CN114014900B - Method for extracting and separating linarin from chrysanthemum indicum stems - Google Patents
Method for extracting and separating linarin from chrysanthemum indicum stems Download PDFInfo
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- YFVGIJBUXMQFOF-PJOVQGMDSA-N 5-hydroxy-2-(4-methoxyphenyl)-7-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one Chemical compound C1=CC(OC)=CC=C1C(OC1=C2)=CC(=O)C1=C(O)C=C2O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@H](O)[C@@H](O)[C@H](C)O2)O)O1 YFVGIJBUXMQFOF-PJOVQGMDSA-N 0.000 title claims abstract description 99
- YFVGIJBUXMQFOF-SAXLCNSLSA-N Linarin Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@H](Oc2cc(O)c3C(=O)C=C(c4ccc(OC)cc4)Oc3c2)O1)[C@H]1[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1 YFVGIJBUXMQFOF-SAXLCNSLSA-N 0.000 title claims abstract description 99
- 238000000034 method Methods 0.000 title claims abstract description 34
- 240000005250 Chrysanthemum indicum Species 0.000 title claims description 14
- 235000018959 Chrysanthemum indicum Nutrition 0.000 title claims description 9
- 235000008495 Chrysanthemum leucanthemum Nutrition 0.000 claims abstract description 44
- 244000035851 Chrysanthemum leucanthemum Species 0.000 claims abstract description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 30
- 238000001556 precipitation Methods 0.000 claims abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 59
- 239000000047 product Substances 0.000 claims description 32
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- 238000001914 filtration Methods 0.000 claims description 19
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- 238000001816 cooling Methods 0.000 claims description 10
- 239000002244 precipitate Substances 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- 238000001953 recrystallisation Methods 0.000 claims description 8
- 238000010992 reflux Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 4
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 claims description 2
- 238000000746 purification Methods 0.000 abstract description 10
- 239000002994 raw material Substances 0.000 abstract description 6
- 238000001640 fractional crystallisation Methods 0.000 abstract description 5
- 238000000926 separation method Methods 0.000 abstract description 5
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- 238000004440 column chromatography Methods 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 238000002481 ethanol extraction Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 29
- 238000000605 extraction Methods 0.000 description 18
- 238000004811 liquid chromatography Methods 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000012535 impurity Substances 0.000 description 9
- 238000010575 fractional recrystallization Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 239000012528 membrane Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 229930182470 glycoside Natural products 0.000 description 6
- 150000002338 glycosides Chemical class 0.000 description 6
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- 235000007516 Chrysanthemum Nutrition 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 5
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- 239000012043 crude product Substances 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 241000628997 Flos Species 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
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- 238000009835 boiling Methods 0.000 description 1
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- 239000003814 drug Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
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- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 229930182486 flavonoid glycoside Natural products 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/06—Benzopyran radicals
- C07H17/065—Benzo[b]pyrans
- C07H17/07—Benzo[b]pyran-4-ones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
Abstract
The invention relates to the technical field of separation and purification of medicinal compounds, in particular to a method for extracting and separating linarin from wild chrysanthemum stems. The method uses the stems remained after picking the head inflorescences of the wild chrysanthemum flowers as raw materials, and adopts ethanol extraction, water precipitation to separate the primary product and fractional crystallization to purify the primary product, thus obtaining the linarin with the purity higher than 95 percent. The method comprises the following steps: the method has the advantages of wide raw material sources, low cost, no acid-base treatment and column chromatography in the whole process to obtain the linarin, environmental friendliness, high yield, high purity and low cost, and is suitable for industrial production of high-purity linarin.
Description
Technical Field
The invention relates to the technical field of separation and purification of medicinal compounds, in particular to a method for extracting and separating linarin from wild chrysanthemum stems.
Background
The linarin is the main component in wild chrysanthemum, belongs to flavonoid glycoside compounds, and has the effects of resisting inflammation, resisting bacteria, resisting oxidation, preventing cardiovascular and cerebrovascular diseases and the like. The medicine application part of the wild chrysanthemum is the head-shaped inflorescence of the wild chrysanthemum, and most of picking is performed manually, so that the yield is low, the cost is high, the price is high, and the acquisition of linarin is greatly limited. The detection shows that the stems of the wild chrysanthemum also contain linarin, and the content can reach more than 1 percent. However, the stalks are mainly used as veterinary drugs and feed additives at present, and are not used for extraction and purification to obtain pure linarin.
The method mainly relates to a reflux extraction method, an alkali extraction and alcohol precipitation method and other methods, and the purification method is extraction, column separation, recrystallization and other methods. For example, patent application 201310491359.9 adopts ethanol micropressure extraction, but the concentration step is to concentrate the extract to a certain volume, then add new ethanol to adjust the solution to a certain alcohol degree, and filter the solution to obtain a precipitate. The linarin is easy to dissolve in ethanol, so that the linarin extraction rate is not high. The subsequent purification of patent application 201210577503.6 adopts liquid phase preparation to obtain the linarin monomer, which is not beneficial to large-scale industrialized production. Patent application 201110195580.0 and patent application 200910057065.9 adopt acid or alkali to adjust the pH value of the solution to achieve the purpose of precipitating linarin, industrial wastewater is easy to generate, industrial production is not facilitated, and the subsequent purification step adopts macroporous resin column separation, so that the cost is high. The subsequent purification of the patent application CN104628801A adopts petroleum ether and ethyl acetate for extraction, and the extractant is a volatile and explosive solvent, so that the requirement on workshop explosion prevention is high. In the prior art, most of the raw materials for extracting and separating the linarin are wild chrysanthemum, namely the head inflorescence of the wild chrysanthemum plant, and the extracting and separating process needs to be further improved to meet the requirement of large-scale industrial production.
Disclosure of Invention
In view of the shortcomings of the prior art, the invention aims to provide a method for extracting and separating linarin from stems of wild chrysanthemum. The method uses the stems remained after picking the head inflorescences of the wild chrysanthemum flowers as raw materials, and adopts ethanol extraction, water precipitation to separate the primary product and fractional crystallization to purify the primary product, thus obtaining the linarin with the purity higher than 95 percent. The method comprises the following steps: the method has the advantages of wide raw material sources, low cost, no acid-base treatment and column chromatography in the whole process to obtain the linarin, environmental friendliness, high yield, high purity and low cost, and is suitable for industrial production of high-purity linarin.
In order to achieve the aim, the invention utilizes the principle that the solubility of the linarin in cold water is smaller to extract and separate, and adopts a fractional recrystallization method for purification. The technical scheme adopted by the invention is as follows:
A method for extracting and separating linarin from stems of wild chrysanthemum comprises the following steps:
(1) Taking dried chrysanthemum indicum stems, and mixing the dried chrysanthemum indicum stems with the following weight of (5-30) mL: adding ethanol with the volume fraction of 40% -100% into 1g, reflux extracting for at least 2 times, each for 1-3 hours, filtering, collecting and mixing filtrates to obtain extractive solution, and concentrating the extractive solution under reduced pressure to 1/30-1/10 of the original extractive solution volume;
(2) Adding hot water with the volume of 4-10 times of the concentrated solution and the temperature of 80-90 ℃ into the concentrated solution obtained in the step (1), stirring to fully mix the solution, filtering and clarifying, standing the filtrate at 0-10 ℃ for 8-16h, centrifuging to obtain a precipitate, washing the precipitate with cold water, and drying to obtain a linarin primary product;
(3) According to 20mL: adding a solution with the volume ratio of water to absolute ethyl alcohol of 1:4-4:1 into the initial product of the linarin obtained in the step (2) according to the proportion of 1g, heating to enable the temperature of the solution to be 80 ℃, cooling to 4 ℃ for crystallization for 8 hours after the solution is completely dissolved, filtering, concentrating filtrate at 80 ℃ under reduced pressure to 1/4-1/3 of the volume of the original filtrate, cooling the concentrated filtrate to 4 ℃ for recrystallization for 8 hours, filtering, taking crystal precipitation, washing with water and absolute ethyl alcohol in sequence, and drying to obtain the linarin with the content of more than 90%.
Further, in the step (1): the stem of flos Chrysanthemi Indici is the overground part left after picking flos Chrysanthemi Indici head inflorescence in flos Chrysanthemi Indici flowering phase; the wild chrysanthemum flower (Chrysanthemum indicum L.) is from the county of the filial city of Hubei province.
Further, in the step (1): the dried wild chrysanthemum stem is obtained by taking fresh wild chrysanthemum stem and drying at 80 ℃ for 24 hours.
Preferably, in the step (1): reflux-extracting with ethanol for 2 times, each for 1-2 hr.
Preferably, in the step (3): the volume ratio of water to absolute ethanol is 1:1, obtaining the linarin with the content of more than 95 percent after fractional crystallization and purification.
Compared with the prior art, the invention has the following outstanding advantages and beneficial effects:
1. Compared with the chrysanthemum indicum flower head inflorescence, the chrysanthemum indicum flower stem has fewer water-soluble impurities and oil-soluble impurities, the subsequent purification steps are simple, and the high-purity linarin finished product is easier to obtain. In addition, the stalks can be harvested by a machine, the sources are wide, the cost is low, the stalks are ideal raw materials for extracting linarin, and the comprehensive utilization of resources can be realized.
2. The aim of purifying and separating the linarin is achieved by utilizing the property that the linarin is easy to dissolve in ethanol and hot water and has low solubility in cold water. Further purifying the initial product of the linarin by adopting a fractional recrystallization method, wherein impurities with smaller solubility are separated out by recrystallization in the first step, and the linarin cannot be separated out by recrystallization in the first step; and the second step of recrystallization further concentrates the first crystallization liquid to separate out the linarin, and impurities with higher solubility still exist in the crystallization liquid, so that the aim of purifying the linarin is fulfilled. The method is free from methods such as column chromatography, extraction and the like, is simple to operate, low in production cost, low in workshop explosion-proof requirement, free from acid-base wastewater to be treated, and suitable for large-scale production of high-purity (more than or equal to 95%) and high-yield linarin.
Drawings
Fig. 1A is a chromatogram of a wild chrysanthemum flower head-like inflorescence in example 1, fig. 1B is a chromatogram of a wild chrysanthemum flower stalk in example 1, and fig. 1C is a chromatogram of a frosting glycoside primary product obtained by extracting and separating a wild chrysanthemum flower stalk and a wild chrysanthemum flower head-like inflorescence in example 1.
FIG. 2 is a chromatogram of a sample of linarin after fractional recrystallization in example 2.
FIG. 3 is a chromatogram of a sample of linarin after fractional recrystallization in example 3.
Detailed Description
The following is a further description of the technical solution of the present invention by the applicant in conjunction with specific embodiments and the accompanying drawings, but the scope of the invention claimed is not limited to these embodiments.
Example 1
(1) Comparing the influence of different extraction materials on the linarin extraction process
The experimental steps are as follows: 10g of each of dried chrysanthemum stem and chrysanthemum flower head inflorescence (all from the Shaoxing county of the filial city of Hubei province) is taken, 100mL of absolute ethyl alcohol is added into each of the chrysanthemum stem and the chrysanthemum flower head inflorescence, reflux extraction is carried out for 1.5h, 1mL of extract is taken, a 0.45 μm filter membrane is adopted, liquid chromatography is carried out, and the linarin content in each sample is measured, and the results are shown in figures 1A and 1B.
The chromatographic conditions were as follows: liquid phase column: AGLIENT TC-C 18 (250 x 4.6mm 5 μm); the flow rate was 1.0mL/min, the detection wavelength λ=334 nm, and the mobile phase conditions were as shown in Table 1 below.
TABLE 1
(2) Extracting and separating the different extracting materials to obtain the linarin primary product
50G of each of dried stems and heads of wild chrysanthemum (both from the county of filial sense of Hubei province) is extracted according to the following steps: adding 500mL of ethanol with volume fraction of 70% into the stems and the heads of the wild chrysanthemum respectively, reflux-extracting for 1.5h, filtering, collecting filtrate, extracting twice, and combining the filtrates to obtain an extracting solution. Concentrating the extractive solution under reduced pressure until the volume of the extractive solution is 50mL (no alcohol smell), adding 80deg.C hot water 400mL, stirring for 5min, vacuum filtering with filter paper, standing at 4deg.C for 8 hr, and observing a large amount of precipitate. Centrifuging to obtain precipitate, washing with cold water (4deg.C), and drying to obtain two different linarin primary products.
Respectively taking two kinds of linarin primary products, preparing into 0.2mg/mL concentration with methanol, filtering with 0.45 μm filter membrane, performing liquid chromatography, and measuring linarin content in sample, and the result is shown in figure 1C. The chromatographic mobile phase conditions are shown in Table 2 below, and the other chromatographic conditions are the same as those of the linarin content measurement method described above.
TABLE 2
Fig. 1A is a wild chrysanthemum flower head-like inflorescence chromatogram, fig. 1B is a wild chrysanthemum flower stem chromatogram, and the retention time of the montan glycoside is about 47 min. The content of linarin in the chrysanthemum flower head is 1.36%, the content of linarin in the stem is 1.27%, and the two contents are basically equivalent. However, the liquid chromatography of the stems of the wild chrysanthemum flowers and the heads of the wild chrysanthemum flowers can clearly find that compared with the stems of the wild chrysanthemum flowers, the liquid chromatography of the heads of the wild chrysanthemum flowers can find a large number of impurity peaks within the retention time of 0-10min, namely more water-soluble impurities; a large number of peaks were also found within the retention time of 70-105min, i.e. oil soluble impurities were also more compared to the stalks. Likewise, compared with the method using the stem as the extraction material, the method using the head-like inflorescence as the extraction material has the advantages that a large number of impurity peaks, namely more water-soluble impurities, are visible in the chromatogram with the retention time of 0-5min, and meanwhile, the peak area and the peak height of the linarin (with the retention time of about 9.5 min) are lower, namely the content is lower (see fig. 1C). The measured peak area of the linarin primary product using the chrysanthemum indicum head inflorescence as the extraction material is 21.69mAU min, and the linarin content is 16.16%; the initial product peak area of linarin by adopting wild chrysanthemum stems as extraction materials is 80.94mAU min, and the linarin content is 60.30%. The comparison shows that the wild chrysanthemum stem is a better material for extracting linarin.
Example 2: a method for extracting and separating linarin from stems of wild chrysanthemum comprises the following steps:
(1) Preparing a primary product of the montan glycoside:
Taking 100g of dried wild chrysanthemum stems (from the Shangxiang county of the Hubei province, the content of linarin is 1.34 percent), adding ethanol with the volume fraction of 70 percent into the wild chrysanthemum stems, carrying out reflux extraction for 1.5 hours, filtering, collecting filtrate, extracting twice, combining the filtrate twice to obtain an extracting solution, concentrating the extracting solution under reduced pressure until the volume of the extracting solution is 100mL (without alcohol taste), adding 80 ℃ hot water 800mL into the concentrated solution, stirring to fully and uniformly mix the solution, filtering and clarifying by adopting a ceramic microfiltration membrane with the thickness of 0.45 mu m, standing the filtrate at the temperature of 4 ℃ for 8 hours, observing a large amount of sediment, centrifuging to obtain sediment, washing the sediment with cold water (4 ℃) and drying to obtain 1.8075g of linarin primary product.
Preparing the crude product of linarin with methanol to a concentration of 0.2mg/mL, filtering with 0.45 μm filter membrane, and performing liquid chromatography to determine linarin content in the sample. The chromatographic conditions were as follows: liquid phase column: AGLIENT TC-C 18 (250 x 4.6mm5 μm); the flow rate was 1.0mL/min, the detection wavelength λ=334 nm, and the mobile phase conditions were as shown in Table 3 below.
TABLE 3 Table 3
Detecting by liquid chromatography: the content of linarin in the initial linarin product is 60.17%. The crude product of the linarin is obtained from the wild chrysanthemum stem through extraction and separation, and the yield of the linarin is 81.16 percent.
(2) Preparing a pure product of the linarin:
Taking 1g of the initial product of the montan glycoside, and adding absolute ethyl alcohol into the initial product of the montan glycoside: water = 1:1 (volume ratio) of 20mL of solution, heating the solution to 80 ℃, completely dissolving the linarin primary product by ultrasonic, obtaining the solution, cooling to 4 ℃, standing for 8h, and separating out crystals. Filtering, removing crystals, concentrating the filtrate at 80deg.C under reduced pressure until the volume of the filtrate is 5mL, cooling the concentrated filtrate to 4deg.C, standing for 8 hr again, precipitating white crystals, and filtering to obtain crystals. Washing the crystals with a small amount of water and absolute ethyl alcohol for 2 times, drying, and collecting crystals to obtain pure linarin, wherein the total amount of linarin is 0.4811g.
Preparing pure linarin with methanol to 0.2mg/mL, filtering with 0.45 μm filter membrane, and performing liquid chromatography to determine linarin content in sample, and the result is shown in figure 2. The chromatographic conditions were as follows: liquid phase column: AGLIENT TC-C 18 (250 x 4.6mm 5 μm); the flow rate was 1.0mL/min, the detection wavelength λ=334 nm, and the mobile phase conditions were as shown in Table 4 below.
TABLE 4 Table 4
Detecting by liquid chromatography: the content of linarin in pure linarin is 96.11%. Purifying the crude linarin product by fractional crystallization, wherein the yield of linarin is 76.85%.
Example 3: a method for extracting and separating linarin from stems of wild chrysanthemum comprises the following steps:
(1) Preparing a primary product of the montan glycoside:
10kg of dried wild chrysanthemum stems (from the Shangxiang county of the Hubei province, the content of linarin is 1.14%) are taken, 200L of ethanol with the volume fraction of 55% is added into the wild chrysanthemum stems, reflux extraction is carried out for 2h, a duplex filter is adopted for filtering (the filter screen is 100 meshes), and the filtrate is collected. Extracting for 2 times, mixing the two filtrates to obtain an extract, recovering ethanol under reduced pressure until the volume of the extract is 40L (no ethanol smell), adding 80 ℃ hot water to the concentrate until the volume is 200L, stirring uniformly, filtering and clarifying the solution by adopting ceramic microfiltration membrane (membrane aperture of 0.45 μm) equipment, standing the filtrate at 4 ℃ for 8h to obtain a precipitate, washing with cold water, and drying to obtain the linarin primary product of 135.3g.
Subjecting the crude linarin product to liquid chromatography to determine linarin content in the sample, and detecting method as in example 2. Detecting by liquid chromatography: the content of the initial product of the linarin is 68.81 percent, and the yield of the linarin is 81.67 percent.
(2) Preparing a pure product of the linarin:
Placing all the initial linarin products into a small crystallization tank for recrystallization, and adding 2.7L of water into the crystallization tank: the volume ratio of the absolute ethyl alcohol is 1:1, regulating the temperature of the crystallization liquid to 80 ℃, stirring to fully dissolve a sample, cooling the crystallization liquid to 4 ℃ for 8 hours, centrifuging, discarding precipitates, concentrating the crystallization liquid to 0.7L at 80 ℃ under reduced pressure, cooling the concentrated liquid to 4 ℃ for 8 hours for crystallization, centrifuging, taking a crystallization sample, washing the crystal with a small amount of water and absolute ethyl alcohol for 2 times in sequence, and drying to obtain 113.9g of light yellow powder.
The sample is taken and subjected to liquid chromatography, the content of linarin in the sample is measured, and the detection method is the same as that of example 2, and the result is shown in figure 3. Detecting by liquid chromatography: the content of linarin in the sample was 96.10% (fig. 3), and the fractional crystallization recovery rate of linarin was 80.90%.
Example 4: a method for extracting and separating linarin from stems of wild chrysanthemum comprises the following steps:
and (3) designing a single factor experiment, and discussing the influence of different recrystallization solutions on the purity of the linarin.
(1) A crude product of linarin was prepared by the process of step (1) of example 2, and the linarin content in the crude product of linarin was 60.20%.
(2) The method comprises the following steps of carrying out fractional recrystallization on a linarin primary product: taking 1.8g of a primary product of the linarin (the linarin content is 60.20%), dividing into 9 equal parts, adding 4mL of absolute ethyl alcohol/water solution with different volume ratios into each part, heating to above 70 ℃ until the solution reaches a micro-boiling state, carrying out ultrasonic treatment to completely dissolve the primary product of the linarin to obtain a solution, cooling to 4 ℃, standing for 8 hours, and carrying out visible crystallization. Centrifuging, removing crystals, concentrating the filtrate at 80deg.C under reduced pressure until the volume of the filtrate is about 1mL, cooling the concentrated filtrate to 4deg.C, standing for 8 hr again, precipitating white crystals, centrifuging, and collecting crystals. The crystals were washed with a small amount of water and absolute ethanol in this order for 2 times, dried, collected and the content of the crystalline linarin was measured by the detection method of example 2.
Wherein, absolute ethyl alcohol: the volume ratio of water is 1:9,2:8,3:7,4:6,5:5,6:4,7:3,8:2,9:1. the results are shown in Table 5 below. It can be seen that when absolute ethanol: the volume ratio of water is 1:1, the content of the linarin in the product after fractional recrystallization is the highest and can reach more than 95 percent.
Table 5: influence of absolute ethanol/Water crystallization solutions with different volume ratios on the content of linarin after fractional recrystallization
Claims (4)
1. The method for extracting and separating linarin from chrysanthemum indicum stems is characterized by comprising the following steps of:
(1) Taking dried wild chrysanthemum stems, and mixing the dried wild chrysanthemum stems with the following weight ratio of (5-30) mL: adding ethanol with the volume fraction of 40% -100% into 1g, reflux extracting for at least 2 times, each for 1-3 hours, filtering, collecting and mixing filtrates to obtain extractive solution, and concentrating the extractive solution under reduced pressure to 1/30-1/10 of the original extractive solution volume; the stem of the wild chrysanthemum is the overground part left after picking the head inflorescence of the wild chrysanthemum in the flowering phase of the wild chrysanthemum;
(2) Adding hot water with the volume of 4-10 times of the concentrated solution and the temperature of 80-90 ℃ into the concentrated solution obtained in the step (1), stirring to fully mix the solution, filtering and clarifying, standing the filtrate at 0-10 ℃ for 8-16h, centrifuging to obtain a precipitate, washing the precipitate with cold water, and drying to obtain a linarin primary product;
(3) According to 20mL: adding a solution with the volume ratio of water to absolute ethyl alcohol of 1:4-4:1 into the initial product of the linarin obtained in the step (2) according to the proportion of 1g, heating to enable the temperature of the solution to be 80 ℃, cooling to 4 ℃ for crystallization for 8 hours after the solution is completely dissolved, filtering, concentrating filtrate at 80 ℃ under reduced pressure to 1/4-1/3 of the volume of the original filtrate, cooling the concentrated filtrate to 4 ℃ for recrystallization for 8 hours, filtering, taking crystal precipitation, washing with water and absolute ethyl alcohol in sequence, and drying to obtain the linarin with high purity.
2. The method according to claim 1, wherein in the step (3): the volume ratio of water to absolute ethanol is 1:1.
3. The method according to claim 1 or 2, wherein in step (1): the dried wild chrysanthemum stem is obtained by taking fresh wild chrysanthemum stem and drying at 80 ℃ for 24 hours.
4. The method according to claim 1 or 2, wherein in step (1): reflux-extracting with ethanol for 2 times, each for 1-2 hr.
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