma components are important factors influencing sensory evaluation of tobacco leaf aroma type style and quality, and research on the tobacco leaf aroma components is of great significance for revealing formation reasons of tobacco leaf style and quality. The tobacco leaf aroma substances mainly comprise volatile chemical substances with various components, complex compositions and low content, and can be divided into organic acids, phenols, alcohols, aldehyde ketones, esters (including lactones), heterocyclic compounds, amino acids, sugars, alkaloids and the like according to different properties of the chemical substances. The type, amount and quality of the aroma are influenced by the type, content and interaction of the aroma components of the tobacco leaves.
The tobacco leaf aroma component composition is extremely complex, and a method for objectively and effectively analyzing the tobacco leaf aroma component composition is developed to know the correlation between the tobacco leaf aroma component and the aroma style characteristics, so that the method has practical significance and application prospect in secondary development of tobacco leaves.
Disclosure of Invention
The prior art has some common problems in the analysis of the aroma components of the tobacco leaves: 1. most of tobacco leaf analysis samples are tobacco leaf essential oil, purified oil and the like, and are treated by means of simultaneous distillation extraction and the like, the treatment means relate to processing steps of high temperature (100 ℃ and above, such as steam distillation and simultaneous distillation extraction) or alcohol precipitation and the like, and the tobacco leaf material basis is changed correspondingly and cannot represent the chemical component composition contained in the tobacco leaf; 2. the tobacco leaf aroma components are quite complex in composition and extremely low in content, have different specific contributions to the aroma type, are directly and comprehensively analyzed without distinguishing and enriching due to lack of effective separation targets, so that the difficulty of analysis and identification is increased, the identified aroma components are numerous, some components have no key effect on the formation of the aroma type, and the analysis and screening of important aroma components are interfered; 3. in the aspect of processing complex samples and separating and qualitatively analyzing trace components, the gas chromatography (GC-MS) has the defects of insufficient resolution, poor separation effect, peak overlapping, insufficient response signals and the like, and for tobacco leaf samples with complicated chemical components and almost trace aroma components, the qualitative accuracy of the aroma components can be greatly influenced by the defects. Therefore, an analysis sample which represents the aroma component composition of the tobacco leaves as much as possible is selected, a key aroma characteristic component group is obtained through effective separation targeted separation, and a proper analysis means is combined to form the key for objectively knowing the aroma component composition of the tobacco leaves.
The tobacco leaf extract is a form which can basically show the self-aroma style of the tobacco leaves in original juice and original taste, the material basis of the tobacco leaf extract is not changed too much, the tobacco leaf extract is obtained by extracting the tobacco leaves with an ethanol solvent, gel chromatography (GPC) is used for pre-separating the tobacco leaf extract, each fraction is grouped according to aroma characteristics through olfactory characteristic evaluation, key aroma characteristic components are screened out, and the obtained key components are qualitatively analyzed by using a full two-dimensional gas chromatography-time of flight mass spectrum (GC multiplied by GC-TOFMS). The method is not only beneficial to separating and analyzing complex tobacco leaf aroma components, but also can correlate the subdivided key aroma characteristic components with the analyzed aroma components, and is beneficial to further understanding the internal relation between the tobacco leaf aroma style and the aroma components.
Therefore, the application provides a method for separating and analyzing aroma components in tobacco leaves, which comprises the following steps:
(1) Extracting the tobacco leaves by using water, an organic solvent or a mixed solvent of the water and the organic solvent to obtain a tobacco leaf extracting solution, and concentrating the tobacco leaf extracting solution to obtain a tobacco leaf extract;
(2) Separating the tobacco leaf extract in the step (1) by using a chromatography method, grouping according to the aroma characteristics of each fraction, and concentrating respectively to obtain grouped concentrates;
(3) And taking deuterated naphthalene and deuterated anthracene as internal standards, carrying out qualitative analysis on the grouped concentrate by adopting GC-TOF MS, and screening to obtain aroma components in the tobacco leaves.
In some embodiments, the chromatography in step (2) is selected from silica gel column chromatography, gel permeation chromatography, or macroporous resin method.
In some embodiments, the filler for gel permeation chromatography is selected from the group consisting of Sephadex G-10, sephadex G-15, sepharose 6B, sepharose ConA, sephadex G-25 and Sephadex LH-20, preferably Sephadex G-10, sephadex G-15 or Sephadex LH-20, more preferably Sephadex LH-20. In some embodiments, the gel chromatography is performed in a glass chromatography column (2.6 cm × 100 cm) (e.g., buchi, switzerland). In some embodiments, elution is performed with the following solvents as the mobile phase: ethanol, propanol, isopropanol, propylene glycol, water, or a mixed system thereof, preferably ethanol. In some embodiments, the flow rate of the mobile phase is 1-8mL/min, such as 1mL/min,2mL/min,3mL/min,4mL/min,5mL/min,6mL/min,7mL/min, or 8mL/min.
In some embodiments, the macroporous resin process filler is selected from Diaion HP-10, -20, -30, -40, or-50, AB-8, D101, D102, and D402; preferably, elution is carried out with the following solvents as mobile phase: ethanol, methanol, isopropanol, water, or a mixed system thereof, preferably an aqueous ethanol solution, for example, a 20% aqueous ethanol solution.
In some embodiments, the extraction in step (1) is performed as follows: extracting the tobacco leaves with 50% -95% ethanol (such as 50% ethanol, 70% ethanol or 95% ethanol), dichloromethane or water as solvent at 50-80 deg.C or under reflux for 10min-2h (such as 10min, 30min, 1h or 2 h).
In some embodiments, the concentrating in step (1) is concentrating under reduced pressure; preferably, the density of the concentrated solution is controlled to be 1.36-1.42g/cm 3 。
In some embodiments, in step (3), the analysis conditions of the comprehensive two-dimensional gas chromatograph are selected from one or more of the following:
(1) A first chromatographic column: HP-5MS (60 m.times.0.25 mm.times.0.25 μm);
(2) And (2) a second chromatographic column: DB-l7 (2.3 m.times.0.1 mm.times.0.1 μm);
(3) Column temperature procedure: 50 ℃ (1 min) → 3 ℃/min → 220 ℃ → 2 ℃/min → 260 ℃ (10 min);
(4) Carrier gas: he;
(5) The carrier gas control mode is as follows: constant current, 1-1.5mL/min;
(6) Sample inlet temperature: 250-300 ℃;
(7) Modulation period: 3-6 seconds;
(8) Sample introduction amount: 1 mu L of the solution;
(9) Shunting mode: split-flow sample injection, wherein the split-flow ratio is 10-30.
In some embodiments, in step (3), the analysis conditions of the mass spectrum are selected from one or more of the following:
(1) An ionization mode: EI;
(2) Ion source temperature: 220 to 280 ℃;
(3) GC/MS transmission line temperature: 250-260 ℃;
(4) EI ionization energy: 70eV;
(5) Solvent delay time: 7-16min;
(6) The collection mode is as follows: scanning the whole Scan;
(7) The mass range is as follows: 45-450 amu.
In some embodiments, in step (3), the qualitative analysis refers to matching the mass spectrum measured by GC × GC-TOF MS with the standard mass spectrum of the compound in the NIST standard library, and selecting the compound with the forward and reverse matching degree of more than 700 (e.g. more than 800, more than 900 or more than 999), i.e. determining the chemical composition corresponding to the corresponding chromatographic peak.
In some embodiments, the tobacco leaf is trimming yuxi tobacco leaf, for example, redried trimming yuxi tobacco leaf.
Advantageous effects of the invention
The application provides a method for separating and analyzing aroma components of tobacco leaves, which comprises the steps of taking tobacco leaf extract as an analysis sample, primarily separating the sample by adopting a chromatography method, enriching fractions with similar aroma characteristics through olfactory characteristic evaluation, further separating the fractions with the similar aroma characteristics by combining a full-two-dimensional gas chromatography-flight time mass spectrum, and determining the nature of each chemical substance in the fractions, thereby screening the aroma components. The method can effectively separate and qualitatively analyze the aroma components of the tobacco leaves. In addition, key fractions capable of reflecting the flavor style of the tobacco leaves can be quickly positioned by means of olfactory characteristic evaluation, the content of the aroma components in the components is greatly improved, the components are simplified, and the separation and qualitative advantages of the full-two-dimensional gas chromatography-flight time mass spectrum are combined, so that the analyzed aroma components of the key components and the aroma characteristic evaluation of the key components can be well verified mutually, and a basis is provided for further understanding the association between the aroma components of the tobacco leaves and the flavor style characteristics. Therefore, the method has a high application prospect.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Example 1
1. Method for extracting Sanming Youyxi tobacco leaves
200g of Sanming Youxi YLC01-2016 tobacco leaves (redried tobacco leaves) are extracted by 1600g of 95% ethanol solvent, the water bath temperature is controlled at 80 ℃, and the extraction time is 1h. The obtained tobacco leaf alcohol extract is subjected to reduced pressure concentration, and the density of the concentrated solution is controlled to be 1.38g/cm 3 To obtain 64.8g of tobacco leaf extract.
2. Gel chromatography column (GPC) fractionation
Dissolving the 2g of tobacco leaf alcohol extract in 4mL of absolute ethyl alcohol, fully shaking and filtering, and performing GPC separation on a filtered sample, wherein the filler is Sephadex LH-20, the absolute ethyl alcohol is used as a mobile phase, and the flow rate is 4mL/min. One fraction was collected every 3min by the fraction collector, and a total of 30 fractions were collected, labeled F1, F2, F3, F4, F5, F6 … F30, respectively. And 6 incense evaluators perform olfactory characteristic evaluation one by one, and the olfactory characteristics in the evaluation results of the fractions are combined to ensure that chemical components with similar aroma are enriched as much as possible so as to be beneficial to analysis of trace components, and the combined components are subjected to olfactory characteristic evaluation to obtain the final evaluation result (the grouping condition and the corresponding olfactory characteristic evaluation result are shown in table 1).
The olfactory characteristics evaluation is carried out according to the fragrance evaluation method in the field of flavors and fragrances (GB/T14454.2-2008 fragrance evaluation method).
TABLE 1 grouping of the streams according to the charm characteristics
From table 1, it can be seen that: (1) the three mings youxi tobacco leaves are generally more concerned about fresh and sweet and honey aroma, while neglecting other aroma, the burnt sweet aroma presented by the component 4 proves that the burnt sweet aroma is also an essential component for explaining the three mings youxi style characteristics. (2) The olfaction characteristics shown by the components 1-4 are different, and a part of the components show a phenomenon of crossing more than two aroma characteristics. Taking the olfaction characteristic evaluation results of the components 2 to 4 as an example, the middle bottom of the component 2 can distinguish strong dry grass fragrance, the component 3 can still distinguish the dry grass fragrance, but the fragrance is weakened compared with the upper component, and meanwhile, a new fragrance note, namely, a scorched sweet fragrance appears at the bottom, while the component 4 can distinguish an obvious scorched sweet fragrance. Two possible reasons for the intersection of aroma characteristics are that GPC does not completely separate different aroma components, the same compound appears in different components before and after, and the other aroma component which is the dominant aroma is not the same compound, so that the elution time is different. Nevertheless, the main aroma characteristics of different components are different, which shows that the separation and enrichment of aroma components according to the aroma characteristics can be realized under the condition. The aroma characteristics of the component 5 are not obvious and the aroma is weak, and the aroma characteristics of the combined components 1-4 are similar to the aroma characteristics of the original tobacco leaf extract, so that the components 1-4 are used as key aroma characteristic components.
3. Qualitative analysis
The key fractions 1-4 were collected separately and concentrated to 1.5mL under reduced pressure for analysis by GC × GC-TOF MS. And automatically searching each spectral peak by using an NIST standard spectral library, manually checking the search result, selecting the forward and reverse search matching degrees to be more than 800, and determining chemical components.
Chromatographic conditions are as follows:
united states of America LECO company comprehensive two-dimensional gas chromatography-time of flight mass spectrometer (GC x GC-TOF); a first chromatographic column: HP-5MS (60 m.times.0.25 mm.times.0.25 μm); and (2) a second chromatographic column: DB-l7 (2.3 m 0.1mm 0.1 μm); column temperature procedure: 50 ℃ (1 min) → 3 ℃/min → 220 ℃ → 2 ℃/min → 260 ℃ (10 min); carrier gas: he; the carrier gas control mode is as follows: constant current, 1mL/min; sample inlet temperature: 280 ℃; modulation period: 5sec; sample injection amount: 1 mu L of the solution; shunting mode: split-flow sample injection, split-flow ratio 10.
Mass spectrum conditions:
an ionization mode: EI; ion source temperature: 230 ℃; GC/MS transmission line temperature: 260 ℃; EI ionization energy: 70eV. Solvent delay time: 7min; the collection mode is as follows: scanning the whole Scan; the mass range is as follows: 45 to 450amu.
Internal standard substance: deuterated naphthalene and deuterated anthracene.
When each component obtained by GPC separation is analyzed by GC XGC-TOF MS, complex chemical components are well separated, the composition of the chemical components is simplified, qualitative identification and content calculation are facilitated, and the analysis result of GC XGC-TOF MS of the component 1 aroma component is shown in Table 2.
TABLE 2 GC XGC-TOF MS analysis results of component 1 aroma component
By analyzing the chemical components of the components, the following results can be obtained: (1) from the chemical composition, the components 1-2 are similar in chemical composition, mainly aldehydes and ketones formed by terpene degradation, wherein part of the components are partially crossed in each fraction, and the aldehydes and ketones comprise 6-methyl-5-octene-2-one, solanone, beta-damascenone, geranylacetone, beta-ionone, dihydroactinidiolide, megastigmatrienone and other fragrant, flowery scent and hay scent compounds; the main component of the component 3 comprises aldehyde and ketone formed by degrading the terpenes, such as faint scent, floral scent and hay scent compounds such as 4-oxo-isophorone, dihydro-actinidiolide, megastigmatrienone and the like, and also comprises heterocyclic compounds such as caramel, angelica lactone, 5-methylfurfural, 5-hydroxymethyl-furfural, methyl cyclopentene alcohol ester, maltol and other scorch scent compounds; the component 4 mainly contains heterocyclic and phenolic compounds, including burnt or spicy fragrant compounds such as furfural, angelica lactone, 5-methylfurfural, 5-hydroxymethyl furfural, methyl cyclopentenol ester, maltol, 2-acetyl pyrrole, guaiacol, 4-vinyl guaiacol and vanillin. The component analysis results of the components 1 to 4 and the olfaction characteristic evaluation thereof can be mutually verified. (2) From the structure of the chemical components, the chemical components of the components 1-2 are basically chain aldehyde, ketone, alcohol structures or chain-carrying cyclic structures (such as propiophenone, megastigmatrienone and the like); the chemical components of the component 4 are basically cyclic compounds such as furan rings, furanones, benzene rings and the like. From each component, 117 kinds of aroma components were identified, wherein, 32 kinds of aldehydes, 38 kinds of ketones, 12 kinds of esters, 4 kinds of alcohols, 12 kinds of phenols, 2 kinds of ethers, 11 kinds of heterocycles, and 6 kinds of acids.
The tobacco leaf aroma components are separated by GPC, and are grouped according to aroma characteristics by sensory assistance, key aroma characteristic components are further screened, the aroma characteristics of all key components can well reflect the aroma style of the tobacco leaves, and the aroma components of the components 1-4 analyzed by GC x GC-TOF MS and the aroma characteristic evaluation thereof can well carry out mutual evidence, so that the internal relation between the tobacco leaf aroma components and the tobacco leaf aroma style can be deeply understood.
Example 2
GPC fractionation
Taking out 2g of the tobacco leaf extract obtained in the example 1, dissolving the tobacco leaf extract in 4mL of absolute ethyl alcohol, fully shaking, filtering, and carrying out gel permeation chromatography separation on a filtered sample, wherein the filler is Sepharose4B, the absolute ethyl alcohol is used as a mobile phase, and the flow rate is 4mL/min. Fractions were collected by fraction collector every 3min for a total of 30 fractions, labeled F1, F2, F3, F4, F5, F6 … F30. Olfactory characteristic evaluation is carried out by 6 incense evaluators one by one (the specific method is shown in example 1), and the olfactory characteristics in the evaluation results of the fractions are closely merged by discussion and confirmation of the incense evaluators, so that chemical components with similar aroma are enriched as much as possible, and trace components are analyzed, and the merged components are subjected to olfactory characteristic evaluation to obtain final evaluation results (the merged groups and the corresponding olfactory characteristic evaluation results are shown in table 3).
TABLE 3 evaluation results of each fraction grouping and its corresponding olfaction characteristics
From table 3, it can be seen that: (1) component 5 had no obvious olfactive characteristics. This may be due to the fact that the last eluted component does not show a clear olfactive character due to its very low concentration. (2) From components 1 to 4, the olfactory characteristics exhibited show a phenomenon of intersection of more than two aroma characteristics. The aroma of the components 2-3 is similar, and the difference lies in that the aroma of the component 3 is weakened compared with that of the component 2, and the aroma among the components is not regularly distinguished, namely the tobacco aroma components are not regularly separated according to the grouping of aroma characteristics, and the aroma is mutually influenced in a cross way. Fractions 1-5 were further collected separately and concentrated to 1.5mL under reduced pressure for analysis by GC × GC-TOF MS.
2. Qualitative analysis
The analysis conditions were the same as in example 1, and the results are shown in Table 4.
TABLE 4 GC XGC-TOF MS analysis of component 2 fragrance component
By analyzing the chemical compositions of the components, the following results can be obtained: (1) from the chemical composition, the components 2 to 3 are similar in chemical composition and mainly contain aldehydes and ketones formed by terpene degradation, wherein the aldehydes and ketones are representative components of faint scent, flower fragrance and hay fragrance, and also contain Xu Duojiao sweet, spicy and bean fragrance components; the component 4 mainly comprises the following components of furfural, angelica lactone, 5-methylfurfural, 5-hydroxymethyl furfural, methyl cyclopentenol ester, maltol, 2-acetyl pyrrole, guaiacol and other burnt aroma compounds. (2) From the structure of chemical components, the elution time of aldehyde and ketone is similar to that of heterocyclic compounds, and the elution time is not too regular. In the example, the tobacco leaf aroma components are separated by GPC, the regularity of grouping separation according to aroma characteristics is not strong through sensory assistance, aroma is mutually influenced in a cross mode, the composition of each component aroma component analyzed by GC × GC-TOFMS is complex, the aroma components cannot be well associated with the aroma characteristics, and the method does not help greatly to understand the internal relationship between the tobacco leaf aroma components and the aroma style.
Example 3
The concentrated solution obtained in example 1 by fractionation on a gel column was prepared as an analysis sample and analyzed by a gas chromatography-mass spectrometer (GC-MS). And automatically searching each spectral peak by using an NIST standard spectral library, manually checking the search result, selecting the search matching degree to be more than 800, and determining chemical components.
The GC-MS conditions used were: agilent 7890-5975C GC, USA, chromatographic column: DB-17MS (60 m 0.25mm 0.25 u m); column temperature procedure: 60 ℃ (2 min) → 2 ℃/min → 240 ℃ (2 min) → 10 ℃/min → 300 ℃ (50 min); carrier gas: he; the carrier gas control mode is as follows: constant current, 1mL/min; sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L of the solution; shunting mode: split-flow injection, split-flow ratio 3:1.
Conditions of Mass Spectrometry
An ionization mode: EI; ion source temperature: 230 ℃; GC/MS transmission line temperature: 280 ℃; EI ionization energy: 70eV. Solvent delay time: 6.2min; the collection mode is as follows: scanning the whole Scan; the mass range is as follows: 35-650 amu.
The components obtained by GPC are analyzed by GC-MS, and 52 kinds of aroma components are identified, wherein 15 kinds of aldehydes, 16 kinds of ketones, 4 kinds of esters, 4 kinds of alcohols, 4 kinds of phenols, 2 kinds of ethers, 4 kinds of heterocycles, and 3 kinds of acids. From component 1 (as shown in table 2), the detected components are far less than the components analyzed by GC × GC-TOF MS, the number of detected aroma components is greatly reduced, and many trace aroma components cannot be detected, such as beta-damascenone, beta-ionone, 3-penten-2-one, 6-methyl-5-octen-2-one, farnesal and the like, which are known important aroma components in tobacco leaves although the content of the aroma components is low, and the lack of the aroma components has a great influence on the association of aroma characteristics of each key component. This comparison shows that the analytical method in example 1 has great advantages for the trace aroma component detection of each key component after GPC pre-separation.
TABLE 5 GC-MS analysis results of the aroma components in component 1
And (4) conclusion:
the fragrant components of the Sanming Yinxi tobacco leaf extract are analyzed by adopting semi-preparative gel chromatography separation and combining a full two-dimensional gas chromatography-time of flight mass spectrometer (GC multiplied by GC-TOF), and 117 fragrant components are identified in total, wherein 32 aldehydes, 38 ketones, 12 esters, 4 alcohols, 12 phenols, 2 ethers, 11 heterocycles and 6 acids are included. The method comprises the steps of extracting tobacco leaves by using 95% ethanol, separating an obtained ethanol extract concentrated solution by using a gel chromatographic column for pre-separation, obtaining 5 components through sensory auxiliary screening, selecting 4 key aroma characteristic components from the 5 components, and detecting each key component by using a full two-dimensional gas chromatography-time of flight mass spectrometer (GC x GC-TOF). The result shows that each key component subjected to GPC pre-separation can well reflect the aroma style characteristics of the tobacco leaves, the content of the aroma components of each component is greatly improved, the composition is relatively simple, and the analyzed aroma components and the sensory evaluation results of each key component can be associated and verified. The research results provide theoretical basis for further understanding the association between the aroma components and the flavor style characteristics of the tobacco leaves.
Although specific embodiments of the invention have been described in detail, it will be appreciated by those skilled in the art that, based upon the overall teachings of the disclosure, various modifications and alternatives to those details could be developed and still be encompassed by the present invention. The full scope of the invention is given by the appended claims and any equivalents thereof.
Claims (10)
1. A method for separating and analyzing aroma components in tobacco leaves comprises the following steps:
(1) Extracting the tobacco leaves for 1 hour by using 95% ethanol water solution under the condition of 80 ℃ water bath to obtain tobacco leaf extracting solution, and concentrating the tobacco leaf extracting solution to obtain tobacco leaf extract;
(2) Separating the tobacco leaf extract in the step (1) by using a gel permeation chromatography, grouping according to aroma characteristics of each fraction, and concentrating respectively to obtain a grouped concentrate, wherein a filler of the gel permeation chromatography is Sephadex LH-20, ethanol is used as a mobile phase for elution, and the flow rate of the mobile phase is 1-8mL/min;
(3) And (3) taking deuterated naphthalene and deuterated anthracene as internal standards, qualitatively analyzing the grouped concentrate obtained in the step (2) by adopting GC × GC-TOF MS, and screening to obtain aroma components in the tobacco leaves, wherein the GC × GC-TOF MS chromatographic analysis conditions are as follows:
1) A first chromatographic column: HP-5MS, 60mX0.25 mm X0.25 μm;
2) And (2) a second chromatographic column: DB-l7,2.3 m.times.0.1 mm.times.0.1 μm;
3) Column temperature procedure: 50 ℃/1min → 3 ℃/min → 220 ℃ → 2 ℃/min → 260 ℃/10min;
4) Carrier gas: he;
5) The carrier gas control mode is as follows: constant current, 1-1.5mL/min;
6) Sample inlet temperature: 250-300 ℃;
7) Modulation period: 3-6 seconds;
8) Sample introduction amount: 1 mu L of the solution;
9) A shunting mode: split-flow sample injection, wherein the split-flow ratio is 10-30;
the GC XGC-TOF MS mass spectrum analysis conditions are as follows:
1) An ionization mode: EI;
2) Ion source temperature: 220 to 280 ℃;
3) GC/MS transmission line temperature: 250-260 ℃;
4) EI ionization energy: 70eV;
5) Solvent delay time: 7-16min;
6) The collection mode is as follows: scanning the whole Scan;
7) The mass range is as follows: 45-450 amu.
2. The method of claim 1, wherein the flow rate of the mobile phase in step (2) is 1mL/min,2mL/min,3mL/min,4mL/min,5mL/min,6mL/min,7mL/min, or 8mL/min.
3. The method of claim 1, wherein in step (1), the concentrating is concentrating under reduced pressure.
4. The method according to claim 1, wherein in the step (1), the density of the concentrate is controlled to be 1.36 to 1.42g/cm 3 。
5. The method of claim 1, wherein in step (3), the qualitative analysis is performed by matching the mass spectrum measured by GC x GC-TOF MS with the standard mass spectrum of the compound in the NIST standard library, and selecting the compound with the forward and reverse matching degree of more than 700, i.e. determining the corresponding chemical composition of the corresponding chromatographic peak.
6. The method of claim 5, selecting a compound having a forward and reverse degree of match of greater than 800.
7. The method of claim 5, selecting a compound having a forward and reverse degree of match of greater than 900.
8. The method of claim 5, selecting a compound having a forward and reverse degree of match of greater than 999.
9. The method of claim 1, wherein the tobacco leaf is trimming yuxi tobacco leaf.
10. The method of claim 1, wherein the tobacco leaf is redried trimming yuxi tobacco leaf.
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| CN102692465A (en) * | 2012-06-25 | 2012-09-26 | 中国烟草总公司郑州烟草研究院 | Sensory omics determination method for tobacco and tobacco products |
| CN104849393A (en) * | 2015-06-09 | 2015-08-19 | 安徽中烟工业有限责任公司 | Method for determining glucosidic bonded fragrance component in tobacco |
| CN106353419B (en) * | 2016-08-16 | 2021-01-22 | 国家烟草质量监督检验中心 | Method for measuring flavor components in cigarette mainstream smoke |
| CN106680408A (en) * | 2016-12-30 | 2017-05-17 | 中国烟草总公司广东省公司 | Method for analyzing aromatic substances of flue-cured tobacco leaves in different tobacco-growing areas |
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