Natural plant-based aerosol-generating substrate for electric heating smoking device

Abstract

A natural plant-based aerosol-generating substrate for an electric heating smoking device. The natural plant-based aerosol-generating substrate for an electric heating smoking device comprises: a natural plant raw material subjected to quantified cell wall disruption treatment, and an exogenous aerosol-generating agent loaded within the natural plant raw material. Cell wall components are decomposed by means of the quantified cell wall disruption treatment, so that a tobacco raw material can be quantitatively and controllably modified, causing localized rupture of a thinner cell wall to form an aerosol carrier, thereby increasing the amount of the aerosol-generating agent loaded onto the tobacco raw material. Furthermore, by monitoring the loading amount, the adjustment of an enzymatic cell wall disruption reaction and the control of the overall reaction mechanism can also be achieved. Moreover, on the premise of maintaining the original processability, more aerosol can be absorbed and more flavor substances can be emitted during heating.

Description

A natural plant aerosol generating substrate for an electrically heated smoking article TECHNICAL FIELD

[0001] The present application relates to the field of novel tobacco technology, in particular to a natural plant aerosol generating substrate for an electrically heated smoking article. BACKGROUND

[0002] In recent years, as the situation of tobacco control becomes increasingly severe and consumers' attention to health gradually increases, novel tobacco products are increasingly favored by the public. Aerosol generating products such as heated cigarettes have achieved good smoking experience after rapid development and optimization in recent years. However, due to the special heating method of heated cigarettes, existing traditional cut tobacco cannot be used as the core material of heated cigarettes, so the core material is mostly reconstituted tobacco, i.e. tobacco sheet, which is mainly composed of tobacco dust, tobacco fragments, tobacco stems or low-grade tobacco leaves, and is mainly prepared by papermaking method, thick pulp method and roller pressing method, which is quite different from traditional cut tobacco and needs to build a new production line. There are also attempts to use traditional cut tobacco for heated cigarettes. In order to release enough flavor substances under heating conditions, high-loss permeabilization treatment such as steam explosion and freeze-drying needs to be performed on traditional tobacco, but direct high-loss permeabilization treatment will seriously reduce the processability of tobacco, resulting in low whole silk rate, and there are also difficulties in rolling and packaging on the cut tobacco production line. SUMMARY

[0003] The present application provides a natural plant aerosol generating substrate for an electrically heated smoking article.

[0004] The present application adopts the following technical solutions:

[0005] In a first aspect, the present application provides a natural plant aerosol generating substrate for an electrically heated smoking article, which comprises: a natural plant raw material subjected to quantitative cell wall breaking treatment, and an exogenous aerosol generating agent loaded in the natural plant raw material.

[0006] Further, the quantitative cell wall breaking treatment comprises 1-5 enzymatic wall breaking steps, each enzymatic wall breaking step comprising: step A: uniformly spraying an active wall breaking enzyme solution on the natural plant raw material; and step B: sealing the sprayed natural plant raw material and performing an enzymatic wall breaking reaction under constant temperature and humidity.

[0007] Further, in step A, the weight ratio of the active wall breaking enzyme to the natural plant raw material is 0.5-1.0%, 1.0-1.5%, 1.5-2.0%, 2.0-2.5% or 2.5-3.0%.

[0008] Further, in step A, the active cell wall-breaking enzyme includes any one or a combination of at least two of pectinase, hemicellulase, cellulase or amylase.

[0009] Further, in step A, the active cell-wall-breaking enzyme is composed of pectinase, hemicellulase and amylase;

[0010] Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, cellulase, and amylase;

[0011] Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, hemicellulase, cellulase, and amylase, in the following weight ratio:

[0012] Pectinase: 15-25%,

[0013] Total hemicellulase and cellulase: 32-48%,

[0014] Amylase: 32-48%.

[0015] Further, in step B, the conditions for each enzymatic hydrolysis and cell wall breaking reaction are independently: temperature of 25-30℃, 30-35℃, 35-40℃, 40-45℃, 45-50℃, 50-55℃ or 55-60℃, relative humidity of 15-20%, 20-25%, 25-30%, 30-35%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 65-70%, 70-75% or 75-80%, and enzymatic hydrolysis and cell wall breaking reaction time of 0.5-1.0h, 1.0-2.0h, 2.0-4.0h, 4.0h-6.0h, 6.0-8.0h, 8.0-12.0h or 12.0-24.0h.

[0016] Further, after step B is completed, step C is performed: the apparent loading and / or micro loading of the natural plant material are detected. When the apparent loading and / or micro loading of the aerosol generating agent of the natural plant material is greater than the target loading, the next enzymatic hydrolysis and cell wall breaking step is not performed.

[0017] Furthermore, based on the difference between the apparent loading and / or microscopic loading of the aerosol generator of the natural plant raw material and the target loading, the enzymatic hydrolysis reaction temperature, relative humidity, and / or reaction time in the next enzymatic hydrolysis step are determined.

[0018] Furthermore, the apparent loading test includes weighing the maximum load of the exogenous aerosol generating agent; the micro loading test includes observing the cell wall and statistically analyzing the cell wall disruption rate.

[0019] Furthermore, after each enzymatic hydrolysis and cell wall breaking reaction is completed, step D is performed: the natural plant raw materials undergo harm reduction treatment.

[0020] Furthermore, the exogenous aerosol generating agent is loaded into the natural plant material that has undergone quantitative cell wall disruption treatment in the following manner: the exogenous aerosol generating agent is sprayed onto the natural plant material that has undergone quantitative cell wall disruption treatment, wherein: the weight of the exogenous aerosol generating agent is less than the weight of the natural plant material × the maximum loading rate × the whole fiber rate; after the exogenous aerosol generating agent has completely penetrated into the natural plant material that has undergone quantitative cell wall disruption treatment, a natural plant aerosol generating matrix is ​​prepared.

[0021] Furthermore, after each enzymatic hydrolysis and cell wall breaking reaction is completed, step E is performed: the whole fiber rate of the natural plant material is detected. If the whole fiber rate of the natural plant material is greater than the whole fiber rate warning value, the next enzymatic hydrolysis and cell wall breaking step will not be performed.

[0022] Furthermore, the warning value for the whole yarn rate is above 60%, above 65%, above 68%, above 70%, above 72%, above 75%, above 78%, or above 80%.

[0023] Secondly, this application provides a method for manufacturing an aerosol-generating matrix as described in the first aspect.

[0024] Thirdly, this application provides an aerosol generating article comprising the natural plant aerosol generating matrix for electric heating smokers as described in the first aspect.

[0025] Fourthly, this application provides an aerosol generation system, which includes an electrically heated aerosol generation device and the aerosol generation article described in the third aspect.

[0026] Compared with the prior art, this application has the following beneficial effects:

[0027] (1) In this application, scanning electron microscopy and thermogravimetric-gas chromatography-mass spectrometry are used to detect and analyze tobacco leaf samples before and after active enzyme treatment, and to explore the influence of active enzyme on the microstructure of tobacco cell wall and heat release pattern. The aim is to enhance the understanding of the properties of tobacco raw materials after active enzyme action and provide technical support for the development of heated cigarette smoke-generating media.

[0028] (2) This application achieves quantitative and controllable modification of tobacco raw materials by quantifying cell wall disruption treatment to decompose cell wall components, thereby locally disrupting thinner cell walls to form aerosol loads and increasing the loading capacity of tobacco raw materials for aerosol generating agents. Furthermore, monitoring the loading capacity allows for adjustment of the enzymatic cell disruption reaction and control of the overall reaction mechanism. Moreover, while maintaining the original processability (especially the whole tobacco yield), it can absorb more aerosols and release more aroma substances during heating, enabling traditional cigarette production lines to be reused with heated cigarettes with only minor modifications. By selecting specific degradation enzyme compositions, enzymatic degradation reactions can be achieved in the existing leaf storage environment without the need for dedicated high-temperature reaction equipment. Since solvent soaking and microwave heating are not required, aroma substance loss is avoided, allowing for full volatilization of the tobacco's natural aroma. The aerosol addition amount determined by the maximum loading rate and whole tobacco yield ensures overall processability. Attached Figure Description

[0029] The above description of this application and the following detailed embodiments will be better understood when read in conjunction with the accompanying drawings. It should be noted that the drawings are merely examples of the claimed technical solutions.

[0030] Figure 1 shows the morphology of tobacco leaf cells treated with pectinase under a transmission electron microscope (scale bar: 2 μm).

[0031] Figure 2 shows the morphology of tobacco leaf cells treated with hemicellulase under a transmission electron microscope (scale bar: 10 μm).

[0032] Figure 3 shows the morphology of tobacco leaf cells treated with hemicellulase under a transmission electron microscope (scale bar: 2 μm).

[0033] Figure 4 is a comparison of the effects of different concentrations of pectinase treatment on the pectin content in the cell walls of tobacco leaves.

[0034] Figure 5 is a comparative diagram of the effects of different concentrations of hemicellulase treatment on the hemicellulose content in the cell wall of tobacco leaves.

[0035] Figure 6 is a comparison of the effects of different concentrations of amylase treatment on the starch content of tobacco leaf cell walls;

[0036] Figure 7 is a schematic diagram of the maximum atomizing agent load testing device of this application, wherein the reference numerals are explained as follows: 1-tobacco, 2-extrusion container, 3-force probe;

[0037] Figure 8A shows the TG curves of tobacco leaves treated with different concentrations of pectinase.

[0038] Figure 8B shows the DSC curves of tobacco leaves treated with different concentrations of pectinase.

[0039] Figure 9A shows the TG curves of tobacco leaves after treatment with different enzymes.

[0040] Figure 9B shows the DSC curves of tobacco leaves after treatment with different enzymes.

[0041] Figure 10A shows the TG curves of tobacco leaves treated with different concentrations of hemicellulase.

[0042] Figure 10B shows the DSC curves of tobacco leaves treated with different concentrations of hemicellulase.

[0043] Figure 11A shows the TG curves of tobacco leaves treated with different concentrations of amylase.

[0044] Figure 11B shows the DSC curves of tobacco leaves treated with different concentrations of amylase.

[0045] Figure 12A shows the TG curves of tobacco leaves treated with different enzyme compositions.

[0046] Figure 12B shows the DSC curves of tobacco leaves treated with different enzyme compositions;

[0047] Figure 13 shows the release of five volatile substances in tobacco leaves after treatment with different concentrations of pectinase;

[0048] Figure 14 shows the detection of aroma components in tobacco leaf samples treated under different storage conditions.

[0049] Figure 15 shows the cell wall rupture rate after treatment with different concentrations of pectinase;

[0050] Figure 16A shows the TG curves after adding different amounts of atomizing agent;

[0051] Figure 16B shows the DSC curves after adding different amounts of atomizing agent;

[0052] Figure 17 is a schematic diagram of the manufacturing process of the natural plant aerosol generation matrix described in this application;

[0053] Figure 18 is a schematic diagram of the main equipment deployment of the production line of this application. Detailed Implementation

[0054] To further illustrate the technical means and effects adopted in this application, the following description, in conjunction with embodiments and accompanying drawings, will provide further details. It is understood that the specific embodiments described herein are merely for explaining this application and not for limiting it.

[0055] The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not limited to those elements and may also include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0056] "Optional" or "any one" means that the matter or event described thereafter may or may not occur, and the description includes both the possibility that the event will occur and the possibility that the event will not occur.

[0057] The indefinite articles “a” and “an” preceding an element or component in this application do not impose any limitation on the quantity (i.e., number of occurrences) of the element or component. Therefore, “an” or “a” should be interpreted as including one or at least one, and the singular form of an element or component also includes the plural form, unless the quantity clearly refers only to the singular form.

[0058] The terms "one embodiment," "some embodiments," "exemplary," "specific example," or "some examples," etc., described in this application refer to a specific feature, structure, material, or characteristic described in connection with that embodiment or example, which is included in at least one embodiment or example of this application. In this document, the illustrative expressions of the above terms are not necessarily directed at the same embodiment or example.

[0059] I. Raw Materials

[0060] Unless otherwise specified below, the tobacco used is 2021 Henan Sanmenxia Mianchi C3F Qinyan 96 flue-cured tobacco.

[0061] The applicant also used the following flue-cured tobacco leaves for experiments: Chenzhou Guiyang B2F, C2F, X2F; Yongzhou Lanshan B3F, C2F, X2F; Sanmenxia Mianchi B2F, C3F, X3F; Guizhou Zunyi B2F, C2F, X2F; Bijie Dafang B3F, C3F, X2F; Guizhou Bijie B2F, C2F, X3F; Yunnan Yuxi B3F, C3F, X2F; Yunnan Kunming B2F, C3F, X2F; Yunnan Dali B2F, C3F, X2F, including tobacco seed varieties such as Honghua Dajinyuan, Cuibi No. 1, and Qinyan 96. In addition, the applicant also used burley tobacco, aromatic tobacco, cigar tobacco, sun-cured tobacco, and cloves for experiments. Unless otherwise specified below, this application also applies to the above-mentioned raw materials.

[0062] II. Enzyme Preparations

[0063] 1. Pectinase: Activity 60000U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., the main components are protopectinase, polygalacturonase, pectin lyase and pectin esterase.

[0064] 2. Hemicellulase: Activity 50000U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., the main component is mannanase, its function is to degrade hemicellulose. The applicant found that after it acts on the tobacco cell wall, the products are mainly mannooligosaccharides and a small amount of mannose, etc.

[0065] 3. Amylase: Activity 50000U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., contains multiple bioactive components such as cellulase, β-glucanase and xylanase. The applicant found that it can degrade non-starch polysaccharides in tobacco, rapidly disintegrate the structure of fiber, protein and starch that are intertwined, promote the separation of each component, and improve the content of residual starch in fiber.

[0066] 4. Cellulase: Activity 50000U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd., Xiasheng Cellulase (CEL-01 type), mainly contains endonuclease (EG), and contains a small amount of exonuclease and a small amount of β-1,4 glucosidase.

[0067] 5. Flavor protease: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd. The applicant discovered that adding suitable flavor proteases to tobacco can hydrolyze flavor precursors and other existing enzymes in tobacco, or the aforementioned added enzymes, thereby releasing flavor substances and enhancing and improving the flavor of the food; it can also control the bitterness of peptides. The principle is that endopeptides cleave the peptide bonds within polypeptides to form short-chain peptides, some of which contain hydrophobic amino acids, thus becoming bitter peptides. Exopeptides then cleave the polypeptide chain one amino acid at a time from the end, thereby completely degrading the bitter peptides into amino acids.

[0068] 6. Neutral protease: Activity 50000 U / g, purchased from Beijing Xiasheng Biotechnology Development Co., Ltd. Neutral proteases belong to the class of hydrolases. Based on the location of the hydrolytic site on the peptide chain, they can be further subdivided into aminopeptidases, carboxypeptidases, and endopeptidases. Aminopeptidases and carboxypeptidases hydrolyze peptide bonds closest to the N-terminus or C-terminus of the protein substrate, while endopeptidases act on the interior of the protein, far from its N-terminus and C-terminus. These enzymes work together to hydrolyze the peptide bonds in the protein molecule, breaking down the protein into oligopeptides and amino acids. The applicant discovered that the neutral protease used mainly contains endopeptidase activity, with small amounts of aminopeptidase and carboxypeptidase activity. This can be used to hydrolyze residual biological enzymes in tobacco leaves after enzyme treatment, avoiding the generation of burnt feather odor during heating.

[0069] III. Other Major Materials, Reagents and Instruments

[0070] Experimental water: Grade I water (resistivity > 18.2 MΩ·cm) as specified in GB / T 6682-2008.

[0071] Instruments: ME204 electronic balance (sensitivity 0.0001g, Mettler Toledo, USA); A33 continuous flow analyzer (BRAN LUEBBE, Germany); Fibertec 2010 fiber analyzer (FOSS, Denmark); Helios NanoLab 600i dual-beam focused ion beam scanning electron microscope (FEI, USA); Spirit 120kV transmission electron microscope (FEI, USA); ICS-3000 multi-functional ion chromatograph (Dionex, USA); 6890N-5975C gas chromatograph-mass spectrometer (Agilent Technologies, USA); TGA / DSC 3+ thermogravimetric analyzer; IST 16 thermal analysis system (Mettler-Toledo, Switzerland); Milli-Q Reference ultrapure water system (Millipore, USA); KBF 720 constant temperature and humidity chamber (Binder, Germany).

[0072] Blank Example A: Enzymatic hydrolysis without the addition of exogenous biological enzymes

[0073] According to traditional processes, the raw tobacco leaves are rehydrated, cut into shreds to obtain tobacco shreds, and then subjected to low-loss permeability treatment and harm reduction treatment, with each group consisting of 50g.

[0074] Take 50g of high-purity water (without active enzyme composition) and place it in a laboratory sprayer, then spray it evenly onto the tobacco product. Seal the sprayed tobacco product and place it in a constant temperature and humidity chamber (temperature 55℃, relative humidity 25%) for 4 hours. Then place the reacted tobacco in an oven at 80℃ for 20 minutes. Finally, dry the final tobacco in an oven at 50℃ for 20 minutes, and use it as a tobacco sample for later use.

[0075] Examples B1-B4: Pectinase hydrolysis

[0076] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0077] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0078] In Examples B1-B4, pectinase was uniformly sprayed onto the tobacco shreds according to the mass ratio of pectinase to tobacco in Table B-1. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0079] Table B-1: Mass ratio of pectinase and tobacco in each example

[0080] Examples B5-B6: Pectinase hydrolysis (at different storage temperatures and humidity levels)

[0081] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0082] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0083] Pectinase was evenly sprayed onto the tobacco shreds at a mass ratio of 1.0% to 1.0%, corresponding to examples B5-B7. The sprayed tobacco shreds were sealed and placed in a constant temperature and humidity chamber for enzymatic hydrolysis for 4 hours according to the conditions set in Table B-2. The hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20min). The finally hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20min) and used as tobacco samples for later use.

[0084] Table B-2: Setting conditions of the constant temperature and humidity chamber in each embodiment

[0085] Examples B7-B12: Pectinase hydrolysis (different hydrolysis reaction times)

[0086] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0087] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0088] Pectinase was evenly sprayed onto the tobacco shreds at a mass ratio of 1.0% to 1.0%, corresponding to examples B7-B12. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%), and the enzymatic hydrolysis reaction time was set according to Table B-3. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20min). For example B10, the spraying and enzymatic hydrolysis process was repeated three times to ensure enzyme activity; for example B11, the spraying and enzymatic hydrolysis process was repeated four times to ensure enzyme activity. The finally enzymatically hydrolyzed tobacco shreds were then placed in an oven to dry (set temperature 50℃, baking time 20min) and used as tobacco shred samples for later use.

[0089] Table B-3: Enzymatic hydrolysis reaction time for each embodiment

[0090] Examples B13-B15: (Different methods for terminating enzymatic hydrolysis reactions)

[0091] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0092] Dilute the pectinase with ultrapure water to prepare a 200 U / g pectinase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0093] Pectinase was evenly sprayed onto the tobacco shreds at a pectinase to tobacco mass ratio of 1.0%, corresponding to examples B13-B15. The sprayed tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction, with the oven parameters set according to Table B-4. The finally hydrolyzed tobacco shreds were then dried in the oven (set temperature 50℃, drying time 20 minutes) and used as tobacco shred samples for later use.

[0094] Table B-4: Conditions for terminating the enzymatic hydrolysis reaction in each example

[0095] Examples C1-C4: Enzymatic hydrolysis of hemicellulase

[0096] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0097] Dilute the hemicellulase with ultrapure water to prepare a 200 U / g hemicellulase solution. Prepare and use immediately. Place the hemicellulase solution into a laboratory sprayer.

[0098] Hemicellulase was evenly sprayed onto the tobacco shreds according to the ratio of hemicellulase to tobacco in Table C-1, corresponding to examples C1-C4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0099] Table C-1: Mass ratio of hemicellulase to tobacco in each example

[0100] Examples D1-D4: Amylase hydrolysis

[0101] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0102] Dilute the amylase with ultrapure water to prepare a 200 U / g amylase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0103] According to the amylase-to-tobacco ratio in Table D-1, amylase was evenly sprayed onto the tobacco shreds, corresponding to examples D1-D4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven for enzymatic hydrolysis (set temperature 80℃, baking time 20 min). Finally, the enzymatically hydrolyzed tobacco shreds were dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0104] Table D-1: Mass ratio of amylase and tobacco in each example

[0105] Examples E1-E4: Cellulase hydrolysis

[0106] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0107] Dilute the cellulase with ultrapure water to prepare a 200 U / g cellulase solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0108] Cellulase was evenly sprayed onto the tobacco shreds according to the ratio of cellulase to tobacco in Table E-1, corresponding to examples E1-E4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0109] Table E-1: Mass ratio of cellulase and tobacco in each example

[0110] Examples F1-F4: Enzymatic hydrolysis with flavor proteases

[0111] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0112] The flavor protease was diluted with ultrapure water to prepare a 200 U / g flavor protease solution. The solution was prepared and used immediately. The flavor protease solution was placed in a laboratory sprayer.

[0113] Flavor protease was evenly sprayed onto the tobacco shreds according to the ratio of flavor protease to tobacco as specified in Table F-1, corresponding to examples F1-F4 respectively. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0114] Table F-1: Mass ratio of flavor protease and tobacco in each example

[0115] Examples F5-F8: Enzymatic hydrolysis with neutral protease

[0116] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0117] Dilute the neutral protease with ultrapure water to prepare a 200 U / g neutral protease solution. Prepare the solution immediately before use and place it in a laboratory sprayer.

[0118] Neutral protease was evenly sprayed onto the tobacco shreds according to the ratio of neutral protease to tobacco in Table F-2, corresponding to examples F1-F4 respectively. The sprayed tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0119] Table F-2: Mass ratio of neutral protease and tobacco in each example

[0120] Examples G1-G6: Enzymatic hydrolysis of enzyme compositions

[0121] According to traditional processes, the raw tobacco leaves are rehydrated and then shredded to obtain tobacco shreds. Each batch of tobacco shreds weighs 50g and undergoes low-loss permeability treatment and harm reduction treatment.

[0122] Prepare the enzyme composition according to the following weight ratio, dilute the enzyme composition with ultrapure water to prepare an enzyme composition solution of 200 U / g, and use it immediately. Put the enzyme composition solution into a laboratory sprayer.

[0123] The enzyme composition formulations are shown in Table G-1, corresponding to examples G1-G6 of the tobacco shreds. The enzyme composition was evenly sprayed onto the tobacco shreds at a mass ratio of 1.0% to 1.0%. After spraying, the tobacco shreds were sealed and placed in a constant temperature and humidity chamber (set temperature 55℃, relative humidity 25%) for enzymatic hydrolysis for 4 hours. The enzymatically hydrolyzed tobacco shreds were then placed in an oven to terminate the enzymatic hydrolysis reaction (set temperature 80℃, baking time 20 min). The finally enzymatically hydrolyzed tobacco shreds were then dried in an oven (set temperature 50℃, baking time 20 min) and used as tobacco shred samples for later use.

[0124] Table G-1: Composition Ratio

[0125] Examples H1-H5: Different nebulization doses

[0126] Add 10%, 15%, 17.5%, 20%, and 25% of the tobacco weight of the tobacco shreds to the tobacco shreds obtained in Example B5 respectively, and then use them as tobacco shreds samples for Examples H1-H5 respectively.

[0127] Examples H6-H9: Different atomizing agent formulations

[0128] Add 15% by weight of atomizing agent, consisting of glycerol and propylene glycol, to the tobacco shreds obtained in Example B5. The formulation of the atomizing agent is shown in Table H-1. Then, use these as tobacco shreds samples for Examples H6-H9.

[0129] Table H-1: Proportions of atomizing agents in each embodiment

[0130] To further demonstrate the beneficial effects of this application, the tobacco samples obtained from blank example A and each of the examples B to H were tested using the following test examples.

[0131] Experimental Example 1: Routine Chemical Composition Detection

[0132] The routine chemical composition of Comparative Example A and Samples B1-B4, C1-C4, and D1-D4 was determined using industry-standard methods, including the following assays:

[0133] YC / T 159―2019 Determination of water-soluble sugars in tobacco and tobacco products - Continuous flow method;

[0134] YC / T 160-2002 Determination of total alkaloids in tobacco and tobacco products by continuous flow method;

[0135] YC / T 161-2002 Determination of total nitrogen in tobacco and tobacco products by continuous flow method;

[0136] YC / T 162-2011 Determination of chlorine in tobacco and tobacco products - Continuous flow method;

[0137] YC / T 217-2007 Determination of potassium in tobacco and tobacco products by continuous flow method.

[0138] All experimental results in this application were repeated three times, and the data are expressed as mean ± standard deviation. SPSS software (SPSS 13.0, SPSS Inc., USA) was used for statistical analysis of differences between groups. One-way analysis of variance (ANOVA) was used to perform the least significant difference (LSD) test. p < 0.05 indicates a statistically significant difference, indicated by *. The results are shown in Tables 1–3.

[0139] Table 1: Effects of pectinase treatment on conventional chemical components of tobacco leaves

[0140] Table 1 shows that pectinase treatment mainly produces sugar compounds. The higher the content of sugar compounds, the better the aroma of heated cigarettes and the smoother and more delicate the smoke.

[0141] Table 2: Effects of hemicellulase treatment on conventional chemical components of tobacco leaves

[0142] Table 2 shows that hemicellulase treatment mainly produces carbohydrate compounds.

[0143] Table 3: Effects of amylase treatment on conventional chemical components of tobacco leaves

[0144] Table 3 shows that amylase treatment mainly produces carbohydrate compounds and affects chlorides.

[0145] Experimental Example 2: Detection of Major Components of the Cell Wall

[0146] The main chemical components of the cell wall in Comparative Example A and Examples B1-B4, C1-C4, and D1-D4 were determined using industry-standard methods:

[0147] YC / T 347-2010 Determination of neutral detergent fiber, acid detergent fiber and acid-washed lignin in tobacco and tobacco products - Detergent method;

[0148] YC / T 346-2010 Determination of pectin in tobacco and tobacco products by ion chromatography.

[0149] Figure 4 shows the test results of pectin content in cell walls after pectinase treatment in Examples B1-B4. Pectinase decomposes pectin with a significant dose-response relationship. When the amount of pectinase added is 1.5%, a statistically significant difference occurs.

[0150] Figure 5 shows the test results of hemicellulose content in the cell wall after hemicellulase treatment. Hemicellulase decomposes hemicellulose with a significant dose-response relationship. When the amount of hemicellulase added is 1.0%, a statistically significant difference is observed.

[0151] Figure 6 shows the results of the starch content test of the cell wall after amylase treatment. The amylase decomposition of starch has a significant dose-response relationship. When the amount of amylase added is 1.0%, a statistically significant difference is observed.

[0152] Test Example 3: Maximum Atomizing Agent Load Test

[0153] Based on the products of Comparative Examples A and Examples B1-B19, C1-C4, D1-D4, E1-E4, F1-F8, and G1-G6, a certain mass of tobacco samples was weighed and placed in a constant temperature and humidity chamber, where they were equilibrated for 4 hours at 25°C and 20% humidity. Using a tobacco filling value measuring instrument, 20g of the equilibrated tobacco was accurately weighed, and 1% (by weight of the tobacco) of an atomizing agent was evenly sprayed onto the tobacco. The atomizing agent could be 100% glycerol. Simultaneously, tests were conducted for Examples A and B1-B4 using 100% propylene glycol as the atomizing agent. Alternatively, 100% propylene glycol or a combination of two or more polyols could be used as the aerosol for the same experiment.

[0154] As shown in Figure 7, tobacco shreds 1 with 1% atomizing agent (relative to the initial tobacco shred mass) are placed in a compression container 2. The compression container 2 is a cylindrical barrel with a diameter of (60.0±0.1) mm and a height of not less than 100 mm. A force-applying probe 3 with a diameter of (55.0±0.1) mm can be applied to the surface of the tobacco shreds, generating a uniform pressure of (29.4±0.5) N on the sample. The pressure application speed of the force-applying probe is (19.5±0.5) mm / s, and the pressure application time is (30.0±0.5) s. After the pressure is applied, the tobacco shreds are poured out onto a flat piece of white paper and gently shaken. If the tobacco shreds can be dispersed well without obvious clumping, then 1% atomizing agent (relative to the initial tobacco shred mass) is applied again, and all other conditions remain unchanged, and the above experiment is repeated. The experiment was terminated when the tobacco shreds were poured onto a flat sheet of white paper and gently shaken. If the tobacco shreds could not disperse well and showed obvious clumping, it indicated that the atomizing agent had not been extracted. The amount of atomizing agent used at this point was recorded as B%. The maximum load of tobacco substances on the atomizing agent under these conditions was (B-1)%. The test results are shown in Tables 4-6.

[0155] Table 4: Maximum loading after different enzyme treatments

[0156] Table 5: Maximum glycerol loading after treatment with different enzyme compositions

[0157] Table 6: Maximum glycerol loading after different total enzymatic hydrolysis times

[0158] Based on scanning electron microscopy experiments and maximum nebulizer loading experiments, it can be deduced that the maximum loading is determined by the cell wall rupture rate, intercellular space, and the viscosity of the nebulizer. Table 7 shows the results of intercellular space, cell wall rupture rate, and maximum nebulizer loading tests for Examples A and B1-B4.

[0159] Table 7: Relationship between intercellular space, cell wall rupture rate and maximum atomizing agent load

[0160] Experimental Example 4: Thermogravimetric Analysis of Tobacco Shreds

[0161] This application conducted thermogravimetric analysis on tobacco leaves treated with 0–2.0% enzyme to investigate the effect of different cell wall rupture degrees on the heat release performance of tobacco leaves.

[0162] Approximately 5 mg of tobacco sample treated with enzyme solution was weighed, and 18% atomizing agent (100% glycerol) was added. The sample was then placed in an alumina crucible and subjected to thermogravimetric analysis (TGA). Since the heating temperature of the cigarette core section did not exceed 400℃, the heating program was selected to increase the temperature from 30℃ to 400℃ at a rate of 20℃ / min; gas flow rate: 40 mL / min; ambient gas: air. Each parallel sample was repeated three times.

[0163] The results of tobacco leaves treated with different concentrations of pectinase in Examples B1-B4 are shown in Figures 8A and 8B. When the temperature is between 30 and 125°C, the weight loss ratio of the tobacco leaves treated with different concentrations of pectinase is relatively small, ranging from 5% to 10%, and the corresponding weight loss rate of the DTG curve is also low. Notably, the temperature at which the tobacco leaves treated with pectinase exhibit the maximum weight loss rate in this temperature range is 25°C lower than that of the blank example A, suggesting that cell wall rupture in this temperature range promotes the evaporation and premature release of water and some small molecule chemical substances. When the temperature is between 125 and 250°C, the TG curve gradually becomes the stage of maximum weight loss for the tobacco leaves with increasing pectinase concentration, accounting for 25% to 35% of the weight loss. The corresponding weight loss rate of the DTG curve is relatively large, with the temperature at which the maximum weight loss rate occurs being 210°C. When the pectinase concentration exceeds 50 U / g, this stage represents the range of maximum weight loss rate, and the weight loss percentage increases with increasing pectinase concentration. The applicant believes this stage may be the release range of major small-molecule aroma compounds. When the temperature is between 250 and 350℃, the TG curve shows that the weight loss percentage of tobacco leaves is between the first two temperature ranges, ranging from 15% to 25%. The corresponding DTG curve shows the maximum weight loss rate at 310℃, and the weight loss percentage decreases with increasing pectinase concentration. The applicant believes that the weight loss at this stage may be due to the decomposition of large-molecule non-volatile substances in tobacco. The enzymatic hydrolysis of pectin reduces the substrate in this stage, leading to a slight decrease in the weight loss percentage. As the amount of enzyme added increased, the proportion of weight loss due to heat in the tobacco-type smoking medium continued to increase. Compared with blank example A, the total weight loss of the experimental groups with enzyme addition of 0.5%, 1%, 1.5% and 2% increased by 4.09%, 5.03%, 7.08% and 10.06% respectively, and the weight loss range was mainly between 170-220℃.

[0164] Taking 1% of different enzyme treatments as an example, thermogravimetric analysis was performed on Examples B2, C2, D2, F2 and F6. The results are shown in Figures 9A and 9B. The TG / DTG results further confirmed the sensory evaluation results. Pectinase, hemicellulase and amylase can significantly increase the ratio of thermal weight loss, which is speculated to be because they destroy the cell wall structure and increase the release of aroma substances. Figures 10-12 further verify this conclusion.

[0165] Thermogravimetric analysis of tobacco samples treated with 0–2% hemicellulase in Examples C1–C4 is shown in Figures 10A and 10B.

[0166] Thermogravimetric analysis of tobacco samples treated with 0-2% amylase in Examples D1-D4 is shown in Figures 11A and 11B.

[0167] Thermogravimetric analysis of tobacco samples treated with the enzyme compositions in Examples G1-G6 is shown in Figures 12A and 12B.

[0168] Experimental Example 5: Thermogravimetric-Gas Mechanism Analysis of Tobacco Shreds

[0169] Approximately 5 mg of enzyme-treated tobacco sample was weighed and placed in a crucible. The thermogravimetric analysis (TGA) temperature program was as follows: temperature increased from 30 °C to 400 °C at a rate of 100 °C / min; gas flow rate was 40 mL / min; and the ambient gas was air. An IST 16 coupled system was used to connect the TGA analyzer and the gas chromatography-mass spectrometer (GC-MS), employing a single-loop injection mode. The valve opening time was set according to the experimental conditions; sampling time: 10 s; loop volume: 120 μL.

[0170] The gas chromatography conditions are as follows:

[0171] The injection mode was external direct injection; the column was a DB-5MS capillary column (30m × 0.25mm, 0.25μm) (Agilent Technologies, USA); the carrier gas was helium; the carrier gas flow rate was 1.0mL / min; the injection port temperature was 260℃; the injection volume was 10μL; and the split ratio was 10:1. The temperature program was: 60℃ for 1 min, then increased to 280℃ at a rate of 15℃ / min, and held at 280℃ for 15 min; the mass spectrometry ionization mode was EI; the ion source temperature was 230℃; the ionization energy was 70eV; the quadrupole temperature was 150℃; the scan range was 30–450 amu; and the solvent delay was 2 min.

[0172] Thermogravimetric analysis (TGA) and gas chromatography-mass spectrometry (GC-MS) were performed on tobacco samples treated with different concentrations of pectinase in Examples B1-B4. Since the tobacco leaves treated with different concentrations of pectinase showed significant differences in thermal release at 210°C, and the heating temperature range of most heated cigarette core segments included 210°C, the quantitative loop valve was set to open at 660 seconds, corresponding to a reaction temperature of 210°C. Gas samples released during thermogravimetric analysis were collected for GC-MS analysis, and some of the results are shown in Figure 13. A total of 18 tobacco-related compounds were identified in the released substances from the tobacco leaves, mainly including nicotine, furfural, furfuryl alcohol, solanone, cyclopentanone, 5-hydroxymethylfurfural, and neophytadiene. Among them, five compounds showed a significant dose-response effect with the amount of pectinase used. As the pectinase treatment concentration increased, the peak area per unit weight of furfural, furfuryl alcohol, 5-hydroxymethylfurfural, and solanone also increased to varying degrees, with the first three showing the most significant increase; the peak area per unit weight of neophytadiene did not change significantly. The results indicate that using pectinase to degrade pectin in tobacco leaves promotes the formation and release of browning reaction products, thereby further enriching the aroma of heated cigarettes. Studies have shown that pectinase can effectively degrade pectin in cell walls, increasing the amount of neutral aroma compounds, with nicotine and some high-molecular-weight aroma compounds being released concentratedly within the heating temperature range of 120℃ to 250℃. Therefore, this application finds that treating tobacco leaves with pectinase can promote the thermal release of aroma compounds in the tobacco raw material. Other active enzyme compositions, such as hemicellulase, amylase, or enzyme combinations, can also achieve similar effects to pectinase.

[0173] Further thermogravimetric-gas chromatography-mass spectrometry (TGA) analysis was performed on tobacco samples treated under different storage conditions in Examples B2, B5, and B6. As shown in Figure 14, after enzyme treatment, the cell walls of tobacco leaves were disrupted, making it easier for cells to exchange substances with the external environment when heated. If the temperature is too high, there will be a significant loss of aroma compounds in the tobacco. Using representative aroma compounds megastigmatrienone, neophytadiene, and furfuryl alcohol as research subjects, it was demonstrated that the storage environment of 29°C + 70% in Example B5 was the best among the three examples.

[0174] Experimental Example 6: Scanning Electron Microscopy Experiment of Tobacco Slices

[0175] This experimental case was tested at the Protein Science Research Platform of the Institute of Biophysics, Chinese Academy of Sciences.

[0176] The tobacco samples were placed in phosphate buffer (0.1 M PB, pH 7.2-7.4) of 2.5% glutaraldehyde + 2% paraformaldehyde at 4°C for 3 days. They were then washed four times on ice with 0.1 M PB for 10 min each time. The tobacco samples were then fixed at 4°C for 2 h with 1% osmium tetroxide solution (0.1 M PB). They were then washed three times on ice with redistilled water for 10 min each time. Gradient dehydration was performed using ethanol solution according to the conditions in Table 8, and gradient osmosis was performed according to the conditions in Table 9.

[0177] Table 8: Gradient elution conditions

[0178] Table 9: Gradient Permeability Conditions

[0179] The tobacco samples, after being permeated with the enzyme solution, were placed in a mold filled with resin (+0.7% DMAE) and polymerized at 45°C for 12 h and 70°C for 24 h, respectively. The polymerized tobacco samples were then cut into 70 nm thick tobacco sheets using a Leica UC6 slicer, and the tobacco sheet samples were backscattered electron images performed using a scanning electron microscope.

[0180] The cell wall rupture rate of tobacco leaves treated with different concentrations of pectinase in Examples B1-B4 (Figure 15) showed that pectinase disrupted the cell wall in a significant dose-response relationship. When the amount of pectinase added was greater than 1%, the cell wall rupture rate of the experimental group was significantly different from that of the blank example A, indicating that under this concentration, pectinase could significantly disrupt the cell wall structure of tobacco leaves.

[0181] Meanwhile, the cell scanning electron microscopy results of the tobacco samples from Examples B1-B4 using a Spirit 120kV transmission electron microscope are shown in Figure 1, where Figure 1-A is blank example A; Figure 1-B is Example B1 (treated with 0.5% pectinase); Figure 1-C is Example B2 (treated with 1.0% pectinase); Figure 1-D is Example B3 (treated with 1.5% pectinase); and Figure 1-E is Example B4 (treated with 2.0% pectinase).

[0182] As shown in the figures, in blank example A (Figure 1-A), the tobacco cells have an intact overall structure, with complete organelles and clear boundaries. However, with increasing pectinase concentration, cell wall rupture, organelle dissolution, and cell wall swelling occur in Figures 1-C, D, and E, respectively. The decomposition and rupture of the cell wall may be the mechanism by which heated cigarette tobacco-type smoke-generating media effectively adsorbs atomizing agents and releases flavor substances upon heating.

[0183] As shown in Figures 2 and 3, where Figure 2-A is blank example A; Figure 2-B is example C1 (treated with 0.5% hemicellulase); Figure 2-C is example C2 (treated with 1.0% hemicellulase); Figure 2-D is example C3 (treated with 1.5% hemicellulase); and Figure 2-E is example C4 (treated with 2.0% hemicellulase).

[0184] Figure 3-A is blank example A; Figure 3-B is example C1 (treated with 0.5% hemicellulase); Figure 3-C is example C2 (treated with 1.0% hemicellulase); Figure 3-D is example C3 (treated with 1.5% hemicellulase); Figure 3-E is example C4 (treated with 2.0% hemicellulase).

[0185] Transmission electron microscopy (TEM) results of 10 μm and 2 μm intercellular gaps in tobacco samples treated with hemicellulase in Examples C1-C4 show that in blank example A (Fig. 2-A) and the low-concentration treatment group (Fig. 2-B), tobacco cells are arranged in an orderly and compact manner with clear cell wall boundaries. However, as the concentration of the gradient treatment increases, in Fig. 2-C, disordered cell arrangement, widening of intercellular gaps, cell wall swelling, and further widening of intercellular gaps leading to cell detachment are observed. With increasing hemicellulase treatment concentration, no obvious rupture is observed on the cell wall surface, and the overall cell morphology changes from round to irregular. Organelles within the cells gradually dissolve, indicating that although hemicellulase treatment does not cause cell wall rupture, it achieves cell penetration and organelle destruction by disrupting the microstructure of the cell wall, thus effectively adsorbing atomizing agents and releasing flavor substances upon heating.

[0186] Amylase or enzyme combinations can also achieve similar effects in adsorbing atomizing agents and effectively releasing flavoring substances upon heating.

[0187] Experiment Example 7: Sensory Evaluation Experiment

[0188] The tobacco samples from Examples H1-H5 were made into cigarettes, and the cigarette tasters used F5202-JS3 heating devices to heat and smoke them. Referring to GB5606.4-2005 Cigarettes Part 4 Sensory Technical Requirements, YC / T 564-2018 Sensory Evaluation Method for Chinese Cigarettes Based on Consumer Experience, and YC / T 497-2014 Sensory Evaluation Method for Chinese Cigarette Style, and combined with the characteristics of heated cigarette smoke, key evaluation indicators were formulated, sensory evaluation was conducted, and the evaluation scores were recorded according to Table 10.

[0189] Table 10: Sensory Evaluation Record Form for Tobacco-Based Heated Cigarettes

[0190] The sensory evaluation results of Examples H1-H5 are shown in Table 11:

[0191] Table 11: Sensory Evaluation Record Table for Examples H1-H5

[0192] Based on the sensory evaluation in Table 11, adding 15% (by weight of tobacco) of atomizing agent to the tobacco sample to produce cigarettes is preferred. This amount of atomizing agent ensures sufficient smoke output while maintaining a relatively balanced distribution of other indicators, thus achieving a satisfactory smoke production effect. Furthermore, in actual production, thermogravimetric analysis (TGA) of the cigarettes produced in Examples H1-H5 was conducted as shown in Experiment 4. The applicant found that as the amount of atomizing agent added increased, the weight loss of the tobacco-type smoke-generating medium gradually increased. When the addition amount reached 25%, the weight loss became significantly greater, but this also led to severe tobacco adhesion, affecting large-scale production. Therefore, considering the sensory evaluation indicators in Table 12, a 15% atomizing agent addition amount is preferred.

[0193] Cigarettes were made by adding 15% (by weight) of atomizing agent to the tobacco samples from Examples B13-B15. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 12.

[0194] Table 12: Sensory Evaluation Record Table for Examples B13-B15

[0195] As can be seen from the sensory evaluation records of Examples B13-B15, among the drying conditions of low temperature long drying (90℃, 10min), medium temperature short drying (170℃, 1-3min), and high temperature fast drying (235℃, 3-5s), high temperature fast drying is more suitable for tobacco-type smoking medium. High temperature fast drying can quickly inactivate biological enzymes while avoiding the volatilization of aroma substances.

[0196] Cigarettes were made by adding 15% (by weight) of atomizing agent to the tobacco samples from Examples B13-B15. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 13.

[0197] Table 13: Sensory evaluation of different dosages of pectinase preparations

[0198] Sensory evaluations after adding different doses of enzyme preparation showed that the overall sensory quality first increased and then decreased with increasing dose, with 1% or 1.5% enzyme preparation being the preferred addition amount.

[0199] Cigarettes were made by adding 15% (by weight) of atomizing agent to tobacco samples from different tobacco storage times. These samples were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 14.

[0200] Table 14: Sensory evaluation of different leaf storage times

[0201] Sensory evaluations of different leaf storage times show that the optimal storage time is 4 hours, with a storage temperature of 29°C and a humidity of 70%.

[0202] Cigarettes were made by adding 15% (by weight) of atomizing agent to tobacco samples from different enzyme preparation examples. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 15.

[0203] Table 15: Sensory evaluation of different enzyme preparations

[0204] Based on the sensory evaluation results of different enzyme preparations in Table 15, different types of enzyme preparations were screened, and pectinase, hemicellulase and amylase were selected as the preferred enzyme preparation formulations for treating tobacco leaves.

[0205] Glycerin and propylene glycol, two of the most common atomizing agents, were used. Glycerin provides a better smoky sensation, while propylene glycol provides sweetness and improves smoke characteristics. Cigarettes were made by adding 15% of the tobacco weight of different atomizing agents to tobacco samples. These cigarettes were then heated and smoked by tobacco tasters using an F5202-JS3 heating device. Sensory evaluations were conducted, and scores were recorded according to Table 10. The sensory evaluation results are shown in Table 16.

[0206] Table 16: Sensory evaluation of different atomizing agent formulations

[0207] Based on the sensory evaluation in Table 16, the ratio of the two atomizing agents was adjusted, and the preferred atomizing agent formulation was 4:6.

[0208] To further demonstrate the application of this application, the following uses single-origin tobacco from nine core production areas, configured into light aroma, medium aroma, and strong aroma modules with distinct style characteristics.

[0209] Strongly aromatic modules: Chenzhou Guiyang C2F, Yongzhou Lanshan C2F, Sanmenxia Mianchi C3F, with a material ratio of 1:1:1;

[0210] Intermediate fragrance modules: Guizhou Zunyi C2F, Bijie Dafang C3F, and Guizhou Bijie C2F, with a material ratio of 1:1:1.

[0211] Lightly scented modules: Yunnan Yuxi C3F, Yunnan Kunming C3F, Yunnan Dali C3F, with a material ratio of 1:1:1.

[0212] The tobacco raw materials of the light aroma module, intermediate aroma module and strong aroma module were respectively made into tobacco shreds samples by the method of Example G2. 15% of the tobacco shred weight of the atomizing agent was added to the tobacco shreds samples to make cigarettes. The atomizing agent was composed of glycerin and propylene glycol in a ratio of 4:6. The cigarettes were subjected to sensory evaluation tests, and the results are shown in Table 17.

[0213] Table 17: Sensory evaluation of different aroma modules

[0214] Based on the sensory evaluation results in Table 17, all three aroma types have a good taste. Among them, the strong aroma type has the best overall sensory quality. The tobacco raw materials have significant differences in style characteristics between shredded heated cigarettes and sheet heated cigarettes, and have a similar style performance to traditional cigarettes.

[0215] Sanmenxia Mianchi B2F was used as the upper tobacco leaf raw material, Sanmenxia Mianchi C3F as the middle tobacco leaf raw material, and Sanmenxia Mianchi X2F as the lower tobacco leaf raw material. Tobacco samples were prepared according to the method of Example G2. Cigarettes were made by adding 15% of the tobacco weight of atomizing agent to the tobacco samples. The atomizing agent was composed of glycerol and propylene glycol in a ratio of 4:6. Sensory evaluation tests were conducted on the prepared cigarettes, and the results are shown in Table 18.

[0216] Table 18: Sensory evaluation of different parts of tobacco leaves

[0217] Sensory quality evaluation was conducted using tobacco leaves from different parts of the same production area. The results showed that the upper tobacco was superior to the middle and lower tobacco. This result is different from that of traditional cigarettes and can provide support for the comprehensive utilization of upper tobacco.

[0218] Test Example 8: Whole Filament Rate Test

[0219] The whole-shred rate of tobacco shreds in some of the above embodiments was determined according to the "Determination Method of Whole Shred Rate and Broken Shred Rate of Tobacco Shreds" (YC / T 178-2003). For other forms of plant materials such as tobacco, tobacco sheets, and cloves, the whole-shred rate can also be measured after shredding in a conventional manner. The whole-shred rate of each embodiment is shown in Table 19.

[0220] Table 19: Whole tobacco yield after different treatment methods

[0221] Test Example 9: Production Line Test

[0222] Referring to Figure 17, the process steps of this application from natural plant raw materials to the formation of aerosol matrix can be combined in various ways. The essential steps include: pretreatment step, low-damage permeable treatment step, aerosol permeation step, harm reduction treatment step and morphological change step. However, the order can be adjusted, and some steps can be carried out in separate steps or in combination with other steps and carried out simultaneously.

[0223] In a specific embodiment, such as Example I1, the process steps are as follows: pretreatment step (rehydration) → first morphological change step (slicing, shredding) → low-damage permeable treatment step (enzymatic hydrolysis) → first harm reduction treatment step (high temperature rapid drying of smoke sheets) → second aerosol permeation step → formation of aerosol generation matrix.

[0224] In one specific embodiment, the process steps are as follows: pretreatment step (rehydration) → first morphological change step (slicing) → low-damage permeable treatment step (enzymatic hydrolysis) → first aerosol permeation step (conducted simultaneously with enzymatic hydrolysis) → first harm reduction treatment step (high-temperature rapid drying of smoked sheets) → second morphological change step (shredding) → second harm reduction treatment step (adding protease) → second aerosol permeation step (simultaneously with adding protease) → formation of aerosol matrix.

[0225] In one specific embodiment, the process steps are as follows: pretreatment step (rehydration) → first morphological change step (slicing) → low-damage permeability treatment step (enzymatic hydrolysis) → first aerosol permeation step (conducted simultaneously with enzymatic hydrolysis) → first harm reduction treatment step (high-temperature rapid drying of smoked sheets) → second morphological change step (shredding) → second harm reduction treatment step (addition of protease) → formation of aerosol matrix. Specifically, as follows:

[0226] As shown in Figure 18, the tobacco shreds and materials of this application can be produced and processed using a traditional cigarette production line. The production line includes: a tobacco sheet processing section 100, a tobacco shred processing section 200, a blending and flavoring device 250, a cigarette rolling section 300, and a production line control system. The production line control system includes one or more processors (CPUs), input / output devices or interfaces, wired or wireless network interfaces, and memory. Input devices include human-machine interface devices to receive control information from operators, including cigarette type and formula. The memory stores computer programs and / or computer instructions, which, when executed by the processor, control the production line.

[0227] The tobacco leaf processing section 100 includes an unpacking device, a metering device, a slicing device, a loosening and rehydration device, a tobacco leaf pre-mixing device, a screening device, a feeding device, and a leaf storage device. After being threshed, re-dried, and aged, the tobacco leaves are boxed and sent to the reuse production line, where they undergo unpacking and metering, slicing, loosening and rehydration, tobacco leaf pre-mixing, screening and feeding, and leaf mixing and storage. First, according to the product's leaf group formula requirements, the tobacco leaves are prepared for feeding. The unpacking device 110 then unpacks the tobacco leaves through the unpacking process. After removing the outer packaging, the tobacco sheets are cut into several pieces of a specified thickness using a slicing device 130. Then, they are loosened and rehydrated using a loosening and rehydration device 150 and a tobacco sheet pre-mixing device to improve the processing resistance of the tobacco sheets and to initially and evenly mix the tobacco sheets. Then, a screening device is used to remove fragments, and tobacco slurry is sprayed onto the tobacco sheets using a feeding device 170. Finally, the selected tobacco sheets are evenly mixed and stored in a leaf storage device 180 for use in the next process stage.

[0228] The feeding device 170 includes a liquid source containing an enzyme combination (with the same proportions as in Example G2) and an atomizing agent composed of glycerol, propylene glycol, and water as the solvent. The feeding device 170 has a nozzle connected to the liquid source, thereby allowing the liquid to be selectively sprayed onto the tobacco sheet according to instructions issued by the production line control system. Because the aerosol is added before the enzymatic hydrolysis reaction, the aerosol can quickly fill the interior of the tobacco cell walls after enzymatic hydrolysis, allowing the tobacco to fully absorb the aerosol and avoiding the problem of the tobacco not being able to disperse or clumping when it is cut into shreds by the shredder 210. It is best to use a single enzymatic hydrolysis reaction on the production line, but multiple enzymatic hydrolysis reactions can also be used. The aerosol can be sprayed before the first or last enzymatic hydrolysis reaction, or the aerosol can be sprayed evenly before each enzymatic hydrolysis reaction. Almost no aerosol precipitation on the tobacco surface can avoid contaminating the production line, especially avoiding the inability to roll the cigarettes in the cigarette rolling section 300. Since the glycerol and propylene glycol in the cigarette products are absorbed in the cell walls, the probability of contact with moisture in the air is reduced, the apparent water absorption is reduced, and thus the shelf life is improved.

[0229] The leaf storage device 180 can be a leaf storage cabinet. The tobacco leaves sprayed with liquid will be sent into the leaf storage cabinet for enzymatic degradation reaction. The temperature of the leaf storage cabinet is 28-33℃ and the relative humidity is 67-75%. The leaf storage time can be selected as needed.

[0230] Because the enzymatically hydrolyzed tobacco sheets of this application have high processability, they can be directly processed into heated tobacco shreds through the tobacco processing section 200 of a traditional tobacco production line, including shredding and other processes. The tobacco processing section 200 includes a shredding device, a drying device, and a blending and flavoring device. First, the shredding device shreds the tobacco sheets 21 to form heated tobacco shreds of a certain width. Then, the drying device dries the tobacco shreds. This step, in addition to drying the heated tobacco shreds to remove some of the moisture from the leaves (to achieve a specified moisture content, such as 11.5-14.0%), thereby improving the filling capacity and sensory quality of the leaves, also includes a high-temperature rapid drying process (temperature 200-250℃, preferably 235℃; time 1-10s, preferably 3-5s) to inactivate the residual enzyme combination. Finally, the blending and flavoring device further adds flavorings and additives suitable for heating cigarettes to the tobacco, including proteases used to degrade biological enzymes to remove impurities, and then sends it to the cigarette rolling section 300.

[0231] The cigarette rolling section 300 includes a cigarette rolling device and a feeding device. The cigarette rolling device rolls heated cigarette tobacco into heated cigarette tobacco segments, and the feeding device feeds heated cigarette tobacco segments and heated cigarette filter segments together to form heated cigarettes.

[0232] This application utilizes cell wall-degrading enzymes to quantitatively and controllably modify traditional cigarette tobacco, enabling it to absorb more aerosols and release more aroma compounds during heating while maintaining its original processability (especially the whole tobacco yield). This allows traditional cigarette production lines to be reused with heated tobacco with only minor modifications. By selecting specific enzyme compositions, the enzymatic degradation reaction can be carried out in the existing leaf storage environment without the need for additional high-temperature reaction equipment.

[0233] The terminology and expressions used herein are for descriptive purposes only and should not be construed as limiting of this application. The use of these terms and expressions does not preclude any illustrative and descriptive equivalents (or parts thereof), and it should be recognized that various modifications that may exist should also be included within the scope of the claims. Other modifications, variations, and substitutions may also exist. Accordingly, the claims should be considered to cover all such equivalents.

[0234] Similarly, it should be noted that although this application has been described with reference to specific embodiments, this application is not limited to the detailed methods described above, that is, it does not mean that this application must rely on the detailed methods described above to be implemented. Those skilled in the art should understand that any improvements to this application, equivalent substitutions of the raw materials of the product of this application, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this application.

Claims

1. A natural plant aerosol generating matrix for electric heating smoke appliances, comprising: Natural plant materials that have undergone quantified cell wall disruption treatment and exogenous aerosol generators loaded within the natural plant materials.

2. The natural plant aerosol generating matrix according to claim 1, wherein, The quantitative cell wall disruption treatment includes 1 to 5 enzymatic hydrolysis steps, each of which includes: Step A: Spray the active cell wall breaking enzyme solution evenly onto the natural plant material; Step B: Seal the sprayed natural plant material and carry out enzymatic hydrolysis and cell wall breaking reaction under constant temperature and humidity conditions.

3. The natural plant aerosol generating matrix according to claim 2, wherein, In step A, the weight ratio of the active cell wall-breaking enzyme to the natural plant raw material is 0.5-1.0%, 1.0-1.5%, 1.5-2.0%, 2.0-2.5%, or 2.5-3.0%.

4. The natural plant aerosol generating matrix according to claim 2, wherein, In step A, the active cell wall-breaking enzyme includes any one or a combination of at least two of pectinase, hemicellulase, cellulase or amylase.

5. The natural plant aerosol generating matrix according to claim 2, wherein, In step A, the active cell-wall-breaking enzyme is composed of pectinase, hemicellulase and amylase; Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, cellulase, and amylase; Alternatively, the active cell-wall-breaking enzyme may be composed of pectinase, hemicellulase, cellulase, and amylase, in the following weight ratio: Pectinase: 15-25%, Total hemicellulase and cellulase: 32-48%, Amylase: 32-48%.

6. The natural plant aerosol generating matrix according to claim 2, wherein, In step B, the conditions for each enzymatic hydrolysis and cell wall breaking reaction are independent: temperature of 25-30℃, 30-35℃, 35-40℃, 40-45℃, 45-50℃, 50-55℃ or 55-60℃, relative humidity of 15-20%, 20-25%, 25-30%, 30-35%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%, 60-65%, 65-70%, 70-75% or 75-80%, and enzymatic hydrolysis and cell wall breaking reaction time of 0.5-1.0h, 1.0-2.0h, 2.0-4.0h, 4.0h-6.0h, 6.0-8.0h, 8.0-12.0h or 12.0-24.0h.

7. The natural plant aerosol generating matrix according to claim 2, wherein, After step B is completed, step C is performed: the apparent loading and / or micro loading of the natural plant material are detected. When the apparent loading and / or micro loading of the aerosol generating agent of the natural plant material is greater than the target loading, the next enzymatic hydrolysis and cell wall breaking step is not performed.

8. The natural plant aerosol generating matrix according to claim 7, wherein, Based on the difference between the apparent loading and / or microscopic loading of the aerosol generator of the natural plant material and the target loading, the enzymatic hydrolysis reaction temperature, relative humidity, and / or reaction time in the next enzymatic hydrolysis step are determined.

9. The natural plant aerosol generating matrix according to claim 7, wherein, The apparent loading test includes weighing the maximum load of the exogenous aerosol generator; the microscopic loading test includes observing the cell wall and statistically analyzing the cell wall disruption rate.

10. The natural plant aerosol generating matrix according to claim 7, wherein, After each enzymatic hydrolysis and cell wall breaking reaction is completed, step D is performed: the natural plant raw material is subjected to a harm reduction treatment.

11. The natural plant aerosol generating matrix according to claim 7, wherein, The exogenous aerosol generator is loaded into the natural plant material that has undergone quantitative cell wall disruption treatment in the following manner: The exogenous aerosol generating agent is sprayed onto the natural plant raw material that has undergone quantified cell wall disruption treatment, wherein: the weight of the exogenous aerosol generating agent is less than the weight of the natural plant raw material × maximum loading rate × whole fiber rate; Once the exogenous aerosol generating agent has completely penetrated into the natural plant raw material that has undergone quantified cell wall disruption treatment, the natural plant aerosol generating matrix is ​​prepared.

12. The natural plant aerosol generating matrix according to claim 7, wherein, After each enzymatic hydrolysis and cell wall breaking reaction is completed, step E is performed: the whole fiber rate of the natural plant material is detected. If the whole fiber rate of the natural plant material is greater than the whole fiber rate warning value, the next enzymatic hydrolysis and cell wall breaking step will not be performed.

13. The natural plant aerosol generating matrix according to claim 12, wherein, The warning value for the whole yarn rate is above 60%, above 65%, above 68%, above 70%, above 72%, above 75%, above 78%, or above 80%.

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