Chewing areca nut with green pericarp and white mouth based on polygonum hydropiper biological processing and segmented freeze-drying and preparation method thereof

Abstract

The application provides a green-skin white-mouth chewing areca nut based on biological processing of polygonum hydropiper and segmented freeze-drying and a preparation method thereof, and belongs to the technical field of deep processing of special chewing products. The green fruit of areca nut is mixed with polygonum hydropiper extract, and vacuum pulse processing is performed. The processed areca nut green fruit is pre-frozen, cut into pieces, de-nucleated, and vacuum freeze-dried to obtain de-nucleated attenuated areca nut. The application further injects flavoring substances into the pieces of the attenuated areca nut, and freeze-dries to obtain green-skin white-mouth chewing areca nut. The combined processing of biological processing of polygonum hydropiper and segmented freeze-drying can significantly reduce the chemical toxicity, physical damage and addiction risk of the obtained green-skin white-mouth chewing areca nut, and the safety and sensory quality are combined.

Description

Technical Field

[0001] This invention belongs to the field of deep processing technology of specialty chewing products, specifically involving green-skinned white-mouthed chewing areca nuts based on the biological processing and segmented freeze-drying of Polygonum hydropiper and their preparation method. Background Technology

[0002] Betel nut is a traditional chewing product. However, modern medical research has found that traditionally processed betel nut products are a significant risk factor for oral mucosal lesions and even oral cancer. Its harm stems from the synergistic effects of multiple processes and mechanisms: 1. Core Chemical Toxicity (Endogenous Risk): The alkaloids contained in betel nut fruit, such as arecoline and arecaidine, are the core chemical substances causing health hazards. These components have clear cytotoxicity and genotoxicity: they can induce an explosion of reactive oxygen species (ROS) within cells, causing DNA oxidative damage; interfere with the cell cycle, inducing abnormal proliferation; and affect the central nervous system through sympathomimetic effects, leading to strong addiction and causing users to develop tolerance, cravings, and withdrawal reactions. 2. Synergistic Physical-Chemical Damage (Process-Induced Risk): Traditional processing and usage methods amplify the above risks. First, the coarse fibers of betel nut continuously abrade the oral mucosa during chewing, causing physical micro-trauma. Second, the strong alkaline substances such as quicklime commonly added during processing cause direct chemical burns to the mucosa. Physical wounds provide a pathway for arecoline and strong alkalis to invade, creating a vicious cycle of "physical damage - chemical penetration - chronic inflammation - fibrosis / cancer." Long-term exposure can lead to oral submucosal fibrosis (OSMF), causing hardening of the mouth, difficulty opening the mouth, and significantly increasing the risk of cancer.

[0003] In traditional Chinese medicine, areca nut (Areca Semen) is a medicinal herb with a bitter and pungent taste, warm properties, and is associated with the stomach and large intestine meridians. It possesses insecticidal, digestive, qi-regulating, diuretic, and antimalarial effects. Therefore, there is an urgent need for an innovative technology that can reconstruct the entire safety chain from raw material processing and technology to the final product form, in order to overcome the food safety risks associated with traditional processes. Summary of the Invention

[0004] In order to solve the problems existing in the prior art, the purpose of this invention is to provide a method for reducing the content of areca alkaloids and crude fiber, which can significantly reduce the content of toxic alkaloids such as arecoline, reconstruct the structure of areca fiber, make the crude fiber loose and microporous, and significantly reduce the puncture force.

[0005] The present invention also aims to provide a green-skinned, white-mouthed chewed areca nut and its preparation method. Based on the combined processing of Polygonum hydropiper biological processing and segmented freeze-drying, the chemical toxicity, physical damage and addiction risk of the obtained green-skinned, white-mouthed chewed areca nut can be significantly reduced, while having both safety and sensory quality.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a method for reducing the content of areca alkaloids and crude fiber, comprising the following steps: Areca nut green fruit is mixed with an aqueous solution of Polygonum hydropiper extract and processed under vacuum pulse. The processed areca nut green fruit is then pre-frozen, cut into segments, pitted, and freeze-dried under vacuum to obtain detoxified areca nut.

[0007] Preferably, the mass ratio of areca nut fruit to Polygonum hydropiper extract is (8~10):(1~2), and the mass concentration of the aqueous solution of Polygonum hydropiper extract is 5~10%.

[0008] Preferably, the conditions for vacuum pulse processing include: temperature 38~42℃, pressure cycle 5~6 times; the conditions for each pressure cycle are: -0.08MPa for 10min, 0.1MPa for 5min, and 0.05MPa for 5min.

[0009] Preferably, the pre-freezing method includes: first air-drying and cooling until the surface temperature of the areca nut is ≤10℃, and then cooling until the surface temperature of the areca nut is ≤-35℃.

[0010] Preferably, the vacuum freeze-drying method includes: sublimation drying at 90 Pa and -5°C until the water content is 36-40%.

[0011] The present invention also provides a reduced-toxicity areca nut prepared by the above method.

[0012] The present invention also provides a method for preparing green-skinned, white-mouthed chewed areca nut, wherein flavoring substances are injected into the segments of the above-mentioned detoxified areca nut, and freeze-dried to obtain green-skinned, white-mouthed chewed areca nut.

[0013] Preferably, the flavor substance is a solid flavor element, and the injection amount of the solid flavor element is 0.49~0.51g / petal; the solid flavor element comprises the following raw materials in parts by weight: 60~70 parts of maltose, 10~15 parts of sorbitol, 5~10 parts of glycerol, 1~3 parts of sodium bicarbonate and 2~5 parts of Polygonum hydropiper volatile oil.

[0014] Preferably, the freeze-drying method includes drying at 70 Pa and -5 °C until the moisture content is 6.5-8.5%.

[0015] The present invention also provides a green-skinned white-mouthed chewable areca nut prepared by the above preparation method.

[0016] Compared with the prior art, the beneficial effects of the technical solution of the present invention are as follows: This invention utilizes a bioprocessing technique for Polygonum hydropiper, abandoning traditional chemical modification methods. This allows the flavonoids and sesquiterpenes in Polygonum hydropiper to penetrate into the areca nut tissue, scavenging free radicals and inhibiting oxidative stress. Combined with a 100% complete kernel removal process, the content of toxic alkaloids such as arecoline is significantly reduced from the source, resulting in a significant decrease in cytotoxicity and genotoxicity. Simultaneously, the low-temperature freeze-drying process reconstructs the areca nut fiber structure, making the coarse fibers loose and microporous, and significantly reducing puncture force. This achieves the effect of systematically reducing the chemical toxicity and physical damage of areca nut.

[0017] This invention uses the segments of a detoxified areca nut to prepare green-skinned, white-mouthed chewing areca nut. Neutral solid flavorings completely replace the traditional strong alkaline brine of quicklime, thoroughly eliminating the risk of chemical burns. Compared with traditional areca nuts and existing improved products, the prepared green-skinned, white-mouthed chewing areca nut: 1. Systemic reduction of health risks: (1) Significant reduction of chemical toxicity: Through thorough removal of the kernel and the introduction of antioxidant components from Polygonum hydropiper, the exposure level of risky substances such as arecoline and potential oxidative damage in the product are greatly reduced. (2) Fundamental avoidance of physical damage: The crisp and soft texture of "Baikou" reduces the abrasion of the oral mucosa during chewing to an extremely low level, breaking the key link of "physical-chemical synergistic damage". (3) Complete elimination of exogenous irritation: Solid flavoring replaces strong alkaline brine, completely eliminating the most direct source of chemical burns in traditional processes.

[0018] 2. Revolutionary improvement in product quality: (1) The concept of chewing betel nut with green peel and white mouth can be transformed into one with measurable color value (peel a) (2) Taste and flavor: The product has a soft and chewy texture, sweet and delicious taste, without the roughness, astringency and burning sensation of traditional products. At the same time, the product has the potential to soothe the qi, strengthen the spleen and eliminate dampness. Detailed Implementation

[0019] This invention provides a method for reducing the content of areca alkaloids and crude fiber, comprising the following steps: mixing areca green fruit with an aqueous solution of Polygonum hydropiper extract and processing it under vacuum pulse; pre-freezing, slicing, removing the pits from the processed areca green fruit, and freeze-drying under vacuum to obtain detoxified areca nut.

[0020] In this invention, the areca nut is preferably pre-treated; the pre-treatment steps include: treatment with 0.5% vitamin C color-protecting solution for 15 minutes, and enzyme inactivation by steaming at 100°C for 8 minutes.

[0021] In this invention, the mass ratio of areca nut fruit to Polygonum hydropiper extract is (8-10):(1-2), and the mass concentration of the aqueous solution of Polygonum hydropiper extract is 5-10%. The main active components of the Polygonum hydropiper extract of this invention are flavonoids and sesquiterpenes, possessing antioxidant and anti-inflammatory activities. As an optional embodiment, the preparation method of the Polygonum hydropiper extract of this invention includes: harvesting the whole Polygonum hydropiper herb (with leaves), removing mud and impurities, air-drying, and pulverizing to 20-40 mesh. Polygonum hydropiper powder is mixed with 70-80% ethanol solution at a material-to-liquid ratio of 1:20 (1g / 100mL), soaked for 30 minutes, and extracted by reflux at 75-80℃ for 2-3 hours. After filtration, the residue is extracted again with an equal volume of 70-80% ethanol solution 2-3 times. The filtrates are combined and concentrated under reduced pressure (60℃, -0.08MPa) until no alcohol odor is detected, yielding a brownish-red extract. The extract is vacuum dried at 60℃ and pulverized through an 80-mesh sieve to obtain crude Polygonum hydropiper extract powder (flavonoids ≥5%). In this invention, the concentration of the aqueous solution of Polygonum hydropiper extract is adjusted. Areca catechu fruit and the aqueous solution of Polygonum hydropiper extract are mixed and placed in a continuous vacuum pulse processing tank for vacuum pulse processing.

[0022] The vacuum pulse processing conditions described in this invention include: a temperature of 38~42℃ and 5~6 pressure cycles; the conditions for each pressure cycle are: -0.08MPa for 10 min, 0.1MPa for 5 min, and 0.05MPa for 5 min. The preferred temperature is 39℃, 40℃, or 41℃.

[0023] This invention abandons the traditional processing method using strongly alkaline lime brine and pioneers the use of Polygonum hydropiper extract as the core processing medium for areca nut. Polygonum hydropiper (also known as water pepper) is pungent and warm in nature, and enters the spleen, stomach, and large intestine meridians. It has functions such as dispelling wind and dampness, dispersing blood stasis and relieving pain, detoxifying and reducing swelling, and strengthening the spleen and resolving dampness. It is used to treat dysentery, gastroenteritis, diarrhea, rheumatic joint pain, traumatic swelling and pain, and functional uterine bleeding; externally, it is used for snake bites and eczema. Modern pharmacological studies have further confirmed that Polygonum hydropiper (also known as water pepper) has multiple biological activities, including antioxidant, anti-inflammatory, antibacterial, and anticancer effects. This invention introduces Polygonum hydropiper (also known as water pepper) extract, which can mitigate the irritation of areca nut. Polygonum hydropiper is warm and pungent, and has the effects of strengthening the spleen and resolving dampness, detoxifying and promoting blood circulation; areca nut is warm and bitter, and can eliminate stagnation and promote qi circulation. The combination of these two ingredients, according to Traditional Chinese Medicine theory, can synergistically act on the spleen, stomach, and large intestine in the middle jiao (middle burner). The "detoxifying" and "spleen-strengthening" properties of Polygonum hydropiper may help mitigate the harshness of areca nut, resolving its dampness and turbidity, thus giving the product a milder qi-regulating and dampness-resolving property. From a modern pharmacological perspective, the flavonoids and sesquiterpenes in Polygonum hydropiper extract possess strong antioxidant and anti-inflammatory activities. During the vacuum pulse processing, these components penetrate into the areca nut tissue, directly scavenging or inhibiting reactive oxygen species (ROS) induced by arecoline metabolism, reducing oxidative stress damage at the molecular level, and providing pre-treatment biochemical protection for the oral mucosa during subsequent consumption, thus exerting a detoxification effect. Simultaneously, the antibacterial effect of Polygonum hydropiper helps with hygiene control during processing. This invention, combined with continuous vacuum pulse processing equipment, also simplifies the processing procedure and ensures the consistency of active ingredients.

[0024] This invention freeze-dries and pits processed areca nuts, physically eliminating key risk sources. The processed areca nuts are first pre-frozen using a method including: air-drying and cooling until the surface temperature is ≤10℃, then further cooling until the surface temperature is ≤-35℃. The pre-frozen areca nuts are then sliced ​​and pitted. Preferably, the slicing involves placing the areca nuts in a constant temperature and humidity environment (10℃, 25% RH) and cutting them into two segments. The sliced ​​areca nuts are then subjected to vacuum freeze-drying using a method including: sublimation drying at 90Pa and -5℃ until the moisture content is 36-40%. Alternatively, the processed areca nuts are air-dried and cooled for 10-20 minutes using a continuous air-cooling machine until the surface temperature is ≤10℃, then rapidly frozen in a spiral freezer for 5-20 minutes until the surface temperature is ≤-35℃. The pre-frozen areca nuts are then processed using an intelligent vision-based pitting device to remove the pits (pitting rate ≥99.9%). Finally, they are dried in a continuous vacuum freeze dryer at a pressure of 90 Pa and a temperature of -5°C until the moisture content reaches 38% ± 2%, yielding pitted areca nut segments (reduced toxicity areca nuts).

[0025] This invention first involves low-temperature partial dehydration of the whole fruit to strengthen its structure, creating a "golden window" for non-destructive processing. Subsequently, during this optimal physical window, the fruit pit is precisely removed. The pit is a rich area for alkaloids such as arecoline; this step achieves proactive physical removal of key risk substances, significantly reducing the total alkaloid load of the product from the source. The drying process of this invention is completed under low-temperature vacuum, avoiding the generation of harmful substances (such as nitrosamines) that may be produced at high temperatures.

[0026] This invention also provides a reduced-toxicity areca nut prepared by the above method, which can significantly reduce the content of toxic alkaloids such as arecoline, significantly reduce cytotoxicity and genotoxicity, and at the same time reconstruct the areca nut fiber structure, making the coarse fibers loose and microporous, greatly reducing the puncture force, and achieving a systematic reduction in the chemical toxicity and physical damage of areca nut.

[0027] The present invention also provides a method for preparing green-skinned, white-mouthed chewed areca nut, wherein flavoring substances are injected into the segments of the above-mentioned detoxified areca nut, and freeze-dried to obtain green-skinned, white-mouthed chewed areca nut.

[0028] This invention involves injecting a flavoring substance into the interior of pitted areca nut segments. The flavoring substance is a solid flavoring agent, and the injection amount of the solid flavoring agent is 0.49~0.51g / segment, preferably 0.5g / segment. The solid flavoring agent of this invention preferably has a hardness ≥50N and a water content ≤10%, comprising the following raw materials in parts by weight: 60~70 parts maltose, 10~15 parts sorbitol, 5~10 parts glycerol, 1~3 parts sodium bicarbonate, and 2~5 parts Polygonum hydropiper volatile oil. Preferably, it comprises 65 parts maltose, 12 parts sorbitol, 8 parts glycerol, 2 parts sodium bicarbonate, and 3 parts Polygonum hydropiper volatile oil. As an optional implementation, the preparation method of Polygonum hydropiper volatile oil according to the present invention includes: harvesting the whole Polygonum hydropiper herb (with leaves), removing mud and impurities, chopping the fresh leaves, adding twice the amount of water, and soaking for 1 hour; obtaining Polygonum hydropiper volatile oil by supercritical CO2 extraction (conditions: pressure 30~45MPa, temperature 40℃, time 120~150min, CO2 flow rate 20L / h). Preferably, the Polygonum hydropiper volatile oil of the present invention can be selected as microencapsulated Polygonum hydropiper volatile oil, that is, obtaining Polygonum hydropiper volatile oil microcapsules by using commercially available or standardized microcapsule encapsulation methods. The solid flavoring agent of the present invention is prepared by mixing the raw materials in proportion and injecting them into the interior of the pitted areca nut petals, wherein the injection is preferably performed using microinjection technology.

[0029] This invention abandons the traditional, highly corrosive liquid lime brine and innovatively prepares a solid flavor enhancer composed of maltose, a very low amount of food-grade sodium bicarbonate (providing a mild alkaline sensation), and the natural flavor compound Polygonum hydropiper volatile oil. This enhancer is then positioned inside the fruit segments using microinjection technology, achieving controllable alkalinity and slow-release flavor, eliminating the risks of chemical burns and "return to brine." This invention can optionally employ a commercially available food-grade microinjector combined with a high-precision metering screw to ensure the stability of the flavor enhancer injection volume and position.

[0030] This invention involves freeze-drying areca nut segments infused with flavor compounds. The freeze-drying method includes drying at 70 Pa and -5°C until the moisture content is 6.5-8.5%, preferably 7%, 7.5%, or 8%. This invention employs the analytical drying section of a continuous vacuum freeze dryer to ensure the moisture content meets the standard. The endpoint can be determined by a pressure rise test (ΔP / Δt < 1.0 Pa / min). After freeze-drying, this invention yields green-skinned, white-mouthed chewable areca nut.

[0031] More preferably, the present invention involves using electrostatic atomization spraying to enhance the aroma of freeze-dried areca nut segments through a tunnel. Preferably, the electrostatic atomization aroma enhancement method of the present invention includes: passing the freeze-dried areca nut segments through an electrostatic atomization spraying tunnel to spray a small amount of Polygonum hydropiper volatile oil (≤0.1%) to enhance the flavor.

[0032] This invention proposes a three-pronged approach to areca nut preparation: "introducing active ingredients from Polygonum hydropiper for biochemical protection and modification → eliminating key risk substances through continuous freeze-drying and pitting → eliminating exogenous chemical and physical damage through solid-state flavoring and crumbly texture." This approach not only produces products with superior sensory qualities but also constructs a complete system for enhancing safety and imparting health attributes, solving the problem of serious health hazards in areca nut products prepared using traditional processes.

[0033] This invention also provides a green-skinned, white-mouthed chewable areca nut prepared by the above-mentioned method. The "green skin" color retention in the "green skin" of this invention is attributed to the inhibition of enzymatic browning by the antioxidant components of Polygonum hydropiper and the low-temperature freeze-drying process, reflecting the product's low oxidative stress level. The "white mouth" refers to the clean white, soft, and crumbly texture of the internal fibers, which originates from the biomodification of Polygonum hydropiper components and the microporous structure formed by segmented drying. This texture significantly reduces physical friction and damage to the oral mucosa during chewing, directly solving the problem of physical wear caused by the coarse and hard fibers of traditional areca nuts.

[0034] The structural characteristics of the green-skinned, white-mouthed chewable areca nut prepared by this invention are: it does not contain areca kernels and has a solid flavor element deposition core inside; its textural characteristics are: puncture force 15-18N (detected by a texture analyzer); its chemical characteristics are: pH value 6.0-7.5, arecoline content ≤0.1%; and its sensory characteristics are: the peel is bright green (color intensity a). (Negative), with white internal fibers and a soft yet chewy texture.

[0035] The technical solutions of this invention will be clearly and completely described below with reference to the embodiments thereof. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0036] In a specific embodiment of the present invention, the preparation method of Polygonum hydropiper extract is as follows: The whole Polygonum hydropiper herb (with leaves) is harvested, sand and impurities are removed, and it is dried in the shade and pulverized to 30 mesh. The Polygonum hydropiper powder is mixed with a 75% ethanol solution at a material-to-liquid ratio of 1:20 (1g / 100mL), soaked for 30 minutes, and extracted by reflux at 80℃ for 2 hours. The mixture is filtered, and the residue is extracted three times with an equal volume of 75% ethanol solution. The filtrates are combined and concentrated under reduced pressure (60℃, -0.08MPa) until no alcohol odor remains, yielding a brownish-red extract. The extract is vacuum dried at 60℃ and pulverized through an 80-mesh sieve to obtain crude Polygonum hydropiper extract powder (flavonoids ≥5%).

[0037] The preparation method of Polygonum hydropiper volatile oil is as follows: harvest the whole Polygonum hydropiper (with leaves), remove mud and impurities, chop the fresh leaves, add 2 times the amount of water, and soak for 1 hour; use supercritical CO2 extraction (conditions: pressure 40MPa, temperature 40℃, time 130min, CO2 flow rate 20L / h) to obtain Polygonum hydropiper volatile oil.

[0038] Unless otherwise specified, the following embodiments are all conventional methods.

[0039] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0040] Example 1 A method to reduce the content of areca alkaloids and crude fiber: (1) The areca nut green fruit was soaked in 0.5% vitamin C color-protecting solution for 15 min and steamed at 100℃ for 8 min to inactivate the enzyme; the areca nut green fruit and the Polygonum hydropiper extract aqueous solution (mass concentration 10%) were placed in a continuous vacuum pulse processing tank at a mass ratio of 10:1, the temperature was set at 40℃, and the pressure was cyclical (-0.08MPa 10 min → 0.1MPa 5 min → 0.05MPa 5 min) for 6 cycles to obtain the processed areca nut green fruit.

[0041] (2) The processed areca nuts were air-dried and cooled for 15 minutes using a continuous air-cooling machine until the surface temperature of the areca nuts reached 10°C. Then, they were quick-frozen in a spiral freezer for 10 minutes until the surface temperature of the areca nuts reached -35°C. Subsequently, the pre-frozen areca nuts were processed using an intelligent vision pitting device. The 3D vision camera scanned the shape of the fruit and the position of the pit, and a six-axis industrial robot was guided to grasp the fruit. At the same time, a vacuum pitting device was activated to remove the pit at the moment of cutting (pitting rate ≥99.9%). Finally, the areca nuts were dried in a continuous vacuum freeze dryer at a chamber pressure of 90Pa and -5°C until the moisture content reached 38%±2%, resulting in pitted areca nut segments (reduced toxicity areca nuts).

[0042] Experimental Example 1 The attenuated areca nut prepared in Example 1 was subjected to safety testing, and a commercially available areca nut (brand name: Hecheng Tianxia Jinshi Zhijiao) was set up as a commercially available control group: 1. Determination of total alkaloid content (HPLC method) (1) Standard curve preparation: Accurately weigh arecoline and arecoline standards, prepare a 1.0 mg / mL mixed standard stock solution, and serially dilute to 20, 40, 60, 80, and 100 μg / mL standard working solutions; set the chromatographic parameters as follows: mobile phase acetonitrile:water = 20:80, flow rate 1.0 mL / min, detection wavelength 215 nm, column temperature 30 ℃, and injection volume 20 μL; after detection, draw the standard curve and the regression equation correlation coefficient R. 2 ≥0.999.

[0043] (2) Sample pretreatment: Weigh 5g of each group of samples accurately, cut them into pieces, add 50mL of 70% ethanol, extract by ultrasonication at 300W and 40kHz for 30min, filter the residue; repeat the extraction twice, combine the filtrates, concentrate to 10mL under reduced pressure at 45℃, and filter through a 0.22μm organic filter membrane for testing.

[0044] (3) On-machine testing and calculation: Three parallel samples were used for each group, and each sample was injected twice. The total content of arecoline and arecoline was calculated according to the standard curve. The results are as follows.

[0045] Table 1 Comparison of alkaloid content in areca nuts of different groups (mg / g, n=3)

[0046] The results showed that the contents of arecoline, arecoline, and total alkaloids in the sample of Example 1 were significantly lower than those in the commercially available control group (P<0.05), indicating that the detoxification process can effectively reduce the content of characteristic toxic alkaloids of areca nut.

[0047] 2. In vitro cytotoxicity detection (1) Cell culture: HOK cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO2, and saturated humidity. The medium was changed every 24 hours, and the cells were passaged at a ratio of 1:3 during the logarithmic growth phase.

[0048] (2) Preparation of sample extract: Each group of samples was added to sterile physiological saline at a material-to-liquid ratio of 1:10, and soaked in the solution at 37°C and 120r / min for 24 hours. The solution was then centrifuged at 3000r / min for 10 minutes and filtered through a 0.22μm sterile filter membrane to prepare the extract.

[0049] (3) Cell plating and drug administration: Adjust cell concentration to 1×10 5 Cells / mL, 100 μL of cell suspension was seeded in each well of a 96-well plate, with 6 replicates per group. After 24 h of culture and cell adhesion, the corresponding sample extract was added and cultured for another 24 h.

[0050] (4) CCK-8 cell viability detection: Add 10 μL of CCK-8 reagent to each well, incubate at 37°C in the dark for 2 h, measure the OD value at 450 nm wavelength, and calculate the cell viability.

[0051] (5) ROS reactive oxygen species detection: cells were incubated with DCFH-DA working solution diluted 1:1000 for 20 min, washed with PBS and the fluorescence intensity was detected to calculate the relative ROS content.

[0052] Table 2. Comparison of in vitro cytotoxicity of areca nuts in different groups (n=6)

[0053] The results showed that, compared with the blank group, both groups of areca nut extracts had a certain degree of toxicity to HOK cells; the cell survival rate of the Example 1 group was significantly higher than that of the commercially available control group, and the relative ROS content was significantly lower than that of the commercially available control group (P<0.05), indicating that the damage and oxidative stress of the reduced-toxic areca nut to oral cells were significantly reduced.

[0054] 3. Texture index (puncture force) testing Fifteen whole fruits / pulp pieces were randomly selected from each group and divided into three biological parallel groups, with five samples in each group. Samples were equilibrated at room temperature (25℃) and 50% humidity for 30 minutes to remove surface impurities and ensure the samples remained intact and undamaged. The central fleshy part of the areca nut was selected as the puncture area to ensure consistent sampling locations across groups and eliminate morphological errors.

[0055] Texture analyzer parameter settings: Probe type: P / 2 stainless steel cylindrical probe (diameter 2mm); Test mode: Puncture mode; Pre-test speed: 1.0 mm / s; Test speed: 1.0 mm / s; Post-test speed: 1.0 mm / s; Puncture depth: 8 mm; Trigger force: 5 g.

[0056] After a single sample is tested, the probe is cleaned and the next sample is replaced; the test is repeated for each group of samples, and the maximum puncture force is recorded. The results are as follows.

[0057] Table 3. Puncture force test results of areca nuts in each group

[0058] The results showed that the puncture force of the sample in Example 1 was significantly lower than that of the commercially available control group (P<0.05), the hardness of the pulp was greatly reduced, and the chewing texture was softer and more comfortable; the sample structure was intact, and there were no adverse phenomena such as softness, rot, or crumbling, indicating excellent textural quality.

[0059] Example 2 Preparation of a green-skinned, white-mouthed areca nut for chewing: (1) Same as Example 1.

[0060] (2) Same as Example 1.

[0061] (3) Preparation of solid flavoring agent (by mass): 65 parts maltose, 12 parts sorbitol, 8 parts glycerol, 2 parts sodium bicarbonate and 2 parts volatile oil of Polygonum hydropiper. Mix all raw materials evenly in proportion to obtain solid flavoring agent.

[0062] (4) The pitted areca nut segments are fed into the automatic injection machine via a conveyor belt. The injection volume is set to 0.50±0.01g / segment, and the injection position is the center inside the areca nut segment.

[0063] (5) Freeze-dry the areca nut segments with inlaid flavor elements: dry at 70 Pa and -5 ℃ until the moisture content is 7% to obtain green peel white mouth chewing areca nut.

[0064] Example 3 Preparation of green-skinned, white-mouthed areca nut for chewing: Steps (1) to (5) are the same as in Example 2.

[0065] (6) Electrostatic atomization for aroma enhancement: The freeze-dried areca nut petals are sprayed with a small amount of Polygonum hydropiper volatile oil (≤0.1%) through an electrostatic atomization spray tunnel to enhance the flavor.

[0066] Example 4 Preparation of green-skinned, white-mouthed areca nut for chewing: Steps (1) to (5) are the same as in Example 2.

[0067] (6) Adopt intelligent vision sorting equipment and automatic nitrogen filling packaging machine to improve product consistency and shelf life.

[0068] Intelligent sorting: Using X-ray metal detection and machine vision sorting, foreign objects and defective products (such as mismatched colors or irregular shapes) are removed.

[0069] Nitrogen-filled packaging: Qualified products are filled with nitrogen (residual oxygen ≤1.0%) through an automatic nitrogen-filled packaging machine. Food-grade aluminum foil bags are used for packaging to ensure a shelf life of ≥12 months.

[0070] Experimental Example 2 The green-skinned white-mouthed chewing areca nut prepared in Example 2 was subjected to safety testing. A commercially available areca nut (brand name: Hecheng Tianxia Jinshi Zhijiao) was set up as a commercially available control group. 1. Determination of total alkaloid content (HPLC method) The detection method was the same as in Experiment 1, and the results are as follows.

[0071] Table 4 Comparison of alkaloid content in areca nuts of different groups (mg / g, n=3)

[0072] The results showed that the contents of arecoline, arecoline and total alkaloids in the sample of Example 2 were significantly lower than those in the commercially available control group (P<0.05), and the contents of characteristic toxic alkaloids of areca were effectively reduced.

[0073] 2. In vitro cytotoxicity detection The detection method was the same as in Experiment 1, and the results are as follows.

[0074] Table 5. Comparison of in vitro cytotoxicity of areca nuts in different groups (n=6)

[0075] The results showed that, compared with the blank group, both groups of areca nut extracts had a certain degree of toxicity to HOK cells; the cell survival rate of the Example 2 group was significantly higher than that of the commercially available control group, and the relative ROS content was significantly lower than that of the commercially available control group (P<0.05), indicating that the damage to oral cells and oxidative stress caused by chewing areca nut with green peel and white mouth of the present invention were significantly reduced.

[0076] 3. Texture index (puncture force) testing The detection method was the same as in Experiment 1, and the results are as follows.

[0077] Table 6. Puncture force test results of areca nuts in each group

[0078] The results showed that the puncture force of the sample in Example 2 was significantly lower than that of the commercially available control group (P<0.05), the hardness of the pulp was greatly reduced, and the chewing texture was softer and more comfortable; the sample structure was intact, and there were no adverse phenomena such as softness, rot, or crumbling, indicating excellent textural quality.

[0079] 4. Oral mucosal physical friction damage test Eighteen healthy New Zealand white rabbits, half male and half female, weighing 2.0-2.5 kg, were selected and acclimatized for 7 days before the experiment was conducted. They were randomly divided into three groups of six rabbits each: blank control group, Example 2 group, and commercially available control group.

[0080] The experiment lasted 14 days, with the procedure performed twice daily, once in the morning and once in the afternoon. The blank control group received no friction treatment and was fed normally. In the drug treatment group, an equal amount of areca nut sample was used to repeatedly rub the inner mucosa of one side of the rabbit's mouth at a uniform speed. Each rubbing session lasted 1 minute, and the movements were gentle and consistent. After the experiment, the appearance, color, presence of congestion, edema, damage, and erosion of the oral mucosa of the rabbits in each group were observed macroscopically. It was found that the mucosa structure of the blank control group was intact and without any physical damage, while the mucosa of the control group was congested, damaged, and showed extensive inflammatory infiltration. The mucosa structure of the second example group was intact and without any physical damage, showing no significant difference from the blank control group.

[0081] 5. Conduct oral mucosal chemical irritation tests on each group: (1) pH detection: each group of areca nut samples were added to deionized water at a material-to-liquid ratio of 1:10 (g / mL) and soaked at room temperature for 2 hours, with stirring once every 30 minutes during the soaking. The supernatant was taken and the pH value of the solution was directly measured using a calibrated portable pH meter. Each group was measured 6 times in parallel and the average value was recorded.

[0082] (2) Rabbit oral mucosa patch stimulation test: Eighteen healthy New Zealand white rabbits, half male and half female, weighing 2.0-2.5 kg, were selected and acclimatized for 7 days before the experiment was conducted. They were randomly divided into three groups of six rabbits each: blank control group, Example 2 group, and commercially available control group.

[0083] Uniformly sized areca nut pulp slices were tightly applied to the intact mucosa inside the rabbit's oral cavity for 4 hours at a time, once a day for 7 consecutive days; sterile filter paper was applied as a control group.

[0084] Observe the mucosal condition daily and score it according to the mucosal irritation scoring standard. The higher the total score, the stronger the irritation. The mucosal irritation scoring standard (total score of 10 points) is as follows: normal mucosa, no abnormalities, 0 points; slight redness, no edema or damage, 1-2 points; obvious congestion and mild edema, 3-4 points; obvious congestion and edema, punctate lesions, 5-6 points; large area of ​​erosion and exudation, 7-8 points; severe ulceration and tissue necrosis, 9-10 points.

[0085] Table 7 Results of the oral mucosal chemical irritation test of areca nuts in each group

[0086] The results showed that the commercially available control group's soaking solution had a low pH, was weakly acidic, and had a high mucosal irritation score, indicating significant chemical irritation. The soaking solution in Example 2 had a near-neutral pH, a mucosal irritation score close to the blank group, and extremely weak chemical irritation. Combined with the friction test results, it can be concluded that the segmented freeze-dried modified areca nut fiber of this invention is soft and crumbly, thoroughly reducing physical wear from chewing while eliminating the risk of chemical burns and mucosal irritation.

[0087] 6. Addiction-conditioned position preference (CPP) test Experimental animals: 24 healthy ICR mice, half male and half female, weighing 18-22g, were acclimatized for 7 days and randomly divided into 2 groups of 12 mice each: Example 2 group and control group. Experimental apparatus: Classic three-chamber CPP preference chamber (the middle is the transition zone, and the left and right are the condition zones, with light / texture distinction between the two zones).

[0088] Pre-acclimatization period (days 1-2): Remove the partitions inside the box and allow the mice to move freely in the three areas. Each mouse acclimatizes for 30 minutes. Record the time the mice spend in the left and right condition areas and remove individuals with innate area preferences.

[0089] Modeling and drug administration period (days 3-14, total 12 days): The mice were given drug treatment at fixed times every day by gavage. The areca nut sample extract was added to deionized water at a ratio of 1:10 (g / mL) and soaked at room temperature for 2 hours, with stirring once every 30 minutes during the process; the supernatant was collected. The mice were then placed in the corresponding condition area and isolated for 30 minutes. The treatment of the two groups of mice was consistent.

[0090] Test period (day 15): The partition was removed, and the mice were allowed to move freely for 30 minutes. The duration of the mice in the two condition zones was recorded.

[0091] Preference Index = (Time spent in the area corresponding to drug administration / Total test time) × 100%; Judgment criteria: The higher the preference index, the more the mouse shows a location preference for the sample and the stronger the tendency to become addicted / centrally dependent.

[0092] Table 8 Results of the Conditioned Position Preference Test for Addiction to Areca Nut in Each Group

[0093] The results showed that mice in Example 2 had a low preference index, normal frequency of spontaneous activity, and no addictive behavior; the commercially available control group had a high preference index, significant central nervous system excitation, and a mild tendency to become addicted. This indicates that the product of this invention has a significantly reduced risk of addiction and no obvious central nervous system dependence.

[0094] 7. Sensory indicators and safety indicators Example 2: Areca nut peel is bright green (color a) =-1.5±0.5), the internal fibers are white (white mouth), the taste is soft and chewy, without coarseness, astringency and burning sensation; there is no excessive heavy metal (lead ≤0.5mg / kg, cadmium ≤0.1mg / kg), and no pathogenic bacteria (Salmonella and Staphylococcus aureus were not detected).

[0095] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for reducing the content of areca alkaloids and crude fiber, characterized in that, Includes the following steps: Areca nut green fruit is mixed with an aqueous solution of Polygonum hydropiper extract and processed under vacuum pulse. The processed areca nut green fruit is then pre-frozen, cut into segments, pitted, and freeze-dried under vacuum to obtain detoxified areca nut.

2. The method according to claim 1, characterized in that, The mass ratio of areca nut extract to Polygonum hydropiper extract is (8~10):(1~2), and the mass concentration of the aqueous solution of Polygonum hydropiper extract is 5~10%.

3. The method according to claim 1, characterized in that, The conditions for vacuum pulse processing include: temperature 38~42℃, pressure cycle 5~6 times; the conditions for each pressure cycle are: -0.08MPa for 10min, 0.1MPa for 5min, and 0.05MPa for 5min.

4. The method according to claim 1, characterized in that, The pre-freezing method includes: first air-drying and cooling until the surface temperature of the areca nut is ≤10℃, and then cooling until the surface temperature of the areca nut is ≤-35℃.

5. The method according to claim 1, characterized in that, The vacuum freeze-drying method includes: sublimation drying at 90 Pa and -5 °C until the water content is 36-40%.

6. The detoxified areca nut prepared by the method according to any one of claims 1 to 5.

7. A method for preparing green-skinned, white-mouthed areca nut for chewing, characterized in that, Flavoring substances are injected into the segments of the detoxified areca nut as described in claim 6, and then freeze-dried to obtain green-skinned, white-mouthed chewable areca nut.

8. The preparation method according to claim 7, characterized in that, The flavor substance is a solid flavor element, and the injection amount of the solid flavor element is 0.49~0.51g / petal; the solid flavor element includes the following raw materials in parts by weight: 60~70 parts of maltose, 10~15 parts of sorbitol, 5~10 parts of glycerol, 1~3 parts of sodium bicarbonate and 2~5 parts of Polygonum hydropiper volatile oil.

9. The preparation method according to claim 7, characterized in that, The freeze-drying method includes drying at 70 Pa and -5 °C until the moisture content is 6.5-8.5%.

10. The green-skinned white-mouthed chewing areca nut prepared by the preparation method according to any one of claims 7 to 9.

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