Method for extracting amide components in Chinese prickly ash by using superfine grinding assisted natural eutectic solvent

Abstract

The invention belongs to the field of comprehensive utilization of plant resources, and relates to a method for extracting amide components in Chinese prickly ash through a natural eutectic solvent assisted by superfine grinding. The method comprises the following steps: firstly, collecting a pepper raw material, drying the pepper raw material by a heat pump, and performing high-speed crushing and low-temperature superfine crushing to obtain pepper superfine powder; preparing a natural eutectic solvent from a hydrogen bond acceptor and a hydrogen bond donor, mixing the natural eutectic solvent with the pepper superfine powder, performing microwave-assisted extraction, centrifugally collecting supernate after extraction, and completing primary extraction; adding NDES into the remaining precipitate, and repeatedly extracting according to the primary extraction step; and repeatedly extracting for several times, combining the supernate, and finally concentrating and filtering with a filter membrane to obtain an extracting solution, thereby realizing the extraction of the amide components in the Chinese prickly ash. The method provided by the invention overcomes the defects of low extraction efficiency, environmental pollution caused by an extraction reagent, high equipment cost and the like in the existing technical means for extracting the amide components in the Chinese prickly ash, realizes efficient and green extraction of the amide components in the Chinese prickly ash, and has a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the field of comprehensive utilization of plant resources, and specifically relates to a method for extracting amide components in Zanthoxylum bungeanum by using ultrafine grinding to assist natural low eutectic solvent. Background Art

[0002] Sichuan pepper, ranked first among the "thirteen spices" of seasonings, is a traditional Chinese medicinal and edible plant with rich color, fresh and spicy flavor, and is widely favored by people. Sichuan pepper is rich in a variety of active ingredients, such as alkaloids, volatile oils, amides, lignin, polyphenols, etc. Among them, amide substances, as characteristic components of Zanthoxylum plants, are the material basis for causing the numbing feeling of Sichuan pepper. Specifically, they are a series of chain polyunsaturated fatty acid amides. Hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-γ-sanshool, hydroxy-ε-sanshool, etc. are the most representative amide substances, accounting for more than 98% of the total amide content. In addition, Sichuan pepper amide also has a variety of physiological functions such as anesthesia, analgesia, anti-inflammatory, anti-aging and anti-cancer.

[0003] Ultrafine grinding technology is a high-tech technology that uses special grinding equipment to crush material particles into fine particles. It uses grinding, impact, shearing and other methods to instantly break solid materials into fine particles with uniform particle size and good fluidity at low temperature. The loss of effective functional components of the raw materials is small, and it belongs to pure physical technology processing, so as to maximize the retention of biologically active ingredients. It has been widely used in food, health care products and other fields. As the particles develop in the direction of micronization, the surface area and porosity of the particles are greatly increased, so that their adsorption and solubility properties are greatly improved, and the release of plant active ingredients is promoted, which can improve the extraction efficiency of plant active ingredients.

[0004] The extraction of amide components in Zanthoxylum bungeanum mainly includes organic solvent extraction and supercritical CO2 extraction. Although the organic solvent extraction method is simple to operate, it has problems such as large demand for organic reagents, high energy consumption, and environmental pollution; while the supercritical CO2 extraction method has the advantages of no solvent pollution and high extraction efficiency, but the operation requirements are high and the equipment is expensive. These limitations hinder the large-scale application of this method in the industry. Deep eutectic solvent (DES) is an economical and new green solvent, usually composed of a hydrogen bond acceptor (HBA) and one or two or more hydrogen bond donors (HBD), forming a uniform and stable transparent liquid by establishing strong hydrogen bond interactions. Natural deep eutectic solvent (NDES) came into being after DES. It is composed of natural products (organic acids, sugars, sugar alcohols, amino acids, organic amines). It has the advantages of non-toxicity, simple preparation, low raw material cost, biodegradability, high thermal stability, etc. It is more suitable for food, daily chemicals, medicines and other products with strict toxicity requirements. Therefore, it has been widely used in extracting natural active ingredients of plants (such as proteins, polysaccharides, phenols, etc.). However, the use of NDES to extract amide components in Zanthoxylum bungeanum is still in its initial stage, and most of them use organic solvents or synthetic reagents, which not only has environmental safety risks, but also the extraction rate needs to be improved. Therefore, establishing an efficient, safe and green extraction process for amide components of Zanthoxylum bungeanum is conducive to the further development and application of Zanthoxylum bungeanum. Summary of the invention

[0005] The purpose of the present invention is to provide a method for extracting amide components in Zanthoxylum bungeanum using ultrafine grinding assisted by natural low eutectic solvent in view of the above-mentioned shortcomings of the existing Zanthoxylum bungeanum amide extraction technology. The extraction method has the advantages of high extraction efficiency, simple operation, safety and non-toxicity.

[0006] To achieve the above purpose, the present invention adopts the following technical means:

[0007] A method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent, the method comprising the following steps:

[0008] (1) Preparing superfine pepper powder: freshly picked mature peppers are selected to remove wilted or brown peppers, and to remove debris, branches, leaves, and fruit stalks to obtain pepper raw materials; the pepper raw materials are subjected to heat pump drying treatment to dry to a final moisture content of ≤12%; the dried peppers are collected and crushed; the crushing treatment is carried out in steps, firstly, the dried peppers are removed from the seeds to obtain pepper shells, the pepper shells are crushed in a high-speed crusher, and after sieving, primary crushed pepper powder is obtained; then, the primary crushed pepper powder is subjected to low-temperature ultrafine crushing, and the temperature is controlled to be 10-13° C. After crushing, the pepper shells are sieved to obtain superfine pepper powder;

[0009] (2) preparing an extract: weighing the Zanthoxylum bungeanum ultrafine powder obtained in step (1), adding a natural low eutectic solvent, shaking and evenly mixing, and then performing microwave-assisted extraction. After the extraction is completed, centrifuging, and collecting the supernatant to complete one extraction; and adding the natural low eutectic solvent to the remaining precipitate again, and repeating the extraction according to the steps of the first extraction to obtain a second extraction supernatant; after repeating the extraction several times, combining the supernatants, and finally concentrating and filtering with a filter membrane to obtain a Zanthoxylum bungeanum NDES extract, thereby achieving the extraction of amide components in Zanthoxylum bungeanum;

[0010] The preparation steps of the natural low eutectic solvent (NDES) are as follows: a hydrogen bond acceptor and a hydrogen bond donor are mixed in a certain molar ratio, and stirred under certain temperature conditions until a uniform, transparent, viscous liquid is formed, and pure water is added to dilute the mixture to a concentration of 10% to 80% (v / v) to obtain a natural low eutectic solvent, which is recorded as NDES; the hydrogen bond acceptor is one of betaine or choline chloride; the hydrogen bond donor is any one of malic acid, lactic acid, citric acid, oxalic acid, tartaric acid, glycerol, propylene glycol, and 1,3-butanediol; and the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:(1 to 10).

[0011] Furthermore, the conditions of the heat pump drying treatment in step (1) are: drying temperature of 35-65° C., relative humidity of 15%-45%, and wind speed of 3-7 m / s.

[0012] Furthermore, the mesh size of the sieve through which the initially crushed pepper powder is sieved in step (1) is 40 to 80 meshes.

[0013] Furthermore, in step (1), the low-temperature ultrafine grinding is performed using a low-temperature ultrafine grinding machine.

[0014] Furthermore, the superfine powder of Sichuan pepper obtained in step (1) is sieved through a mesh number of 200 to 800 meshes, and the particle size is 5 to 75 μm.

[0015] Furthermore, the conditions for microwave-assisted extraction in step (2) are: microwave power of 200-400 W, microwave temperature of 40-80° C., and microwave time of 3-20 min.

[0016] Furthermore, in step (2), the ratio of the ultrafine powder of Zanthoxylum bungeanum to the natural deep eutectic solvent (NDES) is 1 g: (20-80) mL;

[0017] Furthermore, the centrifugal conditions in step (2) are: rotation speed 1500-4000r / min, time 5-20min.

[0018] Furthermore, in step (2), the ratio of the precipitate to NDES is 1 g: (20-80) mL; and the extraction is repeated 1-3 times.

[0019] Furthermore, the concentration in step (2) is to evaporate the supernatant under reduced pressure to 1 / 5 to 1 / 3 of the original volume; the filter membrane is a 0.22 μm nylon organic filter membrane; and the stirring temperature under certain temperature conditions is 60 to 90°C.

[0020] Beneficial effects:

[0021] The present invention provides a method for extracting amide components in Zanthoxylum bungeanum with the aid of ultrafine grinding and natural low eutectic solvent, which mainly solves the shortcomings of existing technical means for extracting amide components in Zanthoxylum bungeanum, such as low extraction efficiency, environmental pollution caused by extraction reagents, and high equipment cost, and realizes efficient and green extraction of amide components in Zanthoxylum bungeanum. The specific beneficial effects are as follows.

[0022] (1) The present invention utilizes heat pump drying technology to dry the pepper. Compared with traditional drying methods such as natural air drying and hot air drying, the technology has the advantages of energy saving, easy control, good drying quality and environmental friendliness.

[0023] (2) The present invention uses a two-step ultrafine grinding process to improve the uniformity of the crushing of the pepper material, and after the pepper is ultrafinely ground, the surface area is greatly increased, the release rate of the pepper amide components is accelerated, and the extraction speed and extraction efficiency of the amide components in the pepper are significantly improved. In addition, low-temperature ultrafine grinding can effectively prevent the heat-sensitive substances in the pepper from being inactivated or lost due to high temperature, which helps to maintain the original flavor and nutritional active ingredients of the pepper.

[0024] (3) The present invention applies natural deep eutectic solvent (NDES) to the extraction of amide components in Zanthoxylum bungeanum, overcoming the shortcomings of insufficient extraction of water as a solvent and high toxicity and high pollution of organic solvents; compared with traditional deep eutectic solvents, the natural deep eutectic solvent used in the present invention is not only conducive to the extraction of amide components, but also has lower toxicity, better biodegradability, and is safer and more environmentally friendly. In addition, the present invention further organically combines microwave extraction with the extraction of amide components in Zanthoxylum bungeanum, accelerates the dissolution of amide components, and greatly improves the extraction efficiency.

[0025] (4) The present invention is simple to operate, does not require complicated processes, and does not require operators to have a strong professional background. The reagents used are low in cost and have high extraction efficiency, providing a technical reference for the green and efficient extraction of prickly ash amide components. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the standard curve of hydroxy-α-sanshool.

[0027] Figure 2 This is a representative diagram of the liquid phase of Zanthoxylum bungeanum NDES extract. DETAILED DESCRIPTION

[0028] In order to more clearly illustrate the content of the present invention, the present invention is further described below in conjunction with specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention and should not be construed as all the embodiments of the present invention.

[0029] 1. Experimental samples, main reagents and instruments

[0030] Samples: Fresh pepper purchased from Sichuan Maowen pepper base was used as the research object.

[0031] Reagents: Hydroxy-α-sanshool standard was purchased from Chengdu Medson Technology Co., Ltd., with a purity greater than 98%. All water used in the experiment was deionized water, and all experimental reagents were analytical grade.

[0032] Instruments: high-speed pulverizer (FW-100, Tianjin Huaxin Instrument Factory); ultra-low temperature ultrafine pulverizer (KC-701, Beijing Kaichuang Tonghe Technology Development Co., Ltd.); heat pump dryer (YGKRK, Dongguan Yonggan Energy Saving Technology Co., Ltd.); high performance liquid chromatograph (Agilent 1260, Agilent Technologies Co., Ltd.); Xianghu computer microwave ultrasonic combined synthesis / extraction instrument (XH-300A, Beijing Xianghu Technology Development Co., Ltd.).

[0033] Embodiment 1:

[0034] S1. Preparation of superfine powder of Sichuan pepper: Pick out the wilted or browned Sichuan pepper from the freshly picked mature Sichuan pepper, remove the debris, branches, leaves, and fruit stems, and weigh 100g to put on a plate. Preheat the heat pump dryer to the set parameters (temperature 50°C, relative humidity 35%, wind speed 5m / s), then put in the Sichuan pepper for drying, and the final moisture content is ≤12%. Collect the dried Sichuan pepper and crush it step by step. First, remove the seeds from the Sichuan pepper to obtain the Sichuan pepper shell, and crush the Sichuan pepper shell in a high-speed grinder. After passing through a 60-mesh sieve, obtain the crushed Sichuan pepper powder; then in the second step, put the crushed Sichuan pepper powder into an ultra-low temperature superfine grinder (the temperature of the crushing chamber is 12°C), pass through a 600-mesh sieve, collect the Sichuan pepper superfine powder, seal it and refrigerate it (4°C) away from light for later use.

[0035] S2. Preparation of NDES: betaine (hydrogen bond acceptor) and citric acid (hydrogen bond donor) were mixed in a molar ratio of 1:5, and magnetically stirred at 85°C until a uniform, transparent, viscous liquid was formed, and diluted with pure water to a concentration of 70% (v / v). The diluted solution was NDES and was set aside.

[0036] S3. Prepare the extract: weigh 5 g of superfine powder of Zanthoxylum bungeanum, add it to 200 mL of NDES (concentration 70%), shake it well, and perform microwave-assisted extraction (microwave power is 300 W, microwave temperature is 50°C, and microwave time is 10 min). After the extraction is completed, centrifuge it at 2500 r / min for 10 min, collect the supernatant, and complete one extraction; then add 200 mL of NDES (concentration 70%) to the precipitate again, perform a second extraction according to the steps of the first extraction, and combine the supernatants collected from the two extractions, and finally concentrate it to 100 mL by reduced pressure evaporation, and filter it through a 0.22 μm organic filter membrane to obtain the prickly ash amide NDES extract.

[0037] Embodiment 2:

[0038] S1. Preparation of superfine powder of Sichuan pepper: Pick out the wilted or browned Sichuan pepper from the freshly picked mature Sichuan pepper, remove the debris, branches, leaves, and fruit stems, and weigh 100g to plate. The heat pump dryer is first preheated to the set parameters (temperature 35°C, relative humidity 15%, wind speed 3m / s), and then put in the Sichuan pepper for drying, and the final moisture content is ≤12%. The dried Sichuan pepper is collected and crushed step by step. First, the Sichuan pepper seeds are removed to obtain the Sichuan pepper shell, and the Sichuan pepper shell is initially crushed in a high-speed crusher. After passing through a 40-mesh sieve, the initially crushed Sichuan pepper powder is placed in an ultra-low temperature superfine pulverizer (the temperature of the crushing chamber is 12°C), and after passing through a 300-mesh sieve, the Sichuan pepper superfine powder is collected, sealed and refrigerated in a dark place for standby use.

[0039] S2. Preparation of NDES: Mix betaine (hydrogen bond acceptor) and glycerol (hydrogen bond donor) in a molar ratio of 1:7, and stir magnetically at 85°C until a uniform, transparent, viscous liquid is formed. Dilute with pure water to a concentration of 60% (v / v) before use. The diluted solution is NDES and is ready for use.

[0040] S3. Prepare the extract: weigh 5 g of superfine powder of Zanthoxylum bungeanum, add 200 mL NDES (concentration 60%), shake well, perform microwave-assisted extraction (microwave power is 200 W, microwave temperature is 40° C., microwave time is 5 min), centrifuge at 2500 r / min for 10 min after the extraction is completed, collect the supernatant, and complete the first extraction; then add 200 mL NDES (concentration 60%) to the precipitate again, perform the second extraction according to the steps of the first extraction, and combine the supernatants collected from the two extractions, finally concentrate to 100 mL by reduced pressure evaporation, and obtain the prickly ash amide NDES extract after filtering through a 0.22 μm organic filter membrane.

[0041] Embodiment 3:

[0042] S1. Preparation of superfine powder of Sichuan pepper: Pick out the wilted or browned Sichuan pepper from the freshly picked mature Sichuan pepper, remove the debris, branches, leaves, and fruit stems, and weigh 100g to plate. The heat pump dryer is first preheated to the set parameters (temperature 65°C, relative humidity 45%, wind speed 7m / s), and then the Sichuan pepper is put in for drying, and the final moisture content is ≤12%. The dried Sichuan pepper is collected and crushed step by step. First, the Sichuan pepper seeds are removed to obtain the Sichuan pepper shell, and the Sichuan pepper shell is initially crushed in a high-speed crusher. After passing through an 80-mesh sieve, the initially crushed Sichuan pepper powder is placed in an ultra-low temperature superfine pulverizer (the temperature of the crushing chamber is 12°C), and after passing through an 800-mesh sieve, the Sichuan pepper superfine powder is collected, sealed and refrigerated in a dark place for standby use.

[0043] S2. Preparation of NDES: Mix choline chloride (hydrogen bond acceptor) and citric acid (hydrogen bond donor) in a molar ratio of 1:5, and stir magnetically at 85°C until a uniform, transparent, viscous liquid is formed. Dilute with pure water to a concentration of 80% (v / v) before use. The diluted solution is NDES and is ready for use.

[0044] S3. Prepare the extract: weigh 5 g of superfine powder of Zanthoxylum bungeanum, add 150 mL of NDES (concentration 80%), shake and evenly shake, perform microwave-assisted extraction (microwave power of 400 W, microwave temperature of 60° C., microwave time of 20 min), centrifuge at 2500 r / min for 10 min after the extraction, collect the supernatant, and complete the first extraction; then add 150 mL of NDES (concentration 80%) to the precipitate again, perform the second extraction according to the steps of the first extraction, and combine the supernatants collected from the two extractions, finally concentrate to 100 mL by evaporation under reduced pressure, and obtain the prickly ash amide NDES extract after filtering through a 0.22 μm organic filter membrane.

[0045] Comparative Example 1:

[0046] Compared with Example 1, the two-step ultrafine grinding process is not used, that is, S1 is operated as follows, and the remaining steps are the same.

[0047] S1. Preparation of pepper powder: Pick out the wilted or browned pepper from the freshly picked mature pepper, remove the debris, branches, leaves, and fruit stems, and weigh 100g to put on a plate. Preheat the heat pump dryer to the set parameters (temperature 50°C, relative humidity 35%, wind speed 5m / s), then put the pepper in for drying, and the final moisture content is ≤12%. Collect the dried pepper, remove the seeds to obtain the pepper shell, crush the pepper shell in a high-speed grinder, pass through a 60-mesh sieve, collect the pepper powder, seal it and refrigerate it in a dark place for later use.

[0048] Comparative Example 2:

[0049] Compared with Example 1, the low-temperature ultrafine grinding process is not used, that is, S1 is operated as follows, and the remaining steps are the same.

[0050] S1. Preparation of pepper powder: Pick out the wilted or browned pepper from the freshly picked mature pepper, remove the debris, branches, leaves, and fruit stems, and weigh 100g to put on a plate. Preheat the heat pump dryer to the set parameters (temperature 50°C, relative humidity 35%, wind speed 5m / s), then put the pepper in for drying, and the final moisture content is ≤12%. Collect the dried pepper and crush it step by step. First, remove the seeds from the pepper to obtain the pepper shell, and crush the pepper shell in a high-speed grinder. After passing through a 60-mesh sieve, obtain the crushed pepper powder; then in the second step, put the crushed pepper powder into an ultrafine grinder (the temperature of the grinding chamber is 65°C), pass through a 600-mesh sieve, collect the pepper ultrafine powder, seal it and refrigerate it (4°C) away from light for later use.

[0051] Comparative Example 3:

[0052] Compared with Example 1, NDES reagent was not used, but 70% methanol was used as the extraction reagent; that is, S2 was operated as follows, and the remaining steps were the same.

[0053] S2. Prepare the extract: weigh 5 g of superfine powder of Zanthoxylum bungeanum, add 200 mL of 70% methanol, shake and evenly mix, perform microwave-assisted extraction (microwave power is 300 W, microwave temperature is 50° C., microwave time is 10 min), centrifuge at 2500 r / min for 10 min after the extraction, collect the supernatant, and complete the first extraction; add 200 mL of 70% methanol to the precipitate again, perform the second extraction according to the steps of the first extraction, combine the supernatants collected from the two extractions, evaporate and concentrate to 100 mL under reduced pressure, and filter through a 0.22 μm organic filter membrane to obtain the methanol extract of Zanthoxylum bungeanum.

[0054] Comparative Example 4:

[0055] Compared with Example 1, microwave-assisted extraction was not used, but water bath heating extraction was used; that is, S3 was operated as follows, and the remaining steps were the same.

[0056] S3. Prepare the extract: weigh 5 g of Zanthoxylum bungeanum ultrafine powder, add 200 mL NDES (concentration 70%), shake well, perform water bath heating extraction (temperature is 50°C, extraction time is 10 min), centrifuge at 2500 r / min for 10 min after the extraction is completed, collect the supernatant, and complete one extraction; then add 200 mL NDES (concentration 70%) to the precipitate again, perform a second extraction according to the steps of the first extraction, and combine the supernatants collected from the two extractions, finally concentrate to 100 mL by reduced pressure evaporation, and obtain the Zanthoxylum bungeanum NDES extract after filtering through a 0.22 μm organic filter membrane.

[0057] Comparative Example 5:

[0058] Compared with Example 1, the two-step ultrafine grinding process was not used; NDES reagent was not used, but 70% methanol was used as the extraction reagent; microwave-assisted extraction was not used, but water bath heating extraction was used.

[0059] S1. Preparation of pepper powder: Pick out the wilted or browned pepper from the freshly picked mature pepper, remove the debris, branches, leaves, and fruit stems, and weigh 100g to put on a plate. Preheat the heat pump dryer to the set parameters (temperature 50°C, relative humidity 35%, wind speed 5m / s), then put the pepper in for drying, and the final moisture content is ≤12%. Collect the dried pepper, remove the seeds to obtain the pepper shell, crush the pepper shell in a high-speed grinder, pass through a 60-mesh sieve, collect the pepper powder, seal it and refrigerate it in a dark place for later use.

[0060] S2. Prepare the extract: weigh 5 g of Zanthoxylum bungeanum powder, add 200 mL of 70% methanol, shake well, and perform water bath heating extraction (temperature is 50°C, extraction time is 10 min). After the extraction is completed, centrifuge at 2500 r / min for 10 min, collect the supernatant, and complete one extraction; add 200 mL of 70% methanol to the precipitate again, perform a second extraction with the same extraction steps, combine the supernatants collected from the two extractions, concentrate to 100 mL by reduced pressure evaporation, and filter through a 0.22 μm organic filter membrane to obtain the pepper amide methanol extract.

[0061] Comparative Example 6:

[0062] Same as Example 1, except that urea is used as the hydrogen bond donor.

[0063] Comparative Example 7:

[0064] Same as Example 1, except that sucrose is used as a hydrogen bond donor.

[0065] Test Example 1: Extraction yield of prickly ash amide components in NDES extract

[0066] The HPLC-MS method was used to determine and calculate the extraction yield of the sanshoamide components in the NDES extract. The liquid phase detection conditions are: the mobile phase is acetonitrile (40%): 1% glacial acetic acid aqueous solution (60%) (v / v), the flow rate is 1.5mL / min, the ultraviolet detection wavelength is 270nm, the injection volume is 20μL, the detection time is 30min, the column temperature is 35℃, and the chromatographic column specifications are Agilent (XDBC18column, 4.6mm×250mm, 5μm). Using the hydroxy-α-sanshool standard as the standard sample, dissolve and dilute the hydroxy-α-sanshool with methanol to prepare a series of standard solutions with different concentrations for liquid chromatography detection, and draw a standard curve with the hydroxy-α-sanshool concentration as the horizontal coordinate and the peak area as the vertical coordinate, as shown in the figure. Figure 1 Take 10 μL of NDES extract and dilute to 2 mL with methanol, take 1.5 mL and filter through 0.22 μm filter membrane for liquid chromatography detection. Figure 2 It is a representative diagram of the liquid phase of the NDES extract; the peak with a retention time of 14-19min is taken as the target peak, and the calculation is performed based on the sum of the calculated peak areas and the linear regression equation corresponding to the standard curve, and then the extraction yield of the prickly ash amide components in the NDES extract is converted according to the following formula.

[0067]

[0068] Where: m1, the mass of the prickly ash amide components in the NDES extract, g; m2, the mass of the weighed prickly ash ultrafine powder, g.

[0069] The results are shown in Table 1:

[0070] Table 1 Extraction yield of prickly ash amide components in NDES extract

[0071] Yield of pepper amide components / % Example 1 15.84±0.66 Example 2 13.25±0.34 Example 3 14.79±0.51 Comparative Example 1 9.65±0.24 Comparative Example 2 10.85±0.31 Comparative Example 3 12.01±0.23 Comparative Example 4 8.81±0.46 Comparative Example 5 5.27±0.43 Comparative Example 6 11.68±0.27 Comparative Example 7 10.57±0.38

[0072] From the results in Table 1, it can be seen that the extraction effects of amide components in Zanthoxylum bungeanum in Examples 1, 2, and 3 using low-temperature ultrafine grinding technology combined with NDES are significantly better than those in Comparative Example 1 which does not use ultrafine grinding technology, wherein the effect of Example 1 is improved by more than 64% compared with Comparative Example 1; and although Comparative Example 2 uses a two-step ultrafine grinding process, low temperature setting is not performed during the ultrafine grinding process, and the extraction effect is reduced by more than 30% compared with Example 1. The seemingly simple adjustment actually produces unexpected effects, indicating that low-temperature ultrafine grinding is more conducive to improving the extraction yield. Compared with Examples 1, 2, and 3 using NDES as the extraction reagent, the extraction yield of Comparative Example 3 using 70% methanol as the extraction reagent is lower than that of Examples 1, 2, and 3 using NDES as the extraction reagent, and methanol as an organic reagent is toxic and easily pollutes the environment, and is not suitable for the fields of food, daily chemicals, and medicines; Compared with Examples 1, 2, and 3 using microwave-assisted extraction, the extraction yield of Comparative Example 4 using ordinary water bath heating is lower than that of Examples 1, 2, and 3 using microwave-assisted extraction, indicating that microwave-assisted extraction can effectively promote the dissolution of prickly ash amide components; Comparative Example 5 further illustrates that low-temperature ultrafine grinding technology, NDES reagent, and microwave-assisted extraction play a vital role in the extraction of prickly ash amide components; and Comparative Example 6 using urea as a hydrogen bond donor and Comparative Example 7 using sucrose as a hydrogen bond donor, their extraction yields are also lower than those of Examples 1, 2, and 3, wherein urea is a non-natural product and its safety needs to be considered, and urea itself has a certain taste and smell, which is easy to affect the flavor of the final product, and the NDES formed by sucrose has a large viscosity, which hinders the mass transfer process from prickly ash powder to the solution, thereby affecting the extraction efficiency of amide components.

[0073] In summary, the present invention is based on ultrafine grinding assisted by natural low eutectic solvent to extract amide components in Zanthoxylum bungeanum, and has achieved very significant technical effects (the existing technology is basically unable to break through the yield of 13%, and achieving 12% is already a very good effect). The present invention is based on a two-step ultrafine grinding process (low temperature setting during ultrafine grinding), a natural low eutectic solvent and microwave-assisted operation, which realizes green and efficient extraction of amide components in Zanthoxylum bungeanum, not only breaking through the yield of 13%, but also Example 1 reaches a yield of more than 15%, achieving unexpected technical effects. At the same time, the method is simple to operate, low in cost, efficient in extraction, green and environmentally friendly, non-toxic and harmless, and provides technical support for the actual production and processing application of amide components in Zanthoxylum bungeanum.

[0074] Note: The above embodiments are only used to illustrate the present invention and are not intended to limit the technical solutions described in the present invention; therefore, although this specification has described the present invention in detail with reference to the above embodiments, a person skilled in the art should understand that the present invention can still be modified or replaced by equivalents; and all technical solutions and improvements thereof that do not depart from the spirit and scope of the present invention should be included in the scope of the claims of the present invention.

Claims

1. A method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent, characterized in that: The following steps are involved: (1) Preparing superfine pepper powder: freshly picked mature peppers are selected to remove wilted or brown peppers, and to remove debris, branches, leaves, and fruit stalks to obtain pepper raw materials; the pepper raw materials are subjected to heat pump drying treatment to dry to a final moisture content of ≤12%; the dried peppers are collected and crushed; the crushing treatment is carried out in steps, firstly, the dried peppers are removed from the seeds to obtain pepper shells, the pepper shells are crushed in a high-speed crusher, and after sieving, primary crushed pepper powder is obtained; then, the primary crushed pepper powder is subjected to low-temperature ultrafine crushing, and the temperature is controlled to be 10-13° C. After crushing, the pepper shells are sieved to obtain superfine pepper powder; (2) preparing an extract: weighing the Zanthoxylum bungeanum ultrafine powder obtained in step (1), adding a natural low eutectic solvent, shaking and evenly mixing, and then performing microwave-assisted extraction. After the extraction is completed, centrifuging, and collecting the supernatant to complete one extraction; and adding the natural low eutectic solvent to the remaining precipitate again, and repeating the extraction according to the steps of the first extraction to obtain a second extraction supernatant; after repeating the extraction several times, combining the supernatants, and finally concentrating and filtering with a filter membrane to obtain a Zanthoxylum bungeanum NDES extract, thereby achieving the extraction of amide components in Zanthoxylum bungeanum; The preparation steps of the natural low eutectic solvent are as follows: a hydrogen bond acceptor and a hydrogen bond donor are mixed in a certain molar ratio, and stirred under certain temperature conditions until a uniform, transparent, viscous liquid is formed, and pure water is added to dilute the mixture to a concentration of 10% to 80% (v / v) to obtain a natural low eutectic solvent, which is recorded as NDES; the hydrogen bond acceptor is one of betaine or choline chloride; the hydrogen bond donor is any one of malic acid, lactic acid, citric acid, oxalic acid, tartaric acid, glycerol, propylene glycol, and 1,3-butanediol; and the molar ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:(1 to 10).

2. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The conditions of the heat pump drying treatment in step (1) are: drying temperature of 35-65° C., relative humidity of 15%-45%, and wind speed of 3-7 m / s.

3. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The mesh number of the sieve through which the initially crushed pepper powder is sieved in step (1) is 40 to 80 meshes.

4. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: In step (1), low-temperature ultrafine grinding is carried out using a low-temperature ultrafine grinding machine.

5. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The superfine powder of Sichuan pepper obtained in step (1) is sieved through a mesh number of 200 to 800 meshes, and the particle size is 5 to 75 μm.

6. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The conditions for microwave-assisted extraction in step (2) are: microwave power of 200-400 W, microwave temperature of 40-80° C., and microwave time of 3-20 min.

7. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The ratio of the superfine Zanthoxylum bungeanum powder to NDES in step (2) is 1 g: (20-80) mL.

8. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The centrifugal conditions in step (2) are: rotation speed 1500-4000r / min, time 5-20min.

9. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The ratio of the precipitate to NDES in step (2) is 1 g: (20-80) mL; the extraction is repeated 1-3 times.

10. The method for extracting amide components from Zanthoxylum bungeanum using ultrafine grinding assisted natural deep eutectic solvent according to claim 1, characterized in that: The concentration in step (2) is to evaporate the supernatant under reduced pressure to 1 / 5 to 1 / 3 of the original volume; the filter membrane is a 0.22 μm nylon organic filter membrane; The stirring temperature under certain temperature conditions is 60-90°C.

Images