Tobacco flavoring using waste tobacco resources and preparation method and application thereof

Abstract

The application provides a tobacco flavor using waste tobacco resources and a preparation method and application thereof. The tobacco flavor is prepared by dissolving a petroleum ether extract of the waste tobacco resources in a co-distilled essential oil of the waste tobacco resources and calamus. The application belongs to the technical field of tobacco flavor essence, and the tobacco flavor provided by the application realizes sufficient extraction and synergistic use of volatile aroma components and non-volatile aroma components, and the sensory evaluation quality is improved in tobacco flavoring.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of tobacco flavor essence, and particularly relates to a tobacco flavor essence using waste tobacco resources and a preparation method and application thereof. BACKGROUND

[0002] Tobacco is an important economic crop in China, and a large amount of field waste tobacco leaves, tobacco flowers, post-curing waste tobacco leaves, tobacco stems, and re-drying broken tobacco leaves are produced every year. With the further improvement of brand concentration in the tobacco industry, the demand for high-quality tobacco leaves is in short supply, which generates more waste tobacco resources. If these waste tobacco resources are not utilized, not only will it cause great waste of resources, but also will cause environmental pollution. High-value utilization of waste tobacco resources has become one of the problems that need to be urgently solved in the tobacco industry. Nationwide, tobacco research and industrial enterprises have widely verified the research of tobacco multi-purpose in the fields of tobacco, energy, medicine and health, functional feed, food, and new materials, and it is believed that the research of tobacco multi-purpose in different fields has different research focuses: the research represented by tobacco essence development, energy, and feed focuses on high-value application; the research represented by medicine and health and new materials focuses on basic research technology reserve. Waste tobacco resources contain a large amount of bioactive substances and a large amount of tobacco aroma components, and recycling these aroma components to prepare tobacco flavor or essence can generate great economic value and realize the recycling of waste resources. Through effective conversion of these waste tobacco resources into valuable products, both economic and environmental benefits can be achieved.

[0003] From waste tobacco resources, aroma extracts can be obtained by methods such as steam distillation, solvent extraction, and supercritical fluid extraction, and the forms can be essential oil, extract, pure oil, balm, and tincture. Essential oil is an oily liquid with aroma characteristics, which is an important form of natural flavor. It is commonly extracted from spice plants by steam distillation, and its liquid characteristics facilitate separation and product perfuming application, and it is widely used in many fields such as perfume, cosmetics, food, and medicine.

[0004] The aroma extracts obtained by solvent extraction and supercritical fluid extraction generally contain other low-grade non-volatile components, and need further refining treatment, and in addition, solvent extraction may cause solvent residues in the aroma extract. Chinese Patent Application CN 114534361 A discloses a method for jointly extracting essential oil, nicotine and extract from waste tobacco resources. After the raw material is crushed and alkalinized by adding alkali water, essential oil, nicotine and extract are obtained by using steam distillation method, and essential oil is obtained by using two-phase solvent extraction method. For waste tobacco resources, literature research and preliminary experimental results show that the volatile aroma components contained therein have a relatively high boiling point. When directly extracted by steam distillation method, the collected essential oil has a very high viscosity, and sometimes the collected essential oil becomes solid after cooling, which is difficult to collect from the essential oil extraction device. The collected viscous essential oil or solid material is also not conducive to flavoring application. If the viscous or solid essential oil is dissolved with an organic solvent, there will be a problem of solvent residue. If refined by molecular distillation or other technologies, the cost is high and the process is complex. It can be seen that the existing technology for recovering aroma components from waste tobacco resources has a relatively complex preparation process (such as steam distillation + solvent extraction, flash extraction + molecular distillation), and is prone to solvent residue problems. Therefore, there is an urgent need for a method for recovering aroma components from waste tobacco resources with relatively simple operation and effectiveness. SUMMARY

[0005] To solve the problems in the prior art, the present application provides a tobacco flavoring agent using waste tobacco resources and a preparation method and application thereof. The volatile aroma components and non-volatile aroma components are fully extracted and synergistically utilized by co-distilling the waste tobacco resources and the calamus at a certain ratio by steam, collecting the co-distilled essential oil, and then dissolving the non-volatile aroma components extracted from the waste tobacco resources in the co-distilled essential oil. The solvent residue problem is avoided, and the sensory evaluation quality is improved in tobacco flavoring.

[0006] The object of the present application will be further illustrated by the following detailed description.

[0007] The present application provides a tobacco flavoring agent using waste tobacco resources, which is prepared by dissolving the petroleum ether extract of waste tobacco resources in the co-distilled essential oil of waste tobacco resources and calamus;

[0008] The preparation of the petroleum ether extract includes the following steps: taking waste tobacco resources, crushing them, and then placing them in a round-bottom flask, adding petroleum ether, heating and refluxing to extract; separating the extract by suction filtration, adding petroleum ether to the filter residue, and continuing to heat and reflux to extract; separating the extract by suction filtration, adding petroleum ether to the filter residue, and continuing to heat and reflux to extract; filtering, combining the extracts, removing the solvent by rotary evaporation, and vacuum drying to obtain the petroleum ether extract;

[0009] The preparation of the co-distilled essential oil comprises the following steps: taking the discarded tobacco resources, crushing, and placing in a flask, adding powder of Acorus calamus and water, heating for water vapor co-distillation, and collecting the co-distilled essential oil.

[0010] The tobacco flavor provided by the application realizes the full extraction and synergistic utilization of volatile and non-volatile aroma components, avoids the solvent residue problem, and realizes the maximum utilization of aroma components.

[0011] Preferably, in the preparation step of the co-distilled essential oil, the mass ratio of the discarded tobacco resources to Acorus calamus is (6-15):1.

[0012] More preferably, in the preparation step of the co-distilled essential oil, the mass ratio of the discarded tobacco resources to Acorus calamus is (8-12):1.

[0013] Preferably, the mass ratio of the petroleum ether extract to the co-distilled essential oil is (1-3):(4-6).

[0014] More preferably, the mass ratio of the petroleum ether extract to the co-distilled essential oil is 1:(2-3).

[0015] Preferably, the discarded tobacco resources include field discarded tobacco leaves, post-cured discarded tobacco leaves, tobacco flower buds, tobacco stems, re-dried tobacco leaves, and discarded tobacco leaves.

[0016] Preferably, the dissolving adopts ultrasonic-assisted dissolving.

[0017] Correspondingly, the application further provides a preparation method of the tobacco flavor using discarded tobacco resources, comprising the following steps: dissolving the petroleum ether extract of discarded tobacco resources in the co-distilled essential oil of discarded tobacco resources and Acorus calamus to obtain;

[0018] The preparation of the petroleum ether extract comprises the following steps: taking discarded tobacco resources, crushing, and placing in a round-bottom flask, adding petroleum ether, and heating for reflux extraction; separating the extract liquid by suction filtration, and adding petroleum ether to the filter residue for continuous heating and reflux extraction; separating the extract liquid by suction filtration, and adding petroleum ether to the filter residue for continuous heating and reflux extraction; suction filtration, combining the extract liquids, removing the solvent by rotary evaporation, and vacuum drying to obtain the petroleum ether extract;

[0019] The preparation of the co-distilled essential oil comprises the following steps: taking the discarded tobacco resources, crushing, and placing in a flask, adding powder of Acorus calamus and water, heating for water vapor co-distillation, and collecting the co-distilled essential oil.

[0020] In addition, the application further provides a tobacco flavor using discarded tobacco resources in the preparation of tobacco products.

[0021] Preferably, the tobacco product includes cigarettes, tobacco shreds, and cigars.

[0022] Compared with the prior art, the beneficial effects of the present application include:

[0023] (1) The present application co-distills the waste tobacco resources and the pinenut tree according to a certain ratio, so that the volatile components with high boiling points in the waste tobacco resources and the pinenut tree essential oil components are co-boiled out with water. After the oil and water are separated, the co-distilled essential oil obtained is an oily liquid without solvent addition. The co-distillation method is relatively simple to operate, avoids the solvent residue caused by the use of organic solvents to dissolve the waste tobacco resources, realizes the compounding of the volatile aroma components (including a-cyperone, neophytadiene, and macobutenone) in the waste tobacco resources and the pinenut tree essential oil, and makes the product liquid, which is convenient for subsequent flavoring application.

[0024] (2) The present application first extracts the non-volatile aroma components from the waste tobacco resources by using the petroleum ether solvent extraction method. The petroleum ether extract obtained can be dissolved in the co-distilled essential oil of the waste tobacco resources and the pinenut tree under ultrasonic assistance. Finally, the liquid tobacco flavor with enhanced aroma is obtained, realizing the full extraction and collaborative use of the volatile aroma components and the non-volatile aroma components. The liquid tobacco flavor has no solvent addition, and the sensory evaluation quality is improved in tobacco flavoring. DETAILED DESCRIPTION

[0025] The present application will be further described in detail through specific examples.

[0026] In the present application, the raw materials involved are all conventional commercially available products, or can be obtained by conventional technical means in the art.

[0027] Example 1: A tobacco flavor using waste tobacco resources

[0028] A tobacco flavor using waste tobacco resources (field waste tobacco leaves) is prepared by dissolving 0.2 g of petroleum ether extract of waste tobacco resources in 0.5 g of co-distilled essential oil of waste tobacco resources and pinenut tree under ultrasonic assistance;

[0029] The preparation of the petroleum ether extract includes the following steps: 100 g of air-dried field waste tobacco leaves are taken, crushed, and then placed in a round-bottom flask. 1 L of petroleum ether is added for heating reflux extraction for 2 h. The extract is separated by suction filtration, and the filter residue is added with 0.5 L of petroleum ether for continued heating reflux extraction for 1 h. The extract is separated by suction filtration, and the filter residue is added with 0.5 L of petroleum ether for continued heating reflux extraction for 1 h. Filtration is performed, and the three times of extract are combined. The solvent is removed by rotary evaporation, and vacuum drying is performed to obtain the petroleum ether extract;

[0030] The preparation of the co-distilled essential oil includes the following steps: taking 300 g of dried field waste tobacco leaves, crushing them and placing them in a flask, adding 30 g of powder of Acorus calamus and 5 L of water, heating and performing steam co-distillation for 5 h, and collecting the co-distilled essential oil.

[0031] Example 2: A tobacco flavor using waste tobacco resources

[0032] A tobacco flavor using waste tobacco resources (waste tobacco leaves after curing) is prepared by dissolving 0.2 g of a petroleum ether extract of the waste tobacco resources in 0.5 g of a co-distilled essential oil of the waste tobacco resources and Acorus calamus under ultrasonic assistance.

[0033] The preparation of the petroleum ether extract includes the following steps: taking 100 g of dried waste tobacco leaves after curing, crushing them and placing them in a round-bottom flask, adding 1 L of petroleum ether, heating and refluxing for 2 h; separating the extract by suction filtration, adding 0.5 L of petroleum ether to the residue, continuing to heat and reflux for 1 h; separating the extract by suction filtration, adding 0.5 L of petroleum ether to the residue, continuing to heat and reflux for 1 h; suction filtering, combining the three extracts, removing the solvent by rotary evaporation, and performing vacuum drying to obtain the petroleum ether extract.

[0034] The preparation of the co-distilled essential oil includes the following steps: taking 300 g of dried waste tobacco leaves after curing, crushing them and placing them in a flask, adding 30 g of powder of Acorus calamus and 5 L of water, heating and performing steam co-distillation for 5 h, and collecting the co-distilled essential oil.

[0035] Example 3: Detection of the co-distilled essential oil of waste tobacco resources and Acorus calamus

[0036] 1. GC-MS analysis of essential oil

[0037] The essential oil of Acorus calamus and the co-distilled essential oils of Example 1 and Example 2 were diluted with n-hexane at 1 :50 (v / v) respectively. GC-MS analysis was performed on a Shimadzu gas chromatograph-mass spectrometer (TQ8040NX) equipped with a TG-5MS capillary column (30 m x 0.25 mm x 0.25 μιη) and an AOC-6000 autosampler. The injector temperature was 250 °C, the column temperature was initially set at 50 °C for 3 min, then ramped at 3 °C / min to 180 °C, and then at 8 °C / min to 250 °C. Helium was used as the carrier gas at a flow rate of 1 mL / min, and the injection volume was 1 μί (split ratio 1 : 100). The mass spectrometry parameters were: mass scan range 45-550, ion source temperature 250 °C, interface temperature 250 °C. The retention indices (Rl) of the components in the essential oils were determined using n-alkanes (C7-C30) under the same operating conditions. The components in the essential oils were qualitatively identified based on the comparison of the mass spectra and the retention indices, and quantitatively determined by calculating the relative percentage content (%) of each component using the area normalization method of the total ion chromatogram. The results are shown in Tables 1 and 2.

[0038] Table 1 Chemical composition of the co-distilled essential oil of Example 1

[0039]

[0040] Table 2 Chemical composition of the co-distilled essential oil of Example 2

[0041]

[0042]

[0043] 2. Antioxidant activity of the essential oils

[0044] The antioxidant activity of the essential oil of Acorus calamus, the co-distilled essential oil of Example 1 and the co-distilled essential oil of Example 2 was evaluated by three different mechanisms, including the DPPH radical scavenging test, the ABTS cation radical scavenging test and the FRAP antioxidant / reducing power assay.

[0045] (1) DPPH radical scavenging test

[0046] A 20 mg / mL solution of the essential oil was prepared in methanol or water. A DPPH methanol solution was prepared (0.0070 g + 250 mL methanol) and stored in the dark until use. The absorbance of the DPPH solution was measured before use and should be in the range of 0.7-0.8. The test was performed by first adding 100 μL of the sample solution to a cuvette, followed by 4 mL of the DPPH solution. The mixture was then left to stand in the dark for 30 min. During the test, the absorbance of the methanol, the sample solution and the DPPH solution was measured at the start of the test (A0) and after 30 min (A1). The DPPH radical scavenging activity was calculated as (A0-A1) / A0*100%. A calibration curve was prepared using Trolox (100, 200, 300, 400, 500, 750 and 1000 μmol / L) in methanol. The radical scavenging activity of the essential oil was expressed as the equivalent concentration of Trolox (μmol TE / g essential oil, TE: Trolox Equivalent).

[0047] (2) ABTS cation radical scavenging test

[0048] A 20 mg / mL solution of the essential oil was prepared in methanol or water. A 7 mmol / L ABTS solution was prepared in phosphate buffer (pH 7.4). The test was performed by first adding 10 mL of the ABTS solution to a cuvette, followed by 176 μL of the potassium persulfate solution. The mixture was then left to stand in the dark for 16 h. Before use, the ABTS cation radical solution was diluted in methanol to an absorbance of 0.70 ± 0.02 at 734 nm. The test was performed by adding 50 μL of the sample solution to a cuvette, followed by 4 mL of the diluted ABTS cation radical solution. The absorbance was measured at the start of the test (A0) and after 6 min (A1). The ABTS cation radical scavenging activity was calculated as (A0-A1) / A0*100%. A calibration curve was prepared using Trolox in methanol. The radical scavenging activity of the essential oil was expressed as the equivalent concentration of Trolox.

[0049] (3) FRAP antioxidant / reducing power test

[0050] Prepare a 20 mg / mL solution of the essential oil in methanol or water. Prepare the FRAP working solution by mixing 50 mL acetate buffer (300 mmol / L, pH 3.6), 5 mL TPTZ solution (10 mmol / L) and 5 mL FeCl3-6H2O solution (20 mmol / L) and leave for 30 min. Before the assay, heat the FRAP working solution to 37°C and keep it ready; during the assay, zero with 150 μL purified water + 450 μL methanol + 4.5 mL purified water. Mix 150 μL of the essential oil solution with 450 μL methanol and then add 4.5 mL of the FRAP working solution. Keep the mixture at 37°C for 30 min and measure the absorbance at 593 nm. Repeat the experiment four times and perform a blank assay. Prepare a calibration curve with Trolox in methanol at known concentrations and express the antioxidant capacity of the essential oil as the equivalent concentration of Trolox.

[0051] The results are shown in Table 3. The co-distilled essential oils of Example 1 and Example 2 have better antioxidant activity than the essential oil of Acorus gramineus, which allows the tobacco flavor provided by the present application to absorb free radicals in the smoke generated by tobacco combustion and to provide certain health benefits.

[0052] Table 3. Antioxidant activity of the co-distilled essential oils of Example 1 and 2

[0053]

[0054] 3. Antimicrobial activity of the essential oils

[0055] The strains used in the antimicrobial assay include three bacteria and one fungus. The bacteria include Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, and the fungus is Candida albicans. The minimum inhibitory concentration MIC (mg / mL) and the minimum microbial concentration MMC (mg / mL) of the essential oils are determined by the double dilution method.

[0056] The essential oils were prepared as 100 mg / mL solutions. A small amount of bacterial biofilm was taken from the slant and the bacteria were dissolved in MHB broth and incubated at 37°C for 20 hours. Fungal samples were dissolved in YMB broth and incubated at 28°C for 48 hours. After the initial incubation, the bacteria were streaked onto MH agar plates and incubated at 37°C for 24 hours to allow the individual colonies to become visible. For the fungi, a similar procedure was used to streak the samples onto YM agar plates and incubate at 28°C for 48 hours. Next, the desired colonies were selected from the grown plates using a sterile loop and added to sterile saline. The bacteria were diluted to an optical density of 0.08-0.11 at 600 nm, which corresponds to a concentration of approximately 10 8 CFU / mL. The bacteria were diluted to 10 6 CFU / mL with saline and then to 10 5 CFU / mL with broth for use in the assay. The fungi were diluted to a 0.5 McFarland turbidity, which corresponds to a concentration of approximately 10 6 CFU / mL. The fungi were diluted to 10 4 CFU / mL with saline and then to 10 3 CFU / mL with broth for use in the assay. The minimum inhibitory concentration (MIC) was determined using a two-fold dilution method in 96-well plates. The first column was filled with 120 μL of the sample solution, and 60 μL was transferred to the second column, which was then serially diluted with broth. After the dilution was complete, 60 μL of the bacterial solution was added to the sample wells, and the bacteria were incubated at 37°C for 20 hours (fungi were incubated at 28°C for 48 hours). The concentration corresponding to the wells in which no growth was observed was the minimum inhibitory concentration (MIC). The minimum bactericidal concentration (MBC) was determined based on the MIC assay by removing 60 μL from the wells corresponding to the MIC, 2x MIC, and 4x MIC and spreading evenly onto plate media. The bacteria were incubated at 37°C for 20-24 hours (fungi were incubated at 28°C for 48 hours), and the concentration corresponding to the wells in which no growth was observed was the minimum bactericidal concentration (MBC). The results are shown in Table 4. As can be seen, the co-distilled essential oils of Examples 1 and 2 both have significant antimicrobial activity, but there is no significant improvement over the essential oil of Acorus calamus.

[0057] Table 4. Antimicrobial activity of the co-distilled essential oils of Examples 1 and 2

[0058]

[0059] Example 4. Evaluation of tobacco flavor

[0060] After the tobacco flavor of example 1 and example 2 is added into cigarettes respectively, a professional person carries out smoking evaluation, and the comprehensive evaluation is carried out from multiple dimensions such as aroma quality, aroma amount, eating taste, miscellaneous gas, irritability and strength, and the results are shown in table 5. It can be seen that the tobacco flavor provided by the present application has better effect in tobacco flavoring, and compared with simple aniseed calamus essential oil, the aroma quality and aroma amount are improved.

[0061] Table 5: smoking evaluation results of the tobacco flavor of example 1 and 2 after being added into cigarettes

[0062]

[0063] The above is a further detailed description of the present application in combination with specific preferred embodiments, and the specific implementation of the present application cannot be limited to these descriptions. For ordinary skilled persons in the technical field to which the present application belongs, a number of simple deductions or substitutions can be made without departing from the concept of the present application, and all of them should be regarded as falling within the protection scope of the present application.

Claims

1. A method for preparing a tobacco flavoring using a discarded tobacco resource, characterized by: The method comprises the following steps: preparing a petroleum ether extract of waste tobacco resources, and dissolving the petroleum ether extract in a co-distilled essential oil of the waste tobacco resources and Acorus calamus Linné; the mass ratio of the petroleum ether extract to the co-distilled essential oil is (1-3):(4-6); The preparation of the petroleum ether extract comprises the following steps: crushing the waste tobacco resources, and placing the crushed waste tobacco resources in a round-bottom flask; adding petroleum ether and heating the mixture to reflux and extract; separating the extract by suction filtration; adding petroleum ether to the residue, and continuing to heat the mixture to reflux and extract; separating the extract by suction filtration; adding petroleum ether to the residue, and continuing to heat the mixture to reflux and extract; separating the extract by suction filtration; combining the extracts, removing the solvent by rotary evaporation, and performing vacuum drying to obtain the petroleum ether extract; The preparation of the co-distilled essential oil comprises the following steps: crushing the waste tobacco resources, and placing the crushed waste tobacco resources in a flask; adding Acorus calamus Linné powder and water, and heating the mixture to perform steam co-distillation to obtain the co-distilled essential oil; in the preparation of the co-distilled essential oil, the mass ratio of the waste tobacco resources to Acorus calamus Linné is (8-12):1; The dissolving is performed by ultrasonic-assisted dissolving.

2. Application of the tobacco flavor prepared by the method for preparing a tobacco flavor from waste tobacco resources according to claim 1 in preparing a tobacco product.

3. Use of a tobacco flavour prepared according to the process for the preparation of a tobacco flavour from waste tobacco resources according to claim 2 for the preparation of a tobacco product, characterized in that: The tobacco product comprises cigarettes, tobacco shreds and cigars.

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