CN107095893B - Extraction method and application of active substances of panax pseudoginseng - Google Patents
Extraction method and application of active substances of panax pseudoginseng Download PDFInfo
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Abstract
The invention discloses an extraction method of active substances of panax pseudoginseng, which comprises the following steps: pretreating raw materials, preparing a Taibai pseudo-ginseng extracting solution, pretreating the Taibai pseudo-ginseng extracting solution, performing high-speed countercurrent extraction, and performing reduced pressure liquid chromatography separation to finally obtain high-purity Taibai pseudo-ginseng total saponins. In addition, the invention also provides application of the active substance of the panax notoginseng in preparing a medicament for treating cardiovascular diseases. According to the invention, through twice ethanol soaking and ultrasonic extraction, active substances in the gynura divaricata are extracted to the maximum extent, then high-speed countercurrent extraction and reduced pressure liquid chromatography separation are adopted, and finally the purity of the extracted gynura divaricata total saponins reaches more than 95%. The total saponins extracted by the invention have high content, stable quality and good color, and the traditional Chinese medicine preparation prepared by taking the total saponins as the raw material completely meets the requirements of modern traditional Chinese medicines.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to an extraction method and application of active substances of panax notoginseng.
Background
Pseudo-ginseng is one of the famous and precious medicinal materials which are special products in China and is the dry root of pseudo-ginseng which belongs to the family of araliaceae. The Taibaishan Chinese herbal medicine is famous for a long time, and especially the Taibaisan pseudo-ginseng has extremely high medical value and is a precious national cultural heritage. The Taibai pseudo-ginseng is the root of the dicotyledonous plant medicine Umbelliferae plant carthamus flower celery, is mainly produced in Yunnan, Shaanxi and other places, is loaded in compendium of materia medica, is a traditional and famous Chinese medicinal material in China, has the effects of promoting blood circulation to remove blood stasis, relieving swelling and pain, stopping bleeding and removing blood stasis and the like, different growing parts of the pseudo-ginseng contain more than twenty saponin components, the main active component of the pseudo-ginseng is total saponins, and the pseudo-ginseng total saponins have the effects of increasing coronary blood flow, protecting brain tissues, expanding blood.
The existing extraction process of panax notoginseng saponins mainly comprises a water decoction and alcohol precipitation method, an ethanol reflux method and a percolation method. Wherein, the water decoction and alcohol precipitation method is easy to gelatinize because the notoginseng contains a large amount of starch and polysaccharide in the water decoction process; after alcohol precipitation, the saponin loss is large due to the adsorption effect of the precipitate, the extraction rate is low, and the content of the product components is not stable, so that the method is mainly used for developing foods; the percolation method has high extraction rate and simple operation, and the extract contains less impurities but consumes too much solvent and is mainly used for medicines; the ethanol reflux method is simple and convenient, the process conditions are easy to control, but the extraction rate is low, and the extraction methods all require that the raw materials are extracted by dried pseudo-ginseng rhizomes (cut openings) and processed leftover materials, so that the extraction period is obviously prolonged.
Disclosure of Invention
The invention provides an extraction method of active substances of gynura divaricata, which solves the problems of low extraction efficiency, high impurity content in the extracted active substances and large loss of active ingredients of the gynura divaricata in the prior art.
The first purpose of the invention is to provide a method for extracting active substances of panax notoginseng, which comprises the following steps:
step 1, pretreatment of raw materials: drying and pulverizing radix Tongoloae Dunnii, and sieving to obtain radix Tongoloae Dunnii powder;
step 2, preparing a Taibai pseudo-ginseng extracting solution, namely adding an ethanol solution with the mass concentration of 75-85% into Taibai pseudo-ginseng powder according to the proportion of 1g to 10-30 m L, soaking for 12-24 h, then carrying out ultrasonic treatment at 60 ℃ for 40-60 min, and filtering after the ultrasonic treatment is finished to obtain a first filtrate and filter residues;
adding 75-85% ethanol solution into the filter residue according to the proportion of 1g to 10-20 m L, soaking for 10-12 h, then performing ultrasonic treatment at 60 ℃ for 10-30 min, and filtering after the ultrasonic treatment is finished to obtain a second filtrate;
mixing the first filtrate and the second filtrate to obtain mixed extractive solution, concentrating the mixed extractive solution, and recovering ethanol to obtain Notoginseng radix extract;
step 3, pretreatment of the gynura segetum extract: adding deionized water which is 5-10 times of the weight of the Taibai pseudo-ginseng extract into the Taibai pseudo-ginseng extract in the step 2, stirring at 40-70 ℃ until the Taibai pseudo-ginseng extract is completely dissolved, then adding an ammonia water solution with the concentration of 10%, and adjusting the pH of the system to 8-9 to obtain a pretreated Taibai pseudo-ginseng extract;
and 4, high-speed countercurrent extraction: pumping the pretreated radix Tongoloae Dunnii extract into an extraction tower, and then mixing the radix Tongoloae Dunnii extract and n-butanol according to the volume ratio of 1: 1-3, uniformly introducing n-butyl alcohol into the extraction tower for circulating high-speed countercurrent extraction, controlling the extraction temperature to be 40-60 ℃, performing circulating extraction for 3-5 times to obtain an extract, concentrating the extract and recovering the n-butyl alcohol to obtain a crude product of the active substance of the panax notoginseng;
and 5, carrying out reduced pressure liquid chromatography separation, namely mixing and dissolving the crude product of the pseudo-ginseng taibai active substance with deionized water according to the proportion of 1g to 3-5 m L to obtain an aqueous solution of the crude product of the pseudo-ginseng tai bai active substance, adding silica gel which is 1.5-2 times of the weight of the crude product of the pseudo-ginseng tai active substance into the aqueous solution of the crude product of the pseudo-ginseng tai bai active substance for sample mixing, simultaneously carrying out wet column packing by using the silica gel which is 20-30 times of the weight of the crude product of the pseudo-ginseng tai bai active substance, wherein the diameter-height ratio is 1: 4-6, controlling the column pressure to be-0.3-0.1 Mpa after the column packing is finished, carrying out sample loading, carrying out silica gel column chromatography, eluting by using a mixed solution of n-butanol and.
Preferably, the active substance of the gynura segetum is gynura segetum total saponin.
Preferably, the gynura segetum in the step 1 is dried and crushed and then sieved by a 100-mesh sieve.
Preferably, the ultrasonic frequency in the step 2 is 120-180 Hz.
Preferably, the specification of the silica gel used in the step 5 is 200-300 meshes.
The second purpose of the invention is to provide the application of the active substance of the panax notoginseng taibai in the medicine for treating cardiovascular diseases.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a method for extracting active substance total saponins from gynura segetum, which comprises the steps of soaking the gynura segetum in ethanol twice, extracting the active substance from the gynura segetum to the maximum extent by combining ultrasonic extraction, then adopting high-speed countercurrent extraction, reduced-pressure liquid chromatography separation and recrystallization, and finally improving the purity of the total saponins to more than 95%. The total saponins extracted by the invention have high content, stable quality and good color, and the traditional Chinese medicine preparation prepared by taking the total saponins as the raw material completely meets the requirements of modern traditional Chinese medicines.
Detailed Description
In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.
The experimental methods described in the embodiments of the present invention are all conventional methods unless otherwise specified; the reagents used, unless otherwise specified, are conventional.
Example 1
A method for extracting active substances of Panax pseudoginseng C.H.Chen comprises the following steps:
step 1, pretreatment of raw materials: drying 1000g of Taibai pseudo-ginseng, crushing, and sieving with a 100-mesh sieve to obtain Taibai pseudo-ginseng powder;
step 2, preparing the gynura segetum extract, namely adding an ethanol solution with the mass concentration of 15L being 75% into the gynura segetum powder, soaking for 12 hours, then carrying out ultrasonic treatment at the temperature of 60 ℃ for 40 minutes, and filtering after the ultrasonic treatment is finished to obtain a first filtrate and filter residues;
adding 10L ethanol solution with mass concentration of 75% into the filter residue, soaking for 10h, then performing ultrasonic treatment at 60 deg.C for 10min, and filtering after the ultrasonic treatment to obtain second filtrate;
mixing the first filtrate and the second filtrate to obtain mixed extractive solution, concentrating the mixed extractive solution, and recovering ethanol to obtain 172g of Notoginseng radix extract;
step 3, pretreating the Taibai pseudo-ginseng extracting solution, namely adding 1.2L deionized water into 172g of the Taibai pseudo-ginseng extract obtained in the step 2, stirring at 70 ℃ until the Taibai pseudo-ginseng extracting solution is completely dissolved, then adding an ammonia water solution with the concentration of 10%, and adjusting the pH value of the system to be 8 to obtain the pretreated Taibai pseudo-ginseng extracting solution;
step 4, high-speed countercurrent extraction, namely pumping the pretreated gynura segetum extract into an extraction tower, then uniformly introducing n-butyl alcohol of 2L into the extraction tower to perform circulating high-speed countercurrent extraction, controlling the extraction temperature to be 40 ℃, performing circulating extraction for 5 times to obtain an extract, concentrating the extract and recovering the n-butyl alcohol to obtain 121g of gynura segetum total saponin crude product;
and 5, carrying out reduced pressure liquid chromatography separation, namely dissolving 121g of the crude product of the panax notoginseng saponins by using 450m L deionized water to obtain an aqueous solution of the crude product of the panax notoginseng saponins, adding 200g of silica gel into the aqueous solution of the crude product of the panax notoginseng saponins, mixing the mixture with a sample, simultaneously carrying out wet column packing by using 2500g of silica gel with the diameter-height ratio of 1: 4, controlling the column pressure to be-0.3 Mpa after the column packing is finished, loading the sample, carrying out silica gel column chromatography, eluting by using a mixed solution of n-butanol and ethanol as an eluent, collecting fractions, concentrating the fractions to obtain 92g of the panax notoginseng saponins, namely the yield is 9.2%, and the purity is 95.37% by liquid chromatography analysis.
Example 2
A method for extracting active substances of Panax pseudoginseng C.H.Chen comprises the following steps:
step 1, pretreatment of raw materials: drying 1200g of radix Tongoloae Dunnii, pulverizing, and sieving with 100 mesh sieve to obtain radix Tongoloae Dunnii powder;
step 2, preparing the gynura segetum extract, namely adding an ethanol solution with the mass concentration of 20L being 80% into gynura segetum powder, soaking for 18h, then carrying out ultrasonic treatment at the temperature of 60 ℃ for 50min, and filtering after the ultrasonic treatment is finished to obtain a first filtrate and filter residues;
adding an ethanol solution with the mass concentration of 15L of 80% into the filter residue, soaking for 11h, then performing ultrasonic treatment at 60 ℃ for 20min, and filtering after the ultrasonic treatment is finished to obtain a second filtrate;
mixing the first filtrate and the second filtrate to obtain mixed extractive solution, concentrating the mixed extractive solution, and recovering ethanol to obtain 210g of Notoginseng radix extract;
step 3, pretreating the Taibai pseudo-ginseng extract, namely adding 1.5L deionized water into 210g of the Taibai pseudo-ginseng extract in the step 2, stirring at 50 ℃ until the Taibai pseudo-ginseng extract is completely dissolved, then adding an ammonia water solution with the concentration of 10%, and adjusting the pH of the system to 8.5 to obtain the pretreated Taibai pseudo-ginseng extract;
step 4, high-speed countercurrent extraction, namely pumping the pretreated gynura segetum extract into an extraction tower, then uniformly introducing 3.5L n-butyl alcohol into the extraction tower to perform circulating high-speed countercurrent extraction, controlling the extraction temperature to be 50 ℃, performing circulating extraction for 4 times to obtain an extract, concentrating the extract and recovering the n-butyl alcohol to obtain 145g of gynura segetum total saponin crude product;
and 5, carrying out reduced pressure liquid chromatography separation, namely dissolving 145g of the crude product of the panax notoginseng saponins by 600m L deionized water to obtain an aqueous solution of the crude product of the panax notoginseng saponins, adding 220g of silica gel into the aqueous solution of the crude product of the panax notoginseng saponins for sample mixing, simultaneously carrying out wet column packing by 3600g of silica gel with the diameter-height ratio of 1: 5, controlling the column pressure to be-0.2 Mpa after the column packing is finished, carrying out sample loading, carrying out silica gel column chromatography, eluting by using a mixed solution of n-butanol and ethanol as an eluent, collecting fractions, concentrating the fractions to obtain 110g of the panax notoginseng saponins, wherein the yield is 9.17%, and the purity is 96.08% by liquid chromatography analysis.
Example 3
A method for extracting active substances of Panax pseudoginseng C.H.Chen comprises the following steps:
step 1, pretreatment of raw materials: drying 1500g of Taibai pseudo-ginseng, crushing, and sieving with a 100-mesh sieve to obtain Taibai pseudo-ginseng powder;
step 2, preparing the gynura segetum extract, namely adding an ethanol solution with the mass concentration of 30L being 85% into gynura segetum powder, soaking for 24 hours, then carrying out ultrasonic treatment at the temperature of 60 ℃ for 60 minutes, and filtering after the ultrasonic treatment is finished to obtain a first filtrate and filter residues;
adding 20L ethanol solution with mass concentration of 85% into the filter residue, soaking for 12h, then performing ultrasonic treatment at 60 deg.C for 10min, and filtering after the ultrasonic treatment to obtain a second filtrate;
mixing the first filtrate and the second filtrate to obtain mixed extractive solution, concentrating the mixed extractive solution, and recovering ethanol to obtain 258g of radix Tongoloae Dunnii extractive solution;
step 3, pretreating the Taibai pseudo-ginseng extracting solution, namely adding 1.8L deionized water into 258g of the Taibai pseudo-ginseng extracting solution in the step 2, stirring at 40 ℃ until the Taibai pseudo-ginseng extracting solution is completely dissolved, then adding an ammonia water solution with the concentration of 10%, and adjusting the pH of the system to be 9 to obtain the pretreated Taibai pseudo-ginseng extracting solution;
step 4, high-speed countercurrent extraction, namely pumping the pretreated gynura segetum extract into an extraction tower, then uniformly introducing 4.5L n-butyl alcohol into the extraction tower to perform circulating high-speed countercurrent extraction, controlling the extraction temperature to be 60 ℃, performing circulating extraction for 3 times to obtain an extract, concentrating the extract and recovering the n-butyl alcohol to obtain 178g of gynura segetum total saponin crude product;
and 5, carrying out reduced pressure liquid chromatography separation, namely dissolving 178g of the crude product of the panax notoginseng saponins by 600m L deionized water to obtain an aqueous solution of the crude product of the panax notoginseng saponins, adding 360g of silica gel into the aqueous solution of the crude product of the panax notoginseng saponins for sample mixing, simultaneously carrying out wet column packing by 5300g of silica gel with the diameter-height ratio of 1: 6, controlling the column pressure to be-0.1 Mpa after the column packing is finished, loading the sample, carrying out silica gel column chromatography, eluting by using a mixed solution of n-butyl alcohol and ethanol as an eluent, collecting fractions, concentrating the fractions to obtain 135g of the panax notoginseng saponins, wherein the yield is 9.0%, and the purity is 96.21% by liquid chromatography analysis.
The total saponins of panax notoginseng taibai with the purity of more than 95% are extracted in the embodiments 1 to 3, and the total saponins of panax notoginseng taibai extracted in the embodiments 1 to 3 have basically the same performance and purity, so the application of the total saponins of panax notoginseng taibai extracted in the embodiment 2 in the medicine for treating cardiovascular diseases is only taken as an illustration.
1. Experimental materials:
1.1 Experimental animals
The weight of Kunming mice is 18-22 g, and the weight of the Kunming mice is half of that of male and female mice; wistar rats, weighing 200-230 g, half male and half female, were provided by the animal experimental center of Heilongjiang university of traditional Chinese medicine.
1.2 test drugs
The total saponins of Panax notoginseng of example 2, pituitrin, aconitine, MDA kit, HD L kit, SOD kit, and urethane.
2. Experimental methods and results
2.1 Effect on the hypoxic survival time of Normal mice
30 mice are taken and randomly divided into 3 groups, each group comprises 10 mice, (1) a control group comprising iv 10m L/kg physiological saline, (2) a group with low dose of total saponins of panax notoginseng taibai comprising iv 5mg/kg, (3) a group with high dose of total saponins of panax notoginseng taibai comprising iv 30mg/kg, the mice are placed into 500m L ground bottles after 30min of injection, and the death time of the mice is observed, and the results are shown in table 1.
TABLE 1 comparative mouse hypoxia survival time test
| Group of | Survival time (min) |
| Control group | 45.38±12.39 |
| Low-dose group of total saponins of panax notoginseng | 73.26±16.25* |
| High-dose group of total saponins of panax notoginseng | 86.46±18.17* |
Note: p < 0.05 compared to control.
As can be seen from Table 1, the mice injected with the Panax notoginsenosides survived for a longer time under the anoxic condition than the mice not injected, which indicates that the Panax notoginsenosides can significantly improve the oxygen supply capacity of the heart.
2.2 Effect on Aconitine-induced arrhythmia in rats
30 Wistar rats are randomly divided into 3 groups, 10 rats in each group are (1) a model group is provided with physiological saline with equal volume iv, (2) a group with high and low doses of total saponins of panax notoginseng of Taibai, iv is 5mg/kg, and (3) a group with high doses of total saponins of panax notoginseng of Taibai, iv is 30 mg/kg.30min, then 20% urethane (5mg/kg) is used for carrying out intraperitoneal injection anesthesia, the animals are fixed on a mouse platform in a supine position and are connected with a B L-420 biological function experiment system, electrocardiogram is traced, after the electrocardiogram of the rats is stabilized, physiological saline is injected into the sublingual veins of a normal control group, and the other two groups are injected with 0.04% aconitine 1m L/kg (40 mu g/kg) into the sublingual veins of 5s, the occurrence time of ventricular premature beats (VP) is observed, and the occurrence rate of Ventricular Tachycardia (VT), Ventricular Fibrillation (VF) and the sinus rhythm recovery rate are counted, and the results are shown in Table 2.
TABLE 2 comparative experimental table of aconitine-induced arrhythmia in rats
Note: p < 0.05 compared to model group.
As can be seen from Table 2, the time of occurrence of the rats injected with the Panax notoginsenosides under the condition of aconitine-induced arrhythmia is obviously later than that of the rats not injected with the aconitine-induced arrhythmia, which indicates that the Panax notoginsenosides have obvious treatment effect on the arrhythmia.
2.3 Effect on chloroform induced ventricular fibrillation in mice
30 mice are taken and randomly divided into 3 groups, 10 mice in each group are (1) a model group is given with physiological saline with equal volume, (2) a group with low dose of total saponins of panax notoginseng taibai is given with iv 5mg/kg, (3) a group with high dose of total saponins of panax notoginseng taibai is given with iv 30 mg/kg., the mice are put into an inverted beaker containing 5m L chloroform cotton balls one by one within 2-3 min after being given, 0.5m L chloroform is needed to be added to each mouse, the mouse is enabled to suck the chloroform until the mouse stops breathing, the mouse is taken out immediately after the breathing stops and is connected with a B L-420 biological function experiment system, electrocardiogram is recorded, and the ventricular fibrillation incidence rate is calculated, and the result is shown in a table 3.
TABLE 3 comparative chloroform induced ventricular fibrillation in mice
| Group of | Number of chamber fluttering objects (only) | Ventricular fibrillation rate (%) |
| Model set | 9 | 90 |
| Low-dose group of total saponins of panax notoginseng | 2 | 20 |
| High-dose group of total saponins of panax notoginseng | 0 | 0 |
Note: p < 0.05 compared to model group.
As can be seen from Table 3, the ventricular fibrillation rate of the mice injected with the Panax notoginseng saponins is obviously reduced, which indicates that the Panax notoginseng saponins have obvious inhibition effect on ventricular fibrillation.
2.4 protective action against Pituitrin-induced myocardial ischemia in rats
The method comprises the steps of injecting 1U/kg of hypophysin into a sublingual vein of a rat, observing the change of an electrocardiogram, selecting a rat sensitive to the hypophysin for an experiment (the T wave is obviously increased, the ST section is increased by more than 0.1mV), using the screened sensitive rat for the model group and the total saponin group, starting the experiment after 24 hours, using each group for lavage with 1m L/100 g of stomach, injecting 20% of uratan (5m L/kg) into an abdominal cavity after 1 hour, fixing the animal on a mouse table in a supine position, connecting with a L-420 biological function experiment system, connecting with a 20% of uratan (5m L/kg) for anesthesia, connecting the animal with the L-420 biological function experiment system, recording the change of the animal serum content, and the normal blood serum superoxide dismutase content after 1 minute, and performing intravenous injection of the intravenous vein, and measuring the change of the serum superoxide dismutase content of the serum after 1 minute, and performing the serokalimetric disproportionation of the normal blood serum dehydrogenase after the test, wherein the normal serum dehydrogenase is determined by immediately after the test, the change of the serum dehydrogenase and the normal blood serum dehydrogenase after the test, the normal blood serum is judged by the intravenous vein, and the normal blood serum dehydrogenase after the normal blood serum is separated by the test, and the normal blood serum is separated by the normal blood serum dehydrogenase after the test, the normal blood serum is separated by the normal blood serum dehydrogenase after the normal blood serum is separated by the normal blood serum dehydrogenase after the.
TABLE 4 comparative test of the Effect on myocardial ischemia rats L DH, MDA and SOD
| Group of | LDH(U/mL) | MDA(nmol/mL) | SOD(U/mL) |
| Control group | 8.61±1.19** | 2.76±0.87** | 96.88±9.15** |
| Model set | 9.92±1.25* | 4.83±1.56* | 73.56±8.90* |
| Low-dose group of total saponins of panax notoginseng | 9.20±1.59** | 3.15±0.64** | 79.73±9.05** |
| High-dose group of total saponins of panax notoginseng | 9.01±1.58** | 2.82±0.83** | 90.86±9.16** |
Note: p < 0.05 compared to control; p < 0.05 compared to model group.
As can be seen from Table 4, the Wistar rat injected with the Panax notoginsenosides has low myocardial ischemia degree, which indicates that the Panax notoginsenosides have significant therapeutic effect on myocardial ischemia.
When the extract of the panax notoginseng is prepared, the panax notoginseng powder is soaked in ethanol and then is extracted by ultrasound, and the diffusion of the extracted components is accelerated and the extracted components are fully contacted with a solvent by utilizing the cavitation action and the secondary effect of the ultrasound, so that the effective components in the panax notoginseng are extracted to the maximum extent, the loss is reduced, and the extraction rate is greatly improved.
After the active substances are extracted, water and ammonia water are added into the extracting solution containing the active substances for further treatment, so that the flavonoids, phenols and other substances in the extracting solution are removed, and the subsequent purification is facilitated. The pretreated Taibai pseudo-ginseng extract after ammonia water treatment is subjected to high-speed countercurrent extraction, and the total saponins in the pretreated Taibai pseudo-ginseng extract can be basically extracted by using n-butyl alcohol as a solvent.
After high-speed countercurrent extraction, a crude product of the total saponins of panax notoginseng is obtained, the crude product contains a small amount of flavone, polysaccharide and the like, the total saponins of panax notoginseng is purified by adopting reduced pressure liquid chromatography, because the chromatographic separation of the reduced pressure column can accelerate the chromatographic speed and shorten the elution time, simultaneously one solvent gradient is developed, and the solvent gradient can be developed again by using another gradient solution after being dried, the mutual crossing and influence among all components are effectively prevented, and the separation efficiency is greatly improved, so the high-purity total saponins of panax notoginseng can be obtained after the reduced pressure liquid chromatography separation. In addition, compared with the conventional normal pressure column chromatography and rapid column chromatography, the method has the advantages of simple equipment, simple and convenient operation, high separation speed, high resolution, large separation capacity and less solvent consumption.
It should be noted that when the following claims refer to numerical ranges, it should be understood that two endpoints of each numerical range and any numerical value between the two endpoints can be selected, and since the steps and methods adopted are the same as those in embodiments 1 to 3, the preferred embodiments are described in the present invention for preventing redundancy, but once a person skilled in the art knows the basic inventive concept, other changes and modifications can be made to the embodiments. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (5)
1. The extraction method of the active substances of the panax notoginseng is characterized by comprising the following steps:
step 1, pretreatment of raw materials: drying and pulverizing radix Tongoloae Dunnii, and sieving to obtain radix Tongoloae Dunnii powder;
step 2, preparing a Taibai pseudo-ginseng extracting solution, namely adding 75-85% ethanol solution into Taibai pseudo-ginseng powder according to the proportion of 1g to 10-30 m L, soaking for 12-24 h, then carrying out ultrasonic treatment at 60 ℃ for 40-60 min, and filtering after the ultrasonic treatment is finished to obtain a first filtrate and filter residue;
adding 75-85% ethanol solution into the filter residue according to the proportion of 1g to 10-20 m L, soaking for 10-12 h, then performing ultrasonic treatment at 60 ℃ for 10-30 min, and filtering after the ultrasonic treatment is finished to obtain a second filtrate;
mixing the first filtrate and the second filtrate to obtain mixed extractive solution, concentrating the mixed extractive solution, and recovering ethanol to obtain Notoginseng radix extract;
step 3, pretreatment of the gynura segetum extract: adding deionized water which is 5-10 times of the weight of the Taibai pseudo-ginseng extract into the Taibai pseudo-ginseng extract in the step 2, stirring at 40-70 ℃ until the Taibai pseudo-ginseng extract is completely dissolved, then adding an ammonia water solution with the concentration of 10%, and adjusting the pH of the system to be 8-9 to obtain a pretreated Taibai pseudo-ginseng extract;
and 4, high-speed countercurrent extraction: pumping the pretreated radix Tongoloae Dunnii extract into an extraction tower, and then mixing the radix Tongoloae Dunnii extract and n-butanol according to the volume ratio of 1: 1-3, uniformly introducing n-butyl alcohol into the extraction tower for circulating high-speed countercurrent extraction, controlling the extraction temperature to be 40-60 ℃, performing circulating extraction for 3-5 times to obtain an extract, concentrating the extract and recovering the n-butyl alcohol to obtain a crude product of the active substance of the panax notoginseng;
step 5, carrying out reduced pressure liquid chromatography separation, namely mixing and dissolving the crude product of the pseudo-ginseng taibaiensis active substance with deionized water according to the proportion of 1 g: 3-5 m L to obtain an aqueous solution of the crude product of the pseudo-ginseng taibaiensis active substance, adding silica gel which is 1.5-2 times of the weight of the crude product of the pseudo-ginseng taibaiensis active substance into the aqueous solution of the crude product of the pseudo-ginseng taibaiensis active substance for sample mixing, simultaneously carrying out wet column packing by using the silica gel which is 20-30 times of the weight of the crude product of the pseudo-ginseng taibaiensis active substance, wherein the diameter-height ratio is 1: 4-6, controlling the column pressure to be-0.3-0.1 Mpa after the column packing is finished, carrying out sample loading, carrying out silica gel column chromatography, eluting by using a mixed solution of n-butanol and ethanol as;
the active substance of the gynura divaricata is gynura divaricata total saponin.
2. The method for extracting active substances of gynura segetum according to claim 1, wherein the gynura segetum in step 1 is dried, crushed and sieved by a 100-mesh sieve.
3. The method for extracting active substances of panax notoginseng taibai according to claim 1, wherein the ultrasonic frequency in the step 2 is 120 to 180 Hz.
4. The method for extracting active substances of panax notoginseng taibai according to claim 1, wherein the silica gel used in the step 5 has a size of 200-300 meshes.
5. Use of the active substance of Panax pseudoginseng C.H. Chen extracted according to the method of claim 1 in the preparation of a medicament for the treatment of cardiovascular diseases.
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