CN113252819A - Method for judging and detecting Maillard reaction product in burley tobacco baking process - Google Patents
Method for judging and detecting Maillard reaction product in burley tobacco baking process Download PDFInfo
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Abstract
The invention provides a method for judging and detecting Maillard reaction products in a burley tobacco baking process. The determination method includes: crushing burley tobacco leaf samples and extracting aroma components, analyzing the extraction solution by adopting gas chromatography-triple quadrupole tandem mass spectrometry, analyzing the aroma components of the extraction solution, determining the aroma components of the baked samples which are increased compared with the reference samples, and judging the characteristic aroma products of Maillard reaction in the burley tobacco baking process by adopting a multiple-of-variation method. The invention also provides a detection method of Maillard reaction products in the burley tobacco baking process. The method provided by the invention has the advantages of simplicity, high efficiency, high sensitivity, good selectivity, high accuracy and the like, and is suitable for detecting and judging the characteristic aroma product of the Maillard reaction in the baking process of burley tobacco.
Description
Technical Field
The invention relates to the field of detection of aroma components, in particular to a method for judging and detecting Maillard reaction products in a burley tobacco baking process.
Background
Blended cigarettes are an important cigarette product type, and have a high market share and a large consumer group. The burley tobacco processing technology is a key link of mixed type cigarette production, and the main technology is feeding and high-temperature baking. The maillard reaction is the predominant reaction carried out during the burley tobacco charge roasting process. The reaction is a complex reaction of a series of reactions such as condensation, rearrangement, cracking, polymerization and the like of amino compounds (such as amino acid, amine, polypeptide or protein and the like) in the tobacco leaves and carbohydrate. Maillard reaction characteristic products in the baking process of burley tobacco comprise various aroma components such as pyrazine, furan, pyrrole, pyridine, aldehyde, alcohol, ester, ether and the like, form a material basis of special aroma and taste of the blended cigarette, and have obvious influence on the sensory quality of the blended cigarette. The judgment and accurate determination of Maillard reaction characteristic products play an important role in the research of burley tobacco processing technology and the quality improvement of blended type cigarette products.
At present, the reported identification and detection method of the Maillard reaction aroma products is mostly a method for measuring the Maillard reaction products of sugar and amino acid in a model system, burley tobacco contains a plurality of sugar and amino compounds, the Maillard reaction generated in the baking process is more complicated than the model system, tobacco is one of the most complicated chemical systems recognized all over the world, the identified components reach 9825, the substrate is extremely complicated, and the identification and detection method of the Maillard reaction aroma products in the reported model system is obviously not suitable for the identification and detection of the Maillard reaction characteristic products in the baking process of the burley tobacco. At present, the identification and detection method specially aiming at Maillard reaction characteristic products in the baking process of burley tobacco is rarely seen. Therefore, it is necessary to establish an effective method for determining Maillard reaction characteristic products and a method for rapidly and accurately detecting Maillard reaction characteristic products in the burley tobacco baking process.
Disclosure of Invention
The embodiment of the invention provides a method for judging Maillard reaction products in a burley tobacco baking process. The method for judging the Maillard reaction product in the burley tobacco baking process fills the blank of the prior art, has the advantages of simplicity, high efficiency, high sensitivity, good selectivity, high accuracy and the like, and is suitable for detecting and judging the Maillard reaction characteristic aroma product in the burley tobacco baking process.
The embodiment of the invention provides a method for judging Maillard reaction products in a burley tobacco baking process, which comprises the following steps: crushing burley tobacco leaf samples and extracting aroma components, adopting gas chromatography-triple quadrupole tandem mass spectrometry (GC-MS/MS) to analyze the extraction solution, analyzing the aroma components of the extraction solution, determining the aroma components of the baked samples which are increased compared with the reference samples, and judging the Maillard reaction characteristic aroma products in the burley tobacco baking process by adopting a multiple-of-variation method.
According to the method for judging the Maillard reaction product in the burley tobacco baking process, provided by the embodiment of the invention, the extraction solvent is methyl tert-butyl ether, dichloromethane, cyclohexane or methanol; preferably, the extraction solvent is methanol; the internal standard substance is phenethyl acetate, and the concentration is 1-2 mg/mL. In the invention, the extraction solvent is adopted, so that the aroma components in the tobacco can be effectively extracted. In particular, the above-mentioned preferable extraction solvent can further improve the effective extraction and extraction efficiency of the aroma component.
According to the method for judging the Maillard reaction product in the baking process of the burley tobacco provided by the embodiment of the invention, the method further comprises the following steps: taking 3-6 g of the tobacco powder sample, adding 20-50 mL of extraction solvent and 40-100 mu L of internal standard substance, carrying out ultrasonic extraction for 1-2 h, standing for 5 +/-1 min, and then carrying out gas chromatography-triple quadrupole tandem mass spectrometry; preferably, the qualitative analysis is performed by retention time confirmation and standard addition, and the detection results of the two groups of samples are compared by an internal standard method.
According to the method for judging the Maillard reaction product in the baking process of the burley tobacco provided by the embodiment of the invention, the method further comprises the following steps: drying and crushing burley tobacco leaf samples, balancing for 12-24 h under the conditions that the relative humidity is 60 +/-3% and the temperature is 20 +/-1 ℃, then drying the balanced burley tobacco leaf samples for 1-2 h at the temperature of 40 +/-1 ℃, crushing, and sieving by a 40-mesh sieve to obtain tobacco powder samples; the burley tobacco leaf samples included both the cured samples treated by curing and the control samples not treated by curing. In the invention, the moisture among the tobacco powder samples obtained by the processing method is kept consistent under the condition that the aroma components are not volatilized.
According to the method for judging the Maillard reaction product in the baking process of the burley tobacco, provided by the embodiment of the invention, the gas chromatography conditions are as follows: the chromatographic column is DB-WAX 30m × 0.25mm × 0.25 μm, HP-INNOWAX 30m × 0.25mm × 0.25 μm or DB-530 m × 0.25mm × 0.25 μm; the temperature of a sample inlet is 230-260 ℃; temperature rising procedure: keeping the initial temperature at 45-60 ℃ for 2-5 min, raising the temperature to 200 ℃ at 1-4 ℃/min, raising the temperature to 240 ℃ at 3-10 ℃/min, and keeping the temperature for 5-15 min; carrier gas: he, 1 mL/min; sample introduction amount: 1 mu L of the solution; sample introduction mode: no shunt sampling; and/or, the mass spectrometry conditions: an ion source: electron bombardment EI +; electron energy: 70 eV; ion source temperature: 230 ℃; transmission line temperature: 250 ℃; multiplication voltage: 500V; a mass analyzer: a four-stage lever; scanning mode: full scan, scan range: 35 to 350 amu.
According to the method for judging the Maillard reaction product in the baking process of the burley tobacco, provided by the embodiment of the invention, the gas chromatography conditions are as follows: a chromatography column, DB-WAX 30m × 0.25mm × 0.25 μm; the temperature of a sample inlet is 250 ℃; temperature rising procedure: the initial temperature is maintained at 50 deg.C for 3min, increased to 200 deg.C at 2 deg.C/min, increased to 240 deg.C at 5 deg.C/min, and maintained for 10 min. Carrier gas: he, 1 mL/min; sample introduction amount: 1 mu L of the solution; sample introduction mode: no split-flow sample introduction.
In the method, in the judging process of the Maillard reaction product in the baking process of the burley tobacco, the gas chromatography and mass spectrum conditions are adopted, so that various aroma components can be effectively separated, and the analysis accuracy is improved.
According to the determination method of the Maillard reaction product in the baking process of the burley tobacco, provided by the embodiment of the invention, the aroma component analysis comprises the following steps: determining target aroma components through NIST spectral library matching, then comparing the consistency of the retention time of the target aroma components in the standard sample and the sample to be detected, and separating and identifying the target aroma components; and analyzing the sample by adopting the ratio of the peak area of the target aroma component to the peak area of the internal standard.
According to the method for judging the Maillard reaction product in the baking process of the burley tobacco, provided by the embodiment of the invention, the multiple change method comprises the following steps: calculating the ratio of the content of the aroma components of the burley tobacco after charging and baking to the unbaked control sample, and selecting a threshold value of 2 times; and if the change multiple is more than or equal to 2, the change of the aroma components after baking is considered to be obvious, and the aroma products with Maillard characteristics are judged.
The embodiment of the invention also provides a detection method of Maillard reaction products in the baking process of burley tobacco, wherein the Maillard reaction products are determined by the determination method, and the detection method comprises the following steps: and (3) carrying out gas chromatography-triple quadrupole tandem mass spectrometry on the extract liquor of the burley tobacco leaf sample, and calculating the content of the Maillard characteristic aroma product in the burley tobacco baking process.
According to the embodiment of the invention, the detection method of Maillard reaction products in the baking process of burley tobacco comprises the following steps: taking a cured burley tobacco leaf sample, drying at the temperature of 40 +/-1 ℃ for 1-2 h, crushing, and sieving with a 40-mesh sieve; accurately weighing 1-3 g of tobacco powder sample, adding 20-60 mL of methanol and 20-60 mu L of phenethyl acetate internal standard, performing ultrasonic extraction for 1-2 h, standing for 5min, filtering the upper layer of extract liquor by 0.45 mu m, performing gas chromatography-triple quadrupole tandem mass spectrometry analysis and determination, and calculating the content of Maillard characteristic aroma products in the baking process of burley tobacco.
According to the detection method of the Maillard reaction product in the baking process of burley tobacco provided by the embodiment of the invention, the gas chromatography conditions are as follows: a chromatographic column: DB-WAX capillary column (30m × 0.25mm × 0.25 μm); temperature rising procedure: the initial temperature is 40-50 ℃, the temperature is kept for 2-5 min, the temperature is raised to 55-80 ℃ at the speed of 3-6 ℃/min, then the temperature is raised to 140-160 ℃ at the speed of 2-5 ℃/min, the temperature is kept for 1-3 min, the temperature is raised to 190-210 ℃ at the speed of 5-10 ℃/min, the temperature is kept for 2-4 min, then the temperature is raised to 230-245 ℃ at the speed of 10-25 ℃/min, and the temperature is kept for 4-6 min; sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L, no shunt sample introduction; in a constant flow mode, the column flow is 1 mL/min; and/or, mass spectrometry conditions: EI ionization mode, ionization energy 70 ev; ion source temperature: 230 ℃; transmission line temperature: 250 ℃; q1 and Q3 temperatures: 150 ℃; collision gas: n is a radical of2The flow rate is 1.5 mL/min; quenching gas: he, flow rate 2.25 mL/min; the scanning mode is as follows: multiple reactions monitor MRM mode.
According to the detection method of the Maillard reaction product in the baking process of burley tobacco, provided by the embodiment of the invention, the gas chromatography conditions are as follows: DB-WAX capillary column 30m × 0.25mm × 0.25 μm; temperature rising procedure: the initial temperature is 50 ℃, the temperature is kept for 3min, the temperature is increased to 65 ℃ at the speed of 5 ℃/min, then the temperature is increased to 150 ℃ at the speed of 3 ℃/min, the temperature is kept for 1min, the temperature is increased to 200 ℃ at the speed of 8 ℃/min, the temperature is kept for 3min, then the temperature is increased to 240 ℃ at the speed of 20 ℃/min, and the temperature is kept for 5 min. Sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L, no shunt sample introduction; constant flow mode, column flow 1 mL/min.
According to the invention, by adopting the specific chromatographic conditions, the measured Maillard characteristic aroma product can be effectively separated, the analysis time is shortened, and the detection efficiency is improved.
The invention has the beneficial effects that: the method provided by the invention fills the blank of the prior art, and adopts a GC-MS/MS combined multiple-of-change method to establish an effective method for identifying and detecting Maillard reaction characteristic products in the baking process of burley tobacco. The method has the advantages of simplicity, high efficiency, high sensitivity, good selectivity, high accuracy and the like, and is suitable for detecting and judging the characteristic aroma product of the Maillard reaction in the baking process of burley tobacco.
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In order to more clearly illustrate the technical solutions of the present invention or the prior art, the drawings needed for the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and those skilled in the art can also obtain other drawings according to the drawings without creative efforts.
FIG. 1 is a MRM total ion flux chromatogram of Maillard reaction-characteristic products during the baking of burley tobacco in example 1 (ingredients numbered 1-28, corresponding names in Table 3);
FIG. 2 is a MRM total ion flux chromatogram of Maillard reaction-characteristic products during the baking of burley tobacco in example 1 (ingredients numbered 29-34, corresponding names in Table 3).
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The instruments and the like are conventional products which are purchased by normal distributors and are not indicated by manufacturers. The process is conventional unless otherwise specified, and the starting materials are commercially available from the open literature. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications.
Example 1
The embodiment provides a method for detecting and judging a Maillard reaction characteristic aroma product in a burley tobacco baking process.
1) Judging the Maillard characteristic aroma product:
taking two samples of a burley tobacco leaf group of a certain brand number 1 cigarette before normal lining material baking, taking one sample as a control sample without baking treatment, taking the other sample as a baking sample, and baking according to a normal baking procedure. After the baking treatment of the samples is finished, the two groups of samples are placed in a constant temperature and humidity box and are balanced for 12 hours under the conditions that the relative humidity is 60 +/-3% and the temperature is 20 +/-1 ℃. The balanced sample is placed in an oven, dried for 1h at the temperature of 40 ℃, crushed and sieved by a 40-mesh screen.
Accurately weighing 5g of tobacco powder sample, repeating 5 times for each sample, respectively adding 30mL of methanol and 60 μ L of n-propyl benzoate internal standard (1mg/mL), rapidly covering, performing ultrasonic extraction for 1h, standing for 5min, filtering the upper layer of extract with a 0.45 μm needle filter, and performing GC-MS/MS determination.
Chromatographic conditions are as follows: column, DB-WAX (30 m.times.0.25 mm. times.0.25 μm); the temperature of a sample inlet is 250 ℃; temperature rising procedure: the initial temperature is maintained at 50 deg.C for 3min, increased to 200 deg.C at 2 deg.C/min, increased to 240 deg.C at 5 deg.C/min, and maintained for 10 min. Carrier gas: he, 1 mL/min; sample introduction amount: 1 mu L of the solution; sample introduction mode: no split-flow sample introduction.
Mass spectrum conditions: an ion source: electron bombardment (EI +); electron energy: 70 eV; ion source temperature: 230 ℃; the transmission line temperature is 250 ℃; multiplication voltage: 500V; a mass analyzer: a four-stage lever; scanning mode: full scan, scan range: 35-350 amu.
Analyzing the extraction solution sample in a GC-MS full-scanning mode, determining the target aroma components through NIST spectral library matching, and then comparing the consistency of the retention time of the target aroma components in the standard sample and the sample to be detected. The total separation identified 73 aroma components, see table 1.
And (4) carrying out semi-quantitative analysis on the sample by adopting the ratio (relative peak area) of the peak area of each target aroma component to the peak area of the internal standard. The baked samples were determined to have a significant increase in the aroma components over the control samples for a total of 33 (table 2). And judging the characteristic aroma product of the Maillard reaction in the burley tobacco baking process by adopting a multiple-change method. And (3) calculating the ratio of the content of the aroma components of the burley tobaccos after being charged and baked to the unbaked control sample, selecting a threshold value of 2 times, and judging that the aroma components are remarkably changed after being baked if the change multiple is more than or equal to 2, judging the aroma components as Maillard characteristic aroma products, and determining 25 Maillard characteristic aroma products in total, wherein the Table 2 shows.
TABLE 1 qualitative analysis results of aroma components after baking burley tobacco
TABLE 2 ratio of the content of the aroma components after the burley tobacco charge is baked to the control
2) Maillard feature aroma product determination:
taking a proper amount of baked burley tobacco samples, baking for 1h at the temperature of 40 ℃, crushing, and sieving with a 40-mesh sieve. Accurately weighing 2g of tobacco powder samples, repeating 3 samples of each sample, respectively adding 40mL of methanol and 40 mu L of n-propyl benzoate internal standard, quickly covering a cover, performing ultrasonic extraction for 1h, standing for 5min, filtering the upper-layer extract by a 0.45 mu m needle filter, performing gas chromatography-triple quadrupole tandem mass spectrometry (GC-MS/MS) determination, and calculating the content of Maillard characteristic aroma products in the baking process of burley tobacco.
Chromatographic conditions are as follows: DB-WAX capillary column (30m × 0.25mm × 0.25 μm); temperature rising procedure: the initial temperature is 50 ℃, the temperature is kept for 3min, the temperature is increased to 65 ℃ at the speed of 5 ℃/min, then the temperature is increased to 150 ℃ at the speed of 3 ℃/min, the temperature is kept for 1min, the temperature is increased to 200 ℃ at the speed of 8 ℃/min, the temperature is kept for 3min, then the temperature is increased to 240 ℃ at the speed of 20 ℃/min, and the temperature is kept for 5 min. Sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L, no shunt sample introduction; constant flow mode, column flow 1 mL/min.
Mass spectrometry conditions: EI ionization mode, ionization energy 70 ev; ion source temperature: 230 ℃; transmission line temperature: 250 ℃; q1 and Q3 temperatures: 150 ℃; collision gas: n is a radical of2The purity is 99.999 percent, and the flow rate is 1.5 mL/min; quenching gas: he, flow rate 2.25 mL/min; the scanning mode is as follows: multiple reactions monitor MRM mode. The retention time, qualitative and quantitative ion pairs and collision energy for each analyte are shown in table 3.
TABLE 3 respective analyte and internal standard MRM Mass Spectrometry parameters
The standard solution was prepared as follows
Accurately weighing 2.5g of phenethyl acetate standard substance, and dissolving the standard substance in a 50mL volumetric flask by using methanol to obtain an internal standard stock solution with the mass concentration of 50 mg/mL. And transferring 100 mu L of internal standard stock solution into a 50mL volumetric flask, diluting with methanol and fixing the volume to obtain the internal standard working solution with the mass concentration of 100 mu g/mL. Respectively weighing 0.1g (accurate to 0.1mg) of aroma standard substances, dissolving the aroma standard substances by using methanol, fixing the volume of the solution in a 100mL volumetric flask to prepare a primary mixed standard solution with the mass concentration of 1mg/mL, transferring 500 mu L of the primary mixed standard solution in a 50mL volumetric flask, diluting the solution by using methanol and fixing the volume to obtain a secondary mixed standard solution with the mass concentration of 10 mu g/mL, and respectively adding 50 mu L of internal standard working solution from the secondary mixed standard solution to prepare 7-grade standard working solution with the concentration range of 5-2000 ng/mL.
The method has the following detection limit, the recovery rate and the repeatability of the added standard:
taking the signal-to-noise ratio of 3 times the minimum concentration of the standard working solution as a detection limit, observing the recovery rate (n is 5) of the Maillard characteristic aroma component by adopting a standard addition method, respectively carrying out sample injection analysis (n is 5) at different time points and different treatment days on the sample treatment day, and calculating the precision in and during the day, wherein the results are shown in a table 4.
TABLE 4 detection limits, recovery rates and precision of the analytes
The results of the maillard feature aroma product measurements after baking for a burley leaf group of certain brand number 1 cigarettes are shown in table 5.
TABLE 5 measurement results of Maillard characteristics of the cured burley tobacco of certain brand No. 1 cigarette
Example 2
This example provides a method for detecting and determining a maillard reaction characteristic aroma product during burley tobacco roasting, which differs from example 1 in the detection method of a burley tobacco sample and a burley tobacco roasting maillard reaction characteristic aroma product:
a burley tobacco sample after baking a brand No. 2 blended type cigarette is taken, baked for 1.5h at the temperature of 40 ℃, crushed and screened by a 40-mesh screen. Accurately weighing 3g of tobacco powder samples, repeating 3 times for each sample, respectively adding 50mL of methanol and 50 mu L of phenethyl acetate internal standard, quickly covering a cover, performing ultrasonic extraction for 1.5h, standing for 5min, filtering the upper-layer extract by a 0.45 mu m needle filter, performing GC-MS/MS determination, and calculating the content of Maillard characteristic aroma products in the baking process of burley tobacco.
The gas chromatography conditions were: a chromatographic column: DB-WAX capillary column (30m × 0.25mm × 0.25 μm); temperature rising procedure: the initial temperature is 45 ℃, the temperature is kept for 5min, the temperature is increased to 60 ℃ at the speed of 6 ℃/min, then the temperature is increased to 155 ℃ at the speed of 4 ℃/min, the temperature is kept for 2min, the temperature is increased to 210 ℃ at the speed of 6 ℃/min, the temperature is kept for 2min, then the temperature is increased to 245 ℃ at the speed of 25 ℃/min, and the temperature is kept for 4 min. Sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L, no shunt sample introduction; constant flow mode, column flow 1 mL/min.
The mass spectrometry conditions were: EI ionization mode, ionization energy 70 ev; ion source temperature: 230 ℃; transmission line temperature: 250 ℃; q1 and Q3 temperatures: 150 ℃; collision gas:N2the purity is 99.999 percent, and the flow rate is 1.5 mL/min; quenching gas: he, flow rate 2.25 mL/min; the scanning mode is as follows: multiple reactions monitor MRM mode.
The results of the maillard characteristics aroma product measurements after baking for the burley leaf group of brand 2 cigarettes are shown in table 6.
TABLE 6 determination results of Maillard characteristics of the aroma product of No. 2 cigarette after baking
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A method for determining Maillard reaction products in a burley tobacco baking process is characterized by comprising the following steps: crushing burley tobacco leaf samples and extracting aroma components, analyzing the extraction solution by adopting gas chromatography-triple quadrupole tandem mass spectrometry, analyzing the aroma components of the extraction solution, determining the aroma components of the baked samples which are increased compared with the reference samples, and judging the characteristic aroma products of Maillard reaction in the burley tobacco baking process by adopting a multiple-of-variation method.
2. The method for determining Maillard reaction products during baking of Burley tobacco according to claim 1, wherein the extraction solvent is methyl tert-butyl ether, dichloromethane, cyclohexane or methanol; preferably, the extraction solvent is methanol; the internal standard substance is phenethyl acetate, and the concentration is 1-2 mg/mL.
3. The method of determining Maillard reaction products during baking of burley tobacco according to claim 2, further comprising: taking 3-6 g of the tobacco powder sample, adding 20-50 mL of extraction solvent and 40-100 mu L of internal standard substance, carrying out ultrasonic extraction for 1-2 h, standing for 5 +/-1 min, and then carrying out gas chromatography-triple quadrupole tandem mass spectrometry; preferably, the qualitative analysis is performed by retention time confirmation and standard addition, and the detection results of the two groups of samples are compared by an internal standard method.
4. The method of determining Maillard reaction products during baking of burley tobacco according to claim 1, further comprising: drying and crushing burley tobacco leaf samples, balancing for 12-24 h under the conditions that the relative humidity is 60 +/-3% and the temperature is 20 +/-1 ℃, then drying the balanced burley tobacco leaf samples for 1-2 h at the temperature of 40 +/-1 ℃, crushing, and sieving by a 40-mesh sieve to obtain tobacco powder samples; the burley tobacco leaf samples included both the cured samples treated by curing and the control samples not treated by curing.
5. A method of determining a Maillard-characteristic aroma product during baking of burley tobacco according to any of claims 1 to 4, wherein the gas chromatography conditions are: the chromatographic column is DB-WAX 30m × 0.25mm × 0.25 μm, HP-INNOWAX 30m × 0.25mm × 0.25 μm or DB-530 m × 0.25mm × 0.25 μm; the temperature of a sample inlet is 230-260 ℃; temperature rising procedure: keeping the initial temperature at 45-60 ℃ for 2-5 min, raising the temperature to 200 ℃ at 1-4 ℃/min, raising the temperature to 240 ℃ at 3-10 ℃/min, and keeping the temperature for 5-15 min; carrier gas: he, 1 mL/min; sample introduction amount: 1 mu L of the solution; sample introduction mode: no shunt sampling; and/or, the mass spectrometry conditions: an ion source: electron bombardment EI +; electron energy: 70 eV; ion source temperature: 230 ℃; transmission line temperature: 250 ℃; multiplication voltage: 500V; a mass analyzer: a four-stage lever; scanning mode: full scan, scan range: 35-350 amu;
preferably, the gas chromatography conditions are: a chromatography column, DB-WAX 30m × 0.25mm × 0.25 μm; the temperature of a sample inlet is 250 ℃; temperature rising procedure: the initial temperature is maintained at 50 deg.C for 3min, increased to 200 deg.C at 2 deg.C/min, increased to 240 deg.C at 5 deg.C/min, and maintained for 10 min. Carrier gas: he, 1 mL/min; sample introduction amount: 1 mu L of the solution; sample introduction mode: no split-flow sample introduction.
6. A method of determining Maillard reaction products during baking of burley tobacco according to any of claims 1 to 5, wherein the aroma component analysis comprises: determining target aroma components through NIST spectral library matching, then comparing the consistency of the retention time of the target aroma components in the standard sample and the sample to be detected, and separating and identifying the target aroma components; and analyzing the sample by adopting the ratio of the peak area of the target aroma component to the peak area of the internal standard.
7. A method of determining Maillard reaction products during baking of burley tobacco according to any of claims 1 to 6, wherein the fold change method: calculating the ratio of the content of the aroma components of the burley tobacco after charging and baking to the unbaked control sample, and selecting a threshold value of 2 times; and if the change multiple is more than or equal to 2, the change of the aroma components after baking is considered to be obvious, and the aroma products with Maillard characteristics are judged.
8. A method for detecting maillard reaction products of a burley tobacco roasting process, wherein the maillard reaction products are determined by the determination method of any one of claims 1 to 7, comprising: and (3) carrying out gas chromatography-triple quadrupole tandem mass spectrometry on the extract liquor of the burley tobacco leaf sample, and calculating the content of the Maillard characteristic aroma product in the burley tobacco baking process.
9. The method of claim 8, comprising: taking a cured burley tobacco leaf sample, drying at the temperature of 40 +/-1 ℃ for 1-2 h, crushing, and sieving with a 40-mesh sieve; accurately weighing 1-3 g of tobacco powder sample, adding 20-60 mL of methanol and 20-60 mu L of phenethyl acetate internal standard, performing ultrasonic extraction for 1-2 h, standing for 5min, filtering the upper layer of extract liquor by 0.45 mu m, performing gas chromatography-triple quadrupole tandem mass spectrometry analysis and determination, and calculating the content of Maillard characteristic aroma products in the baking process of burley tobacco.
10. The method for detecting Maillard reaction products during baking of burley tobacco according to claim 8 or 9, wherein the gas chromatography conditions are: a chromatographic column: DB-WAX capillary column (30m × 0.25mm × 0.25 μm); temperature rising procedure: the initial temperature is 40-50 ℃, the temperature is kept for 2-5 min, the temperature is raised to 55-80 ℃ at the speed of 3-6 ℃/min, then the temperature is raised to 140-160 ℃ at the speed of 2-5 ℃/min, the temperature is kept for 1-3 min, the temperature is raised to 190-210 ℃ at the speed of 5-10 ℃/min, the temperature is kept for 2-4 min, then the temperature is raised to 230-245 ℃ at the speed of 10-25 ℃/min, and the temperature is kept for 4-6 min; sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L, no shunt sample introduction; in a constant flow mode, the column flow is 1 mL/min; and/or, mass spectrometry conditions: EI ionization mode, ionization energy 70 ev; ion source temperature: 230 ℃; transmission line temperature: 250 ℃; q1 and Q3 temperatures: 150 ℃; collision gas: n is a radical of2The flow rate is 1.5 mL/min; quenching gas: he, flow rate 2.25 mL/min; the scanning mode is as follows: multiple reaction monitoring, MRM, mode;
preferably, the gas chromatography conditions are: DB-WAX capillary column 30m × 0.25mm × 0.25 μm; temperature rising procedure: the initial temperature is 50 ℃, the temperature is kept for 3min, the temperature is increased to 65 ℃ at the speed of 5 ℃/min, then the temperature is increased to 150 ℃ at the speed of 3 ℃/min, the temperature is kept for 1min, the temperature is increased to 200 ℃ at the speed of 8 ℃/min, the temperature is kept for 3min, then the temperature is increased to 240 ℃ at the speed of 20 ℃/min, and the temperature is kept for 5 min. Sample inlet temperature: 250 ℃; sample introduction amount: 1 mu L, no shunt sample introduction; constant flow mode, column flow 1 mL/min.
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| PENG LI ET AL.: "Changes in Levels of Amino Acids and Basic Components in Burley Tobacco Produced by Roasting", 《BEITRAGE ZUR TABAKFORSCHUNG INTERNATIONAL》 * |
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