Tobacco leaf extract for improving tobacco quality and preparation method and application thereof

Abstract

The invention belongs to the technical field of tobacco additives, and particularly relates to a tobacco leaf extract for improving tobacco quality and a preparation method and application thereof.The preparation method comprises the following steps that tobacco leaves are collected, dried and smashed and then screened through a screen, and tobacco leaf powder is obtained; sterilizing the tobacco leaf powder, adding sterile water, and stirring to obtain a tobacco leaf suspension; adding a bacterial liquid of enterobacter hormaechei C4 into the tobacco suspension, and carrying out oscillation fermentation at 37 DEG C for 96 hours to obtain a tobacco fermentation liquid; centrifuging the tobacco leaf fermentation liquor, and respectively collecting solids and centrifugate; adding an ethanol solution into the solid, heating and refluxing, filtering, and collecting filtrate; mixing the centrifugate and the filtrate into an extracting solution, and performing rotary evaporation to 1 / 5 of the volume of the extracting solution to obtain a tobacco leaf extract; wherein the preservation number of the enterobacter hormaechei C4 is CGMCC (China General Microbiological Culture Collection Center) When the tobacco extract is prepared, only a single microorganism is used for short-time fermentation, the tobacco extract can be endowed with various aroma substances, and the tobacco extract has an excellent tobacco quality improvement effect.

Description

Technical Field

[0001] The present application belongs to the technical field of tobacco additives, and specifically relates to a tobacco leaf extract for improving tobacco quality, and a preparation method and application thereof. Background Art

[0002] Tobacco is a dark brown viscous paste extracted from tobacco leaves. It has the characteristic aroma of tobacco and has many uses in tobacco processing, such as cigarette additives, tobacco sheet coating additives, and preparation of reaction flavors, etc. It can enhance or supplement the natural aroma characteristics of tobacco products.

[0003] The production process of traditional tobacco extract adopts an organic solvent extraction method. For example, in the prior art CN1014114B, ethanol is used to extract sun-dried tobacco paste, and then mixed with nicotine sulfate to obtain sun-dried red tobacco extract. After long-term development, there are currently technologies for preparing tobacco extracts by means of plasma assistance, combined cold soaking and heating extraction, and combined with thermal alcoholization technology and Maillard reaction. Although the above-mentioned physical or chemical technologies can improve the yield or quality of tobacco extracts, there are problems such as high cost, complicated operation, and single aroma components. Prior art CN102119784B uses a variety of microorganisms to ferment tobacco leaves to obtain spice extracts, but the process cycle of this preparation method requires 40 days of fermentation, which is difficult to carry out large-scale production in industry.

[0004] Therefore, a new method for preparing tobacco leaf extract is needed, which can provide tobacco leaf extract with a variety of aroma substances through biological fermentation while simplifying the process flow and shortening the process cycle. Summary of the invention

[0005] The problem that the present application aims to solve is to provide a tobacco extract for improving tobacco quality, and a preparation method and application thereof. When preparing the tobacco extract, only a single microorganism is used for short-term fermentation, which can give the tobacco extract a variety of active substances and have an excellent tobacco quality improvement effect.

[0006] In order to solve the above technical problems, this application adopts the following technical solutions:

[0007] In one aspect, the present application provides a method for preparing a tobacco extract for improving tobacco quality, comprising the following steps:

[0008] Collect tobacco leaves, dry and crush them, and then sieve them with a sieve to obtain tobacco leaf powder; sterilize the tobacco leaf powder, add sterile water and stir to obtain a tobacco leaf suspension; add a bacterial liquid of Enterobacter hallii C4 to the tobacco leaf suspension, and ferment at 37° C. for 96 hours under shaking to obtain a tobacco leaf fermentation liquid;

[0009] After centrifuging the tobacco fermented liquid, respectively collecting the solid matter and the centrifuge liquid; adding an ethanol solution to the solid matter, heating and refluxing, filtering, and collecting the filtrate; and mixing the centrifuge liquid and the filtrate into an extract, rotary evaporating to 1 / 5 of the volume of the extract, to obtain the tobacco extract;

[0010] Among them, the Enterobacter hormaecheiC4 is preserved in the General Microbiology Center of China Microorganism Culture Collection Administration, the preservation address is No. 3, No. 1, Beichen Road West, Chaoyang District, Beijing, the preservation date is January 13, 2023, and the preservation number is CGMCC No.26480.

[0011] Furthermore, the tobacco leaves include Zimbabwe L1M tobacco leaves.

[0012] Furthermore, the specification of the sieve is 40 mesh.

[0013] Furthermore, the moisture content of the tobacco leaf powder is 2% to 5%.

[0014] Furthermore, the concentration of the bacterial solution is 4.15×10 8 ~4.95×10 8 CUF / mL.

[0015] Furthermore, the concentration of the ethanol solution is 75%.

[0016] Furthermore, the solid-liquid ratio of the tobacco leaf powder to the sterile water is 1:(5-10).

[0017] Furthermore, the volume ratio of the bacterial liquid to the tobacco leaf suspension is 1:(100-150).

[0018] Furthermore, the solid-liquid ratio of the solid matter to the ethanol solution is 1:(5-10).

[0019] Furthermore, the drying temperature is 40°C.

[0020] Furthermore, the sterilization temperature is 121° C., and the sterilization time is 15 min.

[0021] Furthermore, the heating reflux temperature is 50° C., and the heating reflux time is 2 h.

[0022] Furthermore, the temperature of the rotary evaporation is 50° C., and the pressure of the rotary evaporation is 0.1 MPa.

[0023] On the other hand, the present application provides tobacco leaf extract prepared by the above method.

[0024] On the other hand, the present application provides the use of the above method or the above tobacco leaf extract in improving tobacco quality.

[0025] Furthermore, the tobacco quality includes aroma quality, aroma quantity, impurities, concentration, stimulation, aftertaste and sweetness.

[0026] This application has the following beneficial effects:

[0027] 1. The preparation method of the present application is simple to operate, has concise steps, involves readily available raw materials, can reduce costs during industrial production, and is convenient for large-scale production.

[0028] 2. The preparation method of the present application has a short process cycle, and tobacco extract rich in active ingredients and aroma substances can be obtained in only 4 days, which can greatly save the production cycle in industrial production.

[0029] 3. The preparation method of the present application uses microorganisms to ferment the components of tobacco leaves themselves, thereby synthesizing active ingredients and aroma substances, without using substances from non-tobacco sources, which can effectively avoid the problems of inconsistent aroma of tobacco leaf extract and residual chemical components. DETAILED DESCRIPTION

[0030] The technical solutions in some embodiments of the present application are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments provided by the present application, all other embodiments obtained by ordinary technicians in this field belong to the scope of protection of the present application.

[0031] Example 1

[0032] Prepare liquid culture medium as follows:

[0033] Weigh 3 g beef extract, 10 g peptone and 5 g sodium chloride into a beaker, add sterile water and heat and stir until completely dissolved, then add sterile water to 1000 mL, adjust the pH to 7.4, and cool to room temperature for use.

[0034] Example 2

[0035] Prepare the bacterial culture of Enterobacter holmesii C4 according to the following steps:

[0036] The Enterobacter C4 stored in the laboratory was taken out, revived and cultured, and then the strain was selected and added to the liquid culture medium prepared in Example 1, and cultured at 37°C for 24 hours to obtain the bacterial solution of the Enterobacter C4. After testing, the effective viable count in the bacterial solution was 4.15×10 8 ~4.95×10 8 CUF / mL.

[0037] Example 3

[0038] Prepare tobacco extract as follows:

[0039] S11. Collect Zimbabwe L1M tobacco leaves, dry them at 40°C to a moisture content of 2%, grind them, pass them through a 40-mesh sieve, collect the sieved powder, and obtain tobacco leaf powder;

[0040] S12. Take 40 g of the tobacco powder and sterilize it at 121° C. for 15 min using a high pressure steam sterilizer, then add 200 mL of sterile water and stir mechanically for 5 min to obtain a tobacco suspension;

[0041] S13. Take 100 mL of the tobacco suspension, add 1 mL of the bacterial solution prepared in Example 2, and ferment at 37 ° C for 96 h to obtain a tobacco fermentation liquid;

[0042] S14. After the tobacco fermentation liquid was centrifuged at a speed of 3000 rpm for 10 min, the solid matter and the centrifuge liquid were collected respectively;

[0043] S15. After the solid matter is dried by cold air, it is weighed.

[0044] S16. The dried solid was placed in a reflux flask, and a 75% ethanol solution was added at a solid-liquid ratio of 1 (g): 5 (mL), and the mixture was heated to reflux and extracted once at 50°C for 2 hours. After the extraction was completed, the mixture was filtered and the filtrate was collected;

[0045] S17. The centrifuge liquid and the filtrate are mixed to form an extract, and the extract is rotary evaporated at 50° C. under an environment of 0.1 MPa to 1 / 5 of the volume of the extract to obtain the tobacco leaf extract.

[0046] Example 4

[0047] Prepare tobacco extract as follows:

[0048] S11. Collect Zimbabwe L1M tobacco leaves, dry them at 40°C to a moisture content of 3%, grind them, pass them through a 40-mesh sieve, collect the sieved powder, and obtain tobacco leaf powder;

[0049] S12. Take 40 g of the tobacco powder and sterilize it at 121° C. for 15 min using a high pressure steam sterilizer, then add 300 mL of sterile water and stir mechanically for 5 min to obtain a tobacco suspension;

[0050] S13. Take 125 mL of the tobacco suspension, add 1 mL of the bacterial solution prepared in Example 2, and ferment at 37 ° C for 96 h to obtain a tobacco fermentation liquid;

[0051] S14. After the tobacco fermentation liquid was centrifuged at a speed of 3000 rpm for 10 min, the solid matter and the centrifuge liquid were collected respectively;

[0052] S15. After the solid matter is dried by cold air, it is weighed.

[0053] S16. The dried solid was placed in a reflux flask, and a 75% ethanol solution was added at a solid-liquid ratio of 1 (g): 7.5 (mL), and the mixture was heated to reflux and extracted once at 50°C for 2 hours. After the extraction was completed, the mixture was filtered and the filtrate was collected;

[0054] S17. The centrifuge liquid and the filtrate are mixed to form an extract, and the extract is rotary evaporated at 50° C. under an environment of 0.1 MPa to 1 / 5 of the volume of the extract to obtain the tobacco leaf extract.

[0055] Example 5

[0056] Prepare tobacco extract as follows:

[0057] S11. Collect Zimbabwe L1M tobacco leaves, dry them at 40°C to a moisture content of 5%, grind them, pass them through a 40-mesh sieve, collect the sieved powder, and obtain tobacco leaf powder;

[0058] S12. Take 40 g of the tobacco powder and sterilize it at 121° C. for 15 min using a high pressure steam sterilizer, then add 400 mL of sterile water and stir mechanically for 5 min to obtain a tobacco suspension;

[0059] S13. Take 150 mL of the tobacco suspension, add 1 mL of the bacterial solution prepared in Example 2, and ferment at 37 ° C for 96 h to obtain a tobacco fermentation liquid;

[0060] S14. After the tobacco fermentation liquid was centrifuged at a speed of 3000 rpm for 10 min, the solid matter and the centrifuge liquid were collected respectively;

[0061] S15. After the solid matter is dried by cold air, it is weighed.

[0062] S16. The dried solid was placed in a reflux flask, and a 75% ethanol solution was added at a solid-liquid ratio of 1 (g): 10 (mL), and the mixture was heated to reflux and extracted once at 50°C for 2 hours. After the extraction was completed, the mixture was filtered and the filtrate was collected;

[0063] S17. The centrifuge liquid and the filtrate are mixed to form an extract, and the extract is rotary evaporated at 50° C. under an environment of 0.1 MPa to 1 / 5 of the volume of the extract to obtain the tobacco leaf extract.

[0064] Experimental Example 1: Effect of tobacco extract on tobacco leaves

[0065] Yunyan 87 tobacco leaves from the tobacco plantation in Huidong County, Liangshan Prefecture, Sichuan Province were selected for drying to make the moisture content 8%. 1‰ of the weight of the tobacco leaves was added to a portion of the tobacco leaves as experimental groups 1 to 3, and 1‰ of the weight of the tobacco leaves was added to the other portion of the dried tobacco leaves as a control group. The tobacco leaves of each experimental group and the control group were placed in an environment with a temperature of 25°C and a relative humidity of 75% for 30 days, dried and made into cigarettes, and then 7 professional cigarette tasters were invited to perform sensory evaluation according to the "Chinese Cigarette Style Sensory Evaluation Method" YC / T 497-2014 and take the average value. The evaluation results are shown in Table 1.

[0066] Table 1 Sensory evaluation table

[0067] Group Fragrance Aroma volume Miscellaneous gas concentration Stimulate Aftertaste Sweetness Score Control group 5 5 5 5.5 5 5 4.93 35.43 Experimental Group 1 5.5 5.5 5.5 5.5 5.5 5.5 5.5 38.5 Experimental Group 2 5.7 5.6 5.5 5.6 5.7 5.7 5.8 39.6 Experimental Group 3 5.8 5.6 5.6 5.7 5.7 5.8 5.8 40

[0068] By analyzing the data in Table 2, it can be seen that the experimental groups 1 to 3 are higher than the control group in many sensory scores, which shows that tobacco leaf extract can significantly improve the aroma quality and aroma quantity of tobacco, reduce the stimulation of foreign gases, improve the aftertaste and increase the sweetness.

[0069] Experimental Example 2: Analysis of Tobacco Extract Composition

[0070] The components of the tobacco extract prepared in Example 3 were detected and analyzed by GC-MS analysis method, and the detection results are shown in Table 2. The specific steps are as follows: 3.00 g of tobacco extract sample was weighed into a 10.0 mL headspace bottle, equilibrated in a 60° C. water bath for 5 min, headspace extracted for 55 min, analyzed on the machine for 5 min, and analyzed by GC-MS.

[0071] Table 2: Composition of tobacco extract

[0072]

[0073]

[0074] It can be seen from Table 2 that the tobacco extract prepared in the present application contains a variety of active ingredients and aroma substances, among which the ingredients with higher content include (S)-3-(1-methyl-2-pyrrolidinyl)pyridine, (E)-6,8-nonadien-2-one-8-methyl-5-(1-methylethyl), (S)-(+)-1,2-propylene glycol, neophytadiene, (E)-2-butene-1-one-1-(2,6,6-trimethyl-1,3-cyclohexadien-1-yl), nicotine, soytrienone, and pentan-2-ol-4-allyloxy-2-methyl, which can make cigarettes have a richer aroma and effectively improve the quality of tobacco.

[0075] In summary, the method for preparing tobacco extract provided by the present application can be prepared in only 4 to 5 days, and the preparation method is simple, and the raw materials involved are common, easy to obtain and harmless, which is very suitable for industrial production. In addition, the tobacco extract prepared by the present application contains a large number of beneficial ingredients, which can effectively improve the quality of cigarettes, thereby reducing the use of other chemical components in tobacco.

[0076] The above is only a preferred embodiment of the present application. It should be understood that the present application is not limited to the form disclosed herein, and should not be regarded as excluding other embodiments, but can be used in various other combinations, modifications and environments, and can be modified within the scope of the concept described herein through the above teachings or the technology or knowledge in the relevant field. The changes and modifications made by those skilled in the art shall not deviate from the spirit and scope of the present application, and shall be within the scope of protection of the claims attached to the present application.

Claims

1. A method for preparing a tobacco extract for improving tobacco quality, characterized in that: The following steps are involved: Collect tobacco leaves, dry and crush them, and then sieve them using a sieve to obtain tobacco leaf powder; After the tobacco leaf powder is sterilized, sterile water is added and stirred to obtain a tobacco leaf suspension; Adding a bacterial liquid of Enterobacter hallii C4 to the tobacco leaf suspension, and fermenting at 37° C. for 96 hours to obtain a tobacco leaf fermentation liquid; After centrifuging the tobacco fermentation liquid, the solid matter and the centrifuge liquid are collected respectively; Add ethanol solution to the solid, heat and reflux, then filter and collect the filtrate; as well as The centrifuged liquid and the filtrate are mixed into an extract, and the extract is rotary evaporated to 1 / 5 of the volume of the extract to obtain the tobacco leaf extract; Wherein, the preservation number of the Enterobacter hallii C4 is CGMCC No.26480.

2. The method according to claim 1, characterized in that The tobacco leaves include Zimbabwe L1M tobacco leaves.

3. The method according to claim 1, characterized in that The specification of the sieve is 40 mesh; and / or, the moisture content of the tobacco leaf powder is 2% to 5%; And / or, the concentration of the bacterial solution is 4.15×10 8 ~4.95×10 8 CUF / mL; And / or, the concentration of the ethanol solution is 75%.

4. The method according to claim 1, characterized in that: The solid-liquid ratio of the tobacco leaf powder to the sterile water is 1:(5-10); And / or, the volume ratio of the bacterial liquid to the tobacco leaf suspension is 1:(100-150); And / or, the solid-liquid ratio of the solid matter to the ethanol solution is 1:(5-10).

5. The method according to claim 1, characterized in that: The drying temperature is 40°C; And / or, the sterilization temperature is 121°C and the sterilization time is 15 minutes; And / or, the temperature of the shaking fermentation is 37° C., and the time of the shaking fermentation is 96 h; And / or, the heating reflux temperature is 50° C., and the heating reflux time is 2 h; And / or, the temperature of the rotary evaporation is 50° C., and the pressure of the rotary evaporation is 0.1 MPa.

6. Tobacco extract obtained by the method according to any one of claims 1 to 5.

7. Use of the method according to any one of claims 1 to 5 or the tobacco extract according to claim 6 in improving tobacco quality.

8. The use according to claim 7, characterized in that The tobacco quality includes aroma quality, aroma quantity, offal, concentration, stimulation, aftertaste and sweetness.

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