Efficient extraction process of yellow bud soup

Abstract

The invention provides a high-efficiency extraction process of yellow bud soup, which is based on a manganese-doped carbon nitride nano-enzyme mediated plant cell wall directional perforation technology and is used for high-efficiency extraction of the yellow bud soup. The manganese-doped carbon nitride nano-enzyme has peroxidase-like (POD) activity, galacturonic acid hexasaccharide is grafted on the surface of the nano-enzyme, the nano-enzyme can be specifically combined with cell wall pectin, targeted positioning is achieved, the cell wall structure is destroyed through a biomimetic catalytic reaction, active ingredients are promoted to be dissolved out, and the extraction efficiency is improved. The manganese-doped carbon nitride nano-enzyme provided by the invention keeps high enzyme activity in a wide temperature range of 40-70 DEG C, the enzyme activity is kept by 80% or above at 70 DEG C, and the enzyme activity still reaches 85% after the manganese-doped carbon nitride nano-enzyme is stored at normal temperature for 6 months. The manganese-doped carbon nitride nano-enzyme is used for efficiently extracting the yellow bud soup, so that the content of active ingredients such as total polysaccharides, total flavonoids and ginsenoside in the yellow bud soup is remarkably increased, meanwhile, heat-sensitive ingredients are effectively protected, and the retention rate of 6-shogaol and the retention rate of poria cocos oligosaccharide are increased by 0.88-2.13 times and 0.5-1.45 times respectively.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a highly efficient extraction process for Huangya Decoction. Background Technology

[0002] Huangya Decoction is a traditional Chinese medicine formula, a basic formula created to strengthen the spleen and warm the earth element, and regulate the middle qi. It originates from *Sisheng Xinyuan* (Four Sacred Sources of the Heart), Volume Four, "Explanation of Labor Injury," by the renowned Qing Dynasty physician Huang Yuanyu. The book states: "The stomach governs the descending of turbid qi, and the spleen governs the ascending of clear qi. When dampness occurs, the middle qi cannot circulate, the ascending and descending functions reversed, clear yang sinks, and turbid yin rises. This is the root cause of aging, illness, and death. Therefore, the primary focus of medicine is on regulating the middle qi. The middle qi resides at the intersection of the two earth elements. Earth is born from fire, but fire dies from water. Excessive fire dries the earth, and excessive water dampens the earth. Draining water and replenishing fire, supporting yang and suppressing yin, regulates the middle qi..." The Qi circulates, and the clear and turbid return to their proper places. In treating the central Qi, ginseng and ginger are suitable for tonifying Yang and replenishing fire, while licorice and poria are suitable for strengthening the earth and draining water. The formula consists of ginseng, poria, dried ginger, and prepared licorice. Ginseng and dried ginger soothe the liver and dry the spleen, tonify fire and assist Yang, dispel cold and dampness, harmonize blood and promote Qi circulation, and treat coldness in the stomach and intestines, abdominal distension and pain. Combined with poria, it drains water and dries the earth, promotes urination and eliminates dampness, strengthens the spleen and calms the mind, and lowers Qi and relieves depression. The combined use of ginseng and licorice enhances their sweet and warm Qi-tonifying effects, promoting the ascending of spleen Qi and the dispersing of essence. When the spleen ascends, the liver also ascends; when the spleen ascends, the stomach descends; when the stomach descends, the lungs also descend. Therefore, the fundamental principle of Huangya Decoction is based on spleen Qi deficiency, with insufficient spleen Yang and impaired spleen function as key factors. Clinically, patients with spleen and stomach weakness, accompanied by imbalances in Qi movement leading to cold and heat, and imbalances in Qi and blood, can all be treated with modifications to the Huangya Decoction as a base formula to circulate the central Qi, promote the flow of Qi within the body, and restore the functions of various organs. Modern physicians widely apply it to people with insufficient Qi, weak spleen and stomach, overwork, prolonged worry, and irregular diet, as well as to the adjunctive treatment of diseases such as palpitations, chest tightness, shortness of breath, and indigestion.

[0003] The most commonly used traditional Chinese medicine extraction methods include decoction, distillation, maceration, reflux extraction, and supercritical fluid extraction. Although these traditional extraction methods are simple to operate and have low cost, they generally have problems such as low concentration of effective ingredients, high impurity content, low bioavailability, easy destruction of heat-sensitive components, and organic solvent residues in the extract. Summary of the Invention

[0004] Technical problem to be solved: In view of the above-mentioned technical problems, the first objective of the present invention is to provide a nanozyme for efficient extraction of Huangyatang, wherein the nanozyme is a manganese-doped carbon nitride nanozyme with galacturonic acid hexasaccharide grafted on its surface.

[0005] A second objective of this invention is to provide a method for preparing the manganese-doped carbon nitride nanozyme.

[0006] The third objective of this invention is to provide a highly efficient extraction method for Huangyatang (a traditional Chinese medicine) based on the manganese-doped carbon nitride nanozyme-mediated plant cell wall directional perforation technology.

[0007] The fourth objective of this invention is to provide a highly efficient extraction process for the Huangya decoction.

[0008] Technical solution:

[0009] A highly efficient extraction process for Huangya Tang (a traditional Chinese medicine formula) is disclosed. This process utilizes a manganese-doped carbon nitride nanozyme-mediated plant cell wall directional perforation technique for extracting Huangya Tang. The manganese-doped carbon nitride nanozyme has a surface grafted with galacturonic acid hexasaccharide, which can specifically bind to cell wall pectin. It exhibits peroxidase-like activity and catalyzes the generation of hydroxyl radicals (·OH) from H2O2 through a biomimetic catalytic reaction, thereby disrupting the cell wall structure and promoting the dissolution of active ingredients.

[0010] The above-mentioned efficient extraction process of Huangya decoction includes the following steps:

[0011] S1. Ginseng, Poria cocos, dried ginger and roasted licorice are dried separately, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder;

[0012] S2. Add the nanozyme with galacturonic acid hexasaccharide grafted on its surface to the citrate buffer, mix well, add the Chinese medicine powder, and then add H2O2 to react. After the reaction is completed, filter the extract through a filter membrane and centrifuge to obtain Huangya decoction.

[0013] Preferably, by weight, the ginseng in step S1 is 3-10 parts, the poria cocos is 2-7 parts, the dried ginger is 1-7 parts, and the prepared licorice root is 1-6 parts.

[0014] Preferably, the drying temperature of Poria cocos in step S1 is 50~55℃, and the drying time is 4~8 h.

[0015] Preferably, the drying temperature of the dried ginger in step S1 is 50~60℃, and the drying time is 5~10 h.

[0016] Preferably, the drying temperature of the roasted licorice in step S1 is 70~80℃, and the drying time is 18~24h.

[0017] Preferably, the mass ratio of the traditional Chinese medicine powder and nanozyme in step S2 is 1 g: 14~24 mg.

[0018] Preferably, the final concentration of H2O2 in the reaction system in step S2 is 1.7~4.3 mmol / L.

[0019] Preferably, the reaction temperature in step S2 is 40~48℃, the reaction speed is 200~300 rpm, and the reaction time is 30~45 min.

[0020] The preparation method of the nanozyme with galacturonic acid hexasaccharide grafted on its surface as described in step S2 includes the following steps:

[0021] S21. Weigh manganese acetylacetone and thiourea, add anhydrous ethanol, and stir until completely dissolved;

[0022] S22. Evaporate ethanol in a water bath, dry under vacuum, and then grind until a uniform powder is formed;

[0023] S23. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, introduce N2 to remove air, then reduce the N2 flow rate to maintain a dynamic atmosphere, connect the tail gas absorption device to the end of the quartz tube, slowly raise the temperature to 300℃, maintain it for 40~75 min, then continue to raise the temperature to 480~650℃, maintain the constant temperature for 3~6 h, after the reaction is completed, cool naturally to room temperature to obtain the pyrolysis product.

[0024] S24. The pyrolysis product was dispersed in dilute hydrochloric acid and ultrasonically treated. Unreacted metals were removed by stirring. The precipitate was collected by centrifugation, washed with deionized water until neutral, and dried under vacuum to obtain manganese-doped carbon nitride nanozyme.

[0025] S25. Manganese-doped carbon nitride nanozymes were dispersed in MES buffer, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide were added and stirred evenly. The nanozymes were activated at room temperature in the dark, centrifuged to recover the nanozymes, and washed with PBS to obtain the activated nanozymes.

[0026] S26. The activated nanozyme and galacturonic acid hexasaccharide were added to PBS containing Tween-20, mixed evenly, incubated, centrifuged to collect the product, and vacuum dried to obtain a nanozyme with galacturonic acid hexasaccharide grafted on its surface.

[0027] Preferably, the mass ratio of manganese acetylacetone and thiourea in step S21 is 1:4 to 7.5.

[0028] Preferably, the stirring temperature in step S21 is 50~65℃, the stirring speed is 200~250rpm, and the stirring time is 1.5~4h.

[0029] Preferably, the temperature of the water bath in step S22 is 70~85℃, and the water bath time is 50~80 min.

[0030] Preferably, the vacuum drying temperature in step S22 is 55~65℃, and the vacuum drying time is 8~13 h.

[0031] Preferably, the N2 flow rate in step S23 is 200~250 mL / min, and the N2 introduction time is 20~30 min.

[0032] Preferably, the reduced N2 flow rate in step S23 is 50~60 mL / min.

[0033] Preferably, the rate of heating in step S23 is 2~5℃ / min, and the duration of heating to 300℃ is 30~40 min.

[0034] Preferably, the frequency of the ultrasonic treatment in step S24 is 28~40 kHz, and the duration of the ultrasonic treatment is 15~30 min.

[0035] Preferably, the stirring temperature in step S24 is 60~85℃, the stirring speed is 400~700rpm, and the stirring time is 8~12h.

[0036] Preferably, the centrifugation speed in step S24 is 6500~8500 rpm, and the centrifugation time is 10~15 min.

[0037] Preferably, the vacuum drying temperature in step S24 is 40~60℃, and the vacuum drying time is 24~30 h.

[0038] Preferably, the mass ratio of manganese-doped carbon nitride nanozyme, EDC and NHS in step S25 is 1:0.02~0.05:0.03~0.07.

[0039] Preferably, the activation time in step S25 is 30-80 min.

[0040] Preferably, the centrifugation speed in step S25 is 4000~7000 rpm, and the centrifugation time is 3~5 min.

[0041] Preferably, the mass ratio of the activated nanozyme to galacturonic acid hexasaccharide in step S26 is 1:0.056~0.078.

[0042] Preferably, the incubation time in step S26 is 8 to 14 hours.

[0043] Preferably, the centrifugation speed in step S26 is 4000~7000 rpm, and the centrifugation time is 3~5 min.

[0044] Preferably, the vacuum drying temperature in step S26 is 35~45℃, and the vacuum drying time is 20~26 h.

[0045] Beneficial effects:

[0046] 1. The manganese-doped carbon nitride nanozyme provided by this invention possesses peroxidase-like (POD) activity. It achieves highly efficient targeted catalysis by mimicking the structure and catalytic mechanism of natural peroxidases. It constructs an Mn-N4 active center that mimics the metalloporphyrin ring structure of natural enzymes, utilizing the valence state change of manganese ions (Mn... 3+ / Mn 2+ It achieves electron transfer, catalyzes the decomposition of H2O2 to generate highly active free radicals, and grafts galacturonic acid hexasaccharide onto the surface of the nanozyme. Through the specific binding of oligosaccharide with cell wall pectin, it achieves targeted localization. The free radicals generated by catalysis effectively destroy the cell wall structure and promote the dissolution of active ingredients.

[0047] 2. The manganese-doped carbon nitride nanozyme provided by the present invention has excellent stability, maintaining high enzyme activity in a wide temperature range of 40~70℃, retaining more than 80% of enzyme activity after treatment at 70℃ for 20 min, and still retaining more than 85% of enzyme activity after storage at room temperature for 6 months.

[0048] 3. The manganese-doped carbon nitride nanoenzyme provided by this invention can be used for the efficient extraction of Huangyatang (a type of herbal decoction), which can significantly improve the extraction efficiency of target components such as total polysaccharides, total flavonoids, and ginsenosides in Huangyatang, while avoiding the destruction of heat-labile components (such as 6-shogaol and poria oligosaccharides) by high temperature. The retention rates of the two components are increased by 0.88~2.13 times and 0.5~1.45 times, respectively. Attached Figure Description

[0049] Figure 1 For the peroxidase-like activity test of Examples 1-8;

[0050] Figure 2 Thermal stability tests were conducted for Example 1 and Comparative Example 1.

[0051] Figure 3 This is a room temperature storage stability test for Example 1 and Comparative Example 1. Detailed Implementation

[0052] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments:

[0053] Example 1

[0054] This embodiment describes the preparation of manganese-doped carbon nitride nanozymes, including the following steps:

[0055] S1. Accurately weigh 2 g of manganese acetylacetone and 9.2 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0056] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0057] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0058] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0059] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 16 mg of EDC and 30 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 35 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0060] Example 2

[0061] The difference between this embodiment and Embodiment 1 is that the amount of thiourea added in this embodiment is 11 g, and the preparation method includes the following steps:

[0062] S1. Accurately weigh 2 g of manganese acetylacetone and 11 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0063] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0064] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0065] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0066] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 16 mg of EDC and 30 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 35 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0067] Example 3

[0068] The difference between this embodiment and Example 1 is that the amount of thiourea added in this embodiment is 13 g, and the preparation method includes the following steps:

[0069] S1. Accurately weigh 2 g of manganese acetylacetone and 13 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0070] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0071] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0072] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0073] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 16 mg of EDC and 30 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 35 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0074] Example 4

[0075] The difference between this embodiment and Example 1 is that the amount of thiourea added in this embodiment is 15 g, and the preparation method includes the following steps:

[0076] S1. Accurately weigh 2 g of manganese acetylacetone and 15 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0077] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0078] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0079] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0080] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 16 mg of EDC and 30 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 35 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0081] Example 5

[0082] The difference between this embodiment and Embodiment 1 is that the pyrolysis temperature in this embodiment is 500℃, the isothermal time is 5.5h, and the preparation method includes the following steps:

[0083] S1. Accurately weigh 2 g of manganese acetylacetone and 9.2 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0084] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0085] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 500℃, hold for 5.5 h, and let it cool naturally to room temperature.

[0086] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0087] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 16 mg of EDC and 30 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 35 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0088] Example 6

[0089] The difference between this embodiment and Embodiment 1 is that the pyrolysis temperature in this embodiment is 630℃, the isothermal time is 4 hours, and the preparation method includes the following steps:

[0090] S1. Accurately weigh 2 g of manganese acetylacetone and 9.2 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0091] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0092] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and introduce N2 at a flow rate of 220 mL / min for 30 min to remove air. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution to absorb H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 630℃, hold for 4 h, and let it cool naturally to room temperature.

[0093] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0094] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 16 mg of EDC and 30 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 35 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0095] Example 7

[0096] The difference between this embodiment and Embodiment 1 is that in this embodiment, the amount of EDC added is 20 mg, the amount of NHS added is 25 mg, and the amount of galacturonic acid hexasaccharide added is 30 mg. The preparation method includes the following steps:

[0097] S1. Accurately weigh 2 g of manganese acetylacetone and 9.2 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0098] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0099] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0100] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0101] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 20 mg of EDC and 25 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 30 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0102] Example 8

[0103] The difference between this embodiment and Embodiment 1 is that in this embodiment, the amount of EDC added is 12.5 mg, the amount of NHS added is 20 mg, and the amount of galacturonic acid hexasaccharide added is 32.5 mg. The preparation method includes the following steps:

[0104] S1. Accurately weigh 2 g of manganese acetylacetone and 9.2 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0105] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0106] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0107] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0108] S5. Disperse 500 mg of nanozyme in 10 mL of MES buffer (0.1 mol / L, pH 6.0), add 12.5 mg of EDC and 20 mg of NHS, stir well, activate at 25°C in the dark for 60 min, centrifuge at 5000 rpm for 3 min to recover the nanozyme, wash 3 times with PBS (pH 7.4), add the activated nanozyme and 32.5 mg of galacturonic acid hexasaccharide to 10 mL of PBS (containing 0.05% Tween-20), mix well and incubate for 12 h, centrifuge at 5000 rpm for 3 min to collect the product, vacuum dry at 40°C for 24 h and store.

[0109] Comparative Example 1

[0110] The difference between this comparative example and Example 1 is that this comparative example uses horseradish peroxidase.

[0111] Table 1. Characterization indicators of manganese-doped carbon nitride nanozymes in Examples 1-8

[0112]

[0113] Table 1 shows the key characterization parameters of the manganese-doped carbon nitride nanozymes prepared in Examples 1-8. In the manganese-doped carbon nitride nanozymes of Examples 1-8, the Mn element is mainly composed of Mn... 2+ and Mn 3+ With the increase of the thiourea ratio (Examples 1-4), the carbon matrix increases, the Mn loading decreases relatively, and more S element is doped, due to the mixed valence state loading. 2-The reducing properties of Mn affect its valence state distribution. With increasing pyrolysis temperature (Examples 1 and 5-6), the organic components decompose more thoroughly, increasing the Mn loading. However, high temperatures lead to sintering, causing Mn agglomeration into large particles. Some Mn detaches from the C3N4 framework, resulting in a decrease in apparent loading. Simultaneously, high temperatures also cause the C3N4 framework to decompose, reducing the carrier mass and thus lowering the Mn loading. The ratio of covalent coupling agents EDC to NHS also affects the characterization of nanozymes. After NHS reacts with the -COOH group on the carrier surface, the introduced amide bonds may cover some Mn active sites, leading to a decrease in Mn loading.

[0114] like Figure 1 As shown, the peroxidase-like activity of the manganese-doped carbon nitride nanozymes prepared in Examples 1-8 was detected by colorimetry. When the manganese-doped carbon nitride nanozymes were added to a solution containing TMB (3,3',5,5'-tetramethylbenzidine) for reaction, a deep blue product was generated, indicating that the manganese-doped carbon nitride nanozymes prepared in Examples 1-8 of this invention have peroxidase-like activity; the higher the absorbance value, the higher the activity.

[0115] like Figure 2 As shown, the optimal enzyme activity temperature for both manganese-doped carbon nitride nanozyme (Example 1) and horseradish peroxidase (Comparative Example 1) is 40°C. With increasing temperature, the relative enzyme activity of both decreased. After treatment at 50°C, 60°C, and 70°C for 20 min, the relative enzyme activity of Comparative Example 1 decreased sharply. In contrast, Example 1 exhibited excellent enzyme activity stability over a wide temperature range of 40–70°C, and maintained over 80% of its relative enzyme activity even after treatment at 70°C for 20 min.

[0116] like Figure 3 As shown, manganese-doped carbon nitride nanozymes (Example 1) and horseradish peroxidase (Comparative Example 1) were stored at room temperature (25±2℃) for 1 to 6 months. Example 1 showed excellent stability, and could still maintain more than 85% of the enzyme activity after 6 months. However, the relative enzyme activity of Comparative Example 1 showed a continuous downward trend, and only 25.13% of the activity was retained after 6 months. Its stability was significantly lower than that of Example 1.

[0117] Example 9

[0118] This embodiment describes the extraction of Huangya Tang (a traditional Chinese medicine formula) using the nanozyme prepared in Example 1, and includes the following steps:

[0119] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0120] S2. Add 459 mg of the nanozyme prepared in Example 1 to 400 mL of 0.3% citrate buffer (pH 5.0), mix well, and obtain a nanozyme mixture;

[0121] S3. Add the Chinese herbal medicine powder to the nanoenzyme mixture, shake at 44℃ and 270 rpm for 40 min, add H2O2 at 0.5 mL / min (final concentration in the solution is 3 mmol / L), and react at 44℃ and 200 rpm for 35 min. After the reaction is completed, filter the extract through a 0.22 μm membrane, centrifuge at 5000 rpm for 10 min to remove impurities, and then centrifuge at 12000 rpm for 20 min to precipitate the nanoenzyme. The supernatant is Huangya decoction.

[0122] Example 10

[0123] The difference between this embodiment and Embodiment 9 is that the amount of nanozyme added in this embodiment is 378 mg, and it includes the following steps:

[0124] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0125] S2. Add 378 mg of the nanozyme prepared in Example 1 to 400 mL of 0.3% citrate buffer (pH 5.0), mix well, and obtain a nanozyme mixture;

[0126] S3. Add the Chinese herbal medicine powder to the nanoenzyme mixture, shake at 44℃ and 270 rpm for 40 min, add H2O2 at 0.5 mL / min (final concentration in the solution is 3 mmol / L), and react at 44℃ and 200 rpm for 35 min. After the reaction is completed, filter the extract through a 0.22 μm membrane, centrifuge at 5000 rpm for 10 min to remove impurities, and then centrifuge at 12000 rpm for 20 min to precipitate the nanoenzyme. The supernatant is Huangya decoction.

[0127] Example 11

[0128] The difference between this embodiment and Embodiment 9 is that the amount of nanozyme added in this embodiment is 594 mg, and it includes the following steps:

[0129] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0130] S2. Add 594 mg of the nanozyme prepared in Example 1 to 400 mL of 0.3% citrate buffer (pH 5.0), mix well, and obtain a nanozyme mixture;

[0131] S3. Add the Chinese herbal medicine powder to the nanoenzyme mixture, shake at 44℃ and 270 rpm for 40 min, add H2O2 at 0.5 mL / min (final concentration in the solution is 3 mmol / L), and react at 44℃ and 200 rpm for 35 min. After the reaction is completed, filter the extract through a 0.22 μm membrane, centrifuge at 5000 rpm for 10 min to remove impurities, and then centrifuge at 12000 rpm for 20 min to precipitate the nanoenzyme. The supernatant is Huangya decoction.

[0132] Example 12

[0133] The difference between this embodiment and Example 9 is that the reaction temperature in this embodiment is 40°C, the reaction speed is 240 rpm, and the reaction time is 40 min. This embodiment uses the nanozyme prepared in Example 1 to extract Huangya Tang (a type of herbal decoction), and includes the following steps:

[0134] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0135] S2. Add 459 mg of the nanozyme prepared in Example 1 to 400 mL of 0.3% citrate buffer (pH 5.0), mix well, and obtain a nanozyme mixture;

[0136] S3. Add the Chinese herbal medicine powder to the nanozyme mixture, shake at 44℃ and 270 rpm for 40 min, add H2O2 at 0.5 mL / min (final concentration in the solution is 3 mmol / L), and react at 40℃ and 240 rpm for 40 min. After the reaction is completed, filter the extract through a 0.22 μm membrane, centrifuge at 5000 rpm for 10 min to remove impurities, and then centrifuge at 12000 rpm for 20 min to precipitate the nanozyme. The supernatant is Huangya decoction.

[0137] Example 13

[0138] The difference between this embodiment and Embodiment 9 is that the final concentration of H2O2 in the reaction system in this embodiment is 2 mmol / L, and it includes the following steps:

[0139] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0140] S2. Add 459 mg of the nanozyme prepared in Example 1 to 400 mL of 0.3% citrate buffer (pH 5.0), mix well, and obtain a nanozyme mixture;

[0141] S3. Add the Chinese herbal medicine powder to the nanozyme mixture, shake at 44℃ and 270 rpm for 40 min, add H2O2 at 0.5 mL / min (final concentration in the solution is 2 mmol / L), and react at 44℃ and 200 rpm for 35 min. After the reaction is completed, filter the extract through a 0.22 μm membrane, centrifuge at 5000 rpm for 10 min to remove impurities, and then centrifuge at 12000 rpm for 20 min to precipitate the nanozyme. The supernatant is Huangya decoction.

[0142] Comparative Example 2

[0143] The difference between this comparative example and Example 9 is that in this comparative example, the nanozyme is replaced with horseradish peroxidase (HRP), the reaction temperature is 45°C, and the steps include:

[0144] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0145] S2. Mix the Chinese herbal powder with 400 mL of 0.3% citrate buffer (pH 5.0), add 0.3 mL of horseradish peroxidase with an enzyme activity of 200 U / mg and H2O2 (final concentration in solution is 3 mmol / L), shake at 45℃ for 1 h, inactivate HRP by boiling in water bath for 5 min, centrifuge at 5000 rpm for 10 min to remove impurities, centrifuge at 12000 rpm for 20 min, and filter the extract through a 0.22 μm membrane to obtain Huangya decoction.

[0146] Comparative Example 3

[0147] The difference between this comparative example and Example 9 is that the extraction method in this comparative example, which does not add nanozymes, is hot water-assisted ultrasonic extraction, including the following steps:

[0148] S1. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts of ginseng, 6 parts of Poria cocos, 6 parts of dried ginger and 6 parts of prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0149] S2. Add 810 mL of 80℃ hot water to the Chinese herbal powder, let stand for 20 min, sonicate at 500W for 40 min, filter, centrifuge at 12000 rpm for 20 min, and filter the extract through a 0.22 μm membrane to obtain Huangya Decoction.

[0150] Comparative Example 4

[0151] The difference between this comparative example and Example 9 is that the manganese-doped carbon nitride nanozyme in this comparative example does not undergo galacturonic acid hexasaccharide surface modification, and includes the following steps:

[0152] S1. Accurately weigh 2 g of manganese acetylacetone and 9.2 g of thiourea into a 100 mL round-bottom flask, add 50 mL of anhydrous ethanol, and stir magnetically at 60 °C and 220 rpm for 2 h until completely dissolved;

[0153] S2. Transfer the solution to an evaporating dish, evaporate the ethanol in an 80°C water bath for 65 min until it becomes a viscous gel, dry it under vacuum at 60°C for 12 h, and grind it in an agate mortar for 30 min until a uniform powder is formed.

[0154] S3. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, and purge the air with N2 at a flow rate of 220 mL / min for 30 min. Then adjust the flow rate to 50 mL / min to maintain a dynamic atmosphere. Connect the tail gas absorption device (NaOH solution absorbs H2S) to the end of the quartz tube, raise the temperature to 300℃ at 5℃ / min, hold for 40 min, continue to raise the temperature to 550℃, hold for 4.5 h, and let it cool naturally to room temperature.

[0155] S4. The pyrolysis product was dispersed in 1 mol / L HCl, sonicated at 30 kHz for 20 min, stirred at 80℃ and 600 rpm for 12 h to remove unreacted metals, centrifuged at 8000 rpm for 10 min to collect the precipitate, washed with deionized water until neutral, and vacuum dried at 55℃ for 26 h to obtain manganese-doped carbon nitride nanozyme.

[0156] S5. Ginseng slices were dried at 45℃ for 22 h, Poria cocos slices were dried at 55℃ for 6 h, dried ginger was dried at 60℃ for 7 h, and prepared licorice root was dried at 80℃ for 20 h. 9 parts ginseng, 6 parts Poria cocos, 6 parts dried ginger and 6 parts prepared licorice root were taken, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder.

[0157] S2. Add 459 mg of manganese-doped carbon nitride nanozyme to 400 mL of 0.3% citrate buffer (pH 5.0), mix well, and prepare nanozyme mixture;

[0158] S3. Add the Chinese herbal medicine powder to the nanoenzyme mixture, shake at 44℃ and 270 rpm for 40 min, add H2O2 at 0.5 mL / min (final concentration in the solution is 3 mmol / L), and react at 44℃ and 200 rpm for 35 min. After the reaction is completed, filter the extract through a 0.22 μm membrane, centrifuge at 5000 rpm for 10 min to remove impurities, and then centrifuge at 12000 rpm for 20 min to precipitate the nanoenzyme. The supernatant is Huangya decoction.

[0159] Table 2. Content of active ingredients in Huangya decoction of Examples 9-13 and Comparative Examples 2-4

[0160] As shown in Table 2, compared with horseradish peroxidase enzymatic hydrolysis extraction (Comparative Example 2), hot water-assisted ultrasonic extraction (Comparative Example 3), and nanozyme extraction without galacturonic acid oligosaccharide modification (Comparative Example 4), the total polysaccharide, total flavonoid, ginseng total saponins, poria cocos oligosaccharide, and 6-shogaol contents of the nanozymes modified with galacturonic acid oligosaccharide (Examples 9-13) were significantly higher than those of the comparative examples. The nanozymes modified with galacturonic acid oligosaccharide enhanced targeting, specifically recognized and degraded pectin components in the cell wall, and destroyed the plant cell wall structure through the breaking of β-1,4-glycosidic bonds, promoting the dissolution of active ingredients. Effective cell wall destruction could be achieved at a low temperature of 40-50℃. Compared with Comparative Example 3, the retention rates of heat-labile components such as 6-shogaol and poria cocos oligosaccharide in Examples 9-13 were improved, with the retention rate of 6-shogaol increasing by 0.88-2.13 times and the retention rate of poria cocos oligosaccharide increasing by 0.5-1.45 times.

[0161] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A highly efficient extraction process for Huangya decoction, characterized in that: The highly efficient extraction process of Huangya Tang is based on the plant cell wall directional perforation technology mediated by manganese-doped carbon nitride nanozymes, which is used for the extraction of Huangya Tang; the surface of the manganese-doped carbon nitride nanozymes is grafted with galacturonic acid hexasaccharide.

2. The efficient extraction process of Huangya Tang according to claim 1, characterized in that, Includes the following steps: S1. Ginseng, Poria cocos, dried ginger and roasted licorice are dried separately, pulverized and mixed evenly, and passed through an 80-mesh sieve to obtain Chinese medicine powder; S2. Add the nanozyme with galacturonic acid hexasaccharide grafted on its surface to the citrate buffer, mix well, add the Chinese medicine powder, and then add H2O2 to react. After the reaction is completed, filter the extract through a filter membrane and centrifuge to obtain Huangya decoction.

3. The efficient extraction process of Huangya decoction according to claim 2, characterized in that: By weight, the ginseng mentioned in step S1 is 3-10 parts, the poria cocos is 2-7 parts, the dried ginger is 1-7 parts, and the prepared licorice root is 1-6 parts.

4. The efficient extraction process of Huangya Tang according to claim 2, characterized in that, The preparation method of the nanozyme with galacturonic acid hexasaccharide grafted on its surface in step S2 is as follows: S21. Weigh manganese acetylacetone and thiourea, add anhydrous ethanol, and stir until completely dissolved; S22. Evaporate ethanol in a water bath, dry under vacuum, and then grind until a uniform powder is formed; S23. Spread the powder evenly in a quartz boat, place it in the middle of a quartz tube, introduce N2 to remove air, then reduce the N2 flow rate to maintain a dynamic atmosphere, connect the tail gas absorption device to the end of the quartz tube, slowly raise the temperature to 300℃, maintain it for 40~75 min, then continue to raise the temperature to 480~650℃, maintain the constant temperature for 3~6 h, after the reaction is completed, cool naturally to room temperature to obtain the pyrolysis product. S24. The pyrolysis product was dispersed in dilute hydrochloric acid and ultrasonically treated. Unreacted metals were removed by stirring. The precipitate was collected by centrifugation, washed with deionized water until neutral, and dried under vacuum to obtain manganese-doped carbon nitride nanozyme. S25. Manganese-doped carbon nitride nanozymes were dispersed in MES buffer, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide were added and stirred evenly. The nanozymes were activated at room temperature in the dark, centrifuged to recover the nanozymes, and washed with PBS to obtain the activated nanozymes. S26. The activated nanozyme and galacturonic acid hexasaccharide were added to PBS containing Tween-20, mixed evenly, incubated, centrifuged to collect the product, and vacuum dried to obtain a nanozyme with galacturonic acid hexasaccharide grafted on its surface.

5. The efficient extraction process for Huangya decoction according to claim 4, characterized in that: The mass ratio of manganese acetylacetone to thiourea in step S21 is 1:4~7.

5.

6. The efficient extraction process of Huangya decoction according to claim 4, characterized in that: In step S25, the mass ratio of manganese-doped carbon nitride nanozyme, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 1:0.02~0.05:0.03~0.

07.

7. The efficient extraction process of Huangya decoction according to claim 4, characterized in that: The mass ratio of the activated nanozyme to galacturonic acid hexasaccharide in step S26 is 1:0.056~0.

078.

8. The efficient extraction process of Huangya Tang according to claim 2, characterized in that: The mass ratio of the traditional Chinese medicine powder and nanozyme in step S2 is 1 g: 14~24 mg.

9. The efficient extraction process of Huangya decoction according to claim 2, characterized in that: The final concentration of H2O2 in the reaction system in step S2 is 1.7~4.3 mmol / L.

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