A method of reconstituting a tobacco flavour formulation

Abstract

The present application belongs to the field of component analysis of tobacco flavor, and particularly relates to a formula recombination method of tobacco flavor. The formula recombination method of tobacco flavor is to construct a natural flavor raw material liquid chromatography-high resolution mass spectrometry dataset through non-targeted extraction and targeted extraction, to determine the types and quantity of potential natural flavor raw materials through comparison and analysis of the targeted extraction results of the liquid chromatography-high resolution mass spectrometry of the tobacco flavor and the dataset, and to complete the determination of the types and ratio of the natural flavor raw materials by constructing different combinations of natural flavor raw materials and comparing the consistency of different combinations with the tobacco flavor to be recombined. The method is based on the information of ten thousand characteristic peaks of the liquid chromatography-high resolution mass spectrometry dataset, and the determination of the types and ratio of the natural flavor raw materials is completed through the characteristic peak information. The method does not depend on the subjective experience of the flavorists in the implementation process, and has the characteristics of high recombination efficiency and good adaptability.

Description

Technical Field

[0001] This invention belongs to the field of component analysis of tobacco flavorings and fragrances, and specifically relates to a method for recombining the formulation of tobacco flavorings and fragrances. Background Technology

[0002] The natural flavoring ingredients (i.e., natural flavoring monomers) listed in the permitted and temporary licensed lists of tobacco additives mainly include extracts, absolutes, and tinctures, such as maple extract, licorice fluid extract, jujube tincture, Paraguayan tea absolute, apple extract, coffee extract, and fresh grape concentrate. These natural flavoring ingredients constitute various styles of tobacco flavorings and fragrances. The formulation analysis of tobacco flavorings and fragrances involves companies' research on imitation and creation of flavors, and is also of great significance for the quality control of tobacco flavorings and fragrances.

[0003] The difficulty in recombining formulations of tobacco flavorings and fragrances lies in the complexity of their components and those of natural fragrance raw materials. These components encompass volatile, semi-volatile, and non-volatile parts, particularly since most natural fragrance raw materials are primarily water-soluble and alcohol-soluble non-volatile components. This presents significant limitations in component analysis methods, as many natural fragrance raw materials cannot be analyzed using traditional analytical techniques. Establishing the formulation relationship between natural fragrance raw materials and flavorings directly impacts the scientific, accurate, and rational analysis of tobacco flavorings and fragrances, and consequently, the company's independent control over its flavoring technology.

[0004] Chinese invention patent application CN 117990835 A, published on May 7, 2024, discloses a method for recombining tobacco flavoring and fragrance formulations. This method involves analyzing the appearance, physical characteristics, aroma quality, and flavor characteristics of the formulation to be recombined, as well as performing a full chemical composition analysis. The solvent composition and the composition of natural flavoring raw materials are then determined. While the solvent composition can be easily determined using conventional methods, the key lies in determining the composition of the natural flavoring raw materials. First, based on the results of the full chemical composition analysis, highly volatile and semi-volatile components, volatile organic acids, non-volatile acids, and water-soluble sugars are listed as potential flavoring raw material monomers. These are then compared with the flavoring raw material monomers obtained from aroma quality and flavor characteristic analysis to preliminarily determine the composition and content of the flavoring raw material monomers.

[0005] The above recombination methods require multiple testing techniques, including gas chromatography, gas chromatography-mass spectrometry, and high-resolution liquid chromatography-mass spectrometry. The correspondence between volatile components and potential flavor raw material monomers, as well as the determination of flavor raw material monomers based on aroma quality and flavor characteristics, all rely on the experience of the flavorist, resulting in a degree of subjectivity in the implementation of the process. In summary, existing recombination methods suffer from shortcomings such as complex testing processes and reliance on the experience of flavorists to determine the composition and content of natural flavor raw material monomers. This leads to problems such as low recombination efficiency and the need to improve the adaptability of the methods in the formulation recombination of tobacco flavorings and fragrances. Summary of the Invention

[0006] The purpose of this invention is to provide a method for recombining the formulation of tobacco flavorings and fragrances, in order to solve the problems of low recombining efficiency and the need to improve the adaptability of existing recombining methods.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] A method for recombining the formulation of tobacco flavorings and fragrances includes the following steps:

[0009] (1) The characteristic peaks of natural fragrance raw materials are extracted by liquid chromatography-high resolution mass spectrometry using a non-targeting method. The characteristic peaks are aligned and then the liquid chromatography-high resolution mass spectrometry analysis results of natural fragrance raw materials are targeted and extracted using the aligned characteristic peak information to construct a liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials.

[0010] (2) Perform liquid chromatography-high resolution mass spectrometry analysis on the recombinant tobacco flavoring and fragrance and perform the targeted extraction. Compare and analyze the targeted extraction results with the liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials to identify potential natural fragrance raw materials.

[0011] (3) Construct different combinations of potential natural fragrance raw materials, and analyze the consistency between the different combinations of potential natural fragrance raw materials and the tobacco flavorings to be reconstituted based on the liquid chromatography-high resolution mass spectrometry dataset of the natural fragrance raw materials, and determine the types and proportions of natural fragrance raw materials in tobacco flavorings.

[0012] This invention is pioneering. First, it constructs a liquid chromatography-high resolution mass spectrometry (LC-HMS) dataset of natural fragrance raw materials through non-targeted and targeted extraction. By comparing the targeted extraction results of tobacco flavorings and fragrances obtained through LC-HMS with the dataset, the types and quantities of potential natural fragrance raw materials are determined. Then, different combinations of natural fragrance raw materials are constructed, and the consistency between these combinations and the tobacco flavorings and fragrances to be reconstituted is compared to determine the types and proportions of natural fragrance raw materials. This method is based on tens of thousands of characteristic peaks in the LC-HMS dataset, using this peak information to determine the types and proportions of natural fragrance raw materials. The method is independent of the subjective experience of flavorists and features high reconstitution efficiency and good adaptability.

[0013] Preferably, in step (3), constructing different combinations of potential natural flavoring raw materials includes: determining the i characteristic peaks with the highest peak intensities of the potential natural flavoring raw materials, and calculating the ratio R of the i natural flavoring raw materials to the corresponding characteristic peaks of the flavorings and fragrances to be pre-reconstituted in tobacco flavorings. iThe mean R is calculated; the basic ratio D of the natural fragrance raw material is set as 1 / R, and the derived ratios are D / 1.5, D / 2, and D / 3; assuming the quantity of natural fragrance raw materials, a linear combination is constructed based on the quantity of natural fragrance raw materials and the basic and derived ratios of each natural fragrance raw material. More preferably, i = 10~15; assuming the quantity of natural fragrance raw materials is 2, 3, 4, 5, 6, 7, and 8 respectively.

[0014] Preferably, in step (3), analyzing the consistency and determining the types and proportions of the natural flavoring raw materials includes: calculating the characteristic peak intensity of the linear combination and comparing it with the corresponding characteristic peak intensity of the flavorings to be reconstituted in the tobacco flavorings, and calculating the residual; statistically analyzing the top k linear combinations with the smallest residuals under each assumed quantity of natural flavoring raw materials, calculating the mean of the residuals, and determining the assumed quantity of natural flavoring raw materials corresponding to the smallest mean of the residuals; under the determined assumed quantity of natural flavoring raw materials, statistically analyzing the types and proportions of the natural flavoring raw materials with the highest frequency of occurrence in the top k corresponding linear combinations, which are the types and proportions of natural flavoring raw materials in the flavorings to be reconstituted in the tobacco flavorings. More preferably, k = 100~120.

[0015] Preferably, the above-mentioned method for recombining tobacco flavorings includes step (4): detecting the types and contents of solvents in the tobacco flavorings to be recombined; based on the determined types and proportions of natural flavoring raw materials, calculating the contents of each type of solvent according to the detected contents, so that the total mass fraction of the recombined formula is 100%. More preferably, the solvents include one, two, three, or four of water, ethanol, propylene glycol, and glycerin.

[0016] Preferably, in step (1), the characteristic peak is a peak not present in the blank sample or a peak with an intensity greater than 10 times that of the blank sample; the alignment processing rule is: retention time difference ≤ 0.2 min, precise molecular weight difference ≤ 5 ppm; the targeted extraction rule is: retention time difference ≤ 0.2 min, precise molecular weight difference ≤ 5 ppm.

[0017] Preferably, in step (2), the comparative analysis includes batch counting of the number of characteristic peaks of different intensity ranges in the flavorings and fragrances to be reconstituted, as well as the number of characteristic peaks of each natural fragrance raw material corresponding to the intensity range, calculating the detection probability of characteristic peaks of different intensity ranges of all natural fragrance raw materials in the flavorings and fragrances to be reconstituted, and determining the types and quantities of potential natural fragrance raw materials based on the detection probability.

[0018] Preferably, in step (1), the types of natural fragrance raw materials in the liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials cover extracts, tinctures, and absolute oils, and the number of natural fragrance raw materials is more than 400. Attached Figure Description

[0019] Figure 1 This is a total ion chromatogram in positive ion mode of the tobacco flavoring (blue) to be reconstituted and the reconstituted sample (red) in an embodiment of the present invention;

[0020] Figure 2 This is a total ion flow chromatogram of the negative ion mode of the tobacco flavoring (blue) to be reconstituted and the reconstituted sample (red) in an embodiment of the present invention. Detailed Implementation

[0021] The technical concept of this invention is to construct a liquid chromatography-high resolution mass spectrometry dataset and make rational use of the characteristic peak information therein, thereby determining the types and proportions of natural fragrance raw materials; further, combined with the determination of solvent composition, the reorganization of tobacco flavoring and fragrance formulations is completed.

[0022] Furthermore, the determination of the types and proportions of natural fragrance raw materials is based on tens of thousands of characteristic peaks in the dataset. Combined with the construction and statistical analysis of different combinations of natural fragrance raw materials based on a large amount of data, the results obtained are statistically significant and can well reflect the objective essence of tobacco flavoring formulations. Therefore, the recombinant formulation has good consistency with the original formulation, mainly reflected in the good overlap of the total ion chromatogram, the high correlation of characteristic peaks, and the high degree of matching of larger characteristic peaks.

[0023] In addition, the method of the present invention also has the following characteristics:

[0024] First, liquid chromatography-high resolution mass spectrometry can achieve full coverage of volatile, semi-volatile and non-volatile components, and can comprehensively characterize the identity information of natural fragrance raw materials; the powerful qualitative capability of high resolution mass spectrometry makes it possible to pre-reconstitute tobacco flavoring and fragrance formulations without the need for standard products, which greatly reduces economic costs and the threshold for use.

[0025] Secondly, this method enables the rapid batch determination of the types and proportions of natural fragrance raw materials in the tobacco flavorings to be pre-reconstituted, greatly reducing the workload of flavoring technicians and improving the efficiency of formula recombination.

[0026] Third, the natural fragrance raw material liquid chromatography-high resolution mass spectrometry dataset has high availability and easy scalability, and can be dynamically expanded at any time according to needs, thereby meeting the ever-growing or changing pre-recombination needs of tobacco flavoring and fragrance formulations.

[0027] The implementation process of the present invention will be described in detail below with reference to specific embodiments. In the following embodiments, unless otherwise specified, "%" represents volume fraction.

[0028] I. Specific Embodiments of the Formula Recombination Method for Tobacco Flavorings and Fragrances of the Present Invention

[0029] Example 1

[0030] The method for recombining the formulation of tobacco flavorings and fragrances in this embodiment includes the following steps:

[0031] (1) The characteristic peaks of natural fragrance raw materials are extracted by liquid chromatography-high resolution mass spectrometry using a non-targeting method. The characteristic peaks are aligned and then the liquid chromatography-high resolution mass spectrometry analysis results of natural fragrance raw materials are targeted and extracted using the aligned characteristic peak information to construct a liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials.

[0032] The liquid chromatography-high resolution mass spectrometry system used was an Agilent 1290 high performance liquid chromatograph combined with an AB 4600 quadrupole time-of-flight mass spectrometer.

[0033] The sample pretreatment and liquid chromatography-high resolution mass spectrometry (LC-HMS) analysis conditions for natural fragrance raw materials and the tobacco flavorings to be reconstituted remained completely consistent. Sample pretreatment aimed to extract the analytes from the samples, using a 40–60% methanol-water solution as the extraction solvent. 20–25 mL of extraction solvent was sufficient for 20 mg of sample.

[0034] Specifically, accurately weigh 20 mg of natural fragrance raw material or the flavoring to be reconstituted for tobacco use into a 50 mL capped centrifuge tube, then accurately add 20 mL of 50% methanol aqueous solution, vortex for 10 min, centrifuge, filter through an organic phase filter membrane, and take 1.5 mL of the filtrate into a 2 mL chromatographic bottle.

[0035] For liquid chromatography-high resolution mass spectrometry analysis, the chromatographic conditions were as follows: column: Atlantis@T3 column (150 mm length, 2.1 mm inner diameter, 3 μm stationary phase particles); column temperature: 40 °C; flow rate: 0.3 mL / min; injection volume: 10 μL; mobile phase A: ammonium acetate (5 mmol / L) solution containing 0.1% formic acid; mobile phase B: acetonitrile; gradient elution conditions: 0–1 min, 97% A; 1–1.5 min, 97%–85% A; 1.5–18 min, 85%–2% A; 18–25 min, 2% A; 25–25.1 min, 2%–97% A; 25.1–35 min, 97% A.

[0036] The specific conditions for high-resolution mass spectrometry are as follows:

[0037] ESI source; positive / negative ion mode scanning separately; ion source temperature: 500℃; cluster cleavage voltage: 40V; collision energy: 10V; spray voltage: +5500V / -4500V; nebulizer gas pressure: 55psi; curtain gas pressure: 35psi; auxiliary gas pressure: 50psi; scan range: m / z 100~1000.

[0038] The raw data of natural fragrance raw materials and blank samples were processed simultaneously using the OS-Q data processing software built into the high performance liquid chromatography-quadrupole time-of-flight mass spectrometer. First, a non-targeted extraction method was established, with key parameters including a moderate peak detection sensitivity (S3), a threshold of 10 for the peak intensity ratio of natural fragrance raw materials to blank samples, and ensuring that the peak detection time range was consistent with the acquisition time range. By subtracting peaks in natural fragrance raw materials whose peak intensities were not more than 10 times greater than the corresponding peak intensities in blank samples, all characteristic peaks of natural fragrance raw materials were obtained. The characteristic peak information included retention time and precise molecular weight.

[0039] Then, a self-developed Matlab program was used to align the characteristic peaks of all natural fragrance raw materials. The alignment principle was that the difference in retention time should be ≤0.2 min and the difference in precise molecular weight (i.e., mass number, m / z) should be ≤5 ppm. Examples of some characteristic peaks of some natural fragrance raw materials are shown in Table 1, and examples after alignment are shown in Table 2.

[0040] Table 1 Examples of some characteristic peaks of natural fragrance raw materials

[0041]

[0042] Examples of characteristic peak alignment in Table 2 and Table 1

[0043]

[0044] Finally, using OS-Q data processing software, a targeted extraction method was established based on the retention times and precise molecular weights of the aligned characteristic peaks. Key parameters included peak detection using pre-set retention times, retention time differences ≤ 0.2 min, and precise molecular weight differences ≤ 5 ppm. This targeted extraction method was used to extract all characteristic peak intensities from the analytical results of all natural fragrance raw materials, forming a natural fragrance raw material liquid chromatography-high resolution mass spectrometry dataset. This dataset includes the names of natural fragrance raw materials and the retention times, precise molecular weights, and peak intensities of all characteristic peaks, involving tens of thousands of peaks. The types of natural fragrance raw materials in the dataset cover extracts, tinctures, and absolutes, with more than 400 natural fragrance raw materials included.

[0045] Examples of datasets for natural fragrance raw materials are shown in Table 3.

[0046] Table 3. Examples of datasets for natural fragrance raw materials

[0047]

[0048] (2) Perform liquid chromatography-high resolution mass spectrometry analysis on the recombinant tobacco flavoring and fragrance and perform the targeted extraction. Compare and analyze the targeted extraction results with the liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials to identify potential natural fragrance raw materials.

[0049] This step involves batch statistically analyzing the number of characteristic peaks in different intensity ranges within the tobacco flavorings to be reconstituted, as well as the number of characteristic peaks in the corresponding intensity ranges of each natural flavoring ingredient. The detection probability of characteristic peaks in different intensity ranges of all natural flavoring ingredients in the tobacco flavorings to be reconstituted is then calculated, and the types and quantities of potential natural flavoring ingredients are determined based on the detection probability. An example dataset after adding the flavorings to be reconstituted is shown in Table 4.

[0050] Table 4 shows an example of the dataset after adding flavorings and fragrances to be recombined.

[0051]

[0052] Based on the above dataset, the number of characteristic peaks of all natural fragrance raw materials in different intensity ranges (>10,000,000, >5,000,000, >1,000,000, >500,000, >100,000, >50,000, and >10,000) and their detection counts in fragrance and flavor samples were statistically analyzed in batches. Based on this, the detection probability of characteristic peaks of all natural fragrance raw materials in different intensity ranges in fragrance and flavor samples was calculated. According to the corresponding characteristic peak detection probability standards (99%, 97%, 95%, 90%, 80%, 70%, and 50%), the number of potential natural fragrance raw materials was confirmed to be m.

[0053] (3) Construct different combinations of potential natural fragrance raw materials, and analyze the consistency between the different combinations of potential natural fragrance raw materials and the tobacco flavorings to be reconstituted based on the liquid chromatography-high resolution mass spectrometry dataset of the natural fragrance raw materials, and determine the types and proportions of natural fragrance raw materials in tobacco flavorings.

[0054] The specific steps are as follows:

[0055] a) For m potential natural fragrance raw materials, determine the i characteristic peaks with the highest peak intensity of the potential natural fragrance raw materials, i = 10;

[0056] b) Calculate the ratio R of the above i characteristic peaks of the potential natural flavor raw material / to-be-reconstituted tobacco flavoring. i Find the mean R;

[0057] c) Based on the mean R, set the basic ratio D of the corresponding natural fragrance raw material, D = 1 / R; set the possible derivative ratios as D / 1.5, D / 2, and D / 3. Thus, for a certain natural fragrance raw material, there are a total of 4 ratio selection cases, namely D, D / 1.5, D / 2, and D / 3.

[0058] d) Among m potential natural fragrance raw materials, construct linear combinations of different quantities of natural fragrance raw materials, and set the ratio of individual natural fragrance raw materials in each combination (select one from 4 ratios), calculate the combination value of each combination, and compare it with the measured value to calculate the residual. The combination value is the peak intensity value of the highest i characteristic peaks, and the measured value is the peak intensity value of the corresponding i characteristic peaks of the fragrance to be reconstituted.

[0059] e) For n = 2, 3, 4, 5, 6, 7, 8, analyze and statistically analyze the information of the top k combinations with the smallest residuals under the corresponding quantities of natural fragrance raw materials, k = 100;

[0060] f) Compare the mean residuals of the first 100 combinations when n = 2, 3, 4, 5, 6, 7, 8 (if less than 100, use the actual value), and select n with the smallest residual as the determined number of natural fragrance raw materials in the fragrance to be recombined.

[0061] Among the linear combinations of n natural fragrance raw materials, the n natural fragrance raw materials with the highest frequency among the top 100 with the smallest residuals and their proportions are the types and proportions of natural fragrance raw materials in the fragrance and flavor to be reconstituted, as shown in Table 5 below.

[0062] Table 5. Types and proportions of natural fragrance ingredients in the flavorings and fragrances to be reconstituted.

[0063] Serial Number Natural fragrance ingredients Inferring proportions Frequency 1 Apricot extract 4.36％ 32 2 Licorice extract 3.39％ 100 3 Fig extract 2.78％ 82 4 Fenugreek extract from flue-cured tobacco 2.41％ 87 5 Plum extract 2.18％ 49 6 Cocoa extract 1.65％ 100

[0064] The detection probability standard in step (2) above can be adjusted appropriately according to specific cases. Combining the detection probability standard above with step (3) based on frequency further screening can effectively avoid false negatives (avoiding missed selection) and improve the accuracy of formula recombination.

[0065] (4) The water, ethanol, propylene glycol and glycerol in the tobacco flavoring to be reconstituted were determined by gas chromatograph equipped with a hydrogen flame ionization detector. The internal standard method was used for quantification. The results are shown in Table 6 below.

[0066] Table 6 Solvent content test results

[0067] Serial Number name Determine content 1 water 53.62％ 2 ethanol 26.19％ 3 Propylene glycol 15.33％ 4 glycerin Not detected

[0068] Step (3) determined that the total proportion of natural fragrance raw materials was 16.77%. The solvent composition was adjusted according to the detected solvent content to make the total of the pre-reconstituted formula 100%. The adjusted solvent composition of the formula is shown in Table 7 below:

[0069] Table 7 Results after solvent conversion

[0070] Serial Number name Determine content 1 water 46.91％ 2 ethanol 22.91％ 3 Propylene glycol 13.41％

[0071] (5) The tobacco flavoring and fragrance formula to be recombined was carried out in steps (3) and (4), and the results are shown in Table 8 below.

[0072] Table 8 Results of the recombination of tobacco flavoring and fragrance formulations to be recombined

[0073] Serial Number Natural fragrance ingredients Speculated proportions 1 Apricot extract 4.36％ 2 Licorice extract 3.39％ 3 Fig extract 2.78％ 4 Fenugreek extract from flue-cured tobacco 2.41％ 5 Plum extract 2.18％ 6 Cocoa extract 1.65％ 7 water 46.91％ 8 ethanol 22.91％ 9 Propylene glycol 13.41％

[0074] II. Experimental Examples

[0075] The recombinant sample obtained by formula recombination according to the method of Example 1 was subjected to a homogenization pretreatment and liquid chromatography-high resolution mass spectrometry analysis (same as Example 1) with the original tobacco flavoring and fragrance. The total ion current was as follows: Figure 1 and Figure 2 As shown. At the same time, the targeted extraction method in Example 1 was used to perform targeted extraction and comparative analysis on the analysis results of the original tobacco flavoring and recombinant samples prepared by the flavorists. The comparative analysis mainly included the overlap of total ion current, the correlation of characteristic peaks, and the matching degree of larger characteristic peaks (e.g., characteristic peaks with peak intensities in the top 20).

[0076] Depend on Figure 1 and Figure 2 It can be seen that the total ion current has good matching. In addition, the correlation coefficient of the characteristic peaks of the two is 0.98, and the matching degree of the larger characteristic peaks is also relatively consistent, all of which indicate that the recombinant sample has a better effect.

[0077] The flavoring and fragrance formulas for raw tobacco prepared by the perfumers were compared with the formulas for the reconstituted samples. The results are shown in Table 9 below.

[0078] Table 9 Comparison of the original tobacco flavoring and fragrance formulation with the reconstituted sample formulation.

[0079] Serial Number Natural fragrance ingredients Speculated proportions Theoretical ratio 1 Apricot extract 4.36％ 4.00％ 2 Licorice extract 3.39％ 3.50％ 3 Fig extract 2.78％ 3.00％ 4 Fenugreek extract from flue-cured tobacco 2.41％ 2.50％ 5 Plum extract 2.18％ 2.00％ 6 Cocoa extract 1.65％ 1.50％ 7 water 46.91％ 50.00％ 8 ethanol 22.91％ 20.00％ 9 Propylene glycol 13.41％ 13.50％

[0080] The results in Table 10 show that the pre-recombined formulation is basically consistent with the theoretical formulation, which fully demonstrates the effectiveness of the formulation recombination method of the present invention.

[0081] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for recombining the formulation of tobacco flavorings and fragrances, characterized in that, Includes the following steps: (1) The characteristic peaks of the natural fragrance raw materials were extracted by liquid chromatography-high resolution mass spectrometry using a non-targeting method. The characteristic peaks were aligned and then the liquid chromatography-high resolution mass spectrometry analysis results of the natural fragrance raw materials were targeted and extracted using the aligned characteristic peak information to construct a liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials. (2) Perform liquid chromatography-high resolution mass spectrometry analysis on the recombinant tobacco flavoring and fragrance and perform the targeted extraction. Compare and analyze the targeted extraction results with the liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials to identify potential natural fragrance raw materials. (3) Construct different combinations of potential natural fragrance raw materials, and analyze the consistency between the different combinations of potential natural fragrance raw materials and the tobacco flavorings to be reconstituted based on the liquid chromatography-high resolution mass spectrometry dataset of the natural fragrance raw materials, and determine the types and proportions of natural fragrance raw materials in tobacco flavorings; Constructing different combinations of potential natural flavor ingredients includes: identifying the i characteristic peaks with the highest intensities of the potential natural flavor ingredients, and calculating the ratio R of the corresponding characteristic peaks of the i natural flavor ingredients to the flavorings and fragrances to be pre-reconstituted in tobacco flavorings. i Find the mean R; set the basic ratio D of the natural fragrance raw material as 1 / R, and the derived ratios as D / 1.5, D / 2, and D / 3; assume the quantity of natural fragrance raw materials, and construct a linear combination based on the quantity of natural fragrance raw materials and the basic and derived ratios of each natural fragrance raw material; The analysis of consistency and determination of the types and proportions of natural flavoring raw materials include: obtaining the characteristic peak intensity of the linear combination and comparing it with the corresponding characteristic peak intensity of the flavorings to be reconstituted in the tobacco flavorings, and calculating the residual; statistically analyzing the top k linear combinations with the smallest residuals under each assumed quantity of natural flavoring raw materials, obtaining the mean of the residuals, and determining the assumed quantity of natural flavoring raw materials corresponding to the smallest mean of the residuals; under the determined assumed quantity of natural flavoring raw materials, statistically analyzing the types and proportions of natural flavoring raw materials with the highest frequency of occurrence in the top k corresponding linear combinations, which are the types and proportions of natural flavoring raw materials in the flavorings to be reconstituted in the tobacco flavorings.

2. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 1, characterized in that, The i = 10~15; assuming the number of natural fragrance raw materials is 2, 3, 4, 5, 6, 7, and 8 respectively.

3. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 1, characterized in that, k=100~120。 4. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 1, characterized in that, Step (4) includes detecting the types and contents of solvents in the flavorings and fragrances to be recombined. Based on the determined types and proportions of natural flavoring raw materials, the contents of each type of solvent are converted according to the detected contents, so that the total mass fraction of the recombined formula is 100%.

5. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 4, characterized in that, The solvent includes one, two, three, or four of the following: water, ethanol, propylene glycol, and glycerol.

6. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 1, characterized in that, In step (1), the characteristic peak is a peak not present in the blank sample or a peak with an intensity greater than 10 times that of the blank sample; the alignment processing rule is: retention time difference ≤ 0.2 min, precise molecular weight difference ≤ 5 ppm; the targeted extraction rule is: retention time difference ≤ 0.2 min, precise molecular weight difference ≤ 5 ppm.

7. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 1 or 6, characterized in that, In step (2), the comparative analysis includes batch statistical analysis of the number of characteristic peaks of different intensity ranges in the flavorings and fragrances to be reconstituted, as well as the number of characteristic peaks of each natural fragrance raw material corresponding to the intensity range, calculating the detection probability of characteristic peaks of different intensity ranges of all natural fragrance raw materials in the flavorings and fragrances to be reconstituted, and determining the types and quantities of potential natural fragrance raw materials based on the detection probability.

8. The method for recombining the formulation of tobacco flavorings and fragrances as described in claim 1, characterized in that, In step (1), the types of natural fragrance raw materials in the liquid chromatography-high resolution mass spectrometry dataset of natural fragrance raw materials cover extracts, tinctures and absolute oils, and the number of natural fragrance raw materials is more than 400.

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