Method for preparing tobacco essence through co-fermentation of inapplicable tobacco leaves and roses

Abstract

The invention discloses a method for preparing tobacco essence through co-fermentation of inapplicable tobacco leaves and roses, and belongs to the field of tobacco fermentation and tobacco essence. The preparation method comprises the following steps: culturing bacillus subtilis in a proper culture medium under proper culture conditions to obtain a bacillus subtilis liquid; spraying the obtained bacillus subtilis liquid on the rosa damascena, and fermenting under a proper condition; blending the fermented Damascus roses with inapplicable tobacco leaves, and performing fermentation and alcoholization under proper conditions; and extracting the Damascus roses after fermentation and alcoholization and inapplicable tobacco leaves by using an organic solvent, and concentrating to obtain the tobacco essence. The essence prepared by the invention not only has typical rose fragrance, but also better retains the original fragrance of tobacco. When the prepared essence is applied to cigarettes, the aroma quality of the cigarettes can be effectively improved, the irritation is remarkably reduced, and smoke is coordinated and has flower and fruit aroma. According to the invention, the source of the tobacco essence is enriched, and the utilization way of inapplicable tobacco leaves is expanded.

Description

Technical Field

[0001] The invention belongs to the technical field of tobacco fermentation and tobacco flavors, and particularly relates to a method for preparing tobacco flavors without co-fermentation of tobacco leaves and roses. Background Art

[0002] Tobacco is one of my country's important cash crops. The aroma quality of cigarettes directly affects consumers' smoking experience, but with the implementation of the "tar reduction and harm reduction" policy, the aroma of cigarettes has been affected to a certain extent. To improve the aroma loss caused by tar reduction, flavors and fragrances are often added to tobacco. Flavors are generally classified according to their source as natural flavors and synthetic flavors. Currently, most flavors and fragrances on the market are synthetic flavors. However, the production process of synthetic flavors may involve chemical reactions, generating waste and causing environmental pollution. In addition, flavors developed from the original tobacco leaf have long been considered the most compatible product with smoke. Therefore, developing tobacco-derived flavors to replace chemically synthesized tobacco flavors is of great significance to the sustainable development of the tobacco flavor industry.

[0003] Damascus rose, a deciduous, upright shrub from the Rosaceae family, is used for both ornamental and medicinal purposes. Native to Asia Minor and long cultivated in Southern Europe, it is widely used in food, cosmetics, and tobacco research and development. Reports have been published on the co-fermentation of natural plants with tobacco leaves and the extraction of their fermentation products. This has been shown to effectively enhance the aroma and taste of tobacco products through chemical transformation, aroma fusion, reduced irritation, and improved quality and stability, resulting in a superior smoking experience.

[0004] Unsuitable tobacco refers to tobacco raw materials that are classified as substandard or fail to meet grading standards during the tobacco leaf evaluation process and are therefore unsuitable for cigarette processing. In recent years, the cigarette industry has experienced excessive inventories of unsuitable tobacco, placing significant pressure on warehousing, maintenance, cash flow, and quality assurance. To reduce waste and lower costs, companies commonly use methods such as converting unsuitable tobacco into tobacco sheets through papermaking, fermentation to improve usability, and digestion.

[0005] In summary, the co-fermentation of Damascus rose with unsuitable tobacco leaves to produce tobacco flavorings can help improve aroma stability and durability, thereby enhancing the consumer smoking experience. It can also reduce the use of chemical synthetic flavorings through more environmentally friendly production methods, thereby promoting the green and sustainable development of the tobacco industry. Summary of the Invention

[0006] The present invention provides a method for producing tobacco flavorings by co-fermenting unsuitable tobacco leaves with roses. The flavoring produced by this method exhibits a typical rose aroma while retaining the natural aroma of tobacco. Application of this flavoring in cigarettes effectively improves the aroma quality of the cigarettes, significantly reducing irritation and imparting a harmonious floral and fruity aroma to the smoke. This invention not only enriches the sources of tobacco flavorings but also expands the utilization of unsuitable tobacco leaves.

[0007] The present invention is not applicable to the method for preparing tobacco flavoring by co-fermentation of tobacco leaves and roses, which comprises the following steps:

[0008] Step 1: Cultivate Bacillus subtilis in a suitable culture medium and under suitable culture conditions to obtain a Bacillus subtilis bacterial liquid.

[0009] Step 2: Spray the Bacillus subtilis liquid obtained in step 1 onto the Damascus rose and ferment under appropriate conditions.

[0010] Step 3: Blend the fermented Damascus rose with unsuitable tobacco leaves and ferment and mellow them under suitable conditions.

[0011] Step 4: After fermentation and alcoholization, the Damascus rose and unsuitable tobacco leaves are extracted and concentrated using an organic solvent to obtain tobacco flavor.

[0012] In step 1, the culture medium is composed of: 1.5 g lactose, 1 g yeast extract powder, 1 g magnesium sulfate, and water is added to make up to 100 ml.

[0013] In step 1, the culture conditions are: temperature 26-30°C, initial pH 5-7, and rotation speed 150-180 r / min. Preferably, the culture temperature is 30°C, the initial pH = 6, and the rotation speed is 180 r / min.

[0014] In step 2, the viable bacterial count of the Bacillus subtilis solution is 1×10 7 ~1×10 8 The spraying amount is 1-3wt% of the weight of the Damascus rose, and the preferred spraying amount is 3wt%.

[0015] In step 2, the fermentation is carried out at 30° C. for 2 days.

[0016] In step 3, when the fermented Damascus rose is blended with unsuitable tobacco leaves, the mass of the Damascus rose is 1-3 wt % of the mass of the unsuitable tobacco leaves, preferably 3 wt %.

[0017] Furthermore, the moisture content of the blended Damascus rose and tobacco mixture is ≤13%.

[0018] In step 3, the fermentation and alcoholization conditions are: fermentation temperature 30-36° C., fermentation time 30-90 days, and relative humidity 55%, preferably 32° C. and 60 days.

[0019] In step 4, the organic solvent used for extraction is 60-80% by volume ethanol solution, and the material ratio is 1:20-1:35 (g / mL), preferably 80% by volume ethanol solution; the material ratio is 1:30.

[0020] The tobacco flavor prepared by the present invention is added in a flavoring manner by injecting 7 μL into a blank cigarette without flavoring, or prepared into flavor granules and added into a filter rod of a blank cigarette.

[0021] The beneficial effects of the present invention are embodied in:

[0022] The present invention extracts the fermentation product from Damascus roses and unsuitable tobacco leaves through co-fermentation to produce tobacco flavoring. Compared to the production of synthetic flavorings, which can involve complex chemical processes and consume more resources, the present invention places a smaller burden on the environment. From an aroma perspective, naturally fermented flavorings typically have a more complex aroma hierarchy because the various compounds produced during the fermentation process interact with each other to form unique aroma combinations. This layered experience is often difficult for synthetic flavorings to fully simulate. Furthermore, this method provides a new approach to flavor development, unlocking the potential of more natural flavors and bringing innovation and development opportunities to the flavoring industry. It also offers new insights into the development and utilization of unsuitable tobacco leaves. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is the volatile component composition of samples under different treatment methods. DETAILED DESCRIPTION

[0024] The technical solution of the present invention is further illustrated below by using specific implementation methods. Those skilled in the art should understand that the given embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0025] Example 1:

[0026] Bacillus subtilis (purchased from the American Type Culture Collection (ATCC), with the accession number ATCC 23857).

[0027] The preserved Bacillus subtilis was activated on LB plates at 28-32°C for 2 days. Then, a 1 cm piece of microporous material was cut from the edge of the colony using a knife in a clean bench. 2The bacterial block was inoculated into 100 mL of liquid culture medium (liquid culture medium (lactose added in an amount of 1.5 g, yeast extract added in an amount of 1 g, magnesium sulfate added in an amount of 1 g, distilled water 100 ml, sterilized at 121°C for 20 min.) at 30°C, initial pH = 6, and cultured in a constant temperature shaker at 180 r / min for 24 h to obtain a Bacillus subtilis bacterial solution.

[0028] The different effects of using Bacillus subtilis in the present invention compared with other strains are shown in the following table:

[0029] Table 1 Sensory scores of fermentation effects of different strains

[0030]

[0031] The data shown in the table above represent the sensory results of fermenting different strains of Damascus rose oil, blending the fermented Damascus rose oil with unsuitable tobacco leaves, and then fermenting and mellowing under appropriate conditions to extract and prepare tobacco flavoring. (CK) represents the mellowing preparation process without the addition of strains. The Bacillus subtilis strain used in this invention offers advantages such as a superior aroma, excellent comfort, and the highest overall sensory score.

[0032] Example 2:

[0033] Selected damask rose petals free of mold and impurities, and evenly sprayed with the Bacillus subtilis solution prepared in Example 1 at a ratio of 3 wt% to the weight of the damask rose. The damask rose was then incubated in a constant temperature incubator at 30°C for 2 days to obtain fermented damask rose.

[0034] Example 3:

[0035] Instead of using shredded tobacco leaves, blend the fermented rose from Example 2 with shredded tobacco at a ratio of 1% by weight. Ferment and alcoholize for 30 days at 55% relative humidity and 32°C to obtain a co-fermentation product. The co-fermentation product was then heated to reflux in a 90°C waterbath for 2 hours using 60% ethanol at a ratio of 1 g:25 mL. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0036] Example 4:

[0037] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a ratio of 1% by weight. Fermentation and aging were carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated to reflux in a 90°C waterbath for 2 hours using 60% ethanol at a ratio of 1 g:25 mL. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0038] Example 5:

[0039] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a ratio of 1% by weight. Fermentation and aging were performed at 55% relative humidity and 32°C for 90 days to obtain a co-fermentation product. The co-fermentation product was then heated to reflux in a 90°C water bath at a ratio of 1 g:25 mL using 60% ethanol for 2 hours. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0040] Example 6:

[0041] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a rate of 2% by weight. Fermentation was carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated under reflux in a 90°C waterbath with 60% ethanol at a ratio of 1 g:25 mL for 2 hours. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0042] Example 7:

[0043] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a rate of 3% by weight. Fermentation was carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated under reflux in a 90°C waterbath at a ratio of 1 g:25 mL using 60% ethanol for 2 hours. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0044] Example 8:

[0045] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a rate of 3% by weight. Fermentation was carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated under reflux in a 90°C waterbath at a ratio of 1 g:30 mL using 60% ethanol for 2 hours. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0046] Example 9:

[0047] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a rate of 3% by weight. Fermentation was carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated under reflux in a 90°C waterbath with 60% ethanol at a ratio of 1 g:35 mL for 2 hours. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0048] Example 10:

[0049] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a rate of 3% by weight. Fermentation was carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated under reflux in a 90°C waterbath for 2 hours using 70% ethanol at a ratio of 1 g:30 mL. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0050] Example 11:

[0051] Instead of using shredded tobacco leaves, the fermented rose from Example 2 was blended with shredded tobacco at a rate of 3% by weight. Fermentation was carried out at 55% relative humidity and 32°C for 60 days to obtain a co-fermentation product. The co-fermentation product was then heated under reflux in a 90°C waterbath at a ratio of 1 g:30 mL using 80% ethanol for 2 hours. After cooling, the mixture was centrifuged, and the supernatant was removed and concentrated to 1 / 10 of its original volume using a rotary evaporator to obtain a tobacco flavoring.

[0052] Example 12:

[0053] The flavor prepared in the above examples was added to unflavored blank cigarettes at a 7 μL injection volume. The cigarettes were then equilibrated in a constant temperature and humidity chamber at (22±1)°C and (60±3)% for 48 hours. Sensory evaluation was conducted in accordance with GB 5606.4-2005, "Cigarettes - Part 4: Sensory Technical Requirements." A seven-member expert panel evaluated the extracted flavors, finding that adding the flavors to cigarettes reduced irritation, softened the smoke, and enhanced the tobacco's natural aroma, with a distinct floral and fruity aroma.

[0054] Example 13:

[0055] 12g of tobacco flavoring, 4g of β-cyclodextrin, and 3g of microcrystalline cellulose were added to a solvent (20mL of pure water and 1mL of propylene glycol) and thoroughly stirred to form a wet material. The wet material was then added to an extruder (E-25) and extruded at 110r / min. The extrudate was then placed in a spheronizer (Mini S) and spheronized at 1000r / min into spherical wet pellets. The prepared spherical wet pellets were dried in a 40°C oven for 2 hours to obtain a solid flavoring. The solid flavoring was then added to filter rods of blank cigarettes for sensory evaluation. Compared to the blank cigarettes, the flavored cigarettes showed a richer smoke, significantly reduced irritation, and a more sweet and mellow taste, with a distinct floral and fruity aroma.

[0056] Example 14:

[0057] The volatile components of the fermentation and alcoholization of unsuitable tobacco leaves alone (treatment 1), the alcoholization of Damascus rose and unsuitable tobacco leaves together (treatment 2), and the fermentation of Damascus rose by spraying Bacillus subtilis first and then co-fermentation and alcoholization with unsuitable tobacco leaves (treatment 3) are shown in Table 2. Figure 1 shown.

[0058] Table 2 Composition and content of volatile substances in three treatments

[0059]

[0060]

[0061]

[0062] The composition and content of volatile components of samples under three treatment methods are as follows Figure 1 As shown in Table 2, overall, Treatment 3 (prefermenting Damascus rose with Bacillus subtilis before co-fermentation with unsuitable tobacco leaves) produced the richest variety and highest content of volatile components, reaching a total of 1381.28 μg / g, significantly higher than Treatment 2 (1155.20 μg / g) and Treatment 1 (1056.89 μg / g), demonstrating the superiority of its synergistic fermentation. Regarding the content of each category of volatile components, Treatment 3 exhibited a clear advantage in alcohols, esters, and ketones, with a richer composition and significantly higher content than the other treatments.

[0063] In terms of alcohols, phenylethanol (29.24 μg / g) in treatment 3 was significantly higher than that in treatment 1 (0.15 μg / g) and treatment 2 (20.85 μg / g), and had a strong rose fragrance; p-hydroxyphenylethanol (10.71 μg / g) was also an aromatic alcohol with a soft sweet fragrance and was also a common ingredient in perfumes; sclareol (18.02 μg / g) gave a fresh herbal floral fragrance, which was significantly higher than that in treatment 2 (15.78 μg / g), while treatment 1 had a In treatment 3, the contents of linalool (1.35), geraniol (1.58), and citronellol (1.03) were higher than those in treatment 2, but not detected in treatment 1. These three substances have a low odor threshold and contribute floral fragrance and mild freshness to the fragrance. In addition, long-chain alcohols such as dehydrogenated linalool (13.84 μg / g) and n-docosanol (11.64 μg / g) unique to treatment 3 have a light fruity aroma, indicating that synergistic fermentation is conducive to the formation of complex alcohols.

[0064] Among esters, di(2-ethylhexyl) adipate (85.97 μg / g) in treatment 3 was significantly higher than that in the other two groups, giving the system a soft, sweet fragrance and a smooth feel; citronellyl oleate (26.81 μg / g) had a fruity and fatty aroma; in addition, dihydroactin (3.55 μg / g) was an important neutral to fragrant component, constituting the aroma base of treatment 3.

[0065] Among ketones, DDMP (6.95 μg / g) was the highest in treatment 3 and is a typical Maillard intermediate with a sweet caramel aroma. 9-hydroxy-4,7-macrostigena-3-one (37.83 μg / g) and 4-hydroxy-β-dihydrodamascone (6.15 μg / g) imparted sweet and fruity aromas, while 4-hydroxy-β-ionone (2.59 μg / g) had a soft purple floral aroma and was also significantly higher than that in treatment 1.

[0066] In summary, treatment 3 can significantly enhance the production of key aroma substances of floral, fruity and sweet types, with rich aroma types, high intensity and good coordination, indicating that the collaborative fermentation strategy significantly optimizes the composition structure of volatile flavor substances and is an effective means to improve the aroma quality of tobacco leaves.

Claims

1. A method for preparing tobacco flavoring by fermenting tobacco leaves and roses, characterized in that The steps include: Step 1: Cultivating Bacillus subtilis in a suitable culture medium and under suitable culture conditions to obtain a Bacillus subtilis bacterial solution; Step 2: Spraying the Bacillus subtilis liquid obtained in step 1 onto Damascus roses and fermenting under appropriate conditions; Step 3: blending the fermented Damascus rose with unsuitable tobacco leaves and fermenting and mellowing under suitable conditions; Step 4: After fermentation and alcoholization, the Damascus rose and unsuitable tobacco leaves are extracted and concentrated using an organic solvent to obtain tobacco flavor.

2. The method according to claim 1, wherein: The deposit number of the Bacillus subtilis is ATCC 23857.

3. The method according to claim 1, wherein: In step 1, the culture medium is composed of: 1.5 g lactose, 1 g yeast extract powder, 1 g magnesium sulfate, and water is added to make up to 100 ml.

4. The method according to claim 3, wherein: In step 1, the culture conditions are: temperature 26-30° C., initial pH 5-7, and rotation speed 150-180 r / min.

5. The method according to claim 1, wherein: In step 2, the viable bacterial count of the Bacillus subtilis solution is 1×10 7 to 1×10 8 The spraying amount is 1-3wt% of the weight of Damascus rose.

6. The method according to claim 5, characterized in that: In step 2, the fermentation is carried out at 30° C. for 2 days.

7. The method according to claim 1, wherein: In step 3, when the fermented Damascus rose is blended with unsuitable tobacco leaves, the mass of the Damascus rose is 1-3 wt % of the mass of the unsuitable tobacco leaves.

8. The method according to claim 1, wherein: In step 3, the fermentation and alcoholization conditions are: fermentation temperature 30-36° C., fermentation time 30-90 days, and relative air humidity 55%.

9. The method according to claim 1, wherein: In step 4, the organic solvent used for extraction is 60-80vt% ethanol solution, and the material ratio is 1g:20mL-35mL.

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