CN112592329B - Method for extracting apigenin from wild celery and application of apigenin - Google Patents
Method for extracting apigenin from wild celery and application of apigenin Download PDFInfo
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Abstract
The invention relates to the technical field of plant extraction, in particular to a method for extracting apigenin from parsley and application thereof, wherein the method comprises the following steps: (1) drying and crushing the wild celery, adding a solvent A for extraction, and concentrating an extracting solution to obtain a substance A; (2) extracting the substance A with a solvent B, and drying the extracted solvent B layer to obtain a crude product; (3) dissolving the crude product with a solvent C, adsorbing with macroporous adsorption resin, and eluting with a solvent D to obtain an eluent; (4) concentrating and crystallizing the eluent to obtain the product. The invention has the advantages of high yield and purity, and obvious effect of reducing uric acid by combining the apigenin and other effective components.
Description
Technical Field
The invention relates to the technical field of plant extraction, and particularly relates to a method for extracting apigenin from wild celery and application of the method.
Background
The wild celery is a perennial herb plant of the family Umbelliferae and the genus wild celery, and has medicinal value and edible value. The pittosporum tobira has the efficacies of mainly treating rheumatic arthralgia, soreness and pain of waist and knees, cold headache, carbuncle, sore and swelling pain and the like. The wild celery is a high-grade precious vegetable in celery. The nutrient components of the wild celery are higher in the wild vegetables, and the wild celery seedlings can be used as the wild vegetables in spring. The nutrition effect is as follows: (1) celery has high iron content, can supplement the loss of menstrual blood of women, can avoid pale, dry and lusterless skin when eaten, and can lead eyes to be vivid and the hair to be black and bright. (2) Celery is a high fiber food which produces a lignin or intestinal lipid substance by intestinal digestion, which is an antioxidant and a carcinogenic substance which inhibits intestinal bacteria at high concentrations. It also accelerates the transit time of feces in the intestine; reduce the contact of carcinogen and colon mucosa to prevent colon cancer.
Apigenin (Apigenin), also known as Apigenin and Apigenin, is a systematic name of 4',5, 7-trihydroxyflavone, english name Apigenin, CAS number: 520-36-5, is a flavonoid compound and is widely distributed in the nature. Mainly exists in plants of Thymelaeaceae, Verbenaceae and Selaginellaceae, and is widely distributed in vegetables and fruits in temperate zone, especially high in celery content. The structural formula is as follows:
apigenin has effects of scavenging free radicals, improving immunity, resisting oxidation, resisting hyperglycemia and hypertension, resisting inflammation, improving immunity, resisting atherosclerosis, and inhibiting cancer cells. However, the prior art has few methods for extracting apigenin, and the yield and purity in the extraction process need to be further improved.
Chinese patent application CN104876901A discloses a method for preparing apigenin from celery, which comprises the following steps: ethanol reflux extraction; hydrolyzing and filtering; recrystallizing; centrifuging; drying; pulverizing and sieving to obtain apigenin finished product. The preparation method has simple route and cheap raw materials. Is prepared from celery with rich sources. Low cost and easy operation. However, the yield and purity of apigenin in the patent application are to be further improved.
Chinese patent application CN110551092A discloses an apigenin extraction method, comprising the following steps: drying and crushing celery, and sieving the celery with a 40-mesh sieve; adding a polyol mixed system of a nonionic surfactant into the raw materials, extracting by adopting ultrasonic waves with the frequency of 60KHz, filtering, and concentrating the filtrate until no alcohol smell exists; loading the obtained concentrated solution onto a DF01 type macroporous adsorption resin column, eluting with chloroform with the volume of 4-6 times of that of the column and being 60-70%, collecting eluent, and concentrating; recovering the reagent to obtain white powder, namely apigenin. The extraction method of the invention is simple and easy to implement, has high yield, does not need a large amount of organic solvents, is safe and nontoxic, and is suitable for industrial production. However, the purity of apigenin in the patent application is low.
Uric acid is the main metabolite of birds and reptiles, and has the chemical formula C5H4N4O3Slightly soluble in water and prone to form crystals. The product in normal human urine is mainly urea and contains a small amount of uric acid. Uric acid is the end product of purine metabolism, is trioxypurine, and has an alcohol formula with weak acidity. Uric acid produced by oxidation of various purines is excreted with urine.
Normally, uric acid in the body is about 1200 mg, which is newly produced to about 600 mg every day, and 600 mg is excreted and kept in equilibrium. However, if too much uric acid is produced in vivo and excretion is delayed or the uric acid excretion mechanism is degraded, the uric acid in vivo is retained too much, and when the blood uric acid concentration is more than 7 mg/dl, the body fluid of a human body turns acid, the normal function of human body cells is affected, and gout can be caused if the normal function of the cells of the human body is influenced. In addition, over-fatigue or under-rest can also lead to relative retardation of metabolism and thus gout development.
Uric acid in blood is filtered from glomerulus, and the production and excretion speeds of uric acid in normal human bodies are basically constant. The change of uric acid content in body fluid can fully reflect the conditions of metabolism, immunity and other functions in human body. There are four main reasons for uric acid accumulation: the first is the excessive consumption of high purine food; second, problems with purine metabolism occur in vivo; thirdly, the excretion is too small; fourth, uric acid cannot be excreted normally.
The prior art does not disclose the report of using the apigenin for reducing uric acid.
Therefore, it is necessary to develop a method for extracting apigenin from parsley and an application thereof, which can solve the above technical problems.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a method for extracting apigenin from wild celery with higher yield and purity, and the effect of reducing uric acid is obvious by combining the wild celery with other effective components.
The invention is realized by the following technical scheme:
a method for extracting apigenin from wild celery comprises the following steps:
(1) drying and crushing the wild celery, adding a solvent A for extraction, and concentrating an extracting solution to obtain a substance A;
(2) extracting the substance A with a solvent B, and drying the extracted solvent B layer to obtain a crude product;
(3) dissolving the crude product with a solvent C, adsorbing with macroporous adsorption resin, and eluting with a solvent D to obtain an eluent;
(4) concentrating and crystallizing the eluent to obtain the product.
Preferably, the solvent A in the step (1) is a mixed solvent of ethylene glycol, isopropanol and water, and the volume ratio of the ethylene glycol, the isopropanol and the water is 2-4:1: 1-3.
Preferably, the liquid-to-material ratio of the solvent A to the parsley in the step (1) is 5-10 mL/g.
Preferably, the temperature of the extraction in the step (1) is 30-40 ℃, and the extraction time is 1-2 h.
Preferably, in the step (1), the enzymolysis is carried out while adding the solvent A into the parsley.
More preferably, the enzyme used for enzymolysis is at least one of xylanase and glucosidase.
More preferably, in the step (1), 3-8% of xylanase is added for enzymolysis for 10-25min and 5-10% of glucosidase is added at the same time of adding the solvent A into the parsley. The dosage percentages of the xylanase and the glucosidase are the mass percentages of the two enzymes in the solvent A.
More preferably, step (1) comprises the steps of:
drying and crushing the wild celery, adding a solvent A in an amount which is 5-10 times that of the wild celery for extraction, simultaneously adding 3-8% of xylanase for enzymolysis for 10-25min, then adding 5-10% of glucosidase, wherein the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the solvent A to the solvent A is 2-4:1:1-3, the extraction temperature is 30-40 ℃, the total extraction time is 1-2h, and the extracting solution is concentrated to 1/5-1/8 of the original volume to obtain a substance A.
Preferably, the solvent B in the step (2) is at least one of petroleum ether, chloroform and ethyl acetate.
More preferably, in the step (2), petroleum ether, chloroform and ethyl acetate are sequentially added for extraction, and the chloroform and ethyl acetate extraction layers are collected, dried respectively and combined to obtain a crude product.
More preferably, the step (2) comprises the steps of:
and sequentially adding petroleum ether, chloroform and ethyl acetate with the same volume into the substance A for extraction, collecting the chloroform and ethyl acetate extraction layers, respectively drying, and combining to obtain a crude product.
Preferably, the solvent C in step (3) is 30-90% ethanol.
Preferably, the macroporous adsorption resin in the step (3) is D101 macroporous adsorption resin.
Preferably, the solvent D in the step (3) is a mixed solvent of chloroform and methanol, and the volume ratio of the chloroform to the methanol is 1-3: 1.
More preferably, the step (3) comprises the steps of:
dissolving the crude product with 30-90% ethanol, adsorbing with D101 macroporous adsorbent resin, and eluting with solvent D, wherein the solvent D is a mixed solvent of chloroform and methanol at a volume ratio of 1-3:1, to obtain eluate.
More preferably, the method comprises the steps of:
(1) drying and crushing the wild celery, adding a solvent A in an amount which is 5-10 times that of the wild celery for extraction, simultaneously adding 3-8% of xylanase for enzymolysis for 10-25min, then adding 5-10% of glucosidase, wherein the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the solvent A to the solvent A is 2-4:1:1-3, the extraction temperature is 30-40 ℃, the total extraction time is 1-2h, and the extracting solution is concentrated to 1/8-1/5 of the original volume to obtain a substance A;
(2) sequentially adding petroleum ether, chloroform and ethyl acetate with the same volume into the substance A for extraction, collecting the extraction layers of chloroform and ethyl acetate, respectively drying, and combining to obtain a crude product;
(3) dissolving the crude product with 30-90% ethanol, adsorbing with D101 macroporous adsorbent resin, and eluting with solvent D, wherein the solvent D is a mixed solvent of chloroform and methanol at a volume ratio of 1-3:1 to obtain an eluate;
(4) heating the eluate, concentrating, cooling, crystallizing, and drying.
The invention also relates to a composition, which comprises the apigenin prepared by the method.
Preferably, the composition comprises the following components in parts by weight: 20-35 parts of apigenin, 15-20 parts of theine, 0.5-2 parts of folic acid and 5-10 parts of dihydrooroselagin.
The invention also relates to application of the composition in preparation of a medicament for reducing uric acid.
The invention has the beneficial effects that:
the invention optimizes the composition of the solvent A, so that apigenin is extracted as much as possible, and particularly, the yield of the prepared apigenin is higher by adding an enzymolysis process and optimizing a specific enzymolysis process.
The invention optimizes the composition and the extraction sequence of the solvent B and improves the yield and the purity of the apigenin.
The invention optimizes the composition of the solvent D and further improves the yield and the purity of the apigenin.
The composition of the invention has synergistic effect among the components, and the effect of reducing uric acid is obviously improved.
Detailed Description
The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
Example 1
A method for extracting apigenin from wild celery comprises the following steps:
(1) drying and crushing the wild celery, adding a solvent A in an amount which is 5 times that of the wild celery for extraction, simultaneously adding 3% of xylanase for enzymolysis for 10min, then adding 5% of glucosidase, wherein the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the solvent A to the solvent A is 2:1:1, the extraction temperature is 30 ℃, the total extraction time is 1h, and concentrating the extracting solution to 1/5 of the original volume to obtain a substance A;
(2) sequentially adding petroleum ether, chloroform and ethyl acetate with the same volume into the substance A for extraction, collecting the extraction layers of chloroform and ethyl acetate, respectively drying, and combining to obtain a crude product;
(3) dissolving the crude product with 3 times of 30% ethanol, adsorbing with D101 macroporous adsorption resin, and eluting with solvent D, wherein the solvent D is a mixed solvent of chloroform and methanol at a volume ratio of 1:1, to obtain an eluate;
(4) heating the eluate, concentrating, cooling, crystallizing, and drying.
Example 2
A method for extracting apigenin from wild celery comprises the following steps:
(1) drying and crushing the wild celery, adding a 10-time solvent A for extraction, simultaneously adding 8% xylanase for enzymolysis for 25min, adding 10% glucosidase, wherein the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the solvent A to the solvent A is 4:1:3, the extraction temperature is 40 ℃, the total extraction time is 2h, and concentrating the extracting solution to 1/8 of the original volume to obtain a substance A;
(2) sequentially adding petroleum ether, chloroform and ethyl acetate with the same volume into the substance A for extraction, collecting the extraction layers of chloroform and ethyl acetate, respectively drying, and combining to obtain a crude product;
(3) dissolving the crude product with 5 times of 90% ethanol, adsorbing with D101 macroporous adsorption resin, and eluting with solvent D, wherein the solvent D is a mixed solvent of chloroform and methanol at a volume ratio of 3:1, to obtain an eluate;
(4) heating the eluate, concentrating, cooling, crystallizing, and drying.
Example 3
A method for extracting apigenin from wild celery comprises the following steps:
(1) drying and crushing the wild celery, adding 8 times of solvent A for extraction, simultaneously adding 5% xylanase for enzymolysis for 15min, adding 7% glucosidase, wherein the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the solvent A to the solvent A is 3:1:2, the extraction temperature is 35 ℃, the total extraction time is 1.5h, and concentrating the extracting solution to 1/6 of the original volume to obtain a substance A;
(2) sequentially adding petroleum ether, chloroform and ethyl acetate with the same volume into the substance A for extraction, collecting the extraction layers of chloroform and ethyl acetate, respectively drying, and combining to obtain a crude product;
(3) dissolving the crude product with 4 times of 60% ethanol, adsorbing with D101 macroporous adsorption resin, and eluting with solvent D, wherein the solvent D is a mixed solvent of chloroform and methanol at a volume ratio of 2:1, to obtain an eluate;
(4) heating the eluate, concentrating, cooling, crystallizing, and drying.
Example 3-1
The only difference from example 3 is that the xylanase in step (1) was replaced with an equal amount of pectinase, and the other conditions were the same.
Examples 3 to 2
The difference from example 3 is only that xylanase and glucosidase in step (1) are added simultaneously, and the rest conditions are the same, specifically as follows:
(1) drying and crushing the wild celery, adding 8 times of solvent A for extraction, simultaneously adding 5% of xylanase and 7% of glucosidase, wherein the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the solvent A to the solvent A is 3:1:2, the extraction temperature is 35 ℃, the total extraction time is 1.5h, and concentrating the extracting solution to 1/6 of the original volume to obtain a substance A.
Examples 3 to 3
The only difference from example 3 is that the composition of solvent A in step (1) is different, solvent A is replaced by an equal volume of 70% ethanol solution, and the rest conditions are the same.
Examples 3 to 4
The difference from the example 3 is only that the composition of the solvent A in the step (1) is different, the solvent A is a mixed solvent of ethylene glycol, isopropanol and water, the volume ratio of the ethylene glycol to the isopropanol to the water is 2:3:1, and the rest conditions are the same.
Examples 3 to 5
The difference from example 3 is only that the composition of the solvent D in step (3) is different, the solvent D is a mixed solvent of chloroform and methanol, the volume ratio of the chloroform to the methanol is 1:2, and the rest conditions are the same.
Example 4
The composition comprises the following components in parts by weight: 20 parts of apigenin (prepared in example 3), 15 parts of theanine, 0.5 part of folic acid and 5 parts of dihydrooroselin.
The components in the composition are mixed uniformly.
Example 5
The composition comprises the following components in parts by weight: 35 parts apigenin (prepared in example 3), 20 parts theanine, 2 parts folic acid and 10 parts dihydrooroselin.
The components in the composition are mixed uniformly.
Example 6
The composition comprises the following components in parts by weight: 28 parts apigenin (prepared in example 3), 16 parts theanine, 1 part folic acid and 7 parts dihydrooroselin.
The components in the composition are mixed uniformly.
Example 6-1
The difference from the example 6 is only that the dosage and the proportion of each component are different, and the rest conditions are the same, and the concrete conditions are as follows:
the composition comprises the following components in parts by weight: 17 parts of apigenin (prepared in example 3), 12 parts of theanine, 3 parts of folic acid and 20 parts of dihydrooroselin.
Example 6 to 2
The difference from example 6 is only that the dihydrooenanthe javanica element is replaced by an equal amount of oenanthe javanica element, and the rest conditions are the same, and are as follows:
the composition comprises the following components in parts by weight: 35 parts of apigenin (prepared in example 3), 16 parts of theanine and 1 part of folic acid.
Examples 6 to 3
The difference from the example 6 is only that the phedrin is replaced by the same amount of dihydroorophedrin, and the rest conditions are the same, and are as follows:
the composition comprises the following components in parts by weight: 35 parts of dihydrooroselagin, 16 parts of theine and 1 part of folic acid.
Test example 1
The yield and purity of the apigenin prepared in examples 1-3 were tested, and the results are shown in table 1.
TABLE 1 yield and purity test of spilanthol
| Yield (mg/g) | Purity/%) | |
| Example 1 | 78.47 | 98.73 |
| Example 2 | 79.25 | 98.97 |
| Example 3 | 78.36 | 99.01 |
| Examples3-1 | 71.58 | 98.98 |
| Examples 3 to 2 | 73.09 | 99.00 |
| Examples 3 to 3 | 69.79 | 97.62 |
| Examples 3 to 4 | 72.43 | 98.26 |
| Examples 3 to 5 | 75.84 | 95.35 |
Test example 2
The effect of reducing uric acid of the compositions prepared in examples 4-6 was tested.
The male Kunming mice are divided into a blank control group (8 mice) and a hyperuricemia group, wherein the hyperuricemia group adopts an intraperitoneal injection oteracil potassium salt method for modeling, the 300mg/kg dose of the hyperuricemia group is administrated by one-time intraperitoneal injection, the hyperuricemia group is randomly divided into a model group (8 mice) and an experimental group after the modeling is successful, the experimental group respectively irrigates the stomach of the composition (8 mice in each embodiment) prepared in the embodiment 4-6, the dose of the composition is 180mg/kg, the stomach is irrigated for 1 time every day for 7 days, and the blank control group and the model group are administrated with physiological saline with the same volume. The results of observing the activity of the blood uric acid and xanthine oxidase of each group of mice (the activity test of the blood uric acid and xanthine oxidase specifically refers to Cao Rui bamboo, Zhang Sanyin, Daoyong, etc.. the cortex Fraxini total coumarin reduces the level of the blood uric acid of the acute hyperuricemia of the mice and the mechanism research [ J ] Liaoning TCM journal 2010(02): 362-.
TABLE 2 mouse blood uric acid and xanthine oxidase Activity test results
The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.
Claims (2)
1. The composition comprises the following components in parts by weight: 20-35 parts of apigenin, 15-20 parts of theine, 0.5-2 parts of folic acid and 5-10 parts of dihydrooroselagin.
2. Use of the composition of claim 1 for the preparation of a medicament for lowering uric acid.
Priority Applications (1)
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| CN106038536A (en) * | 2016-05-30 | 2016-10-26 | 江苏康缘药业股份有限公司 | Application of columbianedin to medicine preparation |
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| CN103641807B (en) * | 2013-12-25 | 2015-12-09 | 南通大学 | A kind of extracting method of apigenin, the pharmaceutical composition being used for the treatment of diabetes and application thereof |
| CN104193711A (en) * | 2014-09-04 | 2014-12-10 | 南京标科生物科技有限公司 | New method for extracting apigenin from celery |
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| CN103109821A (en) * | 2013-01-28 | 2013-05-22 | 江苏省中国科学院植物研究所 | Use of Columbianetin as agricultural pesticide |
| CN106038536A (en) * | 2016-05-30 | 2016-10-26 | 江苏康缘药业股份有限公司 | Application of columbianedin to medicine preparation |
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| Chemical Characterization of 64 Propolis Samples from Hakkari, Turkey;Ecem Bayram et.al.;《Rec. Nat. Prod.》;20181231;第12卷(第6期);第569-581页 * |
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