Fluid extraction method for extracting active ingredients of natural plants

Abstract

The invention relates to a fluid extraction method for extracting active ingredients of natural plants, and belongs to the technical field of plant extraction. The method comprises the following steps: (1) taking fresh materials of natural plants to prepare natural plant slurry, and taking dry materials of the natural plants to prepare natural plant powder; (2) mixing the natural plant powder with the natural plant slurry, and granulating; and (3) putting the granules obtained by granulation into a supercritical extraction kettle, and extracting by adopting supercritical CO2 fluid so as to separate out the active ingredients of the natural plants. According to the extraction method disclosed by the invention, the extraction of the active ingredients can be realized without the help of ultrasonic-assisted extraction; particularly, active ingredients in the dry materials of natural plants can be extracted more sufficiently.

Description

Technical Field

[0001] The present application relates to the technical field of plant extraction, and in particular to a fluid extraction method for extracting active ingredients from natural plants. Background Art

[0002] Supercritical fluid extraction is a new type of extraction and separation technology. It uses supercritical fluid, that is, a fluid in a thermodynamic state with a temperature higher than the critical temperature and a pressure higher than the critical pressure, as an extractant to extract specific components from liquids or solids to achieve separation purposes.

[0003] Compared with traditional solvent extraction, supercritical fluid extraction has the advantages of high yield, short production cycle, no destruction of active ingredients, no solvent residue or very little solvent residue. However, the disadvantage of supercritical fluid extraction is that it requires higher system pressure and increased solvent circulation times to achieve efficient extraction.

[0004] In the prior art, when using supercritical fluid to extract plant active ingredients, in order to reduce the extraction pressure and extraction temperature, shorten the extraction time, and reduce the amount of entrainer, it is often combined with ultrasonic technology to act on the extract at the same time. For example, a method for extracting active ingredients from natural medicinal plants by supercritical fluid using dual-frequency ultrasound alternating enhancement is disclosed in application number CN200310117437.5, which uses a dual-frequency ultrasound alternating enhancement supercritical fluid extraction device to extract and separate the raw materials, thereby reducing the pressure and extraction temperature of supercritical fluid extraction, shortening the extraction time, and reducing the amount of entrainer. However, the disadvantage of this method is that it must rely on dual-frequency ultrasound alternating enhancement, the operation is not convenient, and the energy consumption is high.

[0005] In addition, when extracting active ingredients from natural plants, the prior art often uses dry materials of natural plants for extraction. Although the dry materials can be crushed into powder to improve their extraction rate to a certain extent, after being crushed to a certain degree, the extraction rate is difficult to continue to increase, and the part of active ingredients that cannot be fully extracted is more difficult to extract. Summary of the invention

[0006] In order to solve or partially solve the problems existing in the related art, the present application provides a fluid extraction method for extracting active ingredients from natural plants, which can achieve the extraction of active ingredients without the aid of ultrasonic-assisted extraction; in particular, the active ingredients in natural plant dry materials can be more fully extracted.

[0007] A fluid extraction method for extracting natural plant active ingredients in the present application comprises the following steps: (1) Prepare natural plant slurry from fresh natural plant materials and prepare natural plant powder from dry natural plant materials; (2) mixing the natural plant powder and the natural plant slurry and then granulating; (3) The granules obtained by granulation are placed in a supercritical extraction kettle and extracted using supercritical CO2 fluid to separate the natural plant active ingredients.

[0008] Specifically, in the step (1), the natural plant slurry is prepared by configuring the natural plant slurry in a ratio of the mass of the natural plant to the volume of water of 1:(2-3), and then grinding the slurry with a colloid mill to obtain the natural plant slurry.

[0009] Specifically, when supercritical CO2 fluid is used for extraction in step (3), the extraction temperature is 35-50°C, the extraction pressure is 10-20 MPa, the extraction time is 3-10 hours, the extraction fluid flow rate is 10-50 L / h, and the amount of entrainer ethanol is 30-100 ml ethanol / 100 g CO2.

[0010] Specifically, the particle size of the natural plant powder is 50-100 meshes.

[0011] Specifically, the natural plant is a plant of the genus Toona of the Meliaceae family or a plant of the genus Coix.

[0012] The mechanism of the present application is: first, the natural plant wall is broken to promote the outflow of its effective ingredients, and then the slurry is mixed with the natural plant powder for granulation. By adopting the above operation, the powder made of natural plant dry materials that are difficult to extract is wrapped and granulated. On the one hand, the extraction fluid can better penetrate into the natural plant dry material powder to extract it. On the other hand, since the effective ingredients wrapped around the periphery of the particles can play a role in increasing the active ingredients, it is speculated that it can guide the hard-to-extract active ingredients in the natural plant dry material powder to be better extracted.

[0013] The beneficial effects of this application are: 1. The fluid extraction method in the present application is to prepare natural plant slurry from fresh natural plant materials, mix it with natural plant powder and granulate it, and then directly perform supercritical CO2 fluid extraction on the granulated particles. The whole process abandons the ultrasonic assisted extraction in the prior art, so the energy consumption is lower, the cost is lower, and the operation is more convenient. At the same time, the control conditions of the supercritical CO2 fluid extraction in the present application are relatively mild in terms of temperature and pressure, so that the active ingredients can be efficiently extracted under the conditions of low extraction pressure and extraction temperature.

[0014] 2. The fluid extraction method in this application can more fully extract the active ingredients in natural plant dry materials. DETAILED DESCRIPTION

[0015] The embodiments of the present application will be described in more detail below with reference to examples. Although the embodiments of the present application are shown in the examples, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to make the present application more thorough and complete, and the scope of the present application can be fully communicated to those skilled in the art. Example 1

[0016] The extraction of flavonoids from toona sinensis leaves specifically comprises the following steps: (1) preparing a toon leaf slurry from fresh toon leaves, and preparing a toon leaf powder from dried toon leaves; the fresh toon leaves are toon leaves harvested within 24 hours; When preparing the toona sinensis leaf slurry, the toona sinensis leaf is prepared in a ratio of 1:2 in terms of the mass of the toona sinensis leaf and the volume of water, and then the toona sinensis leaf is ground with a colloid mill to obtain the toona sinensis leaf slurry.

[0017] The particle size of the toon leaf powder is 50-100 meshes.

[0018] (2) mixing the toona sinensis leaf powder and the toona sinensis leaf slurry and then granulating; (3) placing the granules obtained by granulation in a supercritical extraction kettle with a feed amount of 100 g of granules / L of the capacity of the extraction kettle, and extracting with supercritical CO2 fluid to separate the natural plant active ingredients; Among them, when supercritical CO2 fluid is used for extraction, the extraction temperature is 50°C, the extraction pressure is 15MPa, the extraction time is 3 hours, the extraction fluid flow rate is 50L / h, and the amount of entrainer ethanol is 80ml ethanol / 100gCO2.

[0019] According to the extract and raw materials, the extraction rate of flavonoids was statistically analyzed, and the specific results are shown in Table 1.

[0020] Table 1

[0021] In addition, it should be noted that the content of flavonoids in this embodiment is determined by using rutin as a standard control in the prior art, and using aluminum chloride (AlCl3) colorimetry to determine the total flavonoid content. The specific operating steps are as follows: 1. Drawing of standard curve 1. Preparation of Rutin Standard Solution Stock solution (0.1 mg / mL): Accurately weigh 10 mg of rutin standard, dissolve it in 70% ethanol and dilute to 100 mL volumetric flask.

[0022] Gradient dilution: Take 0, 1, 2, 3, 4, and 5 mL of the mother solution respectively into a 10-mL volumetric flask and fill up to the mark with 70% ethanol to obtain 0, 10, 20, 30, 40, and 50 μg / mL rutin standard solutions.

[0023] 2. Color reaction Take 2 mL of each concentration standard solution in a 10 mL stoppered test tube, and add: 0.1 mL 5% AlCl3 solution (prepared with ethanol), 3 mL 70% ethanol (make up the volume to 5 mL of the total system), shake well and let stand in the dark for 30 min (AlCl3 and flavonoids form a yellow complex).

[0024] 3. Determine absorbance The blank solution (without rutin, only ethanol + AlCl3) was used as a reference, and the absorbance (A) was measured at a wavelength of 420 nm.

[0025] With rutin concentration (μg / mL) as the abscissa and absorbance (A) as the ordinate, a standard curve was drawn to obtain the regression equation.

[0026] 2. Determination of total flavonoids content in samples 1. Sample preparation Raw material Toona sinensis: After drying and crushing, weigh 0.5 g of sample, extract with 70% ethanol by ultrasonic wave (50℃, 60 min), centrifuge to obtain the supernatant, and make up to 50 mL.

[0027] Supercritical CO2 extract: Weigh an appropriate amount of extract (e.g. 10 mg), dissolve it in 70% ethanol and make up to 25 mL.

[0028] 2. Color development and determination Take 2 mL of sample solution (if the concentration is too high, dilute it to within the linear range) and follow the standard curve color development steps: Add 0.1 mL of 5% AlCl3 solution → add 70% ethanol to 5 mL → keep aside in the dark for 30 min → measure A value.

[0029] Each sample was measured three times in parallel and the average value was taken.

[0030] 3. Calculation of total flavonoid content According to the standard curve regression equation, the flavonoid concentration in the sample solution (C, μg / mL) can be calculated. Calculation formula for total flavonoid content:

[0031] C: Flavonoid concentration of sample solution (μg / mL) V: Sample solution volume (mL) D: dilution factor W: sample mass (g) Example 2

[0032] Extracting coix seed oil from coix seed specifically comprises the following steps: (1) fresh coix seed kernels are prepared into coix seed slurry, and dry coix seed kernels are prepared into coix seed powder; fresh coix seed kernels are kernels obtained within 24 hours after the coix seed fruit is picked, after the shell, yellow-brown seed coat and impurities are removed; When preparing coix seed slurry, it is configured according to the ratio of the mass of coix seed kernel to the volume of water of 1:3, and then ground with a colloid mill to obtain coix seed slurry.

[0033] The particle size of coix seed powder is 50-100 mesh.

[0034] (2) mixing the coix seed powder and the coix seed slurry and then granulating; (3) placing the granules obtained by granulation in a supercritical extraction kettle with a feed amount of 120 g of granules / L of the capacity of the extraction kettle, and extracting with supercritical CO2 fluid to separate the natural plant active ingredients; When the supercritical CO2 fluid is used for extraction, the extraction temperature is 35°C, the extraction pressure is 20MPa, the extraction time is 6 hours, the extraction fluid flow rate is 10L / h, and the amount of entrainer ethanol is 100ml ethanol / 100g CO2.

[0035] According to the extract and raw materials, the extraction rate of coix seed oil was calculated. The specific results are shown in Table 2 below: Table 2

[0036] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that in Comparative Example 1, only the dried material of Toona sinensis leaves is extracted to prepare flavonoids. The dried Toona sinensis leaves used in Comparative Example 1 are from the same batch as those used in Example 1.

[0037] Specifically, the steps include: (1) Prepare toona sinensis leaf powder from dried toona sinensis leaves; the particle size of the toona sinensis leaf powder is 50-100 mesh.

[0038] (2) Granulating the toon leaf powder, placing the granules obtained by granulation in a supercritical extraction kettle, with the feeding amount being 100 g of granules / L of the capacity of the extraction kettle, and extracting with supercritical CO2 fluid to separate the natural plant active ingredients; (3) When the supercritical CO2 fluid is used for extraction, the extraction temperature is 50°C, the extraction pressure is 15 MPa, the extraction time is 3 hours, the extraction fluid flow rate is 50 L / h, and the amount of entrainer ethanol is 80 ml ethanol / 100 g CO2.

[0039] (4) According to the extract and raw materials, the extraction rate of flavonoids was calculated. The specific results are shown in Table 3 below: Table 3

[0040] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that in Comparative Example 2, ultrasonic-assisted extraction is used, and the toona sinensis leaves are not prepared into toona sinensis leaf slurry. Specifically, the following steps are included: (1) taking fresh toona sinensis leaves and preparing toona sinensis leaf powder from dried toona sinensis leaves; the fresh toona sinensis leaves are toona sinensis leaves harvested within 24 hours (the same batch as in Example 1); The particle size of the toon leaf powder is 50-100 meshes.

[0041] (2) Fresh toona sinensis leaves were crushed and placed together with toona sinensis leaf powder in a supercritical extraction kettle with a feed amount of 100 g of particles / L of the extraction kettle capacity. Supercritical CO2 fluid was used for extraction, and dual-frequency ultrasound was used for enhanced extraction to separate natural plant active ingredients. The extraction frequencies were 20 kHz and 65 kHz, the power was 300 W, and the sound intensity was 20 W / cm 2 , the double-frequency alternating irradiation time is 1 hour each, and the total time is 2 hours; Among them, when supercritical CO2 fluid is used for extraction, the extraction temperature is 50°C, the extraction pressure is 15MPa, the extraction time is 3 hours, the extraction fluid flow rate is 50L / h, and the amount of entrainer ethanol is 80ml ethanol / 100gCO2.

[0042] According to the extract and the raw material, the extraction rate of flavonoids was statistically analyzed, and the specific results are shown in Table 4 below: Table 4

[0043] Comparative Example 3 The difference between Comparative Example 3 and Comparative Example 1 is that, while supercritical CO2 fluid is used for extraction, dual-frequency ultrasound is used for enhanced extraction to separate natural plant active ingredients, and the remaining operations are the same.

[0044] According to the extract and the raw material, the extraction rate of flavonoids was statistically analyzed, and the specific results are shown in Table 5 below: Table 5

[0045] It can be seen from Examples 1-2 that the extraction method in the present application can achieve efficient extraction of active ingredients in plants without using ultrasonic-assisted extraction. Comparing Example 1 with Comparative Example 2, although the extraction rate in Example 1 is slightly lower than that in Comparative Example 2, Example 1 is achieved by completely abandoning ultrasonic assistance, and its extraction rate is also at a relatively high level. Due to lower energy consumption and more convenient operation, it is more convenient to use.

[0046] Further, from the comparison between Example 1 and Comparative Example 1 and Comparative Example 3, it can be seen that in Comparative Example 3, when ultrasound-assisted extraction is added, the extraction effect is significantly higher than that in Comparative Example 1, which is only extracted under supercritical CO2 fluid conditions. At the same time, it can also be seen that for the extraction of the dry material of Toona sinensis leaves, the extraction method in this application is used, that is, the powder made of the dry material of Toona sinensis leaves is wrapped and granulated with fresh Toona sinensis leaves, and then supercritical CO2 fluid extraction is directly performed, and efficient extraction of effective ingredients can also be achieved without the aid of ultrasound-assisted extraction. In other words, the active ingredients in natural plant dry materials can be more fully extracted using the fluid extraction method in this application.

[0047] The embodiments of the present application have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technology in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A fluid extraction method for extracting active ingredients from natural plants, characterized in that: The following steps are involved: (1) Prepare natural plant slurry from fresh natural plant materials and prepare natural plant powder from dry natural plant materials; (2) mixing the natural plant powder and the natural plant slurry and then granulating; (3) The granules obtained by granulation are placed in a supercritical extraction kettle and extracted using supercritical CO2 fluid to separate the natural plant active ingredients.

2. A fluid extraction method for extracting active ingredients from natural plants according to claim 1, characterized in that: When preparing the natural plant slurry in step (1), the natural plant slurry is prepared in a ratio of the mass of the natural plant to the volume of water of 1:(2-3), and then the slurry is ground with a colloid mill to obtain the natural plant slurry.

3. A fluid extraction method for extracting active ingredients from natural plants according to claim 1, characterized in that: When supercritical CO2 fluid is used for extraction in step (3), the extraction temperature is 35-50°C, the extraction pressure is 10-20 MPa, the extraction time is 3-10 hours, the extraction fluid flow rate is 10-50 L / h, and the amount of entrainer ethanol is 30-100 ml ethanol / 100 g CO2.

4. A fluid extraction method for extracting active ingredients from natural plants according to claim 1, characterized in that: The particle size of the natural plant powder is 50-100 meshes.

5. A fluid extraction method for extracting active ingredients from natural plants according to any one of claims 1 to 5, characterized in that: The natural plants are Toona plants of the Meliaceae family or Coix plants of the Coix genus.

Images