Preparation method of tobacco tree orchid extract as well as product and application of tobacco tree orchid extract

Abstract

The invention relates to a preparation method of a tobacco tree orchid extract and a product and application thereof, and the preparation method comprises the following steps: mixing a tree orchid raw material with water, pulping, mixing with enzyme, carrying out enzymolysis treatment, and carrying out solid-liquid separation to obtain a flower pulp extracting solution and flower residues; the obtained flower pulp extracting solution is subjected to low-temperature distillation, condensation and separation, and orchid fragrance liquid and a fragrance-removed solution are obtained; concentrating the obtained cymbidium odoratum essence, and separating to obtain concentrated aroma liquid and aroma-removed clear liquid; combining the obtained flower residues, the aroma-removed solution and the aroma-removed clear liquid, mixing with glycosidase, carrying out enzymolysis treatment, carrying out enzyme deactivation, carrying out alcohol extraction on a product, then carrying out solid-liquid separation, and concentrating filtrate, so as to obtain an orchid-flavor substrate extract; and adding the obtained concentrated aroma liquid back into the obtained orchid flavor base extract, then carrying out embedding treatment, and homogenizing to obtain the orchid extract for cigarettes. The orchid extract for cigarettes completely retains the top aroma, body aroma and bottom aroma of orchid, and improves the storage stability and fidelity of aroma components.

Description

Technical Field

[0001] This invention belongs to the field of tobacco flavoring technology, and relates to a method for preparing tobacco aralia extract, its products, and applications. Background Technology

[0002] The sensory quality of tobacco products is a key factor determining their market acceptance, and flavorings play an irreplaceable role in shaping the characteristic aroma of cigarettes, enhancing smoke comfort, masking off-flavors, and reducing irritation. With increasing consumer health awareness and more refined tastes, "reducing tar and harm" and "enhancing aroma and moisturizing" have become core research directions in the tobacco industry. Against this backdrop, developing high-quality, highly safe natural tobacco flavorings is particularly important. Compared to synthetic flavorings, natural flavorings have complex and layered components, a naturally harmonious aroma, and are derived from natural plants, making them more in line with health-conscious consumption trends. They are an important resource for enhancing the product's appeal and differentiated competitiveness.

[0003] Aglaia odorata Lour., also known as rice orchid, is an evergreen plant belonging to the genus Aglaia in the family Meliaceae. Its flowers possess a delicate and lasting fragrance, with a sweet floral aroma and hints of jasmine, ylang-ylang, and tea. Developing it as a tobacco flavoring agent not only imbues cigarettes with an elegant and harmonious floral and tea aroma, but also effectively improves the taste of smoke, giving it a smooth and moist feel, and reducing the hot and dry sensation of smoke. This has unique value in enhancing the sensory quality and comfort of cigarettes. Therefore, Aglaia odorata extract, as a natural tobacco flavoring agent with great potential, has attracted much attention for its high-fidelity preparation and stabilization application technology.

[0004] In the existing technology, there have been many explorations into the extraction and application of aroma components of Cymbidium goeringii. CN119499133A discloses the extraction of essential oil from the stems and leaves of Cymbidium goeringii and its application in cosmetics. Although this method can retain some active ingredients, the extraction temperature is high and the time is long, which greatly damages the heat-sensitive top notes. The resulting product has insufficient aroma integrity and poor authenticity, and there is no special treatment for the stability of the aroma during tobacco processing and storage.

[0005] CN116987551A uses a specific complex enzymatic hydrolysis combined with supercritical CO2 extraction to prepare jasmine oil. Its technical limitation is that the process mainly targets and efficiently extracts fat-soluble essential oil components, but the extraction efficiency and comprehensive utilization of water-soluble aroma precursors (such as glycosidic aromas) present in large quantities in the raw materials are insufficient. This may result in an incomplete aroma spectrum of the final product and failure to fully utilize the full flavor value of the raw materials.

[0006] CN108103109A discloses a method for preparing tobacco flavorings by fermenting *Bacillus thuringiensis* using *Bacillus thuringiensis*, which produces new flavor substances through microbial metabolic transformation, enriching the aroma. However, this method has a long process cycle, complex product composition, and alters the fidelity (authenticity) of the original *Bacillus thuringiensis* aroma characteristics. Furthermore, it does not address the issue of the physical stability of the aroma substances.

[0007] In summary, existing technologies for the development of orchid fragrances, especially for the preparation of high-quality orchid extracts for cigarette applications, still suffer from problems such as insufficient aroma fidelity, poor aroma stability, and limited resource utilization. Therefore, in response to the urgent need for high-fidelity, high-stability, and high-efficiency natural fragrances in high-end tobacco products, there is an urgent need to develop an innovative preparation method that can fully preserve the characteristic aroma of orchids, significantly improve their storage and application stability, and achieve comprehensive utilization of raw materials. Summary of the Invention

[0008] In view of the problems of insufficient aroma fidelity, poor stability of aroma components in storage and application, and low raw material utilization in existing orchid flavoring preparation technology, this invention aims to provide a method for preparing high-fidelity orchid extract for tobacco, as well as its products and applications, which can fully preserve the natural aroma of orchid, significantly improve product stability, and realize the full value utilization of raw materials.

[0009] To achieve this objective, the present invention adopts the following technical solution:

[0010] In a first aspect, the present invention provides a method for preparing tobacco aralia extract, characterized in that the preparation method includes the following steps:

[0011] (1) Mix the raw material of *Gymnocalycium mihanovichii* with water and pulp it, then mix it with an enzyme for enzymatic hydrolysis, and separate the solid and liquid to obtain flower pulp extract and flower residue; the enzyme is selected from any one or at least a combination of two of cellulase, pectinase, amylase and hemicellulase.

[0012] (2) The flower extract obtained in step (1) is subjected to low-temperature distillation, and after condensation, it is separated to obtain the orchid fragrance dew and the defragrant solution;

[0013] (3) Concentrate the orchid fragrance obtained in step (2) and separate the concentrated fragrance liquid and the defragrant clear liquid;

[0014] (4) Combine the flower residue obtained in step (1), the deodorizing solution obtained in step (2), and the deodorizing liquid obtained in step (3), mix them with glycosidase for enzymatic hydrolysis, inactivate the enzyme, extract the product with alcohol, then separate the solid and liquid, concentrate the filtrate, and obtain the orchid flavor base extract.

[0015] (5) The concentrated aroma liquid obtained in step (3) is added back to the orchid flavor base extract obtained in step (4), and then encapsulated and homogenized to obtain the orchid extract for tobacco.

[0016] This invention creatively employs enzymatic-assisted extraction, low-temperature distillation, and concentration processes sequentially on *Gynostemma pentaphyllum* raw materials. The concentrated product is then backfilled into a *Gynostemma pentaphyllum* flavor base extract prepared through a specific process (enzymatic hydrolysis combined with alcohol extraction). This results in a final tobacco *Gynostemma pentaphyllum* extract that fully preserves the top, middle, and base aromas of *Gynostemma pentaphyllum*, exhibiting a full and rich aroma with high fidelity. When applied to cigarette flavoring, it imparts an elegant and harmonious floral and tea aroma to the product, effectively enhancing the smoothness of the smoke, reducing dryness, and significantly improving the sensory quality of cigarettes.

[0017] Meanwhile, this preparation method transforms potentially wasteful deodorizing solutions, clear liquids, and flower residues into flavorful base extracts, achieving high-value utilization of the entire process of *Gynostemma pentaphyllum* raw materials and improving the overall yield.

[0018] Furthermore, this preparation method incorporates encapsulation technology, which effectively fixes free aroma molecules within the cavity of the inclusion compound, greatly reducing their volatilization and oxidation losses during storage and improving the aroma retention rate during long-term storage.

[0019] Preferably, the enzyme in step (1) is a combination of cellulase and pectinase.

[0020] For the preparation method of the present invention, the enzymes used in the enzymatic hydrolysis of the orchid raw material are more preferably a combination of cellulase and pectinase. Compared with other enzyme combinations, the orchid extract for tobacco has a higher level of aroma fidelity and a better effect on improving the sensory quality of cigarettes.

[0021] Preferably, the mass ratio of cellulase to pectinase is 1:2 to 2:1, such as 1:2, 1:1.8, 1:1.6, 1:1.5, 1:1.4, 1:1.2, 1:1, 1.2:1, 1.4:1, 1.6:1, 1.8:1, 2:1, etc.

[0022] Preferably, the amount of enzyme added in step (1) is 0.05%-0.2% of the mass of the orchid raw material, such as 0.05%, 0.08%, 0.1%, 0.12%, 0.14%, 0.15%, 0.16%, 0.18%, 0.2%, etc.

[0023] Preferably, the enzymatic hydrolysis in step (1) is carried out at 30-50℃ (e.g., 30℃, 35℃, 40℃, 45℃, 50℃, etc.) for 2-5 h (e.g., 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 5 h, etc.).

[0024] The enzymatic hydrolysis is carried out at a lower temperature, which reduces the damage and loss of heat-sensitive aroma components, helps to preserve the top notes, middle notes and base notes of the tree orchid, and improves the authenticity of the aroma.

[0025] Preferably, the raw material for the tree orchid is tree orchid.

[0026] Preferably, the mass ratio of the orchid raw material to water is 1:(5-20), such as 1:5, 1:7, 1:8, 1:10, 1:12, 1:14, 1:15, 1:16, 1:17, 1:18, 1:20, etc.

[0027] Preferably, the low-temperature distillation in step (2) is carried out at a temperature not exceeding 60°C.

[0028] The distillation is carried out at a lower temperature, which reduces the damage and loss of heat-sensitive aroma components, helps to preserve the top notes, middle notes and base notes of the orchid, and improves the authenticity of the aroma.

[0029] Preferably, the low-temperature distillation is carried out using a rotary cone distillation column with a feed temperature of 40-60℃ (e.g., 40℃, 45℃, 50℃, 55℃, 60℃, etc.) and a vacuum degree of -85 kPa to -50 kPa (e.g., -85 kPa, -80 kPa, -75 kPa, -70 kPa, -65 kPa, -60 kPa, -55 kPa, -50 kPa, etc.).

[0030] Preferably, the concentration in step (3) is achieved by freeze concentration or membrane concentration.

[0031] Preferably, the concentration in step (3) is carried out at a temperature not exceeding 40°C.

[0032] The concentration is carried out at a lower temperature, which reduces the damage and loss of heat-sensitive aroma components, helps to preserve the top notes, middle notes and base notes of the tree orchid, and improves the authenticity of the aroma.

[0033] Preferably, the membrane concentration is a reverse osmosis membrane concentration.

[0034] Preferably, the concentration ratio of the concentration in step (3) is (2-6):1, such as 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, 5.5:1, 6:1, etc.

[0035] Preferably, the glycosidase in step (4) includes β-glucosidase.

[0036] Preferably, the amount of glycosidase used is 0.3-0.5% of the enzymatic hydrolysis substrate, such as 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, etc.

[0037] Preferably, the enzymatic hydrolysis process conditions in step (4) are as follows: enzymatic hydrolysis for 1-3 h (e.g., 1 h, 1.5 h, 2 h, 2.5 h, 3 h, etc.) at pH 4.5-5.5 (e.g., 4.5, 4.7, 4.8, 4.9, 5.0, 5.2, 5.3, 5.4, 5.5, etc.) and temperature 45-55℃ (e.g., 45℃, 47℃, 48℃, 50℃, 53℃, 55℃, etc.).

[0038] Preferably, the alcohol extraction in step (4) is performed by using a 20-70% (e.g., 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, etc.) aqueous ethanol solution for 0.5-2 h (e.g., 0.5 h, 0.8 h, 1 h, 1.2 h, 1.5 h, 1.8 h, 2 h, etc.).

[0039] Preferably, the alcohol extraction method is reflux extraction or hot extraction.

[0040] Preferably, the solids content of the orchid-flavored base extract in step (4) is 40-70%, such as 40%, 45%, 50%, 55%, 60%, 65%, 70%, etc.

[0041] Preferably, the ratio of the concentrated aroma liquid to the orchid flavor base extract in step (5) is 1:(1-5) L / kg, such as 1:1 L / kg, 1:2 L / kg, 1:2.5 L / kg, 1:3 L / kg, 1:3.5 L / kg, 1:4 L / kg, 1:5 L / kg, etc.

[0042] Preferably, the embedding material in step (5) is cyclodextrin.

[0043] Preferably, the cyclodextrin is hydroxypropyl-β-cyclodextrin.

[0044] Preferably, the process conditions for the embedding treatment in step (5) are: stirring at 45-55℃ (e.g., 45℃, 47℃, 48℃, 50℃, 53℃, 55℃, etc.) for 0.5-1.5 h (e.g., 0.5 h, 1 h, 1.2 h, 1.5 h, etc.).

[0045] All other point values ​​not specifically listed within the numerical range involved in this invention can be selected and are all within the protection scope of this invention, and will not be elaborated here.

[0046] More preferably, a secondary embedding process is performed after the embedding process described in step (5).

[0047] Preferably, the material for secondary embedding is a liposome.

[0048] Preferably, the liposomes can be prepared by the following method:

[0049] Soybean lecithin and cholesterol were dissolved in anhydrous ethanol, and the organic solvent was removed by rotary evaporation to form a uniform lipid film. Phosphate buffer was added, and after hydration, the film was ultrasonically treated with a probe to obtain a liposome nanodispersion.

[0050] In a second aspect, the present invention provides a tobacco extract prepared according to the preparation method described in the first aspect.

[0051] Thirdly, the present invention provides the use of the tobacco extract according to the second aspect in the preparation of flavoring materials for tobacco products.

[0052] Compared with the prior art, the present invention has the following beneficial effects:

[0053] This invention creatively employs enzymatic-assisted extraction, low-temperature distillation, and concentration processes sequentially on *Gynostemma pentaphyllum* raw materials. The concentrated product is then backfilled into a *Gynostemma pentaphyllum* flavor base extract prepared through a specific process (enzymatic hydrolysis combined with alcohol extraction). This results in a final tobacco *Gynostemma pentaphyllum* extract that fully preserves the top, middle, and base aromas of *Gynostemma pentaphyllum*, exhibiting a full and rich aroma with high fidelity. When applied to cigarette flavoring, it imparts an elegant and harmonious floral and tea aroma to the product, effectively enhancing the smoothness of the smoke, reducing dryness, and significantly improving the sensory quality of cigarettes.

[0054] Meanwhile, this preparation method transforms potentially wasteful deodorizing solutions, clear liquids, and flower residues into flavorful base extracts, achieving high-value utilization of the entire process of *Gynostemma pentaphyllum* raw materials and improving the overall yield.

[0055] Furthermore, this preparation method incorporates encapsulation technology, which effectively fixes free aroma molecules within the cavity of the inclusion compound, greatly reducing their volatilization and oxidation losses during storage and improving the aroma retention rate during long-term storage. Detailed Implementation

[0056] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0057] The source information for some of the raw materials mentioned below is as follows:

[0058]

[0059]

[0060] Example 1

[0061] This embodiment provides a tobacco-grade orchid extract, the preparation method of which is as follows:

[0062] (1) Mix 2.5 kg of freshly washed cymbidium orchids with 10 times the weight of water and pulp them. Then add 2.5 g of cellulase (activity 100,000 U / g) and 1.25 g of pectinase (activity 30,000 U / g) to the pulp. Extract at 48℃ with constant temperature stirring for 3.5 h. After extraction, filter using a horizontal screw centrifuge to separate the pulp extract and wet flower residue for later use.

[0063] (2) Pump the flower extract obtained in step (1) into a rotary cone distillation column and set the key parameters: feed rate 50L / h, feed temperature 48℃, system vacuum maintained at -70kpa, and extraction efficiency set at 10% to obtain tree orchid fragrance and deodorized solution.

[0064] (3) Transfer the orchid fragrance obtained in step (2) to a membrane separation device and concentrate it using a reverse osmosis membrane until the volume is 1L to obtain concentrated fragrance liquid; the clear liquid produced during the concentration process is collected as defragrant clear liquid;

[0065] (4) Combine all the flower residue obtained in step (1), all the deodorized solution obtained in step (2), and all the deodorized clear liquid obtained in step (3), and stir evenly. Adjust the pH of the mixture to 5.0 with citric acid, add 0.5% β-glucosidase (activity 100,000 U / g) of the enzymatic substrate, and enzymatically hydrolyze in a 50℃ water bath shaker for 2 h. After the enzymatic hydrolysis is completed, inactivate the enzyme at 90℃ for 15 minutes, add 1.5 times the mass of 50% ethanol aqueous solution of the system, heat and reflux for 1 h, then cool and separate by horizontal centrifuge. The supernatant is concentrated at -0.095 MPa to a solid content of 55% to obtain a dark brown, richly flavored orchid-flavored base extract;

[0066] (5) Under stirring, the concentrated aroma liquid obtained in step (3) is added dropwise to the orchid flavor base extract obtained in step (4) (ratio of 1 L: 2.5 kg), and then 5.0% of hydroxypropyl-β-cyclodextrin is added according to the mass of the base extract. The mixture is continuously stirred at 52°C for 1.2 h for encapsulation. Finally, the mixture is homogenized three times in a high-pressure homogenizer at a pressure of 60 MPa to obtain the orchid extract for tobacco (referred to as sample E1).

[0067] Example 2

[0068] This embodiment provides a tobacco-grade orchid extract, the preparation method of which is as follows:

[0069] (1) Mix 2.5 kg of cleaned fresh tree flower with 15 times the weight of water and then add 0.1% of cellulase (activity 100,000 U / g) and pectinase (activity 30,000 U / g) of the total mass of tree flower to the flower pulp. The mass ratio of cellulase to pectinase is 2:1. Extract at 35℃ with constant temperature stirring for 5 h. After extraction, filter using a horizontal screw centrifuge to separate the flower pulp extract and wet flower residue for later use.

[0070] (2) Pump the flower extract obtained in step (1) into a rotary cone distillation column and set the key parameters: feed rate 50L / h, feed temperature 54℃, system vacuum maintained at -80kpa, and extraction efficiency set at 10% to obtain tree orchid fragrance and deodorized solution.

[0071] (3) Transfer the orchid fragrance obtained in step (2) to a membrane separation device and concentrate it using a reverse osmosis membrane until the volume is 1.5L to obtain concentrated fragrance liquid; the clear liquid produced during the concentration process is collected as defragrant clear liquid;

[0072] (4) Combine all the flower residue obtained in step (1), all the deodorized solution obtained in step (2), and all the deodorized clear liquid obtained in step (3), and stir evenly. Adjust the pH of the mixture to 5.5 with citric acid, add 0.5% β-glucosidase (activity 100,000 U / g) of the enzymatic substrate, and enzymatically hydrolyze for 1 h in a water bath shaker at 55℃. After enzymatic hydrolysis, inactivate the enzyme at 90℃ for 15 minutes, add 1.5 times the mass of 70% ethanol aqueous solution of the system, heat and reflux for 1.5 h, then cool and separate by horizontal centrifuge. Concentrate the supernatant at -0.095 MPa to a solid content of 55% to obtain a dark brown, richly flavored orchid-flavored base extract;

[0073] (5) Under stirring, the concentrated aroma liquid obtained in step (3) is added dropwise to the orchid flavor base extract obtained in step (4) (ratio of 1 L: 2.8 kg), and then 5.0% of hydroxypropyl-β-cyclodextrin is added according to the mass of the base extract. The mixture is continuously stirred at 55°C for 1 h for encapsulation. Finally, the mixture is homogenized three times in a high-pressure homogenizer at a pressure of 60 MPa to obtain the orchid extract for tobacco (denoted as sample E2).

[0074] Example 3

[0075] This embodiment provides a tobacco-grade orchid extract, the preparation method of which is as follows:

[0076] (1) Mix 2.5 kg of cleaned fresh tree flower with 8 times the weight of water and pulp. Then add 0.2% of the total weight of tree flower to the pulp, including cellulase (100,000 U / g activity) and pectinase (30,000 U / g activity), with a mass ratio of cellulase to pectinase of 1:2. Extract at 45℃ with constant temperature stirring for 2 h. After extraction, filter using a horizontal screw centrifuge to separate the pulp extract and wet flower residue for later use.

[0077] (2) Pump the flower pulp extract obtained in step (1) into a rotary cone distillation column and set the key parameters: feed rate 50L / h, feed temperature 45℃, system vacuum maintained at -65kpa, and extraction efficiency set at 10% to obtain tree orchid fragrance and deodorized solution.

[0078] (3) Transfer the orchid fragrance obtained in step (2) to a membrane separation device and concentrate it using a reverse osmosis membrane until the volume is 0.8L to obtain concentrated fragrance liquid; the clear liquid produced during the concentration process is collected as defragrant clear liquid;

[0079] (4) Combine all the flower residue obtained in step (1), all the deodorized solution obtained in step (2), and all the deodorized clear liquid obtained in step (3), and stir evenly. Adjust the pH of the mixture to 4.5 with citric acid, add 0.3% β-glucosidase (activity 100,000 U / g) of the enzymatic substrate, and enzymatically hydrolyze for 3 h in a water bath shaker at 45℃. After enzymatic hydrolysis, inactivate the enzyme at 90℃ for 15 minutes, add 1.5 times the mass of 30% ethanol aqueous solution of the system, heat and reflux for 2 h, then cool and separate by horizontal centrifuge. Concentrate the supernatant at -0.095 MPa to a solid content of 55% to obtain a dark brown, richly flavored orchid-flavored base extract;

[0080] (5) Under stirring, the concentrated aroma liquid obtained in step (3) is added dropwise to the orchid flavor base extract obtained in step (4) (ratio of 1 L: 2.6 kg), and then 5.0% of hydroxypropyl-β-cyclodextrin is added according to the mass of the base extract. The mixture is continuously stirred at 45°C for 1.5 h for encapsulation. Finally, the mixture is homogenized three times in a high-pressure homogenizer at a pressure of 60 MPa to obtain the orchid extract for tobacco (referred to as sample E3).

[0081] Example 4

[0082] This embodiment provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in that a liposome encapsulation treatment is added, specifically:

[0083] (1) Weigh 6.0 g of soybean lecithin and 1.2 g of cholesterol and dissolve them in 40 mL of anhydrous ethanol. Remove the organic solvent by rotary evaporation at 40 °C to form a uniform lipid film. Add 60 mL of phosphate buffer (pH 7.0), hydrate, and then sonicate with a probe (300 W power, 2 seconds working, 3 seconds intermittent, total duration 5 minutes) to obtain a liposome nanodispersion.

[0084] (2) Prepare 500 g of the mixture before homogenization and complete cyclodextrin encapsulation according to Example 1, mix it with all the liposome dispersions prepared above, and perform 3 "freeze-thaw" cycles (freeze at -20℃ for 12 hours, thaw in a water bath at 25℃ for 2 hours).

[0085] (3) The material that has undergone double encapsulation treatment is homogenized twice under a pressure of 60 MPa to obtain the final product (referred to as sample E4).

[0086] Example 5

[0087] This embodiment provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in that: in step (1), 0.15% of the total mass of the orchid flower is added to the flower pulp, consisting of cellulase (activity 100,000 U / g), hemicellulase (activity 50,000 U / g), and pectinase (activity 30,000 U / g), with the mass ratio of cellulase to hemicellulase to pectinase being 1:1:1, and other conditions remaining unchanged, to obtain the final product (referred to as sample E5).

[0088] Example 6

[0089] This embodiment provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in that: in step (1), 0.15% of the total mass of the orchid flower mass of amylase (activity 10,000 U / g) and pectinase (activity 30,000 U / g) are added to the flower pulp, the mass ratio of amylase to pectinase is 2:1, and other conditions remain unchanged, and the final product (referred to as sample E6) is obtained.

[0090] Comparative Example 1

[0091] This comparative example provides a tobacco aralia extract, the preparation method of which differs from that of Example 1 only in that: in step (4), all the deodorized solution obtained in step (2) and all the deodorized clear liquid obtained in step (3) are combined and stirred evenly as raw materials for the preparation of aralia flavor base extract; and in step (5), hydroxypropyl-β-cyclodextrin is not added and high-pressure homogenization is not performed. The operation and parameters of other steps remain unchanged, and the final product (denoted as sample C1) is obtained.

[0092] Comparative Example 2

[0093] This comparative example provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in that: in step (4), all the flower residue obtained in step (1) and all the deodorized clear liquid obtained in step (3) are combined and stirred evenly as raw materials for the preparation of orchid flavor base extract; and in step (5), hydroxypropyl-β-cyclodextrin is not added and high-pressure homogenization is not performed. The operation and parameters of other steps remain unchanged, and the final product (denoted as sample C2) is obtained.

[0094] Comparative Example 3

[0095] This comparative example provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in that: in step (4), all the flower residue obtained in step (1) and all the deodorized solution obtained in step (2) are combined and stirred evenly as raw materials for the preparation of orchid flavor base extract; and in step (5), hydroxypropyl-β-cyclodextrin is not added and high-pressure homogenization is not performed. The operation and parameters of other steps remain unchanged, and the final product (denoted as sample C3) is obtained.

[0096] Comparative Example 4

[0097] This comparative example provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in the preparation process of the base extract in step (4), as follows:

[0098] All the flower residue obtained in step (1), all the deodorized solution obtained in step (2), and all the deodorized clear liquid obtained in step (3) were combined and stirred evenly. The pH of the mixture was adjusted to 5.0 with citric acid, and 0.5% β-glucosidase (activity 50,000 U / g) of the substrate was added. The mixture was enzymatically hydrolyzed in a 50°C water bath shaker for 3 h. After the enzymatic hydrolysis was completed, the enzyme was inactivated at 90°C for 15 minutes, and then separated by horizontal centrifuge after cooling. The supernatant was concentrated at -0.095 MPa to a solid content of 55% to obtain the base extract. Hydroxypropyl-β-cyclodextrin was not added in step (5), and high-pressure homogenization was not performed. The operation and parameters of other steps remained unchanged to obtain the final product (denoted as sample C4).

[0099] Comparative Example 5

[0100] This comparative example provides a tobacco orchid extract, the preparation method of which differs from that of Example 1 only in the preparation process of the base extract in step (4), as follows:

[0101] All the flower residue obtained in step (1), all the deodorized solution obtained in step (2), and all the deodorized clear liquid obtained in step (3) were combined and stirred evenly. 1.5 times the mass of 50% ethanol aqueous solution was added to the system and heated to a gentle boil under reflux for 2 hours. After cooling, the mixture was separated by horizontal centrifuge. The supernatant was concentrated at -0.095 MPa until the solid content was 55% to obtain the base extract. Hydroxypropyl-β-cyclodextrin was not added in step (5), and high-pressure homogenization was not performed. The operation and parameters of other steps remained unchanged to obtain the final product (denoted as sample C5).

[0102] Comparative Example 6

[0103] This comparative example provides a tobacco-grade orchid extract, the preparation method of which is as follows:

[0104] (1) Mix 2.5 kg of freshly washed tree flower with 10 times the weight of water and then add 0.15% of the total weight of tree flower to the flower pulp. The enzyme is composed of glucose isomerase, non-specific enzyme streptase E and bromelain in a mass ratio of 2:0.5:4. The mixture is stirred at 48℃ for 3.5 h. After extraction, the mixture is filtered by a horizontal screw centrifuge to separate the flower pulp extract and wet flower residue.

[0105] (2) Place the flower pulp extract in a microwave extraction device, set the power to 400W, and perform microwave-assisted steam distillation. When the amount of essential oil in the collection device no longer increases, separate the essential oil.

[0106] (3) The wet flower residue from step (1) was ultrasonically extracted with 95% ethanol (material-liquid ratio 1:3) at 45 kHz for 1 h. The ethanol extract was filtered and the ethanol was recovered under reduced pressure to obtain the concentrate.

[0107] (4) Combine the essential oil from step (2) with the concentrate from step (3) to obtain the final product (referred to as sample C6).

[0108] Comparative Example 7

[0109] This comparative example provides a tobacco-grade orchid extract, the preparation method of which is as follows:

[0110] (1) Mix 2.5 kg of freshly washed tree flower with 10 times the weight of water and then add 0.15% of the total weight of tree flower to the flower pulp. The mixture consists of a complex enzyme composed of hemicellulase, pectinase, xylanase, β-glucanase and subtilisin (enzyme activity ratio 5:4:4:2:1). Extract at 48℃ with constant temperature stirring for 3.5 h.

[0111] (2) The enzymatically hydrolyzed material was subjected to supercritical CO2 fluid extraction under the following conditions: pressure 30 MPa, temperature 45℃, extraction time 2 h, separation pressure 6 MPa, and separation temperature 55℃. The extract was collected to obtain orchid oil (denoted as sample C7).

[0112] Test Example 1

[0113] Sensory descriptive analysis and similarity scoring were used to verify the aroma fidelity of the products prepared in Examples 1-6 and Comparative Examples 1-7.

[0114] Ten specially trained sensory evaluators were selected to dilute the products obtained in Examples 1-6 and Comparative Examples 1-7 to a concentration of 1% (w / w) using an equal volume of propylene glycol / deionized water (1:1) mixed solvent. Fresh jasmine flowers (which released their aroma after being flash-frozen in liquid nitrogen and then thawed at room temperature) were used as a natural aroma reference. During the evaluation, each sample solution (0.1 mL) was dropped onto a standard olfactory strip, and the evaluators conducted an olfactory evaluation.

[0115] First, a descriptive analysis was conducted, recording the intensity (0-5 points) of each sample across eight attributes: "fresh floral," "sweet," "green," "fruity," "tea-like," "waxy," "burnt," and "herbal." Then, a similarity score was calculated, using the natural aroma of jasmine as a benchmark (defined as 10 points), to evaluate the overall closeness (0-10 points) between each sample's aroma and the natural aroma. All samples were presented in random order, and the average score was used to assess the similarity of each product's aroma profile to the natural raw material. The evaluation results are shown in Table 1.

[0116] Table 1

[0117]

[0118] Table 1 shows that Examples 1-4 (especially E4) exhibited high intensity in positive attributes such as fresh floral and sweet aromas, while achieving the best control over negative indicators such as waxy and herbal odors. The aroma profile was closest to that of fresh flowers (similarity 9.1-9.5), verifying the high-fidelity reproduction of the characteristic aroma of *Gymnocalycium mihanovichii* through the synergistic process of "enzymatic hydrolysis-SCC extraction-backfilling-embedding." E5, due to enhanced cell wall breakdown by hemicellulase, released aroma more fully but was slightly less pure; E6, due to amylase mismatch, showed slightly more off-odors. Comparative Examples 1-5, lacking components such as flower residue and deodorizing liquid, or the embedding step, had poor aroma integrity and significantly increased off-odors. Comparative Example 6, due to the high-temperature microwave treatment producing burnt and herbal odors, had the lowest similarity; Comparative Example 7, although prominent in floral aroma, lacked base backfilling during supercritical extraction, resulting in a thin aroma and an overall profile deviating from that of fresh flowers.

[0119] Test Example 2

[0120] Sensory evaluation of cigarette flavoring

[0121] The products obtained in Examples 1-6 and Comparative Examples 1-7 were diluted at an addition rate of 0.1% (w / w) and uniformly sprayed onto the same batch of blank tobacco shreds, then rolled into cigarettes. A tasting panel of nine evaluators evaluated the cigarette samples and blank control samples according to YC / T 497-2014 "Sensory Evaluation Method for Chinese-style Cigarette Style," assessing aroma quality, aroma quantity, harmony, irritation, dryness, and aftertaste. A 9-point scoring system was used (higher scores were better), and the average results were taken. The evaluation results immediately after rolling are shown in Table 2. The evaluation results after the cigarettes were equilibrated in a constant temperature and humidity chamber at 22°C and 60% humidity for three weeks are shown in Table 3.

[0122] Table 2

[0123]

[0124] Table 3

[0125]

[0126] The combined evaluation results in Tables 2 and 3 fully verify the synergistic effect of the present invention on improving the sensory quality and aroma stability of cigarettes. In the immediate evaluation (Table 2), E4, with its double encapsulation technology, leads with a score of 51.2, demonstrating excellent performance in aroma quality, harmony, and irritation. E1-E3, relying on the complete "enzymatic hydrolysis-SCC extraction-backfilling-encapsulation" process, follow closely behind, with scores all exceeding 49.5, significantly better than E5 (48.9) and E6 (46.9).

[0127] After three weeks of accelerated testing (Table 3), E4 showed the smallest score reduction (only 1.0 point), E1-E3 saw reductions controlled at 1.2-1.3 points, while E5 and E6 experienced larger reductions of 2.3-2.9 points. Comparative Examples 1-5, due to the lack of embedding or base components, showed reductions of 4.2-4.8 points; Comparative Examples 6 and 7, due to the lack of embedding protection and process defects, showed reductions as high as 6.7-6.9 points. After three weeks, their total scores were close to the blank control, thoroughly verifying the substantial progress of the synergistic process of this invention in maintaining the long-term sensory quality of cigarettes.

[0128] Test Example 3

[0129] Key aroma component retention analysis (GC-MS):

[0130] Equal amounts of the products from Examples 1, 4, 6, and 7 were placed in a 40°C constant temperature oven for accelerated stability testing. Samples were taken on day 0 and day 7, and the relative contents of five representative heat-sensitive and volatile aroma components were determined by GC-MS. The retention rates relative to the day 0 sample were calculated.

[0131] Sample pretreatment method: Accurately weigh a certain mass of sample, add a known concentration of phenethyl acetate (internal standard) dichloromethane solution, then dilute and bring to volume with chromatographic grade dichloromethane. After vortexing and sonication to completely dissolve and mix the sample, take the dichloromethane phase and filter it through a 0.22 μm organic microporous membrane. Transfer the resulting clear filtrate to a sample vial for GC-MS analysis, which will be used for subsequent qualitative and semi-quantitative analysis of volatile components.

[0132] Instrumental analysis conditions: Instrumental analysis was performed on a gas chromatography-mass spectrometry (GC-MS) system equipped with an autosampler. A DB-5MS capillary column (30 m × 0.25 mm × 0.25 μm) was used for separation. High-purity helium was used as the carrier gas at a constant flow rate of 1.0 mL / min. Samples were injected in split mode (split ratio 10:1), with the injection port temperature set at 250℃. Chromatographic separation employed a programmed temperature ramp: an initial temperature of 50℃ held for 2 minutes, followed by ramping at 5℃ / min to 150℃, and then at 10℃ / min to 280℃ held for 10 minutes. The eluent fractions were transferred to the mass spectrometer ion source via a 280℃ transfer line, where fragment ions were generated under 70 eV electron impact ionization. The mass spectrometry scan range was m / z 35-550, with the ion source and quadrupole temperatures set at 230℃ and 150℃, respectively. Data were acquired using full scan mode for qualitative and semi-quantitative analysis. The results are shown in Table 4.

[0133] Table 4

[0134]

[0135] As shown in Table 4, the retention rate of key aroma components of the orchid extract prepared by the method of the present invention is significantly higher than that of the comparative example under high temperature accelerated conditions. Among them, Example 4 with double encapsulation performed the best, indicating that the product of the present invention has excellent aroma persistence and stability.

[0136] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0137] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0138] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method for preparing tobacco aralia extract, characterized in that, The preparation method includes the following steps: (1) Mix the raw material of *Gymnocalycium mihanovichii* with water and pulp it, then mix it with an enzyme for enzymatic hydrolysis, and separate the solid and liquid to obtain flower pulp extract and flower residue; the enzyme is selected from any one or at least a combination of two of cellulase, pectinase, amylase and hemicellulase. (2) The flower extract obtained in step (1) is subjected to low-temperature distillation, and after condensation, it is separated to obtain the orchid fragrance dew and the defragrant solution; (3) Concentrate the orchid fragrance obtained in step (2) and separate the concentrated fragrance liquid and the defragrant clear liquid; (4) Combine the flower residue obtained in step (1), the deodorizing solution obtained in step (2), and the deodorizing liquid obtained in step (3), mix them with glycosidase for enzymatic hydrolysis, inactivate the enzyme, extract the product with alcohol, then separate the solid and liquid, concentrate the filtrate, and obtain the orchid flavor base extract. (5) The concentrated aroma liquid obtained in step (3) is added back to the orchid flavor base extract obtained in step (4), and then encapsulated and homogenized to obtain the orchid extract for tobacco.

2. The preparation method according to claim 1, characterized in that, The enzyme described in step (1) is a combination of cellulase and pectinase; Preferably, the mass ratio of cellulase to pectinase is 1:2-2:1; Preferably, the amount of enzyme added in step (1) is 0.05%-0.2% of the mass of the orchid raw material; Preferably, the enzymatic hydrolysis in step (1) is carried out at 30-50°C for 2-5 hours.

3. The preparation method according to claim 1 or 2, characterized in that, The raw material for the tree orchid is tree orchid flower; Preferably, the mass ratio of the orchid raw material to water is 1:(5-20).

4. The preparation method according to any one of claims 1-3, characterized in that, The low-temperature distillation in step (2) is carried out at a temperature not exceeding 60°C; Preferably, the low-temperature distillation is carried out using a rotary cone distillation column with a feed temperature of 40-60℃ and a vacuum degree of -85 kPa to -50 kPa.

5. The preparation method according to any one of claims 1-4, characterized in that, The concentration in step (3) is achieved by either freeze concentration or membrane concentration. Preferably, the concentration in step (3) is carried out at a temperature not exceeding 40°C; Preferably, the membrane concentration is a reverse osmosis membrane concentration; Preferably, the concentration ratio of the concentration in step (3) is (2-6):

1.

6. The preparation method according to any one of claims 1-5, characterized in that, The glycosidase mentioned in step (4) includes β-glucosidase; Preferably, the amount of glycosidase used is 0.3-0.5% of the enzymatic hydrolysis substrate; Preferably, the enzymatic hydrolysis process conditions in step (4) are: enzymatic hydrolysis at pH 4.5-5.5 and temperature 45-55℃ for 1-3 h; Preferably, the alcohol extraction in step (4) is performed by using a 20-70% aqueous ethanol solution for 0.5-2 hours; Preferably, the solids content of the orchid-flavored base extract in step (4) is 40-70%.

7. The preparation method according to any one of claims 1-6, characterized in that, The ratio of the concentrated aroma liquid to the orchid flavor base extract in step (5) is 1:(1-5) L / kg; Preferably, the embedding material in step (5) is cyclodextrin; Preferably, the process conditions for the embedding treatment in step (5) are: stirring at 45-55℃ for 0.5-1.5 h.

8. The preparation method according to any one of claims 1-7, characterized in that, After the embedding process described in step (5), a secondary embedding process is performed; Preferably, the material for secondary embedding is a liposome.

9. The tobacco extract prepared by any one of claims 1-8.

10. The application of the tobacco extract according to claim 9 in the preparation of flavoring materials for tobacco products.

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