CN112851621B - Total iridoid extract of caulis et folium fici Tikouae, extraction and purification method and application thereof - Google Patents
Total iridoid extract of caulis et folium fici Tikouae, extraction and purification method and application thereof Download PDFInfo
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Abstract
The invention provides a caulis millettiae speciosae total iridoid extract, an extraction and purification method and application thereof, wherein the extraction and purification method sequentially comprises the following steps: adding ethanol into caulis et folium Hedyotis Hedyotideae, adding chitosan, adjusting pH to 5.8-6.5, extracting for 0.5-8 hr, filtering to obtain extractive solution and residue, heating and reflux-extracting the residue for 2-4 times, mixing extractive solutions, and concentrating to obtain dry extract; and then purifying and purifying the dry paste extract to obtain the total iridoid extract of the caulis spatholobi. The extraction method has high extraction rate and good purification effect, and the obtained caulis et folium Hedyotidis Diffusae total iridoid extract has effect in resisting esophageal cancer.
Description
Technical Field
The invention relates to an active component extracted from traditional Chinese medicine, a preparation method and application thereof, in particular to a total iridoid extract with an anti-tumor effect and taking a caulis sinomenii as a raw material, an extraction and purification method and application thereof.
Background
The iridoid compound is a large class of natural products and comprises nearly thousands of compounds, wherein more than 90 percent of the compounds are iridoid glycosides. The structural parent nucleus can be divided into cyclopentane iridoid and secoiridoid according to whether the structural parent nucleus has complete cyclopentane structural units, wherein the cyclopentane iridoid comprises jasminoidin, loganin, cantaxanthin, aucubin, and spicate vitexin, and the secoiridoid comprises swertiamarin, gentiopicroside, swertisin, picroside, and morroniside.
The traditional Chinese medicine composition can be widely stored in various traditional Chinese medicinal materials, such as gardenia, picrorhiza rhizome, eucommia bark, carpet bugle, honeysuckle and the like, and is one of the effective components of a plurality of common Chinese patent medicines, such as gardenia golden flower pills, honeysuckle antipyretic capsules, lung-heat clearing and fire suppressing tablets, nine-ingredient swertia herb pills, nine-ingredient dispersion and the like.
The common Chinese medicine of the bovine white vine also contains iridoid compounds, the bovine white vine refers to dried stem leaves of the bovine white vine of the genus Erysia of the family Rubiaceae, also called hairy fevervine herb, sweet tea, sparerib and the like, and is mainly produced in areas of Guangdong, guangxi, yunnan, guizhou, fujian, taiwan and the like.
Iridoid compounds have various biological activities such as anti-tumor and anti-inflammatory activities, have great research value, and are receiving more and more attention in recent years.
The iridoid compound is extracted and separated by a solvent method, and the solvent is usually water, methanol, ethanol, dilute acetone, ethyl acetate and the like.
In the prior art, the extraction and separation method of the iridoid has low extraction rate and poor purification effect.
Disclosure of Invention
The invention mainly aims to overcome the defects of the prior art and provides a caulis fici Pumilae total iridoid extract, an extraction and purification method and application thereof. The method has high extraction rate and good purification effect of the caulis et folium fici Tikouae total iridoid extract.
In order to achieve the above objects, in a first aspect, the present invention provides a method for extracting and purifying a total iridoid extract of fevervine, which sequentially comprises the following steps:
(1) Adding ethanol into caulis et folium Hedyotis Hedyotideae, adding chitosan, adjusting pH to 5.8-6.5, extracting for 0.5-8 hr, filtering to obtain extractive solution and residue, heating and reflux-extracting the residue for 2-4 times, mixing extractive solutions, and concentrating to obtain dry extract.
Preferably, the caulis et folium piperis is caulis et folium piperis powder.
Preferably, the concentration of the ethanol in percentage by weight is 55-65%, and the weight ratio of the caulis sinomenii powder to the ethanol is 1 (8-10).
Preferably, the weight ratio of the caulis Sinomenii to the chitosan is (10-20): 1, and the chitosan can release iridoid compounds in the caulis Sinomenii better in the extraction process, thereby improving the yield.
During the extraction process, the chitosan can promote the release of the iridoid compounds and can adsorb part of the iridoid compounds, if the dosage of the chitosan is too high, the adsorption effect of the chitosan exceeds the release promotion effect in the extraction process, and the yield of the total iridoid extract in the extract after filtration is reduced; if the consumption of the fructan is too small, the corresponding effect cannot be achieved.
Preferably, ultrasonic extraction is used, and the extraction time is 0.5-0.6 hours.
Preferably, hydrochloric acid is used for adjusting the pH value.
In the extraction method of the total iridoid extract of the solanum lyratum thunb, if the pH value is lower than 5.8 or higher than 6.5, the yield is reduced and is equivalent to the yield of the traditional extraction method.
Preferably, the method for extracting and purifying the total iridoid extract of the caulis spatholobi further comprises the following steps:
(2) Dissolving the dry extract prepared in the step (1) with hot water, filtering to prepare a crude extract of the iridoid of the bauhinia championii, purifying by adopting an extraction method, and adopting a solid-phase extraction column during extraction.
Wherein the temperature of the hot water is 50-60 ℃, the hot water cannot be completely dissolved due to too low temperature, and the hot water is easy to generate chemical reaction due to too high temperature, thereby influencing the quality of the product.
Wherein the weight ratio of the dry extract to the hot water is 0.1-0.4.
The solid phase extraction column is formed by filling three components A, B and C from top to bottom, and isolation plates are arranged between layers.
Preferably, the volume ratio of A, B and C is 1 (0.8-1.2) to (0.8-1.2).
Preferably, the component A of the filler is neutral Al 2 O 3 200-300 mesh, component B is graphitized carbon black of 120-400 meshC component is C18 alkyl filler bonded by silica gel as matrix, the particle size is 40-63 μm, the average pore diameter is 60A, the carbon content is 16-18%, and the C component seals tail.
Experiments prove that the C8 alkyl filler bonded by taking silica gel as a matrix has overlarge polarity, is not suitable for purifying the total iridoid extract of the solanum lyratum thunb, and has overlow yield; the phenyl filler bonded by taking silica gel as a matrix has too low polarity, is not suitable for purifying the caulis sinomenii total iridoid extract, and has too low yield.
The filling diameter and height ratio of the above fillers is 1: (2-3) column, prepared as a solid phase extraction column, preferably a PVC column; the function is to remove pigment, acidic interference impurities, sugar and fat-soluble impurities in the caulis spatholobi without adsorbing iridoid components.
The extraction method comprises the following steps: activating a solid-phase extraction column by using methanol with 2-6 times of column volume and water with 2-6 times of column volume respectively, taking the crude extract of the bovine caulis spatholobi iridoid with 2-3 times of column volume, passing through the solid-phase extraction column, drying the column by blowing, and collecting the filtrate to obtain the purified crude extract of the total iridoid of the bovine caulis spatholobi.
The method for extracting and purifying the total iridoid extract of the caulis sinomenii further comprises the following steps:
(3) Enriching and purifying total cyclic vinyl ether terpenoid:
and (3) respectively passing the purified crude extract of the total iridoid of the solanum lyratum thunb obtained in the step (2) through a polyamide resin adsorption column and a macroporous resin adsorption column, washing with water, then washing with 30-40% ethanol, and collecting ethanol eluent.
Preferably, the filling method of the polyamide resin adsorption column is as follows: the purified polyamide (200-300 mesh, acid washed, alkali washed, and then water washed to neutral) was packed in a column to give a resin having a length to diameter ratio (1-2): 1.
Preferably, the filling method of the macroporous resin adsorption column comprises the following steps: the purified macroporous resin AMBERLITE XDA-4 (acid washing, alkali washing, and water washing to neutrality) is used for filling the column, so that the length-diameter ratio of the resin is 4-5.
The polyamide resin adsorption column and the macroporous resin adsorption column are connected in series to obtain a series column, and a liquid drainage and liquid adding device is arranged between the two columns.
Taking iridoid compound extract with the same volume as the serial column, passing through the serial column, washing with water, washing with 30-40% ethanol 4-8 times the volume of the serial column, and collecting ethanol eluate; and (3) concentrating the obtained ethanol eluent to dryness to obtain the total iridoid extract of the solanum lyratum thunb.
In a second aspect, the invention also provides the total iridoid extract of the caulis sinomenii obtained by the preparation method of the total iridoid extract of the caulis sinomenii, which comprises the monotropein, asperulosidic acid, deacetylasperulosidic acid, paederosidine, asperuloside and hydroxycrystalloside.
The specific structural formula is as follows:
in a third aspect, the invention also provides application of the total iridoid extract of the solanum lyratum thunb in the prevention of esophageal cancer.
The invention has the following beneficial effects:
the method for extracting the caulis sinomenii total iridoid extract has high extraction rate and good purification effect, and the prepared caulis sinomenii total iridoid extract can be used for resisting esophageal cancer.
Drawings
FIG. 1 shows that the total iridoid extract of caulis Sinomenii prepared in example 1 acts on EC109, KYSE/140, KYSE410 and KYSE510 esophageal cancer cells in logarithmic phase, and CCK8 reagent is adopted to detect the proliferation activity of esophageal cancer cells before and after drug intervention.
Detailed Description
The technical solutions in the embodiments of the present invention are described clearly and completely 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.
It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In the examples, the chitosan was produced by Winhadda marine biologicals Co., ltd, all of the three filler components A, B and C were produced by Aladdin reagent (Shanghai) Co., ltd, and the polyamide resin and AMBERLITE XDA-4 were produced by Aladdin reagent (Shanghai) Co., ltd and Shanghai spectral vibration Biotechnology Co., ltd, respectively.
Example 1
Extracting and purifying iridoid compounds in the caulis spatholobi:
(1) Extraction of iridoid compound
Taking 100g of caulis sinomenii powder, adding 55% ethanol according to a material-liquid ratio of 1.
(2) Purification of crude extract of iridoid from rattan
Taking 1g of the dry extract prepared in the step (1), dissolving with 25ml of hot water at 55-60 ℃, filtering to prepare a crude extract of the iridoid of the bauhinia variegata, and purifying by adopting a solid phase extraction method; the extraction method and conditions are as follows:
a) 3g of filler of the solid-phase extraction column, wherein the filler is formed by sequentially filling three components A, B and C in layers from A to C from top to bottom according to the proportion of 1;
the component A of the filler is neutral Al 2 O 3 200 meshes to 300 meshes, B component is graphitized carbon black of 120 meshes to 400 meshes, C component is C18 filler bonded by silica gel as a substrate, the particle size is 40 to 63 mu m, the average pore diameter is 60A, the carbon content is 17 percent, and the tail is sealed; filling the above fillers into a PVC column with diameter of 1.0cm and height of 2cm to prepare a solid phase extraction column;
b) An elution mode: activating a solid-phase extraction column with 5ml of methanol and 5ml of water respectively, taking 25ml of crude extract of the iridoid of the solanum lyratum, passing through the solid-phase extraction column, eluting with 40% ethanol, drying the column by blowing, and collecting filtrate to obtain purified iridoid extract.
(3) Enriching and purifying the total iridoid:
column 1: the purified polyamide (200-300 mesh, acid-washed, alkali-washed, and then water-washed to neutrality) was taken 40ml and packed in a column so that the resin aspect ratio was 1.
Column 2: 40ml of the purified macroporous resin AMBERLITE XDA-4 (acid washed, alkali washed, and then water washed to neutrality) was packed in a column to give a resin length to diameter ratio of 4.
The column 1 and the column 2 are connected in series to form a funnel shape, and a liquid discharging and adding device is arranged between the two columns. Thus, column 1 is of larger diameter, a thick short column relative to column 2; the post 2 is of a smaller diameter and is an elongated post relative to the post 1. The thick short column (column 1, thin filler, 200-300 meshes) can reduce resistance while realizing adsorption separation; the slender column (column 2, the filler is thicker, 10-20 meshes) can realize effective adsorption and enrichment and is not easy to block resin pores, particularly after passing through the column 1. The clever combination effectively improves the purification effect and efficiency.
And (3) taking 30ml of the iridoid compound extract purified in the step (2), passing through a series column of a column 1 and a column 2, washing with water, washing with 250ml of 40% ethanol for 3 times, and collecting ethanol eluent to obtain the purified total iridoid compound extract.
(4) Preparation of extract of total iridoid of caulis et folium fici Tikouae
And (4) concentrating the purified total iridoid compound extract obtained in the step (3) to be dry to obtain the compound.
(5) Identification of iridoid extract components of caulis et folium fici Tikouae
The total iridoid extract is identified by HPLC-MS method, and comprises 6 iridoid components such as astragaloside, asperulosidic acid, deacetylasperulosidic acid, paederosidic acid, asperuloside and hydroxycysteronine.
(6) Total iridoid content determination
Precisely weighing about 0.05g of total iridoid extract, placing in a 100mL conical flask, adding 10mL of 60% ethanol, weighing, ultrasonically treating for 60min, shaking, filtering to obtain test solution, taking 5mg of gentiopicroside, diluting to 10mL with 60% ethanol to constant volume, and making into reference solution. The ultraviolet absorption values of the reference substance and the test substance are respectively measured, the concentration of the total iridoid can be known through the ratio of the absorption values of the reference substance and the test substance, and the results of the two measurements are shown in Table 1.
TABLE 1 Total iridoid extract content determination
Comparative example 1
Compared with the traditional extraction method:
according to the embodiment 1, 100g of the caulis sinomenii powder is taken, ethanol with the concentration of 65% is added according to the material-liquid ratio of 1. The results are shown in Table 2-1:
TABLE 2-1 comparison of the quality of extracts obtained by different extraction methods
Compared with the traditional extraction method such as hot reflux extraction, an ultrasonic extraction method, supercritical fluid extraction and the like, the method adopted by the invention is simple and convenient, has higher extraction efficiency, and particularly, chitosan is added, so that the iridoid compounds in the caulis sinomenii are better released while impurities are adsorbed.
Adjusting pH to 5.8-6.5 to increase solubility of iridoid such as gentiopicroside in extractive solution and greatly improve extraction efficiency (see Table 2-2), adjusting pH to 7 and keeping the pH unchanged in example 1, or adjusting pH to 5 and keeping the pH unchanged in example 1, and comparing the extraction rate in step (1) with the mass of dry extract accounting for the mass of caulis et folium fici Tikouae powder as extraction rate (%).
TABLE 2-2 comparison of extract quality at different pH values
Compared with the traditional purification method, the purification method of the crude extract of the iridoid of the rattan comprises the following steps:
1g of the dry paste extract prepared in the step (1) in the example 1 is taken, dissolved in 25ml of hot water with the temperature of 55-60 ℃, filtered to prepare a crude extract of the iridoid of the bovine rattan, and the crude extract is purified by adopting a solid-phase extraction method. Then concentrating to dry weight, measuring the content of the total iridoid in the purified dry paste extract, and simultaneously carrying out a standard experiment, and simultaneously taking the content and the recovery rate as the evaluation of the purification efficiency, wherein the higher the content after purification is, the higher the recovery rate is, the higher the purification efficiency is, the packing volume and the column of each filler in the table 3 used singly are consistent with the embodiment 1, and when the three components A, B and C are combined, the packing volume and the column are consistent with the embodiment 1 according to the proportion of 1.
TABLE 3 comparison of purification efficiency of different purification methods
Wherein the component A of the filler is neutral Al 2 O 3 200-300 meshes, B component is graphitized carbon black and 120-400 meshes, C component is C18 filler bonded by silica gel as a matrix, the particle size is 40-63 mu m, the average pore diameter is 60A, and the carbon content is 17%. The extract of caulis et folium nigellae contains more sinapic acid, which can be converted into gentiopicroside via hydrogen bondCompound, etc., where a: al (Al) 2 O 3 Can specifically adsorb erucic acid in the extractive solution, free gentiopicroside, and improve purification effect. The graphitized carbon black can absorb pigment and fatty acid, and the acidic substances are reduced and the purification effect of the graphitized carbon black is improved through the eluent which is used for removing erucic acid and is obtained by the step A. And (3) passing the eluent subjected to the treatment A and the treatment B through a reaction system C: when the C18 is filled, dead adsorption of the C18 can be reduced, and the enrichment and purification effects of the C18 are further improved.
The comparison result shows that the ABC combined purification method has the highest content measurement result which is far higher than other methods, and the recovery rate is high. Therefore, the ABC combined purification method has the best purification effect.
The purification method of the crude extract of the iridoid of the rattan comprises the following steps:
30ml of iridoid compound extract purified in the step (2) of the embodiment 1 is taken, and the content and the recovery rate of the total iridoid in the purified and purified dry extract are used as the evaluation of the purification efficiency by comparing the macroporous resin method, the polyamide method and the combined method of the invention reported in the literature. The higher the purified content, the higher the recovery rate, indicating the higher the purification efficiency, the column volume was the same as in example 1 for polyamide and macroporous resin alone, and the column volume was the same as in column 1 and column 2 of example 1 for polyamide and macroporous resin in combination. The results are shown in Table 4.
TABLE 4 comparison of purification efficiency of different purification methods
The caulis Sinomenii contains various iridoid chemical components, and the content and purity of the obtained product are different by different extraction and purification (including purification) methods. The same extraction method is used for different extracts, and the results are different. The experimental results of the extraction and purification (including purification) of iridoid contained in the caulis sinomenii show that the XDA-4 has better effect in the macroporous resin, and the combination of the polyamide and the macroporous resin obviously improves the purification efficiency compared with the single use of the polyamide or the macroporous resin.
The method adopts a tandem chromatography column method for purifying the terpene components of the total iridoid in the caulis sinomenii for the first time. Compared with the method of passing through a column in a fractional manner (firstly passing through polyamide and then passing through macroporous resin), the method of the series chromatography column method (polyamide and macroporous resin) has the advantages of greatly improving the recovery rate (taking gentiopicroside as a labeling experiment) and being more convenient to operate due to the reduction of experimental steps and the reduction of the used solvent amount. Proved by series connection of polyamide and macroporous resin column XDA-4, the effect is best.
Example 2
The extraction and purification method of example 2 is different from example 1 in that ultrasonic waves are used for heating reflux in step (1) and the reflux extraction time is 0.5 hours, and the method is otherwise the same as example 1.
Example 3
The extraction and purification method of example 3 is different from example 1 in that the amount of the caulis sinomenii powder used in step (1) is 120g, and the other steps are the same as example 1.
Example 4
The extraction and purification method of example 4 is different from example 1 in that the amount of chitosan used in step (1) is 8g, and the method is otherwise the same as example 1.
Example 5
The extraction and purification method of example 5 is different from example 1 in that pH in step (1) is 6.2, and the method is otherwise the same as example 1.
Comparative example 2
The extraction and purification method of comparative example 2 is different from that of example 1 in that chitosan is not added in step (1), and the other steps are the same as those of example 1.
Comparative example 3
The extraction and purification method of comparative example 3 is different from example 1 in that the amount of chitosan added in step (1) is 25g, and the other steps are the same as example 1.
Comparative example 4
The preparation method of comparative example 4 is different from example 1 in that the raw material in step (1) is clove leaf powder instead of bovine fevervine root powder, and the other steps are the same as example 1.
The results of the measurements of the examples and comparative examples after the final extraction and purification are shown in Table 5.
TABLE 5 Total iridoid extract content and extraction yield
As can be seen from comparative examples 2-3, the total iridoid extract obtained by the method is greatly reduced in purity and extraction rate without adding chitosan or with too large amount; as can be seen from comparative example 4, the extraction method of the invention is suitable for the extraction and purification of the total iridoid extract of the caulis sinomenii.
Example 1 application of total iridoid extract of caulis et folium piperis niveae in resisting esophageal cancer:
the invention adopts a pharmacological molecular biology method, and evaluates the pharmacological action of the caulis sinomenii total cycloolefine ether terpene extract on esophageal cancer resistance to esophageal cancer through cell in-vitro experiments. 4 kinds of esophageal cancer cells are taken as research objects, the activity change of tumor cells before and after the intervention of the total iridoid extract of the caulis sinomenii is detected by using a CCK-8 (Cell Counting Kit-8) cytotoxicity activity detection reagent, and the influence of the drug intervention on the migration of the tumor cells is examined by using a Cell scratching experiment. Evaluating the killing effect of the medicament on tumor cells, and researching the influence of the total iridoid extract of the bovine solanum lyratum on the activity and migration of the tumor cells.
1. CCK8 detection is used for researching the influence of the caulis sinomenii total iridoid extract on the survival rate of esophageal cancer cells.
The caulis et folium piperis total iridoid extract diluted into culture medium with different drug concentrations acts on EC109, KYSE/140, KYSE410 and KYSE510 esophageal cancer cells in logarithmic phase, and CCK8 reagent is adopted to detect esophageal cancer cell proliferation activity before and after drug intervention (see figure 1 specifically). The relative cell survival rate and the relative cell inhibition rate before and after intervention of the caulis sinomenii on esophageal cancer cells are measured by taking the same background drug and no cell as a blank group, taking the tumor cells which are not administrated as a control group and taking 100 mug/mL cis-platinum and 100 mug/mL 5-fluorouracil as positive control groups respectively, and the Half inhibition Concentration (IC 50) is calculated (see Table 6 in detail). Experimental results show that in the esophageal cancer cells, the caulis sinomenii with the concentration of more than 50 mug/mL has obvious inhibition effect on the esophageal cancer cells. The half inhibitory concentration IC50 of the bovine rattan iridoid extract on esophageal cancer cells is between 32.81 and 104.9 mu g/mL. Under the action of effective IC50 drug concentration, the bovine rattan iridoid extract can obviously reduce the relative survival rate of esophageal cancer cells and shows certain dose dependence.
TABLE 6 Semithragma nivea total iridoid extract half inhibitory concentration IC50 on esophageal cancer cells
2. The scratch cell is adopted to research the influence of the caulis sinomenii iridoid extract on the movement of esophageal cancer cells:
after the IC50 bovine rattan acts on the serum-free cultured tumor cells, the healing conditions of the scratch at different time points are detected, the area of the scratch is calculated, and the healing rate of the scratch is measured. The experimental result is as follows: after the IC50 medicine intervenes in esophageal cancer cells, along with the prolonging of action time, the tumor cells of the blank control group gradually move to the scratch, the scratch is gradually healed, while the tumor cells of the administration group have slower moving speed, the cell morphology changes at the later stage, and part of the cells are suspended and dead. Compared with a control group, along with the prolonging of the action time of the medicine, the cells of the administration group are not healed, even the scratch area is enlarged, the healing rate of the scratch is far less than that of the control group, which shows that under the action of the medicine concentration of effective IC50, the healing capacity of the tumor cells is obviously inhibited, the cell migration capacity is weakened, and the medicine can inhibit the activity and migration of the tumor cells. According to CCK8 cell activity experiments and scratch experiments, the total iridoid extract of the caulis sinomenii is presumed to realize the anti-tumor effect by inhibiting cell migration/promoting tumor cell death.
TABLE 7 healing rate of esophageal cancer cell scratch before and after administration of total iridoid extract of caulis et folium fici Tikouae
The first discovery shows that the total iridoid extract of the bovine solanum lyratum can inhibit the survival rate of tumor cells and inhibit the migration of the tumor cells, thereby realizing the effect of resisting esophageal cancer.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and all modifications and equivalents of the present invention, which are made by the present specification and directly/indirectly applied to other related technical fields within the spirit of the present invention are included in the scope of the present invention.
Claims (5)
1. A method for extracting and purifying a caulis millettiae speciosae total iridoid extract is characterized by sequentially comprising the following steps of:
(1) Adding ethanol into caulis et folium Hedyotis Hedyotideae, adding chitosan, adjusting pH to 5.8-6.5, extracting with ultrasonic wave for 0.5-0.6 hr, filtering to obtain extractive solution and residue, and concentrating the extractive solution to obtain dry extract; the weight percentage concentration of the ethanol is 55-65%, the weight ratio of the caulis sinomenii to the ethanol is 1 (8-10), and the weight ratio of the caulis sinomenii to the chitosan is (10-20) to 1;
(2) Dissolving the dry extract prepared in the step (1) with hot water, filtering to prepare a crude extract of the iridoid of the bauhinia championii, purifying by adopting an extraction method, and adopting a solid-phase extraction column during extraction; wherein the temperature of the hot water is 50-60 ℃; the weight ratio of the dry extract to the hot water is (0.1-0.4) to 5; the solid phase extraction column comprises three filler components A, B and C from top to bottom, wherein the filler components A, B and C are filled according to the volume ratio of 1 (0.8-1.2) to 0.8-1.2; the component of the filler A is neutral Al 2 O 3 200-300 meshes, the component of the filler B is graphitized carbon black, 120-400 meshes, the component of the filler C is the filler CThe component C is C18 alkyl filler bonded by taking silica gel as a matrix, the particle size is 40-63 mu m, the average pore diameter is 60A, the carbon content is 16-18%, and the component C is tail-sealed; the extraction method comprises the following steps: activating a solid phase extraction column by using methanol with 2-6 times of column volume and water with 2-6 times of column volume respectively, taking the crude extract of the caulis sinomenii iridoid with 2-3 times of column volume, passing the crude extract of the caulis sinomenii iridoid through the solid phase extraction column, drying the column by blowing, and collecting filtrate to obtain the purified crude extract of the total iridoid of the caulis sinomenii.
2. The method for extracting and purifying the total iridoid extract of bauhinia championii as claimed in claim 1, wherein the ratio of the diameter to the height of the solid-phase extraction column is 1: (2-3).
3. The method for extracting and purifying the extract of caulis spatholobi total iridoid as claimed in claim 1, further comprising the following step (3): and (3) respectively passing the purified crude extract of the total iridoid extract of the solanum lyratum thunb obtained in the step (2) through a polyamide resin adsorption column and a macroporous resin adsorption column, washing with water, then washing with 30-40% ethanol, and collecting ethanol eluent.
4. The method for extracting and purifying the total iridoid extract of caulis boehmeriae as claimed in claim 3, wherein the polyamide resin adsorption column is filled by the following steps: filling the purified polyamide into a column, wherein the length-diameter ratio (1-2) of the resin is 1; the filling method of the macroporous resin adsorption column comprises the following steps: the column was packed with the purified macroporous resin AMBERLITE XDA-4, the aspect ratio of the resin (4-5) was 1.
5. The method for extracting and purifying the total iridoid extract of caulis boehmeriae as claimed in claim 3, wherein the polyamide resin adsorption column and the macroporous resin adsorption column are connected in series.
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