Prickly pear refined extract, its preparation method and its use in the field of reconstituted tobacco.

Abstract

This invention provides a refined extract of prickly pear, its preparation method, and its application in reconstituted tobacco. The preparation method of the refined prickly pear extract includes the following steps: raw material pretreatment; extraction and purification; concentration; enzymatic hydrolysis of prickly pear extract; freeze-drying; adsorption purification; Maillard reaction: the supernatant of the enzymatic hydrolysis is added to an alkaline solution to adjust the pH to alkaline, stirred under water bath, and concentrated under reduced pressure to obtain a prickly pear alkali extract; a stabilizer, amino acids, and sugar are added sequentially to the prickly pear alkali extract, and the mixture is stirred and reacted under high temperature conditions. After the reaction is completed, the mixture is cooled to obtain the refined prickly pear extract. This invention uses prickly pear as raw material, and the enzymatic hydrolysis treatment effectively improves the flavor of the prickly pear extract. The product after the Maillard reaction has good quality, high clarity, and a prominent aroma. This refined prickly pear extract, when used in reconstituted tobacco, can, to a certain extent, mask the impurities of the tobacco itself, effectively improve the aroma and overall comfort, and add a specific aroma to tobacco products.

Description

Technical Field

[0001] This invention relates to tobacco technology, and more particularly to a refined extract of prickly pear, its preparation method, and its use in the field of reconstituted tobacco. Background Technology

[0002] Reconstituted tobacco is a recycled tobacco product made from tobacco waste (waste tobacco dust, broken tobacco leaves, tobacco stems, etc.) through processes such as extraction, concentration, separation, pulping, grinding, sheet forming, coating, and drying. Adding an appropriate amount of reconstituted tobacco to cigarettes can reduce the pressure on the use of high-quality tobacco raw materials, significantly reduce cigarette costs, and also help improve the intrinsic quality of cigarettes, reduce tar and harmful substances, and to a certain extent adjust and improve the physical properties and chemical composition of cigarettes.

[0003] prickly pear ( Rosa roxburghii Tratt Rosa chinensis is a perennial deciduous shrub belonging to the genus Rosa in the family Rosaceae. It is a natural plant used for both medicinal and edible purposes. Its fruit is rich in sugar, vitamins, carotene, organic acids, more than 20 kinds of amino acids, and more than 10 kinds of trace elements beneficial to the human body. It is mainly distributed in southwestern my country and is a unique wild resource of the Yunnan-Guizhou Plateau and the Panxi Plateau. Guizhou is the main producer, with the most varieties and the highest annual yield, followed by Yunnan. It is also found in the Liangshan area of ​​western Sichuan.

[0004] Prickly pear extract, due to its unique aroma characteristics, can better harmonize with tobacco aromas, enhancing the quality and quantity of fragrance, masking off-flavors, and improving the aftertaste. It is a safe and effective natural tobacco flavoring. It can effectively improve the aroma's texture and overall comfort and harmony. The Maillard reaction is a chemical reaction that, by reacting sugar compounds with amino acids or amino acid derivatives, can generate compounds with special aromas. Therefore, prickly pear Maillard reaction products typically possess various aromas, including fruity and floral notes.

[0005] Chinese patent literature discloses "A method for preparing natural prickly pear extract," application publication number CN115363153A. This invention uses purified water extraction, concentration, freeze-drying, or spray-drying processes to prepare prickly pear extract. However, the refrigeration time of the extract in this process is too long (24 hours), which is not conducive to continuous production and widespread application. In addition, the spray-drying process of the prickly pear extract is too high, resulting in the loss of effective components and the phenomenon of sticking to the wall, affecting the quality of the prickly pear extract. Chinese patent literature also discloses "Prickly pear extract and its preparation method, application, and low-temperature heating non-combustible product," application publication number CN114209086A. This invention carbonizes the prickly pear, then extracts it with ethanol, and obtains the extract by centrifugation and filtration. The low-temperature heating non-combustible product prepared from the obtained prickly pear extract loses the original flavor of the prickly pear after carbonization in this process. Chinese patent literature also discloses "Maillard products for enhancing tobacco aroma and reducing free radicals in tobacco leaves and their usage method", application publication number CN113397197A. This invention prepares Maillard reaction base liquid by using tobacco extract and prickly pear extract as raw materials, and optimizes Maillard reaction conditions to prepare aroma substances from the Maillard reaction base liquid. However, the initial pH of this process is 5-9 (too low pH is not conducive to the Maillard reaction) and the heating time is 3-7 hours (too long a reaction time will affect the flavor of the reactants). Summary of the Invention

[0006] The purpose of this invention is to address the problems of long extraction cycles and low efficiency in existing prickly pear extracts by proposing a method for preparing refined prickly pear extract through enzymatic hydrolysis. This method uses prickly pear as raw material, and the enzymatic hydrolysis effectively improves the flavor of the extract. The product after Maillard reaction exhibits high quality, clarity, and a prominent aroma. This refined prickly pear extract, when used in reconstituted tobacco, can mask the inherent impurities of tobacco to a certain extent, effectively improve aroma and overall comfort, and add a specific aroma to tobacco products.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is: a method for preparing refined extract of prickly pear by enzymatic hydrolysis, comprising the following steps:

[0008] (1) Raw material pretreatment: Remove impurities from fresh prickly pear fruit / dried prickly pear fruit and then crush the whole tissue. Soak the crushed dried prickly pear fruit in water containing antioxidants; after crushing fresh prickly pear fruit, mix it directly with antioxidants.

[0009] (2) Extraction and purification: The pretreated raw material is added to the solvent for extraction; after extraction, solid-liquid separation and liquid-phase vacuum filtration are performed to obtain prickly pear extract;

[0010] (3) Concentration: The prickly pear extract is mixed and concentrated under vacuum membrane to obtain prickly pear paste, and the solvent is recovered;

[0011] (4) Enzymatic hydrolysis of prickly pear extract: The enzyme preparation is completely dissolved in water to obtain an enzyme solution. After filtering the enzyme solution, the supernatant is added to the prickly pear extract for enzymatic hydrolysis.

[0012] (5) Freezing and standing: Seal the enzymatically hydrolyzed material and place it in a refrigerator to stand;

[0013] (6) Adsorption purification: After standing, take the supernatant and add adsorbent for purification to obtain enzymatic hydrolysis supernatant;

[0014] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant, add alkaline solution to adjust the pH to alkaline, stir under water bath, and concentrate under reduced pressure to a prickly pear alkaloid extract with a solid content of 65-70%; add stabilizer, amino acid and sugar to the prickly pear alkaloid extract in sequence, stir and react under high temperature, cool after the reaction is completed to obtain prickly pear refined extract (also known as prickly pear Maillard reaction product).

[0015] Furthermore, the prickly pear refers to fresh or dried prickly pear fruit, whether artificially cultivated or wild.

[0016] Furthermore, the prickly pear varieties mentioned in step (1) are seeded prickly pear (also known as prickly pear) and seedless prickly pear (golden prickly pear).

[0017] Furthermore, in step (1), the prickly pear raw material only needs to have impurities removed, while retaining the sepals, thorns, and seeds, and the whole tissue is extracted.

[0018] Further, the particle size of the pulverized dried prickly pear fruit in step (1) is 0.20~0.60 mm, preferably 0.25~0.60 mm.

[0019] Further, the mass ratio of the crushed dried prickly pear fruit to water containing antioxidants in step (1) is 1:1~3, preferably 1:1.5~2.

[0020] Furthermore, the immersion temperature in step (1) is 0~30℃ and the immersion time is 3~5 min.

[0021] Furthermore, the fresh prickly pear fruit mentioned in step (1) is crushed using a pulping machine to a degree that allows it to pass through a 40-mesh sieve.

[0022] Furthermore, the antioxidant mentioned in step (1) is one or a combination of phytic acid, propyl gallate, and sodium phytate. The addition of antioxidants can effectively reduce the loss of effective components of prickly pear (vitamins, SOD, flavonoids, etc.) during the extraction process.

[0023] Furthermore, the antioxidant addition ratio in the raw material after pulverization in step (1) is 0.01~0.10%, preferably 0.01~0.05%.

[0024] Further, in step (2), the mass ratio of the pretreated raw material to the solvent is 1:1 to 5, the extraction is performed twice, and the extraction solvent is water. Preferably, the mass ratio of the pretreated raw material to the solvent is 1:3.5 to 5, and more preferably, the mass ratio of the pretreated raw material to the solvent is 1:1.5 to 2.5.

[0025] Further, the extraction method described in step (2) is countercurrent extraction, high-pressure homogenization extraction or high-pressure microfluidic extraction.

[0026] Further, the extraction time in step (2) is 15~45 min, and the extraction temperature is 0~70℃. Preferably, the extraction time is 25~30 min, and the extraction temperature is 50~70℃.

[0027] Further, in step (2), the solvent is recovered for use as a secondary extraction solvent;

[0028] Furthermore, the filtration device in step (2) is a vacuum filter bag filter.

[0029] Further, in step (2), the vacuum degree of filtration is 0.03~0.05Mpa, and the filtration accuracy of the filter bag is 200~800 mesh. Preferably, the filtration accuracy of the filter bag is 200~400 mesh.

[0030] Further, in step (3), the vacuum film concentration temperature is 30~100℃, the vacuum degree is 0.01~0.1Mpa, and the solid content of the prickly pear extract is 65~70wt%. Preferably, the concentration temperature is 50~70℃ and the vacuum degree is 0.02~0.04Mpa.

[0031] Further, the enzyme preparation in step (4) is one or more of β-glucanase (100,000 U), galacturonase (60,000 U), sutilanase (50,000 U), and thioglycoside enzyme (2,000 U). The enzyme preparation is added in a ratio of 0.09~0.27 wt%.

[0032] Further, the enzyme preparation in step (4) comprises β-glucanosidase (100,000 U), galacturonase (60,000 U), sutilanase (50,000 U), and thioglycoside polysaccharide enzyme (2,000 U). The addition ratio of the β-glucanosidase (100,000 U) is 0.05~0.15 wt%, preferably 0.05~0.10 wt%. The addition ratio of the galacturonase (60,000 U) is 0.01~0.03 wt%, preferably 0.01~0.025 wt%. The addition ratio of the sutilanase (50,000 U) is 0.02~0.06 wt%, preferably 0.025~0.04 wt%. The addition ratio of the thioglycoside polysaccharide enzyme (2,000 U) is 0.01%~0.03% wt%, preferably 0.01~0.025 wt%.

[0033] Further, in step (4), the enzymatic hydrolysis temperature is 50~60℃ and the enzymatic hydrolysis time is 0.5~2.0h; preferably, the enzymatic hydrolysis temperature is 55~60℃ and the enzymatic hydrolysis time is 0.5~1.5h.

[0034] Furthermore, the enzyme preparation does not need to be inactivated after enzymatic hydrolysis in step (4).

[0035] Further, in step (5), the refrigerator temperature is -18 to -4°C, and the settling time is 30 to 45 minutes. Preferably, the refrigerator temperature is -18 to 0°C, and the settling time is 30 to 40 minutes.

[0036] Further, the adsorption and purification agent in step (6) is one or more of wood-based activated carbon, lily clay, paint mist flocculant, and adsorption resin. The addition ratio of the adsorption and purification agent is 0.2-0.5% of the upper liquid, and the adsorption and purification time is 10-20 min. Preferably, the addition ratio is 0.2-0.4% of the upper liquid, and the adsorption and purification time is 10-15 min.

[0037] Furthermore, the addition ratios of wood-based activated carbon, lily clay, paint mist flocculant and adsorption resin in step (6) are 20~25%, 40~45%, 25~30% and 10~20%, respectively.

[0038] Further, the alkaline solution in step (7) is an 8-12% KOH aqueous solution and / or an 8-12% NaOH aqueous solution, the alkaline solution addition ratio is 10-20%, and the pH value is 9-11. Preferably, the alkaline solution addition ratio is 12-18%, and the pH range is 10-11.

[0039] Further, the water bath temperature in step (7) is 40~60℃, and the stirring time is 10~30 min. Preferably, the water bath reaction temperature is 45~60℃, and the stirring time is 15~25 min.

[0040] Further, the stabilizer in step (7) is hyaluronic acid and / or triacetin, with an addition ratio of 60-80%, preferably 60-65%;

[0041] And / or, the amino acid is one or a combination of lysine, tryptophan, ornithine, and methionine, and is added in an amount of 0.66~1.43%;

[0042] And / or, the sugar is one or a combination of deoxyribose, threose, lysoose, erythrose and glyceraldehyde, and is added in an amount of 2.15 to 3.9%.

[0043] Further, the preferred amino acids are cyproheptadine, tryptophan, ornithine, and methionine, with addition ratios of 0.01~0.03%, 0.1~0.3%, 0.4~0.7%, and 0.15~0.4%, respectively. Preferably, the addition ratios of cyproheptadine, tryptophan, ornithine, and methionine are 0.01~0.25%, 0.15~0.25%, 0.4~0.65%, and 0.15~0.35%, respectively.

[0044] Further, the preferred sugars are deoxyribose, threose, lysose, erythrose, and glyceraldehyde, with addition ratios of 1.0-1.5%, 0.5-1.0%, 0.1-0.3%, 0.4-0.7%, and 0.15-0.4%, respectively. Preferably, the addition ratios of deoxyribose, threose, lysose, erythrose, and glyceraldehyde are 1.0-1.25%, 0.8-1.0%, 0.1-0.2%, 0.4-0.6%, and 0.15-0.3%, respectively.

[0045] Further, in step (7), the stirring reaction temperature is 90~110℃, the stirring reaction time is 2~4h, and the cooling temperature is 0~4℃. Preferably, the stirring reaction temperature is 95~110℃, the stirring reaction time is 2~3.5h, and the cooling temperature is 0℃.

[0046] Furthermore, the reaction temperature medium in step (7) is a water bath or an oil bath.

[0047] Another object of the present invention discloses a prickly pear Maillard reaction product prepared by the above method.

[0048] Another object of the present invention is to disclose the use of a prickly pear Maillard reaction product in the field of reconstituted tobacco.

[0049] Another object of the present invention discloses a reconstituted tobacco leaf comprising the above-mentioned prickly pear Maillard reaction product.

[0050] Furthermore, the amount of the prickly pear Maillard reaction product added to the reconstituted tobacco is 0.005~0.015wt%, preferably 0.005~0.010wt.

[0051] Furthermore, the reconstituted tobacco leaves are conventional cigarette reconstituted tobacco leaves or special reconstituted tobacco leaves for low-temperature heating and non-combustion.

[0052] Unless otherwise specified, all percentages in this invention refer to mass percentages.

[0053] The method for preparing the Maillard reaction product of prickly pear and its application in reconstituted tobacco, as described in this invention, has the following advantages compared with the prior art:

[0054] (1) The extraction medium for dried and fresh prickly pear fruit in this invention is water, which is convenient and environmentally friendly. At the same time, the prickly pear Maillard reaction products are prepared by combining bio-enzymatic hydrolysis technology with Maillard technology, which can fully explore the value of prickly pear and make it more valuable. The extracted residue can be used as raw material fiber for reconstituted tobacco substrate or can be treated and used as compost, which can avoid resource waste and environmental pollution problems.

[0055] (2) In this invention, the prickly pear extract after enzymatic hydrolysis is pre-cooled at -18~-4℃ before the Maillard reaction, which can inhibit enzyme activity. Then, the uninactivated enzyme can be inactivated through the Maillard reaction process. Therefore, the enzyme preparation does not need to be inactivated after enzymatic hydrolysis in this invention, which reduces the loss of effective ingredients and energy consumption caused by enzyme inactivation, thereby preparing a prickly pear Maillard reaction product with better quality.

[0056] (3) The prickly pear Maillard reaction product prepared by the present invention has good flowability (viscosity range between 80 and 85 cP), pure color and flavor, retains the original aroma of prickly pear, can enrich the aroma of tobacco, can effectively improve the aroma quality and overall comfort, make the aroma of cigarettes elegant, and make the mouth comfortable and clean, and has a good promoting effect on improving the quality of reconstituted tobacco leaves.

[0057] (4) The prickly pear Maillard reaction product prepared by the present invention only requires the removal of impurities from the prickly pear raw material. There is no need to remove the calyx, thorns and seeds. The whole tissue is extracted, which can significantly retain the original aroma of the prickly pear. It can be added to traditional cigarette reconstituted tobacco leaves or heat-not-burn cigarette reconstituted tobacco leaves at a ratio of 0.005~0.010wt%. Due to its good quality, only a very small amount is added to improve the aroma quality of the cigarette without increasing the strength and irritation. It is less harmful to the human body.

[0058] In summary, this invention uses prickly pear as raw material. The pulverization process effectively improves extraction efficiency, and the addition of antioxidants maximizes the retention of active ingredients during extraction. The Maillard reaction products prepared by this method are of high quality. Different processing methods are employed for different types of prickly pear raw material matrices, making the process more feasible and continuous. Furthermore, the combination of bio-enzyme technology and Maillard technology effectively removes large molecules detrimental to sensory quality, increases the content of small molecule volatile components, especially aroma components, while preserving the natural aroma of prickly pear. When added to tobacco, it improves the smoke to a certain extent, effectively enhancing the aroma and overall comfort, further harmonizing the internal chemical composition of cigarettes, and improving their quality. This not only opens up new avenues for enhancing the use value of prickly pear but also reduces the harm of tobacco to the human body. Detailed Implementation

[0059] The present invention will be further described below with reference to the embodiments: Example 1

[0060] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0061] (1) Raw material pretreatment: After removing impurities, artificially cultivated prickly pear dried fruit is crushed to a particle size of 0.25 mm, and water containing 0.01% phytic acid antioxidant is added at a mass ratio of 1:2 to soak and stir evenly.

[0062] (2) Extraction and purification: The soaked raw material was mixed with water at 60℃ at a mass ratio of 1:3.5 and extracted under high pressure at 60℃ for 30 min; after extraction, the extract residue was separated from the liquid; the liquid was vacuum filtered through a 200-mesh filter bag under a vacuum of 0.03 MPa to obtain prickly pear extract.

[0063] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 60℃ and 0.02Mpa to a solid content of 65% to obtain prickly pear paste.

[0064] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.05% β-glucanase (100,000 U), 0.01% galacturonase (60,000 U), 0.04% sutilanase (50,000 U), and 0.01% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 1 h in a 50℃ water bath.

[0065] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -4℃ refrigerator for 30 min.

[0066] (6) Adsorption and purification: After standing, 0.2% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 40%: 25%: 15%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 15 minutes. The supernatant is then taken for later use.

[0067] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 12% alkaline solution (10% KOH: 10% NaOH = 1:1) to adjust the pH to 10. Stir at 45℃ for 25 min. After the reaction, concentrate under reduced pressure to a prickly pear alkaloid extract with a solid content of 70%. Add 60% hyaluronic acid, 0.25% lysine, 0.25% tryptophan, 0.65% ornithine, 0.35% methionine, 1.25% deoxyribose, 1.0% threose, 0.2% lysose, 0.6% erythrose, and 0.3% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir the reaction in an oil bath at 110℃ for 3 h. After the reaction, cool with water at 0℃ to obtain the prickly pear Maillard reaction product. Example 2

[0068] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0069] (1) Raw material pretreatment: After removing impurities, the dried wild prickly pear fruit is crushed to a particle size of 0.35 mm, and water containing 0.01% phytic acid antioxidant is added at a mass ratio of 1:2 to soak and stir evenly.

[0070] (2) Extraction and purification: The soaked raw material is mixed with 70℃ water at a mass ratio of 1:3.5 and extracted countercurrently at 70℃ for 30 min; after extraction, the extract residue is separated from the liquid; the liquid is vacuum filtered through a 200-mesh filter bag under a vacuum degree of 0.03Mpa to obtain prickly pear extract.

[0071] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 70℃ and 0.04Mpa until the solid content was 65% to obtain prickly pear paste.

[0072] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.05% β-glucanase (100,000 U), 0.01% galacturonase (60,000 U), 0.04% sutilanase (50,000 U), and 0.01% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 1 h in a 50℃ water bath.

[0073] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -4℃ refrigerator for 30 min.

[0074] (6) Adsorption and purification: After standing, 0.2% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 40%: 25%: 15%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 15 minutes. The supernatant is then taken for later use.

[0075] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 12% alkaline solution (8% KOH: 12% NaOH = 1:1) to adjust the pH to 11. Stir at 60℃ for 15 min. After the reaction, concentrate under reduced pressure to a prickly pear alkaloid extract with a solid content of 70%. Add 60% hyaluronic acid, 0.25% lysine, 0.25% tryptophan, 0.65% ornithine, 0.35% methionine, 1.25% deoxyribose, 1.0% threose, 0.2% lysose, 0.6% erythrose, and 0.3% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir the reaction in an oil bath at 110℃ for 3 h. After the reaction, cool with water at 0℃ to obtain the prickly pear Maillard reaction product. Example 3

[0076] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0077] (1) Raw material pretreatment: After removing impurities, the artificially cultivated golden prickly pear dried fruit is crushed to a particle size of 0.40 mm, and water containing 0.01% propyl gallate antioxidant is added at a mass ratio of 1:1.5 and stirred evenly.

[0078] (2) Extraction and purification: The soaked raw material was mixed with 50℃ water at a mass ratio of 1:4 and extracted under high pressure at 55℃ for 30 min; after extraction, the extract residue was separated from the liquid; the liquid was vacuum filtered through a 200-mesh filter bag under a vacuum of 0.03Mpa to obtain prickly pear extract.

[0079] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 55℃ and 0.03Mpa until the solid content was 65% to obtain prickly pear paste.

[0080] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.08% β-glucanase (100,000 U), 0.012% galacturonase (60,000 U), 0.025% sutilanase (50,000 U), and 0.01% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 0.5 h in a 55℃ water bath.

[0081] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -4℃ refrigerator for 30 min.

[0082] (6) Adsorption and purification: After standing, 0.2% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 40%: 25%: 15%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 15 minutes. The supernatant is then taken for later use.

[0083] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 14% alkaline solution (9% KOH: 11% NaOH = 1:1) to adjust the pH to 10. Stir at 45℃ for 20 min. After the reaction, concentrate under reduced pressure to a prickly pear alkaloid extract with a solid content of 70%. Add 65% hyaluronic acid, 0.01% lysine, 0.15% tryptophan, 0.4% ornithine, 0.15% methionine, 1.0% deoxyribose, 0.8% threose, 0.1% lysose, 0.4% erythrose, and 0.15% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir and react in a 95℃ water bath for 3.5 h. After the reaction, cool at 4℃ to obtain the prickly pear Maillard reaction product. Example 4

[0084] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0085] (1) Raw material pretreatment: After removing impurities, the dried wild golden prickly pear fruit is crushed to a particle size of 0.60 mm. It is then soaked in water containing 0.02% propyl gallate antioxidant at a mass ratio of 1:1.5 and stirred evenly.

[0086] (2) Extraction and purification: The soaked raw material was mixed with water at 50℃ at a mass ratio of 1:5 and extracted under high pressure at 55℃ for 30 min; after extraction, the residue was separated from the liquid; the liquid was vacuum filtered through a 200-mesh filter bag under a vacuum of 0.03 MPa to obtain prickly pear extract.

[0087] (3) Concentration: The dried prickly pear extract was concentrated under vacuum thin film at 55℃ and 0.03Mpa until the solid content was 65% to obtain prickly pear paste.

[0088] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.08% β-glucanase (100,000 U), 0.025% galacturonase (60,000 U), 0.03% sutilanase (50,000 U), and 0.015% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 0.5 h in a 60℃ water bath.

[0089] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -4℃ refrigerator for 30 min.

[0090] (6) Adsorption and purification: After standing, 0.2% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 40%: 25%: 15%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 15 minutes. The supernatant is then taken for later use.

[0091] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 14% alkaline solution (11% KOH: 9% NaOH = 1:1) to adjust the pH to 10. Stir at 60℃ for 15 min. After the reaction, concentrate under reduced pressure to obtain a prickly pear alkaloid extract with a solid content of 70%. Add 65% hyaluronic acid, 0.01% lysine, 0.15% tryptophan, 0.4% ornithine, 0.15% methionine, 1.0% deoxyribose, 0.8% threose, 0.1% lysose, 0.4% erythrose, and 0.15% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir the reaction in a 100℃ water bath for 3 h. After the reaction, cool at 4℃ to obtain the prickly pear Maillard reaction product. Example 5

[0092] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0093] (1) Raw material pretreatment: After removing impurities, artificially cultivated prickly pear fruits are crushed and passed through a 40-mesh sieve. Phytic acid and propyl gallate antioxidants are added at a mass ratio of 0.05%, and the mixture is stirred evenly.

[0094] (2) Extraction and purification: The raw material was mixed with water at 50℃ at a mass ratio of 1:2.5 and extracted under high pressure microjet at 50℃ for 30 min; after extraction, the extract residue was separated from the liquid; the liquid was vacuum filtered through a 400-mesh filter bag under a vacuum of 0.05 MPa to obtain prickly pear extract.

[0095] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 50℃ and 0.02Mpa to a solid content of 70% to obtain prickly pear paste.

[0096] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.05% β-glucanase (100,000 U), 0.025% galacturonase (60,000 U), 0.04% sutilanase (50,000 U), and 0.025% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 1 h in a 50℃ water bath.

[0097] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -18℃ refrigerator for 25 minutes.

[0098] (6) Adsorption and purification: After standing, 0.4% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 45%: 25%: 10%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 10 minutes. The supernatant is then taken for later use.

[0099] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 18% alkaline solution (8% KOH: 12% NaOH = 1:1) to adjust the pH to 11. Stir at 45℃ for 15 min. After the reaction, concentrate under reduced pressure to obtain a prickly pear alkaloid extract with a solid content of 70%. Add 60% triacetin, 0.05% lysine, 0.20% tryptophan, 0.5% ornithine, 0.20% methionine, 1.2% deoxyribose, 0.9% threose, 0.15% lysose, 0.5% erythrose, and 0.20% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir the reaction in an oil bath at 110℃ for 2.5 h. After the reaction, cool with water at 0℃ to obtain the prickly pear Maillard reaction product. Example 6

[0100] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0101] (1) Raw material pretreatment: Wild prickly pear fresh fruit is pulverized after removing impurities and passed through a 40-mesh sieve. Phytic acid and propyl gallate antioxidant are added at a mass ratio of 0.05% and stirred evenly.

[0102] (2) Extraction and purification: The raw material is mixed with water at 60℃ at a mass ratio of 1:2.5 and extracted countercurrently at 60℃ for 30 min; after extraction, the extract residue is separated from the liquid; the liquid is vacuum filtered through a 400-mesh filter bag under a vacuum degree of 0.05Mpa to obtain prickly pear extract.

[0103] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 60℃ and 0.02Mpa to a solid content of 70% to obtain prickly pear paste.

[0104] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.05% β-glucanase (100,000 U), 0.025% galacturonase (60,000 U), 0.04% sutilanase (50,000 U), and 0.025% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 0.5 h in a 60℃ water bath.

[0105] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -18℃ refrigerator for 25 minutes.

[0106] (6) Adsorption and purification: After standing, 0.4% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 45%: 25%: 10%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 10 minutes. The supernatant is then taken for later use.

[0107] (7) Maillard reaction: The supernatant of the enzymatic hydrolysis was added to 18% alkaline solution (9% KOH: 11% NaOH = 1:1) to adjust the pH to 11. The reaction was stirred at 45℃ for 15 min. After the reaction was completed, the solution was concentrated under reduced pressure to a solid content of 70% prickly pear alkaloid extract. 65% triacetin, 0.05% lysine, 0.20% tryptophan, 0.5% ornithine, 0.20% methionine, 1.2% deoxyribose, 0.9% threose, 0.15% lysose, 0.5% erythrose, and 0.20% glyceraldehyde were added to the prickly pear alkaloid extract in sequence. The mixture was stirred in an oil bath at 110℃ for 2 h. After the reaction was completed, the mixture was cooled at 0℃ to obtain the prickly pear Maillard reaction product. Example 7

[0108] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0109] (1) Raw material pretreatment: After removing impurities, artificially cultivated golden prickly pear fresh fruit is crushed and passed through a 40-mesh sieve. Phytic acid and sodium phytate antioxidants are added at a mass ratio of 0.05% and stirred evenly.

[0110] (2) Extraction and purification: The raw material is mixed with water at 60℃ at a mass ratio of 1:2 and extracted under high pressure at 60℃ for 25 min; after extraction, the residue is separated from the liquid; the liquid is vacuum filtered through a 400-mesh filter bag under a vacuum of 0.05 MPa to obtain prickly pear extract.

[0111] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 60℃ and 0.02Mpa to a solid content of 70% to obtain prickly pear paste.

[0112] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.05% β-glucanase (100,000 U), 0.025% galacturonase (60,000 U), 0.04% sutilanase (50,000 U), and 0.025% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 0.5 h in a 60℃ water bath.

[0113] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -18℃ refrigerator for 25 min.

[0114] (6) Adsorption and purification: After standing, 0.4% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 45%: 25%: 10%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 10 minutes. The supernatant is then taken for later use.

[0115] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 16% alkaline solution (10% KOH: 10% NaOH = 1:1) to adjust the pH to 10. Stir at 50℃ for 20 min. After the reaction, concentrate under reduced pressure to a prickly pear alkaloid extract with a solid content of 70%. Add 65% triacetin, 0.10% lysine, 0.25% tryptophan, 0.60% ornithine, 0.30% methionine, 1.25% deoxyribose, 1.0% threose, 0.2% lysose, 0.40% erythrose, and 0.15% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir the reaction in an oil bath at 110℃ for 3 h. After the reaction, cool with water at 0℃ to obtain the prickly pear Maillard reaction product. Example 8

[0116] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, the specific steps of which are as follows:

[0117] (1) Raw material pretreatment: After removing impurities from the fresh wild golden prickly pear fruit, crush it and pass it through a 40-mesh sieve. Add sodium phytate and propyl gallate antioxidants at a mass ratio of 0.05% and stir evenly.

[0118] (2) Extraction and purification: The raw material was mixed with water at 55℃ at a mass ratio of 1:1.5 and extracted countercurrently at 55℃ for 25 min; after extraction, the residue was separated from the liquid; the liquid was vacuum filtered through a 400-mesh filter bag under a vacuum of 0.05 MPa to obtain prickly pear extract.

[0119] (3) Concentration: The prickly pear extract was concentrated under vacuum thin film at 60℃ and 0.04Mpa to a solid content of 60% to obtain prickly pear paste.

[0120] (4) Enzymatic hydrolysis of prickly pear extract: Take 0.05% β-glucanase (100,000 U), 0.025% galacturonase (60,000 U), 0.04% sutilanase (50,000 U), and 0.025% thiaminase (2,000 U) of prickly pear extract respectively, add water to completely dissolve to obtain enzyme solution, filter, take the supernatant of enzyme solution and add it to dry prickly pear extract, mix well, seal and hydrolyze for 1 h in a 55℃ water bath.

[0121] (5) Freezing and standing: After cooling the enzymatically hydrolyzed prickly pear paste, seal it and place it in a -18℃ refrigerator for 25 min;

[0122] (6) Adsorption and purification: After standing, 0.4% adsorption and purification agent (wood activated carbon: lily soil: paint mist flocculant: adsorption resin = 20%: 45%: 25%: 10%) is sealed with breathable gauze and added to the liquid in step (5) for adsorption and purification for 10 minutes. The supernatant is then taken for later use.

[0123] (7) Maillard reaction: Take the enzymatic hydrolysis supernatant and add 16% alkaline solution (11% KOH: 9% NaOH = 1:1) to adjust the pH to 10. Stir at 55℃ for 15 min. After the reaction, concentrate under reduced pressure to obtain a prickly pear alkaloid extract with a solid content of 70%. Add 60% triacetin, 0.10% lysine, 0.25% tryptophan, 0.50% ornithine, 0.20% methionine, 1.20% deoxyribose, 0.8% threose, 0.2% lysose, 0.40% erythrose, and 0.15% glyceraldehyde to the prickly pear alkaloid extract in sequence. Stir the reaction in an oil bath at 110℃ for 3 h. After the reaction, cool with water at 0℃ to obtain the prickly pear Maillard reaction product. Example 9

[0124] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 1 in that:

[0125] The 12% alkali solution ratio in step (7) is (10% KOH: 10% NaOH = 2:1), the pH is adjusted to 11, and the stabilizer is 60% triacetin; the reaction temperature is 100℃, the reaction medium is a water bath, and the stirring reaction time is 3.5h. Example 10

[0126] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 2 in that:

[0127] The 12% alkali solution ratio in step (7) is (8% KOH: 12% NaOH = 2:1), the pH is adjusted to 11, the stabilizer is 60% triacetin, the reaction temperature is 95℃, the reaction medium is a water bath, and the stirring reaction time is 3.5h. Example 11

[0128] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 3 in that:

[0129] The 12% alkali solution ratio in step (7) is (9% KOH: 11% NaOH = 2:1), the pH is adjusted to 11, the stabilizer is 65% triacetin, the reaction temperature is 105℃, the reaction medium is an oil bath, and the stirring reaction time is 2.5h. Example 12

[0130] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 4 in that:

[0131] The 12% alkali solution ratio in step (7) is (11% KOH: 9% NaOH = 2:1), the pH is adjusted to 11, the stabilizer is 65% triacetin, the reaction temperature is 105℃, the reaction medium is an oil bath, and the stirring reaction time is 2.5h. Example 13

[0132] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 5 in that:

[0133] The 16% alkali solution ratio in step (7) is (8% KOH: 12% NaOH = 2:1), the pH is adjusted to 10, and the stabilizer is 65% hyaluronic acid; the reaction temperature is 100℃, the reaction medium is a water bath, and the stirring reaction time is 2.5h. Example 14

[0134] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 6 in that:

[0135] The 16% alkali solution ratio in step (7) is (9% KOH: 11% NaOH = 2:1), the pH is adjusted to 10, and the stabilizer is 65% hyaluronic acid; the reaction temperature is 100℃, the reaction medium is a water bath, and the stirring reaction time is 2.5h. Example 15

[0136] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 7 in that:

[0137] The 14% alkali solution ratio in step (7) is (10% KOH: 10% NaOH = 2:1), the pH is adjusted to 11, and the stabilizer is 60% hyaluronic acid; the reaction temperature is 100℃, the reaction medium is a water bath, and the stirring reaction time is 3.5h. Example 16

[0138] This embodiment discloses a method for preparing the Maillard reaction product of prickly pear, which differs from Example 8 in that:

[0139] The 18% alkali solution ratio in step (7) is (11% KOH: 9% NaOH = 2:1), the pH is adjusted to 11, and the stabilizer is 65% hyaluronic acid; the reaction temperature is 95℃, the reaction medium is a water bath, and the stirring reaction time is 3.5h.

[0140] Examples 17-32

[0141] Examples 17-32 disclose various reconstituted tobacco leaves. The reconstituted tobacco leaves of Examples 17-32 respectively include the prickly pear Maillard reaction products of Examples 1-16, with an addition amount of 0.005-0.010%. The reconstituted tobacco leaves are used in conventional cigarette reconstituted tobacco leaves or heat-not-burn special reconstituted tobacco leaves, and sensory evaluation is carried out.

[0142] The results showed that the sensory evaluation of reconstituted tobacco leaves with the Maillard reaction product prepared according to this invention was superior to that of the control sample without the Maillard reaction product. Specifically:

[0143] (1) Conventional cigarette reconstituted tobacco leaves with the Maillard reaction product of prickly pear prepared in this invention have prominent prickly pear characteristic aroma, good coordination between prickly pear aroma and tobacco aroma, rich and full tobacco flavor, improved tobacco aftertaste, comfortable and clean mouth, and pure and delicate tobacco aroma.

[0144] (2) The reconstituted tobacco leaves prepared by adding the Maillard reaction product of prickly pear prepared in this invention exhibit good aroma richness, sweet and refreshing taste, smooth and mellow smoke, good consistency and stable quality.

[0145] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing the Maillard reaction product of prickly pear for reconstituted tobacco, characterized in that, Includes the following steps: (1) Raw material pretreatment: After removing impurities, the fresh prickly pear fruit / dried prickly pear fruit is completely crushed, and the crushed dried prickly pear fruit is soaked in water containing antioxidants; the fresh prickly pear fruit is directly mixed with antioxidants after crushing; the antioxidants are one or a combination of phytic acid, propyl gallate and sodium phytate. (2) Extraction and purification: The pretreated raw material is added to the solvent for extraction; after extraction, solid-liquid separation and liquid-phase vacuum filtration are performed to obtain prickly pear extract; (3) Concentration: The prickly pear extract is concentrated by vacuum membrane to obtain prickly pear paste, and the solvent is recovered; (4) Enzymatic hydrolysis of prickly pear extract: The enzyme preparation is completely dissolved in water to obtain an enzyme solution. After filtering the enzyme solution, the supernatant is added to the prickly pear extract for enzymatic hydrolysis. (5) Freezing and standing: The enzymatically hydrolyzed material is sealed and placed in a refrigerator for standing at a temperature of -18 to -4°C for 30 to 45 minutes. (6) Adsorption and purification: After standing, take the supernatant and add adsorption and purification agent to purify it to obtain enzymatic hydrolysis supernatant; (7) Maillard reaction: Take the enzymatic hydrolysis supernatant, add alkaline solution to adjust the pH to alkaline, stir under water bath, concentrate under reduced pressure to obtain prickly pear alkaloid extract with a solid content of 70%; add stabilizer, amino acid and sugar to prickly pear alkaloid extract in sequence, stir and cool after the reaction is completed to obtain prickly pear Maillard reaction product; The stabilizer is hyaluronic acid and / or triacetin; The amino acid is one or a combination of lysine, tryptophan, ornithine, and methionine.

2. The preparation method according to claim 1, characterized in that, The vacuum film concentration temperature in step (3) is 30~100℃, the vacuum degree is 0.01~0.1Mp a, and the solid content of the prickly pear extract is 65~70wt%.

3. The preparation method according to claim 1, characterized in that, The enzyme preparation in step (4) is one or a mixture of β-glucanase, galacturonase, sutilanase and thioglycoside enzyme.

4. The preparation method according to claim 1, characterized in that, The adsorption and purification agent in step (6) is one or more of wood-based activated carbon, lily soil, paint mist flocculant and adsorption resin, and the adsorption and purification time is 10~20 min.

5. A Maillard reaction product of prickly pear, prepared by the method described in any one of claims 1 to 4.

6. Use of the prickly pear Maillard reaction product of claim 5 in the field of reconstituted tobacco.

7. A reconstituted tobacco leaf, characterized in that, Includes the prickly pear Maillard reaction product as described in claim 6.