Method for simultaneous extraction and determination of tobacco ether extract and glandular hair secretion components

Abstract

The invention discloses a method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components, which comprises: extracting tobacco ether extract with petroleum ether and determining the content of petroleum ether extract; dissolving glandular hair secretion components with ethanol solution and removing high molecular fat-soluble impurities; determining the content of diterpenoid aroma components in tobacco glandular hair secretion components: quantitatively taking glandular hair secretion component solution, diluting with mobile phase, and simultaneously determining the content of cephatriene glycol, cephatriene monool and cis-abirol by liquid chromatography; determining the content of sucrose ester in tobacco glandular hair secretion components: dissolving the remaining glandular hair secretion components with liquid nitrogen and blowing to dryness, and determining the content of sucrose ester in tobacco glandular hair secretion components by gas chromatography-mass spectrometry after derivatization. The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components of the invention can simultaneously extract and determine tobacco ether extract and glandular hair secretion main components, is simple to operate, has high detection efficiency, and reduces solvent waste.

Description

Technical Field

[0001] The invention relates to the technical field of tobacco leaf characteristic aroma component detection, in particular to a method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components. Background Art

[0002] Tobacco petroleum ether extract, referred to as ether extract, is a mixture obtained by extracting tobacco leaf samples with petroleum ether as solvent. Its components mainly include resins, waxes, lipids, sterols, oils, volatile oils, fatty acids, pigments, etc. Most of these substances are closely related to the aroma and oil content of tobacco. Tobacco leaves with high ether extract content have full chroma, strong gloss, high oil content, good elasticity and full aroma.

[0003] Tobacco leaves, stems, and flowers are densely covered with glandular hairs. The secretion components of tobacco glandular hairs are characteristic secondary metabolites of tobacco, mainly cepetane diterpenes, cepetane diterpenes, and sucrose ester compounds. At present, 125 types of tobacco cepetane diterpenes have been discovered. The most abundant cepetane diterpenes in tobacco are cepetane triene diol (α-CBT-diol, β-CBT-diol) and cepetane triene monool (CBT-ol), reaching more than 90%; cis-abietanol is the most important cepetane compound in tobacco, which is generally found in oriental tobacco, and generally cultivated flue-cured tobacco does not contain cis-abietanol; cultivated flue-cured tobacco sucrose ester compounds are generally divided into 6 types (SEⅠ-SEⅥ) according to different molecular weights (622, 636, 650, 664, 678, 692).

[0004] During the burning process of cigarettes, most of the cedarene diterpenes undergo pyrolysis to generate flavor components such as solanone and norsolanone. Solanone and oxysolanone give the smoke a ketone flavor; tobacco lysine degradation products such as ambroxan and ambergris lactone have a strong ambergris (cedar aroma) aroma and are considered to be the main compounds that cause the characteristic aroma of oriental tobacco; sugar esters can be pyrolyzed during the smoking process of tobacco to release low-level carboxylic acids that have a positive effect on the taste of smoke, giving the smoke an aromatic flavor and making the smoke delicate and mellow. The secretion components of tobacco glandular hairs are not only important precursors of cigarette aroma, but cedarene diterpenes also have good antibacterial, anti-tumor, and anti-insect activities, and have good development value in the direction of plant-based pesticides and medicines.

[0005] Tobacco ether extracts and glandular hair secretion components have an important impact on the aroma quality of cigarettes and are important indicators for evaluating tobacco aroma quality. Systematic determination of tobacco aroma components and clarifying the effects of different varieties, different parts or different ecological and cultivation conditions on the content of main aroma components are important contents of tobacco quality evaluation.

[0006] At present, the determination of ether extracts generally refers to the industry standard "Determination of petroleum ether extracts of tobacco and tobacco products (YC / T176-2003)". The GC-MS method is currently the industry recommended standard method for the detection of tobacco cephalotriene diol. This method requires a complex post-derivation reaction, the cost of the derivatization reagent is high, and it is easy to cause chromatographic column contamination; in recent years, a method for determining the content of cephalotriene diol (α-CBT-diol, β-CBT-diol) by liquid chromatography has been proposed. This method is simple to operate, uses less solvent, has a short detection cycle, and has high precision and accuracy. At present, there are few reports on the determination of cephalotriene monool (CBT-ol). A method for determining cis-abietanol by liquid chromatography (ZL 201710047317.4) has been proposed. At present, there are no reports on the simultaneous extraction and determination of tobacco cephalotriene diol, cephalotriene monool and cis-abietanol. In addition, a method for analyzing sucrose ester content was also proposed. This method established a method for determining the sucrose ester (SE) content in oriental tobacco by gas chromatography / mass spectrometry using sucrose octaacetate as an internal standard, dichloromethane ultrasonic extraction, N,O-bis(trimethylsilyl)trifluoroacetamide derivatization. In summary, under the current experimental technical conditions, the determination of tobacco ether extracts and the main components of glandular hair secretion (cebutriene diol, cebutriene monool, cis-abirol, sucrose esters) requires a large amount of work, a long working cycle, and a large waste of reagents.

[0007] Therefore, there is an urgent need for a method for simultaneously extracting and determining tobacco ether extracts and glandular hair secretion components. Summary of the invention

[0008] The object of the present invention is to provide a method for simultaneously extracting and determining components of tobacco ether extract and glandular hair secretion, so as to solve the above-mentioned problems in the prior art. The method can simultaneously extract and determine the main components of tobacco ether extract and glandular hair secretion (cephalotriene glycol, cephalotriene monool, cis-abietanol, sucrose esters), has simple operation, high detection efficiency and reduces solvent waste.

[0009] The present invention provides a method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components, which comprises:

[0010] Extracting tobacco ether extract with petroleum ether and determining the content of tobacco petroleum ether extract;

[0011] Using ethanol solution to dissolve the glandular hair secretion components in the tobacco petroleum ether extract and removing high molecular fat-soluble impurities to obtain a glandular hair secretion component solution;

[0012] Determination of the content of diterpenoid aroma components in the glandular hair secretion components of tobacco: quantitatively pipette the glandular hair secretion component solution, dilute it with a mobile phase, and simultaneously determine the contents of cephariol, cephariol, and cis-abieol in the glandular hair secretion components of tobacco by liquid chromatography;

[0013] Determination of sucrose ester content in tobacco glandular hair secretion components: The remaining glandular hair secretion components were dissolved in liquid nitrogen and blown to dryness. After derivatization, the sucrose ester content in the tobacco glandular hair secretion components was determined by gas chromatography-mass spectrometry.

[0014] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components as described above, wherein preferably, the method of extracting tobacco ether extract with petroleum ether and determining the content of tobacco petroleum ether extract specifically comprises:

[0015] Refer to the industry standard YC / T 176-2003 and use petroleum ether at 30℃-60℃ to extract tobacco ether extract;

[0016] After the extraction is completed, the solvent is removed by vacuum rotary evaporation at 35°C-45°C;

[0017] Wipe off the moisture on the outside of the rotary evaporator, put it in a desiccator and dry it to constant weight, weigh it, and calculate the content of tobacco petroleum ether extract.

[0018] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components as described above, wherein preferably, the glandular hair secretion components in the tobacco petroleum ether extract are dissolved by using an ethanol solution and high molecular weight fat-soluble impurities are removed to obtain a glandular hair secretion component solution, specifically comprising:

[0019] After the tobacco petroleum ether extract is weighed and the content is determined, an ethanol solution with a volume concentration of 80%-85% is added;

[0020] Under normal temperature, the tobacco petroleum ether extract was dissolved by ultrasound;

[0021] Transfer the dissolved solution of petroleum ether extract into a volumetric flask, dissolve it 2-4 times continuously, make up to volume, and wait for testing.

[0022] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components as described above, wherein preferably, in determining the content of diterpenoid aroma components in tobacco glandular hair secretion components, the glandular hair secretion component solution is quantitatively pipetted, diluted with a mobile phase, and the contents of cephalotriene glycol, cephalotriene monool, and cis-abietanol in the tobacco glandular hair secretion components are simultaneously determined by liquid chromatography, specifically comprising:

[0023] Quantitatively remove the glandular hair secretion component solution and dilute it with an acetonitrile solution with a volume concentration of 80%-85%;

[0024] The contents of cephariene glycol, cephariene monool and cis-abieol in the secretion components of tobacco glandular hairs were detected using a DAD detector under liquid chromatography gradient elution conditions.

[0025] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components as described above, wherein, preferably, under the liquid chromatography gradient elution conditions, the liquid chromatography conditions are: ultra-high performance liquid chromatography, the analytical column is ACQUITY UPLC BEH C18 chromatographic column, specifications are 2.1mm×50mm, 1.7μm, the mobile phases are acetonitrile as mobile phase A and ultrapure water as mobile phase B, the elution program is 0min, 20% mobile phase A, 80% mobile phase B; 2min, 40% mobile phase A, 60% mobile phase B; 4min, 60% mobile phase A, 40% mobile phase B; 6min, 80% A; 8min, 100% A; 9min, 100% A; 11min, 80% mobile phase A, 20% mobile phase B; 13min, 60% mobile phase A, 40% mobile phase B; 14min, 40% mobile phase A, 60% mobile phase B; 16min, 20% mobile phase A, 80% mobile phase B; the flow rate is 0.3mL / min, the column temperature is 35℃, and the external standard method is used for quantification.

[0026] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components as described above, wherein preferably, in determining the content of sucrose ester in tobacco glandular hair secretion components, the remaining glandular hair secretion components are dissolved in liquid nitrogen and blown to dryness, and the sucrose ester content in the tobacco glandular hair secretion components is determined by gas chromatography-mass spectrometry after derivatization, specifically comprising:

[0027] After determining the content of diterpenoid aroma components in the glandular hair secretion components of tobacco, add a petroleum ether internal standard solution containing sucrose octaacetate to the remaining glandular hair secretion component solution, the concentration is 0.5 mg / mL-0.7 mg / mL, the volume is 80 μL-120 μL, and blown to dryness with nitrogen;

[0028] Add a mixture of dimethylformamide and N,O-bistrimethyltrifluoroacetamide in a volume ratio of 0.8:1-1.2:1, heat at 70°C-80°C for 0.8h-1.2h, derivatize, pass through an organic filter with a pore size of 0.22μm, and perform GC / MS determination;

[0029] Semi-quantitative analysis was performed using sucrose octaacetate as an internal standard.

[0030] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components as described above, wherein, preferably, in the GC / MS determination, the GC / MS chromatographic conditions are: the chromatographic column is HP-MS, the specifications are 30m×250μm×0.25μm, the carrier gas is helium, the flow rate is 0.8mL / min, the injection port temperature is 290°C, the split mode is used for injection, the injection volume 1 is μL, the split ratio is 5:1, the starting temperature of the chromatographic column is 230°C, maintained for 1 min, increased to 270°C at a rate of 5°C / min, maintained for 2 min, increased to 290°C at a rate of 1°C / min, and maintained for 4 min.

[0031] The invention provides a method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components, which can realize the simultaneous extraction and detection of tobacco ether extract and main aroma components of glandular hair secretion, and realizes the simultaneous high-performance liquid chromatography detection of main diterpenoid aroma components of tobacco glandular hair secretion, namely, α-cepinatriene diol, β-cepinatriene diol, cepinatriene monool and cis-abirol for the first time; in the process of detecting tobacco glandular hair secretion components, an ethanol solution with a concentration of 80%-85% can be used to dissolve the tobacco petroleum ether extract, so that the target detection components can be efficiently dissolved, and the pollution of high-molecular fat-soluble components such as wax, solanesol and phytosterol on liquid chromatography and gas chromatography-mass spectrometry chromatograph used in the process of detecting glandular hair secretion components and the interference on the target detection components can be avoided; compared with the currently commonly used detection method for the target components, the method of the invention has simple operation steps, saves solvent, improves the detection efficiency and reduces the detection cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further described below with reference to the accompanying drawings, in which:

[0033] Figure 1 A flow chart of an embodiment of the method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components provided by the present invention;

[0034] Figure 2 Liquid chromatogram of the test sample;

[0035] Figure 3 This is the total ion current of the detected sample. DETAILED DESCRIPTION

[0036] Various exemplary embodiments of the present disclosure will now be described in detail with reference to the accompanying drawings. The description of the exemplary embodiments is merely illustrative and is in no way intended to limit the present disclosure and its application or use. The present disclosure can be implemented in many different forms and is not limited to the embodiments described herein. These embodiments are provided to make the present disclosure thorough and complete and to fully express the scope of the present disclosure to those skilled in the art. It should be noted that unless otherwise specifically stated, the relative arrangement of the components and steps, the composition of the materials, the numerical expressions and the numerical values ​​set forth in these embodiments should be interpreted as being merely exemplary and not as limiting.

[0037] The words "first", "second" and similar words used in this disclosure do not indicate any order, quantity or importance, but are only used to distinguish different parts. The words "include" or "comprises" and similar words mean that the elements before the word include the elements listed after the word, and do not exclude the possibility of including other elements. "Up", "down" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0038] In the present disclosure, when a specific component is described as being located between a first component and a second component, there may or may not be an intermediate component between the specific component and the first component or the second component. When a specific component is described as being connected to other components, the specific component may be directly connected to the other components without an intermediate component, or may not be directly connected to the other components but have an intermediate component.

[0039] All terms (including technical terms or scientific terms) used in the present disclosure have the same meanings as those understood by ordinary technicians in the field to which the present disclosure belongs, unless otherwise specifically defined. It should also be understood that terms defined in general dictionaries, such as general dictionaries, should be interpreted as having meanings consistent with their meanings in the context of the relevant technology, and should not be interpreted in an idealized or extremely formal sense, unless explicitly defined herein.

[0040] Technologies, methods, and equipment known to ordinary technicians in the relevant art may not be discussed in detail, but where appropriate, the technologies, methods, and equipment should be considered part of the specification.

[0041] like Figure 1 As shown, the embodiment of the present invention provides a method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components, which specifically includes the following steps during actual implementation:

[0042] Step S1, extracting tobacco ether extract with petroleum ether, and determining the content of the tobacco petroleum ether extract.

[0043] Wherein, before step S1, a step of preparing a sample is also included. In a specific implementation, fresh tobacco leaves of the oriental tobacco variety Samsun are taken, the tendons are removed, freeze-dried, and crushed. In one embodiment of the method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components of the present invention, the step S1 may specifically include:

[0044] Step S11, extracting tobacco ether extract using petroleum ether at 30° C.-60° C. according to the industry standard YC / T 176-2003.

[0045] Step S12: After the extraction is completed, the solvent is removed by rotary evaporation under reduced pressure at 35° C.-45° C.

[0046] Step S13, wipe off the moisture on the outside of the rotary evaporator, put it into a dryer and dry it to constant weight, weigh it, and calculate the content of the tobacco petroleum ether extract.

[0047] In one embodiment of the present invention, the content of the aromatic tobacco petroleum ether extract measured by the above method is 6.25%.

[0048] Step S2, using an ethanol solution to dissolve the glandular hair secretion components in the tobacco petroleum ether extract and remove high-molecular fat-soluble impurities to obtain a glandular hair secretion component solution.

[0049] In one embodiment of the method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components of the present invention, step S2 may specifically include:

[0050] Step S21: After weighing and determining the content of the tobacco petroleum ether extract, add an ethanol solution with a volume concentration of 80%-85%.

[0051] Step S22: dissolving the tobacco petroleum ether extract by ultrasound at room temperature.

[0052] Step S23, transfer the dissolved solution of the petroleum ether extract into a volumetric flask, dissolve it 2-4 times continuously, fix the volume, and wait for testing.

[0053] Step S3, determination of the content of diterpenoid aroma components in the glandular hair secretion components of tobacco: quantitatively pipette the glandular hair secretion component solution, dilute it with the mobile phase, and use liquid chromatography to simultaneously determine the contents of cedrindiol, cedrinone monool, and cis-abietanol in the glandular hair secretion components of tobacco.

[0054] In one embodiment of the method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components of the present invention, step S3 may specifically include:

[0055] Step S31, quantitatively pipette the glandular hair secretion component solution and dilute it with an acetonitrile solution with a volume concentration of 80%-85%.

[0056] Step S32, using a DAD detector under liquid chromatography gradient elution conditions to detect the contents of cedrindiol, cedrinone monool, and cis-abietanol in the secretion components of tobacco glandular hairs.

[0057] Wherein, under the liquid chromatography gradient elution conditions, the liquid chromatography conditions are: ultra-high performance liquid chromatography, the analytical column is ACQUITY UPLC BEH C18 chromatographic column, specifications are 2.1mm×50mm, 1.7μm, the mobile phases are acetonitrile as mobile phase A and ultrapure water as mobile phase B, the elution program is 0min, 20% mobile phase A, 80% mobile phase B; 2min, 40% mobile phase A, 60% mobile phase B; 4min, 60% mobile phase A, 40% mobile phase B; 6min, 80% A; 8min, 100% A; 9min, 100% A; 11min, 80% mobile phase A, 20% mobile phase B; 13min, 60% mobile phase A, 40% mobile phase B; 14min, 40% mobile phase A, 60% mobile phase B; 16min, 20% mobile phase A, 80% mobile phase B; the flow rate is 0.3mL / min, the column temperature is 35℃, and the external standard method is used for quantification.

[0058] Among them, the liquid chromatogram of the test sample is as follows Figure 2 As shown by Figure 2 It can be obtained that the content determination results of α-cetabtriene diol, β-cetabtriene diol, cetabtriene monool and cis-abieol are 5.86 mg / g, 2.31 mg / g, 1.84 mg / g and 1.33 mg / g respectively.

[0059] Step S4, determination of the content of sucrose esters in the secretory components of tobacco glandular hairs: dissolving the remaining secretory components of tobacco glandular hairs in liquid nitrogen and blowing them to dryness, and determining the content of sucrose esters in the secretory components of tobacco glandular hairs by gas chromatography-mass spectrometry after derivatization.

[0060] In one embodiment of the method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components of the present invention, step S4 may specifically include:

[0061] Step S41, after determining the content of diterpenoid aroma components in the glandular hair secretion components of tobacco, add a petroleum ether internal standard solution containing sucrose octaacetate to the remaining glandular hair secretion component solution, the concentration is 0.5mg / mL-0.7mg / mL, the volume is 80μL-120μL, and blown to dryness with nitrogen.

[0062] Step S42, adding a mixed solution of dimethylformamide and N,O-bistrimethyltrifluoroacetamide in a volume ratio of 0.8:1-1.2:1, heating at 70°C-80°C for 0.8h-1.2h, derivatizing, passing through an organic filter with a pore size of 0.22μm, and performing GC / MS determination.

[0063] Specifically, in the GC / MS determination, the GC / MS chromatographic conditions are as follows: the chromatographic column is HP-MS with specifications of 30m×250μm×0.25μm, the carrier gas is helium, the flow rate is 0.8mL / min, the injection port temperature is 290°C, the split mode is used for injection, the injection volume 1 is μL, the split ratio is 5:1, the starting temperature of the chromatographic column is 230°C, maintained for 1 min, increased to 270°C at a rate of 5°C / min, maintained for 2 min, increased to 290°C at a rate of 1°C / min, and maintained for 4 min.

[0064] Step S43, performing semi-quantitative analysis using sucrose octaacetate as an internal standard.

[0065] Among them, the total ion current diagram of the test sample obtained is as follows Figure 3 As shown, from Figure 3 It can be seen that the sucrose esters of oriental tobacco are composed of types Ⅰ, Ⅱ, Ⅲ, Ⅳ and Ⅴ, and the total sucrose ester content is 460 μg / g.

[0066] In some embodiments of the present invention, the reproducibility of the detection results is also verified. Specifically, the samples were tested for 6 consecutive days to examine the reproducibility of the detection method of the present invention. The results are shown in Table 1. As can be seen from Table 1, the relative standard deviation (RSD) of the target detection component is in the range of 1.88-5.12%, which meets the requirements of quantitative analysis.

[0067] Table 1 Reproducibility results of chemical composition detection

[0068]

[0069]

[0070] In some embodiments of the present invention, the target component detected by the method of the present invention is also compared with the extraction and determination methods of the target component. Specifically, the determination results of the ether extract, α-cephalotriene glycol, and β-cephalotriene glycol obtained by the present invention are compared with the industry standard method. Since there is no industry standard for the determination of cis-abietanol, the determination results of cis-abietanol of the present invention are compared with the determination method in the patent application number ZL 201710047317.4 and the patent name is a method for determining afir alcohol in tobacco and tobacco products. Since there is no industry standard for the determination of sucrose esters, the determination results of sucrose esters of the present invention are compared with the currently commonly used determination methods. The comparison results are shown in Table 2. Furthermore, Table 3 compares the time required to complete the detection of the target components and the reagents used.

[0071] Table 2 Comparison of the test results of this method with those of industry standard methods or conventional methods

[0072]

[0073] Table 3 Comparison of time and reagents required to complete target component detection

[0074]

[0075] It can be seen from Table 2 and Table 3 that the method of the present invention can be used to simultaneously determine tobacco ether extract, α-cetabtriene diol, β-cetabtriene diol, cetabtriene monool, abietol and sucrose ester. Compared with the separate extraction and determination of the above components, the difference in the determination results is not significant, but the method of the present invention can greatly save analysis time and solvent usage.

[0076] The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components provided by the embodiment of the present invention can realize the simultaneous extraction and detection of tobacco ether extract and main aroma components of glandular hair secretion, and realizes the simultaneous high-performance liquid chromatography detection of the main diterpenoid aroma components of tobacco glandular hair secretion, namely, α-cepinatriene diol, β-cepinatriene diol, cepinatriene monool and cis-abirol for the first time; in the process of detecting the glandular hair secretion components of tobacco, the tobacco petroleum ether extract can be dissolved in an ethanol solution with a concentration of 80%-85%, so that the target detection components can be efficiently dissolved, and the contamination of the liquid chromatography and gas chromatography-mass spectrometry chromatograph used in the process of detecting the glandular hair secretion components by high molecular fat-soluble components such as wax, solanesol and phytosterol and the interference with the target detection components can be avoided; compared with the currently commonly used detection method for the target components, the method of the present invention has simple operation steps, saves solvents, improves the detection efficiency and reduces the detection cost.

[0077] So far, various embodiments of the present disclosure have been described in detail. In order to avoid obscuring the concept of the present disclosure, some details known in the art are not described. Based on the above description, those skilled in the art can fully understand how to implement the technical solution disclosed here.

[0078] Although some specific embodiments of the present disclosure have been described in detail by way of examples, it should be understood by those skilled in the art that the above examples are for illustration only and are not intended to limit the scope of the present disclosure. It should be understood by those skilled in the art that the above embodiments may be modified or some technical features may be replaced by equivalents without departing from the scope and spirit of the present disclosure. The scope of the present disclosure is defined by the appended claims.

Claims

1. A method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components, characterized in that: include: Extracting tobacco ether extract with petroleum ether and determining the content of tobacco petroleum ether extract; Using ethanol solution to dissolve the glandular hair secretion components in the tobacco petroleum ether extract and removing high molecular fat-soluble impurities to obtain a glandular hair secretion component solution; Determination of the content of diterpenoid aroma components in tobacco glandular hair secretion components: quantitatively pipette the glandular hair secretion component solution, dilute it with a mobile phase, and use liquid chromatography to simultaneously determine the contents of α-cepatriene diol, β-cepatriene diol, cepatriene monool, and cis-abirol in the tobacco glandular hair secretion components. Specifically, the contents of α-cepatriene diol, β-cepatriene diol, cepatriene monool, and cis-abirol in the tobacco glandular hair secretion components are detected using a DAD detector under liquid chromatography gradient elution conditions, wherein under the liquid chromatography gradient elution conditions, the liquid chromatography conditions are: ultra-high performance liquid chromatograph, the analytical column is an ACQUITY UPLC BEH C18 column with a specification of 2.1 mm×50 mm, 1.7 μm, the mobile phase is acetonitrile as mobile phase A and ultrapure water as mobile phase B, and the elution program is 0 min, 20% mobile phase A, 80% mobile phase B; 2 min, 40% Mobile phase A, 60% mobile phase B; 4 min, 60% mobile phase A, 40% mobile phase B; 6 min, 80% A; 8 min, 100% A; 9 min, 100% A; 11 min, 80% mobile phase A, 20% mobile phase B; 13 min, 60% mobile phase A, 40% mobile phase B; 14 min, 40% mobile phase A, 60% mobile phase B; 16 min, 20% mobile phase A, 80% mobile phase B; flow rate was 0.3 mL / min, column temperature was 35°C, and quantification was performed using the external standard method; Determination of sucrose ester content in tobacco glandular hair secretion components: The remaining glandular hair secretion components were dissolved in liquid nitrogen and blown to dryness. After derivatization, the sucrose ester content in the tobacco glandular hair secretion components was determined by gas chromatography-mass spectrometry.

2. The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components according to claim 1, characterized in that: The method of extracting tobacco ether extract with petroleum ether and determining the content of tobacco petroleum ether extract specifically includes: Refer to the industry standard YC / T 176-2003 and use petroleum ether at 30℃-60℃ to extract tobacco ether extract; After the extraction is completed, the solvent is removed by vacuum rotary evaporation at 35°C-45°C; Wipe off the moisture on the outside of the rotary evaporator, put it in a desiccator and dry it to constant weight, weigh it, and calculate the content of tobacco petroleum ether extract.

3. The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components according to claim 2, characterized in that: The method of using an ethanol solution to dissolve the glandular hair secretion components in the tobacco petroleum ether extract and removing high-molecular fat-soluble impurities to obtain a glandular hair secretion component solution specifically comprises: After weighing and determining the content of the tobacco petroleum ether extract, add an ethanol solution with a volume concentration of 80%-85%; Under normal temperature, the tobacco petroleum ether extract was dissolved by ultrasound; Transfer the dissolved solution of petroleum ether extract into a volumetric flask, dissolve it 2-4 times continuously, make up to volume, and wait for testing.

4. The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components according to claim 1, characterized in that: In the determination of the content of diterpenoid aroma components in the glandular hair secretion components of tobacco, the quantitative removal of the glandular hair secretion component solution, dilution with a mobile phase, and simultaneous determination of the contents of α-cetabtriene diol, β-cetabtriene diol, cetabtriene monool, and cis-abieol in the glandular hair secretion components of tobacco by liquid chromatography specifically includes: Quantitatively pipette the glandular hair secretion component solution, and dilute it with an acetonitrile solution with a volume concentration of 80%-85%; The contents of α-cepintriene glycol, β-cepintriene glycol, cepintriene monool and cis-abieol in the secretion components of tobacco glandular hairs were detected using a DAD detector under liquid chromatography gradient elution conditions.

5. The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components according to claim 4, characterized in that: In the determination of the sucrose ester content in the glandular hair secretion components of tobacco, the remaining glandular hair secretion components are dissolved in liquid nitrogen and blown to dryness, and the sucrose ester content in the glandular hair secretion components of tobacco is determined by gas chromatography-mass spectrometry after derivatization, which specifically includes: After determining the content of diterpenoid aroma components in the glandular hair secretion components of tobacco, add a petroleum ether internal standard solution containing sucrose octaacetate to the remaining glandular hair secretion component solution at a concentration of 0.5 mg / mL-0.7 mg / mL in a volume of 80 μL-120 μL, and blow nitrogen to dryness; Add a mixture of dimethylformamide and N,O-bistrimethyltrifluoroacetamide in a volume ratio of 0.8:1-1.2:1, heat at 70°C-80°C for 0.8h-1.2h, derivatize, pass through an organic filter with a pore size of 0.22 μm, and perform GC / MS determination; Semi-quantitative analysis was performed using sucrose octaacetate as an internal standard.

6. The method for simultaneously extracting and determining tobacco ether extract and glandular hair secretion components according to claim 5, characterized in that: In the GC / MS determination, the GC / MS chromatographic conditions are as follows: the chromatographic column is HP-MS with specifications of 30 m×250μm×0.25μm, the carrier gas is helium, the flow rate is 0.8mL / min, the injection port temperature is 290°C, the injection is in split mode, the injection volume is 1 μL, the split ratio is 5:1, the starting temperature of the chromatographic column is 230°C, maintained for 1 min, increased to 270°C at a rate of 5°C / min, maintained for 2 min, increased to 290°C at a rate of 1°C / min, and maintained for 4 min.

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