阅读说明：本技术 一种刺五加总皂苷、提取方法及应用 (Acanthopanax senticosus total saponin, extraction method and application ) 是由 马佰偏 孙冬云 于 2021-07-30 设计创作，主要内容包括：本发明公开一种刺五加总皂苷、提取方法及应用,涉及生物制药技术领域。取刺五加茎、根茎或根,用水作溶剂提取多次,获得刺五加初级提取液；将获得的刺五加初级提取液采用膜浓缩设备进行膜浓缩,然后进行减压浓缩；获得减压浓缩液；冷却至常温后加乙醇溶液,搅拌均匀,醇沉；取醇沉后的上清液,采用多效浓缩器进行减压浓缩,用水溶解,用大孔吸附树脂进行吸附,用乙醇溶液进行洗脱；加氧化镁进行脱色,过滤,浓缩,喷雾干燥,过筛,即得。本发明采用膜浓缩,可以增加浓缩效率,低温浓缩能减少有效成分损失。本发明操作简单方便,产品收率高,可提高0.5-1％。本发明用YKDH-2大孔吸附树脂进行吸附,效果较好。(The invention discloses a acanthopanax senticosus total saponin, an extraction method and application thereof, and relates to the technical field of biological pharmacy. Extracting stems, rhizomes or roots of acanthopanax for multiple times by using water as a solvent to obtain a primary acanthopanax extracting solution; performing membrane concentration on the obtained acanthopanax primary extract by adopting membrane concentration equipment, and then performing reduced pressure concentration; obtaining a reduced pressure concentrated solution; cooling to normal temperature, adding ethanol solution, stirring, and precipitating with ethanol; collecting supernatant after alcohol precipitation, concentrating under reduced pressure with multiple-effect concentrator, dissolving with water, adsorbing with macroporous adsorbent resin, and eluting with ethanol solution; adding magnesium oxide for decolorizing, filtering, concentrating, spray drying, and sieving. The invention adopts membrane concentration, can increase the concentration efficiency, and can reduce the loss of effective components by low-temperature concentration. The invention has simple and convenient operation and high product yield which can be improved by 0.5 to 1 percent. The invention uses YKDH-2 macroporous adsorption resin for adsorption, and has better effect.)

1. The method for extracting the acanthopanax senticosus total saponins is characterized by comprising the following steps:

step one, taking stems, rhizomes or roots of acanthopanax senticosus, and extracting for multiple times by using water as a solvent to obtain a primary acanthopanax senticosus extracting solution;

step two, performing membrane concentration on the obtained acanthopanax primary extract by adopting membrane concentration equipment, and then performing reduced pressure concentration by adopting a multi-effect concentrator; obtaining a reduced pressure concentrated solution;

step three, cooling the reduced pressure concentrated solution obtained in the step two to normal temperature, adding an ethanol solution, uniformly stirring, and precipitating with ethanol;

step four, taking the supernatant obtained after alcohol precipitation in the step three, carrying out reduced pressure concentration by adopting a multi-effect concentrator, dissolving by using water, adsorbing by using macroporous adsorption resin, and eluting by using an ethanol solution;

and step five, adding magnesium oxide into the eluent obtained in the step four for decoloring, filtering, concentrating, spray-drying and sieving to obtain the acanthopanax senticosus total saponin.

2. The method for extracting acanthopanax senticosus total saponin according to claim 1, wherein in the first step, water is used as a solvent for extraction for 3 times, the first time is decocted for 1 hour by 8-10 times of water, the second time is decocted for 1 hour by 6-8 times of water, and the third time is decocted for 20 minutes by 5-6 times of water.

3. The method of claim 2, wherein the third extraction water is recycled as the first extraction water of the next batch.

4. The method for extracting acanthopanax senticosus total saponin according to claim 1, wherein the vacuum degree in the second step of reduced pressure concentration is-0.07 to-0.09 MPa.

5. The method for extracting acanthopanax senticosus total saponin according to claim 1, wherein in the third step, after cooling to normal temperature, 80% ethanol solution which is 5-6 times of the normal temperature is added, and the mixture is stirred uniformly.

6. The method for extracting acanthopanax senticosus total saponin according to claim 1, wherein in the third step, alcohol precipitation is carried out for 12 hours.

7. The method for extracting acanthopanax senticosus total saponin according to claim 1, wherein in the fourth step, a multi-effect concentrator is adopted for carrying out reduced pressure concentration, and the vacuum degree is-0.07 to-0.09 MPa;

dissolving with water until each liter of liquid is equivalent to each kilogram of medicinal materials;

eluting with 60% ethanol solution at a flow rate of 1.5BV and a dosage of 2 BV.

8. The method for extracting acanthopanax senticosus total saponin according to claim 1, wherein in the step five, the acanthopanax senticosus total saponin is concentrated to a relative density of 1.17-1.25gcm^-1And sieving with 80 mesh sieve.

9. Total saponins of Acanthopanax senticosus harms extracted by the extraction method according to any one of claims 1 to 8.

10. Use of the total saponins of acanthopanax senticosus according to claim 9 in the preparation of immunopotentiator drugs.

Technical Field

The invention belongs to the technical field of biological pharmacy, and particularly relates to acanthopanax senticosus total saponins, an extraction method and application thereof.

Background

At present, acanthopanax is one of acanthopanax of the family araliaceae, has mild medicine property and no toxicity, has the effects of tonifying and strengthening body, tonifying qi and spleen, strengthening body resistance, tranquilizing and allaying excitement and the like, is an immunopotentiator, can regulate the metabolism of a human body and enhance the resistance of the human body to various diseases, is mainly distributed in northeast and north China areas, and has a long history as a medicine in Chinese medicine and pharmacology. Modern medical research proves that acanthopanax has the effects of resisting inflammation, tumors, radiation, aging, immunity and the like. Has wide physiological activity and thus is concerned by scholars at home and abroad.

With the intensive research on acanthopanax senticosus in recent years, it is found that the main active ingredient with pharmacological action is saponin, and is mostly oleanane type triterpene saponin. Eleutheroside I, K, M and eleutheroside A, B, C, D, E, F isolated from leaves of Acanthopanax senticosus in the early 80 th of the 20 th century, and eleutheroside A1, A2, A3, A4, B, C1, C2, C3, C4, D1, D2, D3 and E isolated from Shao Chunjie et al on this basis, were all oleanane type compounds.

The traditional process is a method of extracting acanthopanax root decoction pieces by water or 75% ethanol, or decoloring by macroporous resin after extraction, so that the extraction and separation efficiency is low, and the purity of effective parts or components is not high, therefore, the research on the extraction and separation method of the acanthopanax root saponin with more effective and high purity has important significance.

Although there are many methods for extracting acanthopanax senticosus total saponins so far, the methods all have many disadvantages, such as long extraction time of an extraction method which adopts water or a proportionally mixed organic solvent aqueous solution as an extracting solution according to saponin characteristics; ultrasonic extraction methods that achieve extraction by direct cell disruption can result in increased impurities; the microwave extraction method for extracting saponin by microwave field has high requirements on equipment.

The main methods for separating and purifying the acanthopanax senticosus total saponins at present comprise: macroporous resin adsorption, chemical extraction and ultrafiltration membrane. Although the above methods can achieve the purpose of purification, the single method has disadvantages and shortcomings.

The prior art provides a method for extracting acanthopanax senticosus total saponin from acanthopanax senticosus leaves, which comprises the following steps:

step 1: grinding the dried leaves of radix Acanthopanacis Senticosi into powder, mixing with ethanol water solution, performing ultrasonic treatment for 10min-20min, filtering to obtain first extractive solution, adding ethanol water solution with the same volume as the above residue, repeating the above steps for 1-2 times, and mixing the obtained extractive solution with the first extractive solution;

wherein the volume ratio of the acanthopanax leaf powder to the ethanol water solution is 1 (20-30);

step 2: concentrating the mixed extracting solution obtained in the step 1 to 10-30% of the original volume, adding activated carbon, heating in a water bath at 30-60 ℃ for 10-15 min, and filtering to obtain a filtrate;

and step 3: adding petroleum ether with the same volume as the petroleum ether into the filtrate obtained in the step 2, decoloring for several times until the petroleum ether layer is clear and colorless, retaining the lower-layer extracting solution, adding saturated n-butyl alcohol with the same volume as the petroleum ether layer into the lower-layer extracting solution, extracting for 2-3 times, uniformly mixing the extracted upper-layer n-butyl alcohol solution, then performing rotary evaporation on the mixed n-butyl alcohol solution to remove the n-butyl alcohol, adding deionized water for dissolving, and freeze-drying to obtain a powder;

and 4, step 4: soaking the macroporous resin in 15-20 wt.% of sodium hydroxide aqueous solution for 6-12 h, washing the macroporous resin with deionized water until the discharged cleaning solution is neutral, soaking the macroporous resin in 15-20 wt.% of hydrochloric acid solution for 6-12 h, washing the macroporous resin with deionized water until the discharged cleaning solution is neutral, soaking the macroporous resin with absolute ethyl alcohol for 6-12 h, filling the macroporous resin into a chromatographic column, adding deionized water for washing until the effluent is white and turbid, dissolving the powder obtained in the step 3 in 10ml of deionized water, and filling the solution into the chromatographic column for static adsorption for 30-50 min;

wherein the mass ratio of the macroporous resin to the powder is (10-30) g:1 g;

and 5: and (4) washing the macroporous resin in the chromatographic column obtained in the step (4) by using deionized water until the eluent is clear, eluting the chromatographic column by using 70-90 wt.% of ethanol water solution to obtain eluent, evaporating to remove ethanol in the eluent, and freeze-drying to obtain the acanthopanax senticosus total saponin dry powder.

Through the above analysis, the problems and defects of the prior art are as follows:

(1) in the prior art, the concentration efficiency is low, low-temperature concentration is not adopted, and the loss of effective components is easy to cause.

(2) The preparation process in the prior art has the disadvantages of complex operation and low product yield.

(3) In the preparation process in the prior art, the adsorption effect is poor.

Disclosure of Invention

In order to overcome the problems in the related art, the disclosed embodiment of the invention provides acanthopanax senticosus total saponin, an extraction method and application. The technical scheme is as follows:

according to a first aspect of the disclosed embodiments of the present invention, there is provided a method for extracting total saponins of acanthopanax senticosus, comprising:

step one, taking stems, rhizomes or roots of acanthopanax senticosus, and extracting for multiple times by using water as a solvent to obtain a primary acanthopanax senticosus extracting solution;

step two, performing membrane concentration on the obtained acanthopanax primary extract by adopting membrane concentration equipment, and then performing reduced pressure concentration by adopting a multi-effect concentrator; obtaining a reduced pressure concentrated solution;

step three, cooling the reduced pressure concentrated solution obtained in the step two to normal temperature, adding an ethanol solution, uniformly stirring, and precipitating with ethanol;

step four, taking the supernatant obtained after alcohol precipitation in the step three, carrying out reduced pressure concentration by adopting a multi-effect concentrator, dissolving by using water, adsorbing by using macroporous adsorption resin, and eluting by using an ethanol solution;

and step five, adding magnesium oxide (added according to actual requirements) into the eluent obtained in the step four for decoloring, filtering, concentrating, spray-drying and sieving to obtain the acanthopanax senticosus total saponin.

In one embodiment of the invention, in the first step, water is used as a solvent for extraction for 3 times, the first time is decocted for 1 hour by 8-10 times of water, the second time is decocted for 1 hour by 6-8 times of water, and the third time is decocted for 20 minutes by 5-6 times of water;

in an embodiment of the present invention, the third extraction water is recycled as the first extraction water of the next batch.

In one embodiment of the invention, the vacuum degree in the second step of reduced pressure concentration is-0.07 MPa to-0.09 MPa.

In an embodiment of the invention, in the third step, after cooling to normal temperature, 5-6 times of 80% ethanol solution is added, and the mixture is stirred uniformly.

In one embodiment of the present invention, in the third step, alcohol precipitation is performed for 12 hours.

In one embodiment of the invention, in the fourth step, a multi-effect concentrator is adopted for carrying out reduced pressure concentration, and the vacuum degree is-0.07 MPa to-0.09 MPa;

dissolving with water until each liter of liquid is equivalent to each kilogram of medicinal materials;

eluting with 60% ethanol solution at a flow rate of 1.5BV and a dosage of 2 BV.

In one embodiment of the present invention, in the step five, the mixture is concentrated to a relative density of 1.17-1.25gcm^-1And sieving with 80 mesh sieve.

According to a second aspect of the disclosed embodiments of the invention, there is provided a acanthopanax senticosus total saponin extracted by the extraction method.

According to a third aspect of the disclosed embodiments of the present invention, there is provided a use of the total saponins of acanthopanax senticosus harms in preparing an immunopotentiator medicament.

The technical scheme provided by the embodiment of the invention has the following beneficial effects:

the invention adopts membrane concentration, can increase the concentration efficiency, and can reduce the loss of effective components by low-temperature concentration.

The invention has simple and convenient operation and high product yield which can be improved by 0.5 to 1 percent.

The invention uses YKDH-2 macroporous adsorption resin for adsorption, and has better effect.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as disclosed.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present disclosure and together with the description, serve to explain the principles of the disclosure.

Fig. 1 is a flow chart of the method for extracting total saponins of acanthopanax senticosus provided by the embodiment of the invention.

Detailed Description

Reference will now be made in detail to the exemplary embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, like numbers in different drawings represent the same or similar elements unless otherwise indicated. The implementations described in the exemplary embodiments below are not intended to represent all implementations consistent with the present disclosure. Rather, they are merely examples of apparatus and methods consistent with certain aspects of the present disclosure, as detailed in the appended claims.

As shown in fig. 1, the method for extracting total saponins of acanthopanax senticosus provided by the embodiment of the present disclosure comprises:

s101, extracting stems, rhizomes or roots of acanthopanax senticosus for 3 times by using water as a solvent, decocting the stems, rhizomes or roots of acanthopanax senticosus for 1 hour by using 8-10 times of water for the first time, decocting the stems, rhizomes or roots of acanthopanax senticosus for 1 hour by using 6-8 times of water for the second time, decocting the stems, rhizomes or roots of acanthopanax senticosus for 1 hour by using 5-6 times of water for the third time for 20 minutes, and using the extracted water for the third time as the extracted water for the first time in the next batch.

S102, performing membrane concentration on the liquid medicine by adopting a membrane concentration device, and then performing reduced pressure concentration (the vacuum degree is between-0.06 MPa and-0.09 MPa) to the relative density is between 1.26 and 1.28 (measured at 80 ℃) by adopting a multi-effect concentrator;

s103, cooling to normal temperature, adding 80% ethanol solution which is 5-6 times of the normal temperature, uniformly stirring, and carrying out alcohol precipitation for 12 hours;

s104, carrying out reduced pressure concentration (vacuum degree of-0.06 MPa to-0.09 MPa) on the supernatant by using a multi-effect concentrator until the relative density is 1.30 to 1.32 (measured at 80 ℃), dissolving the supernatant with water until each liter of liquid is equivalent to each kilogram of medicinal material, adsorbing the supernatant with YKDH-2 macroporous adsorption resin, eluting the supernatant with 60 percent ethanol solution, wherein the using amount of eluent is 2BV, and the flow rate is 1.5 BV;

and S105, adding magnesium oxide into the eluent for decoloring, filtering, concentrating until the relative density is 1.17-1.25 (measured at normal temperature), spray-drying, and sieving by using a 80-mesh sieve to obtain the product.

The technical solution of the present invention is further described below with reference to the experimental results.

Experiments show that:

the invention adopts membrane concentration, can increase the concentration efficiency, and can reduce the loss of effective components by low-temperature concentration.

The invention has simple and convenient operation and high product yield which can be improved by 0.5 to 1 percent.

The invention uses YKDH-2 macroporous adsorption resin for adsorption, and has better effect.

Other embodiments of the disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the disclosure disclosed herein. This application is intended to cover any variations, uses, or adaptations of the disclosure following, in general, the principles of the disclosure and including such departures from the present disclosure as come within known or customary practice within the art to which the disclosure pertains. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the disclosure being indicated by the following claims.

It will be understood that the present disclosure is not limited to the precise arrangements described above and shown in the drawings and that various modifications and changes may be made without departing from the scope thereof. The scope of the present disclosure should be limited only by the attached claims.