CN114369026B - Method for extracting chlorogenic acid and rutin from ficus pumila leaves - Google Patents
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Abstract
The invention provides a method for extracting chlorogenic acid and rutin from Ficus pumila leaves, which comprises the following steps: pulverizing Ficus pumila leaves, extracting with ethanol, filtering to obtain extractive solution and residue, concentrating the extractive solution under reduced pressure to obtain concentrated solution, refrigerating overnight, and filtering to obtain crude rutin and filtrate A; adding acid water into the filter residue for extraction and filtration to obtain an acid water extract; mixing the acid water extract with the filtrate A, decolorizing with activated carbon, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate under reduced pressure to obtain concentrated solution A; adding diethyl ether into the concentrated solution A to extract to obtain chlorogenic acid solution, regulating pH to 4, refrigerating overnight, and filtering to obtain chlorogenic acid pure product; dissolving crude rutin with water, filtering while the crude rutin is hot, and extracting the filtrate with ethyl acetate to obtain rutin solution; and (5) carrying out refrigerator overnight on the rutin solution, and filtering to obtain a rutin pure product. The method adopts alcohol extraction and low-temperature crystallization to rapidly separate chlorogenic acid from rutin, avoids the use of column chromatography purification, and effectively improves the utilization rate of functional components in the ficus pumila leaves.
Description
Technical Field
The invention belongs to the technical field of natural products, and particularly relates to a method for extracting chlorogenic acid and rutin from ficus pumila leaves.
Background
The ficus pumila is also called bean jelly, chinese magnolia and the like. Moraceae Ficus plants, climbing or creeping shrubs, leaves, adventitious roots on branches, and leaf egg-shaped hearts. The water washing of the clematis fruit can be used as bean jelly and the vine leaves can be used for medicines. The wild plant resource is widely distributed, the wild plant resource is extremely rich, the growth of the wild plant resource does not select soil or climate, the drought resistance is realized, and the wild plant resource is extremely easy to artificially plant, so the wild plant resource is a novel safe and healthy natural plant resource with development potential.
The ficus pumila fruit serving as a plant for both medicine and food has high medicinal value and comprehensive nutritional ingredients. Pharmacological studies show that the ficus pumila has the main functions of: dispelling wind and removing dampness, promoting blood circulation and dredging collaterals; immunity enhancing and human body function enhancing functions, cardiovascular disease preventing and regulating functions; antibacterial, anti-inflammatory and antitumor functions; has effects of supporting yang, stopping bleeding, promoting lactation, and treating chronic dysentery. Many literature data show that the related research on the ficus pumila is mainly focused on the research on the nutrition components, pectin extraction and drug effects of ficus pumila, the research on the cultivation and propagation of ficus pumila and the comprehensive treatment of stony desertification by using the ficus pumila, but the research on the utilization of ficus pumila leaves is mainly focused on the research on the aspects of tea making and additives, such as the patent on the preparation method 201810885634.8 of ficus pumila leaf fermented tea, the preparation process 201810967946.3 of ficus pumila leaf tea cream, the brewing method 201810868516.6 of ficus pumila leaf health vinegar, the processing method 201610694324.9 of ficus pumila leaf fermented bean curd, the functional component extraction is related to the patent on the ficus pumila leaf polysaccharide and the preparation method thereof and the application 201810474592.9 in the preparation of anticomplement drugs, and the availability is low, and the utilization of flavonoid substances is recently reported. The invention aims to fully utilize the Ficus pumila leaves by adopting the modern extraction technology, separate and extract the functional components such as chlorogenic acid, rutin, ursolic acid and the like which are rich, form a reasonable industry chain for Ficus pumila production and Ficus pumila product development and utilization, and lead the development prospect to be wider.
Disclosure of Invention
The invention aims to provide a method for extracting chlorogenic acid and rutin from Ficus pumila leaves, which aims to solve the problems that Ficus pumila leaves proposed in the background art are not fully utilized, some functional components are not high in extraction rate, and in addition, chlorogenic acid is easy to oxidize, so that the production cost and quality of flavonoid substances of the Ficus pumila leaves are affected.
In order to achieve the above purpose, the present invention provides the following technical solutions:
a method for extracting chlorogenic acid and rutin from Ficus pumila leaves comprises the following steps:
1. pulverizing Ficus pumila leaves, and leaching with 30-50% ethanol solution at room temperature for 10-15 hr, wherein the ratio of weight kg of Ficus pumila leaves to volume L of 30-50% ethanol solution is 1:2-5;
2. filtering to obtain leaching solution and filter residue, adding acid water into the filter residue, stirring at 40-60deg.C, extracting for 3-5 h, cooling, and filtering to obtain acid water extract, wherein the ratio of She Chongliang kg of Ficus pumila to L of acid water is 1:2-5;
3. concentrating the leaching solution at 55-60deg.C under reduced pressure to 1/3-1/10 of the original volume to obtain concentrated solution, placing into a refrigerator at 4deg.C overnight, and filtering to obtain crude rutin and filtrate A;
4. mixing the acid water extract with the filtrate A, adding activated carbon for decolorization for 30-50min, and filtering to obtain decolorized filtrate, wherein the ratio of the weight g of the activated carbon to the volume L of the mixed solution is 10-30:1, a step of;
5. concentrating the decolorized filtrate at 55-60deg.C under reduced pressure to 1/5-1/10 of the original volume to obtain concentrated solution A;
6. extracting the concentrated solution A with diethyl ether of 2-4 times volume, and removing diethyl ether layer to obtain chlorogenic acid solution;
7. adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution to adjust pH to 4, placing into a refrigerator at 4deg.C overnight, and filtering to obtain chlorogenic acid pure product;
8. dissolving crude rutin with 50-80deg.C water, filtering while it is hot, adding ethyl acetate 2-4 times of volume of the filtrate, extracting, removing ethyl acetate layer to obtain rutin solution, wherein the ratio of volume L of water to crude rutin kg is 1-3:1, a step of;
9. placing the rutin solution in a refrigerator at 4deg.C overnight, and filtering to obtain rutin pure product.
Preferably, the 30-50% ethanol solution is prepared by adding water into ethanol, wherein the 30% ethanol solution is prepared by adding 70ml water into 30ml ethanol and mixing.
Preferably, the acid water is hydrochloric acid water solution with pH of 1-2.5, and 0.1mol/L hydrochloric acid solution is used for adjusting.
Preferably, the filtration is performed 1-2 times by using a plate frame.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention adopts the modern separation technology of combining alcohol extraction and low-temperature crystallization phase to rapidly separate chlorogenic acid from rutin, avoids the use of column chromatography purification, and effectively improves the utilization rate of functional components in ficus pumila leaves.
(2) The method comprises the steps of alcohol extraction and acid water high-temperature extraction, so that the risk of hydrolysis in the rutin extraction process is effectively avoided, and the chlorogenic acid extraction rate is improved.
Drawings
Fig. 1: rutin and chlorogenic acid are mixed into a standard sample chromatogram;
fig. 2: rutin pure product chromatogram;
fig. 3: chlorogenic acid pure chromatogram.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
100kg of Ficus pumila leaves are crushed, and 300L of 50% ethanol solution is adopted for room temperature leaching for 15h; filtering to obtain leaching solution and filter residue, adding 350L acid water into the filter residue, stirring at 60 ℃ for extraction for 5h, cooling, and filtering to obtain 300L acid water extract; concentrating the leaching solution at 55-60deg.C under reduced pressure to obtain 40L concentrate, placing into a refrigerator at 4deg.C overnight, and filtering to obtain 1360g of crude rutin and filtrate A; mixing the acid water extract with the filtrate A, adding 3500g of activated carbon for decolorizing for 40min, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate at 55-60deg.C under reduced pressure to obtain 40L concentrate A; extracting the concentrated solution A with 100L diethyl ether, and removing diethyl ether layer to obtain chlorogenic acid solution; adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution to adjust the pH to 4, placing the chlorogenic acid solution into a refrigerator at 4 ℃ for overnight, and filtering to obtain 285g chlorogenic acid pure product; dissolving crude rutin with 60 deg.C water 3L, filtering, adding 10L ethyl acetate into the filtrate, extracting, and removing ethyl acetate layer to obtain rutin solution; the rutin solution is placed in a refrigerator at 4 ℃ for overnight, and 861g of rutin pure product is obtained by filtration.
The detection equipment is an LC-20AT high performance liquid chromatograph, and the chromatographic column is InertSustatin C 18 Column (4.6mm.times.250mm, 5 μm), chlorogenic acid detection conditions: mobile phase of 18% acetonitrile, 82% 0.2% phosphoric acid water solution, detection wavelength of 326nm, flow rate of L ml/min, sample injection amount of 20 μl, column temperature of 35deg.C; liquid chromatography detection conditions of rutin: the mobile phase is 18% acetonitrile, 82% 0.2% phosphoric acid aqueous solution, the detection wavelength is 254nm, the flow rate is L ml/min, the sample injection amount is 20 mu L, and the column temperature is 35 ℃. The purity of chlorogenic acid pure product is 99.57%, and the purity of rutin pure product is 99.24%, see figure 2 and figure 3.
Example 2
200kg of ficus pumila leaves are crushed, and 700L of 40% ethanol solution is adopted for room temperature leaching for 14h; filtering to obtain leaching solution and filter residue, adding 600L acid water into the filter residue, stirring and extracting at 50 ℃ for 4 hours, cooling, and filtering to obtain 550L acid water extract; concentrating the leaching solution at 55-60deg.C under reduced pressure to obtain 75L concentrate, placing into a refrigerator at 4deg.C overnight, and filtering to obtain 2980g of crude rutin and filtrate A; mixing the acid water extract with the filtrate A, decolorizing with 10kg of activated carbon for 30min, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate at 55-60deg.C under reduced pressure to obtain 65L concentrate A; extracting the concentrated solution A with 150L diethyl ether, and removing diethyl ether layer to obtain chlorogenic acid solution; adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution to adjust the pH to 4, placing the chlorogenic acid solution into a refrigerator at 4 ℃ for overnight, and filtering to obtain 584g chlorogenic acid pure product; dissolving crude rutin with 70deg.C water 7L, filtering, adding 20L ethyl acetate into the filtrate, extracting, and removing ethyl acetate layer to obtain rutin solution; placing the rutin solution in a refrigerator at 4deg.C overnight, and filtering to obtain 1725g rutin pure product. The purity of the product was examined by the examination method of example 1, wherein the purity of chlorogenic acid was 99.42% and the purity of rutin was 99.35%.
Comparative example 1
Pulverizing 100kg of Ficus pumila leaves, leaching with 50% ethanol solution 300L at room temperature for 15 hr, and filtering to obtain leaching solution and residue; concentrating the leaching solution at 55-60deg.C under reduced pressure to obtain 40L concentrate, placing into a refrigerator at 4deg.C overnight, and filtering to obtain 1360g of crude rutin and filtrate A; adding 500g of activated carbon into the filtrate A for decoloration for 40min, and filtering to obtain decolored filtrate; vacuum concentrating the decolorized filtrate at 55-60deg.C to obtain 5L concentrate A; adding 15L diethyl ether into the concentrated solution A for extraction, and removing diethyl ether layer to obtain chlorogenic acid solution; adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution to adjust the pH to 4, placing the chlorogenic acid solution into a refrigerator at 4 ℃ for overnight, and filtering to obtain 124g chlorogenic acid pure product; dissolving crude rutin with 60 deg.C water 3L, filtering, adding 10L ethyl acetate into the filtrate, extracting, and removing ethyl acetate layer to obtain rutin solution; the rutin solution is placed in a refrigerator at 4 ℃ for overnight, and 861g of rutin pure product is obtained by filtration. The purity of the product was examined by the examination method of example 1, wherein the purity of chlorogenic acid was 99.48% and the purity of rutin was 99.24%.
Comparative example 2
Pulverizing 100kg of Ficus pumila leaves, adding 350L of acid water, stirring at 60deg.C, extracting for 5 hr, cooling, and filtering to obtain 300L of acid water extract; adding 3500g of active carbon into the acid water extract for decoloring for 40min, and filtering to obtain decolored filtrate; concentrating the decolorized filtrate at 55-60deg.C under reduced pressure to obtain 40L concentrate A; extracting the concentrated solution A with 100L diethyl ether, and removing diethyl ether layer to obtain chlorogenic acid solution; adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution to adjust pH to 4, placing into a refrigerator at 4deg.C overnight, and filtering to obtain 264g chlorogenic acid pure product. The purity of the product was measured by the method of example 1, wherein the purity of chlorogenic acid was 99.23%.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (4)
1. A method for extracting chlorogenic acid and rutin from ficus pumila leaves is characterized by comprising the following steps:
(1) Pulverizing Ficus pumila leaves, and leaching with 30-50% ethanol solution at room temperature for 10-15 hr, wherein the ratio of She Chongliang kg of Ficus pumila to 30-50% ethanol solution at volume L is 1:2-5;
(2) Filtering to obtain leaching solution and filter residue, adding acid water into the filter residue, stirring at 40-60deg.C, extracting for 3-5 h, cooling, and filtering to obtain acid water extract, wherein the ratio of She Chongliang kg of Ficus pumila to L of acid water is 1:2-5;
(3) Concentrating the leaching solution at 55-60deg.C under reduced pressure to 1/3-1/10 of the original volume to obtain concentrated solution, placing into a refrigerator at 4deg.C overnight, and filtering to obtain crude rutin and filtrate A;
(4) Mixing the acid water extract with the filtrate A, adding activated carbon for decolorization for 30-50min, and filtering to obtain decolorized filtrate, wherein the ratio of the weight g of the activated carbon to the volume L of the mixed solution is 10-30:1, a step of;
(5) Concentrating the decolorized filtrate at 55-60deg.C under reduced pressure to 1/5-1/10 of the original volume to obtain concentrated solution A;
(6) Extracting the concentrated solution A with diethyl ether of 2-4 times volume, and removing diethyl ether layer to obtain chlorogenic acid solution;
(7) Adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution to adjust pH to be equal to 4, placing into a refrigerator at 4deg.C overnight, and filtering to obtain chlorogenic acid pure product;
(8) Dissolving crude rutin with 50-80deg.C water, filtering while it is hot, adding ethyl acetate 2-4 times of volume of the filtrate, extracting, removing ethyl acetate layer to obtain rutin solution, wherein the ratio of volume L of water to crude rutin kg is 1-3:1, a step of;
(9) Placing the rutin solution in a refrigerator at 4deg.C overnight, and filtering to obtain rutin pure product.
2. The method for extracting chlorogenic acid and rutin from Ficus pumila leaves according to claim 1, wherein the ethanol solution with volume concentration of 30-50% is prepared by adding water into ethanol.
3. The method for extracting chlorogenic acid and rutin from the leaves of ficus pumila of claim 1, wherein said acid water is aqueous hydrochloric acid solution having a pH of 1-2.5, and is adjusted with 0.1mol/L hydrochloric acid solution.
4. The method for extracting chlorogenic acid and rutin from the leaves of ficus pumila of claim 1, wherein said filtering is performed 1-2 times with a plate frame.
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