CN114369026A - Method for extracting chlorogenic acid and rutin from Ficus pumila leaves - Google Patents
Method for extracting chlorogenic acid and rutin from Ficus pumila leaves Download PDFInfo
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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: crushing climbing fig leaves, extracting with ethanol, filtering to obtain an extract and filter residues, concentrating the extract under reduced pressure to obtain a concentrated solution, standing overnight in a refrigerator, and filtering to obtain crude rutin and a filtrate A; adding acid water into the filter residue, extracting and filtering to obtain an acid water extracting solution; mixing the acid water extract and the filtrate A, and decolorizing and filtering with activated carbon to obtain decolorized filtrate; concentrating the decolorized filtrate under reduced pressure to obtain concentrated solution A; extracting the concentrated solution A with diethyl ether to obtain chlorogenic acid solution, adjusting pH to 4, refrigerating overnight, and filtering to obtain chlorogenic acid pure product; dissolving the crude rutin with water, filtering while hot, and extracting the filtrate with ethyl acetate to obtain a rutin solution; and (4) refrigerating the rutin solution overnight, and filtering to obtain a pure rutin product. The method adopts alcohol extraction and low-temperature crystallization to quickly separate the chlorogenic acid and the rutin, avoids column chromatography purification, and effectively improves the utilization rate of functional components in the climbing fig 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
Ficus pumila is also called bean jelly, and Mulian, etc. The plant of Ficus of Moraceae, climbing or creeping shrub, leaf type, adventitious root on the branch node of the fruiting tree, and leaf-egg-shaped heart shape. The cleome fruit can be used as bean jelly and vine leaf. Wide distribution, extremely rich wild resources, no soil and climate selection for growth, drought resistance and easy artificial planting, and is a safe and healthy novel natural plant resource with development potential.
The ficus pumila fruit serving as a medicine-food dual-purpose plant not only has high medicinal value, but also has comprehensive nutritional ingredients. Pharmacological research shows that the Ficus pumila has the following main functions: functions of dispelling wind and eliminating dampness, promoting blood circulation and removing obstruction in channels; immunity enhancing and human body function enhancing functions, cardiovascular disease preventing and regulating functions; antibacterial, anti-inflammatory and anti-tumor functions; and has effects in supporting yang, stopping nocturnal emission, stopping bleeding, promoting lactation, and treating chronic dysentery and intestinal diseases. Many literature data show that the research on the ficus pumila is mainly focused on the research on the nutrient components, pectin extraction and drug effect of ficus pumila fruits and the research on the comprehensive stony desertification control by using the ficus pumila cultivation and propagation, but the research on the utilization of ficus pumila leaves is also mainly focused on the research on the aspects of tea making and additives, such as the patent 'preparation method 201810885634.8 of uniflower ficus pumila leaf fermented tea, the preparation process 201810967946.3 of uniflower ficus pumila leaf tea paste, the brewing method 201810868516.6 of uniflower ficus pumila leaf health vinegar and the processing method 201610694324.9 of uniflower ficus pumila leaf nutrient preserved beancurd', although the extraction of functional components is involved in the patent 'Ficus pumila leaf polysaccharide and the preparation method thereof and the application 201810474592.9 in the preparation of anticomplement drugs', the utilization degree is low, and particularly, the utilization of flavonoid substances is reported. The invention aims to fully utilize the leaves of the Ficus pumila by adopting the modern extraction technology, separate and extract functional components such as chlorogenic acid, rutin, ursolic acid and the like, form a reasonable industrial chain of Ficus pumila production and Ficus pumila product development and utilization, and have wider development prospect.
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 the Ficus pumila leaves proposed in the background art are not fully utilized, the extraction rate of some functional components is low, and the chlorogenic acid is easily oxidized to influence the production cost and quality of flavonoid substances of the chlorogenic acid.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for extracting chlorogenic acid and rutin from Ficus pumila leaves comprises the following steps:
1. crushing Ficus pumila leaves, and leaching at room temperature for 10-15 h by adopting 30-50% ethanol solution, wherein the volume ratio L of the weight kg of the Ficus pumila leaves to the volume L of the 30-50% ethanol solution is 1: 2-5;
2. filtering to obtain leaching liquor and filter residue, adding acid water into the filter residue, stirring and extracting at 40-60 ℃ for 3-5 h, cooling, and filtering to obtain acid water extracting solution, wherein the ratio of weight kg of ficus pumila leaves to volume L of the acid water is 1: 2-5;
3. concentrating the leaching solution at 55-60 deg.C under reduced pressure to 1/3-1/10 of original volume to obtain concentrated solution, placing in 4 deg.C refrigerator overnight, and filtering to obtain crude rutin and filtrate A;
4. and (3) combining the acid water extracting solution and the filtrate A, adding activated carbon for decoloring for 30-50min, and filtering to obtain a decolored filtrate, wherein the ratio of the weight g of the activated carbon to the volume L of the combined solution is 10-30: 1;
5. concentrating the decolorized filtrate at 55-60 deg.C under reduced pressure to 1/5-1/10 of the original volume to obtain concentrated solution A;
6. adding 2-4 times of diethyl ether into the concentrated solution A for extraction, and removing diethyl ether layer to obtain chlorogenic acid solution;
7. adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution, adjusting pH to 4, placing in refrigerator at 4 deg.C overnight, and filtering to obtain chlorogenic acid pure product;
8. dissolving the crude rutin with water of 50-80 ℃, filtering while hot, adding ethyl acetate of 2-4 times of the volume of the filtrate for extraction, removing an ethyl acetate layer to obtain a rutin solution, wherein the ratio of the volume L of the added water to the kg of the crude rutin is 1-3: 1;
9. and putting the rutin solution in a refrigerator at 4 ℃ overnight, and filtering to obtain a pure rutin 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.
Preferably, the acid water is hydrochloric acid aqueous solution with pH of 1-2.5, and is adjusted by 0.1mol/L hydrochloric acid solution.
Preferably, the filtration is performed for 1-2 times by adopting plate and frame filtration.
Compared with the prior art, the invention has the beneficial effects that:
(1) the method adopts the modern separation technology combining alcohol extraction and low-temperature crystallization phase, so that the chlorogenic acid and the rutin are quickly separated, column chromatography purification is avoided, and the utilization rate of functional components in the climbing fig leaves is effectively improved.
(2) The method firstly extracts alcohol and then extracts acid water at high temperature, thereby effectively avoiding the risk of hydrolysis in the rutin extraction process and improving the extraction rate of chlorogenic acid.
Drawings
FIG. 1: a mixed standard sample chromatogram of rutin and chlorogenic acid;
FIG. 2: a chromatogram of the pure rutin product;
FIG. 3: chromatogram of chlorogenic acid.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Pulverizing Ficus Pumila leaves 100kg, and extracting with 50% ethanol solution 300L at room temperature for 15 hr; filtering to obtain leaching solution and residue, adding 350L acid water into the residue, stirring at 60 deg.C for 5 hr, cooling, and filtering to obtain 300L acid water extractive solution; concentrating the leaching solution at 55-60 deg.C under reduced pressure to obtain 40L concentrated solution, placing in 4 deg.C refrigerator overnight, and filtering to obtain 1360g of crude rutin and filtrate A; mixing the acid water extractive solution and filtrate A, adding 3500g activated carbon for decolorizing for 40min, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate at 55-60 deg.C under reduced pressure to obtain 40L concentrated solution A; adding 100L 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, adjusting pH to 4, placing in refrigerator at 4 deg.C overnight, and filtering to obtain 285g chlorogenic acid pure product; dissolving the crude rutin with 3L of water at 60 deg.C, filtering while hot, adding 10L of ethyl acetate into the filtrate, extracting, and removing ethyl acetate layer to obtain rutin solution; the rutin solution is put into a refrigerator at 4 ℃ overnight, and 861g of rutin pure product is obtained by filtering.
The detection equipment is an LC-20AT high performance liquid chromatograph, and the chromatographic column is Inertsustatin C18Column (4.6mm × 250mm, 5 μm), chlorogenic acid detection condition with mobile phase of 18% acetonitrile and 82% 0.2% phosphoric acid aqueous solution, detection wavelength of 326nm, flow rate of L ml/min, sample injection amount of 20 μ L, and column temperature of 35 deg.C; the liquid chromatography detection condition of rutin is as follows: the mobile phase is 18% acetonitrile and 82% 0.2% phosphoric acid water 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%, shown in fig. 2 and fig. 3.
Example 2
Pulverizing Ficus Pumila leaves 200kg, and extracting with 40% ethanol solution 700L at room temperature for 14 h; filtering to obtain leaching solution and residue, adding 600L acid water into the residue, stirring and extracting at 50 deg.C for 4 hr, cooling, and filtering to obtain 550L acid water extractive solution; concentrating the leaching solution at 55-60 deg.C under reduced pressure to obtain 75L concentrated solution, placing in 4 deg.C refrigerator overnight, and filtering to obtain 2980g crude rutin and filtrate A; mixing the acid water extractive solution and filtrate A, adding 10kg of active carbon for decolorizing for 30min, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate at 55-60 deg.C under reduced pressure to obtain 65L concentrated solution 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 the chlorogenic acid solution to adjust the pH to 4, putting the chlorogenic acid solution into a refrigerator at 4 ℃ for overnight, and filtering to obtain 584g of pure chlorogenic acid; dissolving the crude rutin with water 7L at 70 deg.C, filtering while hot, adding ethyl acetate 20L into the filtrate, extracting, and removing ethyl acetate layer to obtain rutin solution; the rutin solution is put into a refrigerator at 4 ℃ overnight, and filtered to obtain 1725g of pure rutin. And (3) detecting the purity of the product according to the detection method of the embodiment 1, wherein the purity of the pure chlorogenic acid product is 99.42 percent, and the purity of the pure rutin product is 99.35 percent.
Comparative example 1
Crushing 100kg of climbing fig leaves, leaching for 15h at room temperature by adopting 300L of 50% ethanol solution, and filtering to obtain leaching liquor and filter residues; concentrating the leaching solution at 55-60 deg.C under reduced pressure to obtain 40L concentrated solution, placing in 4 deg.C refrigerator overnight, and filtering to obtain 1360g of crude rutin and filtrate A; adding 500g of activated carbon into the filtrate A for decolorization for 40min, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate at 55-60 deg.C under reduced pressure to obtain 5L concentrated solution 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, adjusting pH to 4, placing in refrigerator at 4 deg.C overnight, and filtering to obtain pure chlorogenic acid 124 g; dissolving the crude rutin with 3L of water at 60 deg.C, filtering while hot, adding 10L of ethyl acetate into the filtrate, extracting, and removing ethyl acetate layer to obtain rutin solution; the rutin solution is put into a refrigerator at 4 ℃ overnight, and 861g of rutin pure product is obtained by filtering. And (3) detecting the purity of the product according to the detection method of the embodiment 1, wherein the purity of the pure chlorogenic acid is 99.48%, and the purity of the pure rutin is 99.24%.
Comparative example 2
Pulverizing Ficus Pumila leaves 100kg, adding 350L acid water, stirring at 60 deg.C for 5 hr, cooling, and filtering to obtain 300L acid water extractive solution; adding 3500g activated carbon into the acid water extractive solution, decolorizing for 40min, and filtering to obtain decolorized filtrate; concentrating the decolorized filtrate at 55-60 deg.C under reduced pressure to obtain 40L concentrated solution A; adding 100L 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, adjusting pH to 4, placing in refrigerator at 4 deg.C overnight, and filtering to obtain 264g chlorogenic acid pure product. The purity of the product was tested according to the test method of example 1, wherein the purity of the pure chlorogenic acid was 99.23%.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments 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) crushing Ficus pumila leaves, and leaching at room temperature for 10-15 h by adopting 30-50% ethanol solution, wherein the volume ratio L of the weight kg of the Ficus pumila leaves to the volume L of the 30-50% ethanol solution is 1: 2-5;
(2) filtering to obtain leaching liquor and filter residue, adding acid water into the filter residue, stirring and extracting at 40-60 ℃ for 3-5 h, cooling, and filtering to obtain acid water extracting solution, wherein the ratio of weight kg of ficus pumila leaves to volume L of the acid water is 1: 2-5;
(3) concentrating the leaching solution at 55-60 deg.C under reduced pressure to 1/3-1/10 of original volume to obtain concentrated solution, placing in 4 deg.C refrigerator overnight, and filtering to obtain crude rutin and filtrate A;
(4) and (3) combining the acid water extracting solution and the filtrate A, adding activated carbon for decoloring for 30-50min, and filtering to obtain a decolored filtrate, wherein the ratio of the weight g of the activated carbon to the volume L of the combined solution is 10-30: 1;
(5) concentrating the decolorized filtrate at 55-60 deg.C under reduced pressure to 1/5-1/10 of the original volume to obtain concentrated solution A;
(6) adding 2-4 times of diethyl ether into the concentrated solution A for extraction, and removing diethyl ether layer to obtain chlorogenic acid solution;
(7) adding 0.1mol/L sodium hydroxide solution into chlorogenic acid solution, adjusting pH to 4, placing in refrigerator at 4 deg.C overnight, and filtering to obtain chlorogenic acid pure product;
(8) dissolving the crude rutin with water of 50-80 ℃, filtering while hot, adding ethyl acetate of 2-4 times of the volume of the filtrate for extraction, removing an ethyl acetate layer to obtain a rutin solution, wherein the ratio of the volume L of the added water to the kg of the crude rutin is 1-3: 1;
(9) and putting the rutin solution in a refrigerator at 4 ℃ overnight, and filtering to obtain a pure rutin product.
2. The method of claim 1, wherein the 30-50% ethanol solution is prepared by adding water to ethanol.
3. The process according to claim 1, characterized in that the acid water is an aqueous hydrochloric acid solution having a pH of 1-2.5, adjusted with a 0.1mol/L hydrochloric acid solution.
4. The method according to claim 1, wherein the filtration is performed 1-2 times using plate and frame filtration.
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- 2021-12-06 CN CN202111478686.1A patent/CN114369026B/en active Active
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| CN103204765A (en) * | 2013-01-30 | 2013-07-17 | 湖南中烟工业有限责任公司 | Method for extracting solanesol and chlorogenic acid from discard tobacco leaves |
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