阅读说明：本技术 一种改进的淫羊藿提取物的制备工艺 (Improved preparation process of epimedium extract ) 是由 孙小鹦 廖萍 廖伟 廖忠 于 2020-12-25 设计创作，主要内容包括：本发明公开了一种改进的淫羊藿提取物的制备工艺,其特征在于：包括以下步骤：1)取淫羊藿药材,粉碎成粗粉；2)将淫羊藿粗粉加水低温煎煮浸润；3)向浸润后的淫羊藿中加入70％～80％乙醇,浸渍18～23小时,渗漉提取2次,合并两次提取液；4)将提取液浓缩得浓缩液；5)将淫羊藿浓缩液用大孔吸附树脂柱进行吸附洗脱得洗脱液；6)将洗脱液浓缩成浸膏；7)采用合适的干燥方法对浓缩液进行干燥、粉碎过筛,得淫羊藿提取物。本发明的优点是：工艺简单易行、效率高,适于工业化生产,提高了淫羊藿提物纯度、提取物含量高,有利于下一步淫羊藿有效成分的分离。(The invention discloses an improved preparation process of an epimedium extract, which is characterized by comprising the following steps: the method comprises the following steps: 1) pulverizing herba Epimedii into coarse powder; 2) decocting herba Epimedii coarse powder in water at low temperature; 3) adding 70-80% ethanol into the soaked epimedium, soaking for 18-23 hours, percolating and extracting for 2 times, and combining the two extracting solutions; 4) concentrating the extract to obtain a concentrated solution; 5) adsorbing and eluting herba Epimedii concentrated solution with macroporous adsorbent resin column to obtain eluate; 6) concentrating the eluent into extract; 7) drying the concentrated solution by a proper drying method, pulverizing and sieving to obtain herba Epimedii extract. The invention has the advantages that: the method has the advantages of simple and easy process, high efficiency, suitability for industrial production, improvement of the purity of the epimedium extract, high extract content and contribution to the next step of separation of the effective components of the epimedium.)

1. An improved preparation process of an epimedium extract is characterized by comprising the following steps:

1) raw material treatment: pulverizing herba Epimedii into coarse powder in a pulverizer;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration until the medicinal materials are soaked;

3) extraction: adding 70-80% ethanol into the soaked epimedium, soaking for 18-23 hours, percolating and extracting for 2 times, and combining two percolates;

4) concentration: concentrating the percolate to obtain a concentrated solution, wherein the relative density of the percolate at 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution by using a macroporous adsorption resin column, carrying out flow elution and impurity removal by using purified water with the volume of 3-10 times, carrying out secondary elution by using 50-90% ethanol with the volume of 3-8 times, and collecting epimedium ethanol eluate;

6) concentration: concentrating the eluate to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by a proper drying method, pulverizing, and sieving to obtain herba Epimedii extract.

2. The preparation method according to claim 1, wherein the temperature for low-temperature soaking in water in the step 2) is 50-60 ℃.

3. The method according to claim 2, wherein the temperature for low temperature infiltration with water is 55 ℃.

4. The production method according to claim 1, wherein in the step 3), the dipping time is 21 hours.

5. The preparation method according to claim 1, wherein in the step 3), the mass ratio of the epimedium herb to 70-80% of ethanol in the amount of ethanol extracted for 2 times is 1: 16-20.

6. The preparation method of claim 5, wherein the amount of the ethanol extracted for 2 times is 1:18 of the mass ratio of the epimedium herb to the 75% ethanol.

7. The method according to claim 1, wherein the concentration in the steps 4) and 6) is performed under reduced pressure at a temperature of 40 to 60 ℃ and a pressure of-0.20 to-0.07 MPa.

8. The preparation method according to claim 1, wherein in the step 5), the purified water with 7 times volume is used for removing impurities by flow elution, and then the ethanol with 60% -70% of 6 times volume is used for secondary elution.

9. The method according to claim 1, wherein the drying method of the concentrated solution in the step 7) is vacuum freeze drying, spray drying or evaporation drying.

10. The process according to claim 9, wherein the drying is vacuum freeze-drying at a temperature of-35 ℃ to-45 ℃ under a vacuum of 55pa for 7 hours.

Technical Field

The invention relates to the field of extraction of effective ingredients of medicaments, in particular to a preparation process of an epimedium extract.

Background

Dried stems and leaves of Epimedium brevicornum Maxim, Epimedium sagittatum Maxim, Epimedium pubescens Maxim, or Epimedium koreanum Nakai belonging to the genus berberidaceae. The epimedium is a traditional Chinese medicine tonic, has the effects of tonifying yang and kidney, strengthening the bones and the discs, dispelling wind-damp, is used for impotence and spermatorrhea, flaccidity of the bones and the discs, rheumatic arthralgia, numbness and contracture, and climacteric hypertension, can effectively inhibit staphylococcus, and is anti-aging. Icariin is one of main active ingredients of epimedium, has the activities of immunoregulation, anti-inflammation, anti-aging, anti-tumor and the like, and has certain improvement effect on relevant diseases such as nerve, immunity, endocrine, cardiovascular and bone. The existing epimedium medicinal materials have more extraction processes, but the existing processes have the following problems: the consumption of raw materials and extraction solvents is large, the content of the extract is generally low, the production cost is high, the production period is long, and the process production is not facilitated, such as:

the invention relates to a capsule for treating kidney deficiency and yang deficiency and process for preparation, wherein the capsule comprises the following components: guizhou Jianxing pharmaceutical industry Co., Ltd, discloses a preparation method as follows: weighing herba epimedii, crushing into coarse powder, soaking for 24 hours by using 60-80% ethanol as a solvent, slowly percolating, collecting percolate, recovering ethanol at 50-70 ℃ under reduced pressure, concentrating to obtain clear paste with the relative density of 1.09-1.11, drying in vacuum at 60-80 ℃, and crushing into fine powder to obtain the herba epimedii extract. The process flow is simple, but the content of icariin in the extract is low, and the extract is not purified, so that the subsequent application of the epimedium extract is not facilitated.

The invention of China CN201711368332.5 is a method for extracting icariin from epimedium, and the applicant: the extraction method disclosed by Sichuan Hesheng Biotech limited comprises the following steps: cleaning herba Epimedii, pulverizing, and decocting in water at low temperature for 30min for 2-4 times; adding ethanol into the wetted epimedium, and soaking for 12-24 h; putting the soaked herba Epimedii into a percolation pipe, and continuously pouring ethanol from the top of the percolation pipe for percolation; and (3) pouring ethanol into the percolation pipe for elution, and putting the filtrate and the eluent into a vacuum evaporation pipe for evaporation and purification. The extraction method is simple, but the times of water decoction and infiltration are more, the time is longer, the time cost of a production enterprise is increased, and the industrial production is not facilitated.

The invention relates to a epimedium extract CN202011137960.4 and relative production, wherein the applicant comprises: the preparation technology disclosed by Jiangxi Wulin Xiang biological technology company Limited is as follows: pulverizing dried herba Epimedii leaf, adding warm water, heating and concentrating to obtain concentrated solution; adding diluted ethanol water solution into the concentrated solution, preserving at constant temperature, and heating and concentrating to obtain concentrated solution; filtering the concentrated solution by using a micro-filtration membrane to obtain filtrate; eluting the filtrate with macroporous adsorbent resin to obtain eluate; and evaporating and drying the eluent by using a heating pipe to obtain solid particles. The preparation process adopts water extraction and ethanol extraction, and has the advantages of simple process, low content of the obtained extract, high concentration frequency, increased equipment requirement of the concentration process, increased production cost of enterprises, and no contribution to industrial production.

In order to solve the problems, the applicant improves the original preparation process through long-term grope tests, and solves the technical problems of low extraction efficiency, large raw material consumption, generally low icariin content and high production cost in the prior art.

Disclosure of Invention

The invention aims to provide an improved preparation process of an epimedium extract.

The invention relates to an improved preparation process of an epimedium extract, which comprises the following steps:

1) raw material treatment: pulverizing herba Epimedii into coarse powder in a pulverizer;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration until the medicinal materials are soaked;

3) extraction: adding 70-80% ethanol into the soaked epimedium, soaking for 18-23 hours, percolating and extracting for 2 times, and combining two percolates;

4) concentration: concentrating the percolate to obtain a concentrated solution, wherein the relative density of the percolate at 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution by using a macroporous adsorption resin column, carrying out flow elution and impurity removal by using purified water with the volume of 3-10 times, carrying out secondary elution by using 50-90% ethanol with the volume of 3-8 times, and collecting epimedium ethanol eluate;

6) concentration: concentrating the eluate to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by a proper drying method, pulverizing, and sieving to obtain herba Epimedii extract.

Preferably, the temperature of the low-temperature soaking by adding water is 50-60 ℃.

More preferably, the temperature for low-temperature soaking in water is 55 ℃.

Preferably, in step 3), the immersion time is 21 hours.

Preferably, in the step 3), the mass ratio of the ethanol in 2 times of extraction to the epimedium medicinal material and 70-80% ethanol is 1: 16-20.

Further preferably, in the step 3), the amount of the ethanol in 2 times of extraction is 1:18 of the mass ratio of the epimedium herb to the 75% ethanol.

Preferably, in the steps 4) and 6), the concentration mode is reduced pressure concentration, the temperature of the reduced pressure concentration is 40-60 ℃, and the pressure is-0.20-0.07 MPa;

preferably, in the step 5), the elution mode is that purified water with 7 times of volume is firstly used for removing impurities by flowing elution, and then 60% -70% ethanol with 6 times of volume is used for secondary elution;

preferably, in step 7), the drying method of the concentrated solution is vacuum freeze drying, spray drying or evaporation drying.

More preferably, the drying method is vacuum freeze drying, the drying temperature is-35 ℃ to-45 ℃, the vacuum degree is 55pa, and the drying time is 7 hours.

The invention has the advantages of

Compared with the prior art, the invention provides an improved preparation process of epimedium extract, which has the following beneficial effects:

1. the extraction process of the invention is to carry out two times of percolation extraction after crushing the epimedium medicinal material, the solvent ethanol of the method flows downwards under the action of gravity, the leachate left on the upper layer replaces the solvent position on the lower layer to continuously cause concentration difference, and the medicinal material is immersed for countless times, which is a dynamic process capable of continuous operation, so that the effective components in the epimedium medicinal material are released with high efficiency, and the extraction yield of the icariin is improved.

2. According to the invention, the ethanol extraction solvent is used for 2 times of extraction, and 2 times of concentration are carried out, so that effective components in the epimedium medicinal material can be fully released, and the content of icariin is effectively increased.

3. The infiltration step before extraction softens the medicinal materials, accelerates the fusion time of the solvent and the medicinal materials in the subsequent extraction step, and ensures that the solvent and the medicinal materials are fully fused and the extraction is more full.

4. The invention can remove the impurities in the extracting solution by carrying out adsorption elution through the macroporous adsorption resin column, is beneficial to improving the purity of the epimedium extract and is beneficial to subsequent application.

5. The invention uses water solution and ethanol water solution to elute the extracting solution, which can ensure the purity of epimedium extract.

6. The invention adopts a vacuum freeze-drying method for drying, the properties of the effective components in the extract are protected under the low-temperature condition, the extract keeps the original structure without concentration, the quality of the finished product is ensured, and the long-term storage and application of the finished product are facilitated.

7. The preparation process is simple, the operation is convenient, the requirement on working conditions is not high, the control is easy, and the content of active ingredients such as icariin in the extract is high, so that the preparation method is suitable for industrial production.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Example 1:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 55 ℃ until the medicinal materials are soaked;

3) extraction: adding 180g of 75% ethanol into the soaked epimedium medicinal material, soaking for 20 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at 50 ℃ and under the pressure of-0.20 MPa to obtain a concentrated solution, wherein the relative density of the percolate is 1.10-1.12 at 60 ℃;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 7 times volume of purified water for removing impurities, eluting with 6 times volume of 70% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 50 deg.C and-0.13 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-40 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 2:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 50 ℃ until the medicinal materials are soaked;

3) extraction: adding 160g of 76% ethanol into the soaked epimedium medicinal material, soaking for 18 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 40 ℃ and the pressure of-0.20 MPa to obtain a concentrated solution, wherein the relative density of the percolate at the temperature of 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 3 times volume of purified water for removing impurities, eluting with 3 times volume of 50% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 40 deg.C and-0.20 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-35 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 3:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 51 ℃ until the medicinal materials are soaked;

3) extraction: adding 165g of 71% ethanol into the soaked epimedium medicinal material, soaking for 19 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at 42 ℃ and under the pressure of-0.18 MPa to obtain a concentrated solution, wherein the relative density of the percolate is 1.10-1.12 at 60 ℃;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 4 times volume of purified water to remove impurities, eluting with 4 times volume of 55% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 42 deg.C and-0.18 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-36 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 4:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at 52 ℃ until the medicinal materials are soaked;

3) extraction: adding 170g of 72% ethanol into the soaked epimedium medicinal material, soaking for 20 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at 44 ℃ and under the pressure of-0.16 MPa to obtain a concentrated solution, wherein the relative density of the percolate is 1.10-1.12 at 60 ℃;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 5 times volume of purified water for removing impurities, eluting with 5 times volume of 60% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 44 deg.C and-0.16 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-37 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 5:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 53 ℃ until the medicinal materials are soaked;

3) extraction: adding 175g of 73% ethanol into the soaked epimedium medicinal material, soaking for 21 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 46 ℃ and the pressure of-0.14 MPa to obtain a concentrated solution, wherein the relative density of the percolate at the temperature of 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 6 times volume of purified water to remove impurities, eluting with 6 times volume of 65% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 46 deg.C and-0.14 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-38 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 6:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at 54 ℃ until the medicinal materials are soaked;

3) extraction: adding 180g of 74% ethanol into the soaked epimedium medicinal material, soaking for 22 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 48 ℃ and under the pressure of-0.12 MPa to obtain a concentrated solution, wherein the relative density of the percolate is 1.10-1.12 at the temperature of 60 ℃;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 7 times volume of purified water for removing impurities, eluting with 7 times volume of 70% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 48 deg.C and-0.12 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-39 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 7:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 55 ℃ until the medicinal materials are soaked;

3) extraction: adding 185g of 75% ethanol into the soaked epimedium medicinal material, soaking for 23 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at 50 ℃ and under the pressure of-0.10 MPa to obtain a concentrated solution, wherein the relative density of the percolate at 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 8 times volume of purified water to remove impurities, eluting with 8 times volume of 75% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 50 deg.C and-0.10 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-40 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 8:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 57 ℃ until the medicinal materials are soaked;

3) extraction: adding 190g of 76% ethanol into the soaked epimedium medicinal material, soaking for 23 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 52 ℃ and the pressure of-0.08 MPa to obtain a concentrated solution, wherein the relative density of the percolate is 1.10-1.12 at the temperature of 60 ℃;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 9 times volume of purified water to remove impurities, eluting with 8 times volume of 80% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 52 deg.C and-0.08 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-41 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 9:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 58 ℃ until the medicinal materials are soaked;

3) extraction: adding 195g of 77% ethanol into the soaked epimedium medicinal material, soaking for 23 hours, percolating and extracting for 2 times, and combining percolates of the two times;

4) concentration: concentrating the percolate at 54 ℃ and under the pressure of-0.09 MPa to obtain a concentrated solution, wherein the relative density of the percolate is 1.10-1.12 at 60 ℃;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 10 times volume of purified water to remove impurities, eluting with 8 times volume of 85% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 54 deg.C and-0.09 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-42 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 10:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 59 ℃ until the medicinal materials are soaked;

3) extraction: adding 196g of 78% ethanol into the soaked epimedium medicinal material, soaking for 23 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 56 ℃ and under the pressure of-0.07 MPa to obtain a concentrated solution, wherein the relative density of the percolate at the temperature of 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 10 times volume of purified water to remove impurities, eluting with 8 times volume of 86% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 56 deg.C and-0.07 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-43 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 11:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 60 ℃ until the medicinal materials are soaked;

3) extraction: adding 198g of 79% ethanol into the soaked epimedium medicinal material, soaking for 23 hours, percolating and extracting for 2 times, and combining percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 58 ℃ and under the pressure of-0.07 MPa to obtain a concentrated solution, wherein the relative density of the percolate at the temperature of 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 10 times volume of purified water for removing impurities, eluting with 8 times volume of 88% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 58 deg.C and-0.07 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-44 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

Example 12:

1) raw material treatment: taking 10g of epimedium herb, and putting the epimedium herb into a grinder to be ground into coarse powder;

2) infiltrating: placing the crushed coarse powder of the epimedium herb in an extraction tank, adding water for low-temperature infiltration at the infiltration temperature of 60 ℃ until the medicinal materials are soaked;

3) extraction: adding 200g of 80% ethanol into the soaked epimedium medicinal material, soaking for 23 hours, percolating and extracting for 2 times, and mixing percolates of the two times;

4) concentration: concentrating the percolate at the temperature of 60 ℃ and under the pressure of-0.07 MPa to obtain a concentrated solution, wherein the relative density of the percolate at the temperature of 60 ℃ is 1.10-1.12;

5) and (3) purification: adsorbing and eluting the concentrated solution with macroporous adsorbent resin column, eluting with 10 times volume of purified water for removing impurities, eluting with 8 times volume of 90% ethanol for the second time, and collecting herba Epimedii ethanol eluate;

6) concentration: concentrating the eluate at 60 deg.C and-0.07 MPa to obtain extract with relative density of 1.14-1.21 at 50 deg.C;

7) and (3) drying: drying the concentrated solution by vacuum freeze drying at-45 deg.C under vacuum degree of 55pa for 7 hr, pulverizing, and sieving to obtain herba Epimedii extract.

To further verify the effectiveness of the present invention, the invention was subjected to a series of verification tests, summarized as follows:

the content of icariin described below was measured by high performance liquid chromatography (appendix VI D of the first part of the 2010 edition of the Chinese pharmacopoeia).

Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-water (30: 70) as a mobile phase; the detection wavelength was 270 mn. The number of theoretical plates should not be less than 1500 calculated on icariin peaks.

Preparation of control solutions: taking appropriate amount of icariin reference substance, precisely weighing, and adding methanol to obtain solution containing 0.1mg per 1 ml.

Preparation of a test solution: weighing about 0.1g of the powder, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of dilute ethanol, weighing, ultrasonically treating for 1 hr, weighing again, supplementing the reduced weight with dilute ethanol, shaking, filtering, and collecting the filtrate.

The determination method comprises the following steps: precisely sucking 10ul of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and measuring.

First, wetting Process investigation

1. Content of research

The inventor develops the infiltration process research before extraction aiming at the problems of long fusion time, large solvent dosage, insufficient extraction and the like of the extraction solvent and the epimedium medicinal material in the original preparation process.

2. Technical study of wetting process

The original epimedium extraction technology has no wetting process, insufficient extraction, long extraction time, large solvent consumption and is not beneficial to industrial production, so the research aims at solving the problems of low extract content, insufficient extraction and the like.

The method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 50 deg.C, 55 deg.C, and 60 deg.C at low temperature to soak the medicinal materials, performing the rest steps according to the invention in example 1 to obtain herba Epimedii extract, and measuring icariin content according to high performance liquid chromatography (appendix VI D of 2010 edition).

The second method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 50 deg.C, 55 deg.C, and 60 deg.C at low temperature to soak the medicinal materials, performing the rest steps according to the invention in example 1 to obtain herba Epimedii extract, and measuring icariin content according to high performance liquid chromatography (appendix VI D of 2010 edition).

Table 1 wetting mode examination table

As can be seen from the above table, the icariin content in the first method is higher than that in the second method, so the optimum wetting process in establishing the epimedium extraction process is as follows: the optimal soaking temperature is 55 ℃ by adopting a soaking and wetting method.

Second, investigation of extraction Process

The investigation content is as follows: since the content of icariin extracted by using ethanol solution in one time is low in the original extraction process, the research mainly focuses on the extraction method, the extraction solvent, the solvent dosage, the extraction times, the extraction time and the like.

1. Examination of extraction methods

The method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C for 24 hr, and extracting with water under heating and refluxing to examine icariin content. See table 2.

The second method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C at low temperature, adding 70% ethanol, heating and reflux extracting for 24 hr, and observing icariin content. See table 2.

The third method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C, adding 70% ethanol, percolating and extracting for 24 hr, and observing icariin content. See table 2.

The method four comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C, adding 70% ethanol, percolating for 24 hr, filtering, adding 70% ethanol into the residue, percolating for 24 hr, mixing filtrates, and examining icariin content. See table 2.

Table 2: method of extraction survey table

As can be seen from the above table, ethanol extraction is better than water extraction, percolation extraction is better than heating reflux, and percolation extraction is better than percolation extraction for 2 times, so that the optimal extraction process of the present invention is determined as follows: percolating and extracting for 2 times by using ethanol as an extraction solvent.

2. Investigation of solvent dosage

The method comprises the following steps: taking 10g of epimedium herb, crushing into coarse powder, adding water for low-temperature infiltration at 55 ℃ until the medicinal materials are soaked, respectively adding 160g, 180g and 200g of 70% ethanol for percolation extraction for 2 times, merging filtrates, and inspecting the content of icariin. See table 3.

Table 3: examination table of extraction solvent 70% ethanol and 70% ethanol content

As is clear from the above table, the addition of 160g, 180g and 200g of 70% ethanol icariin was not particularly preferable, and further investigation was required.

The second method comprises the following steps: taking 10g of epimedium herb, crushing into coarse powder, adding water for low-temperature infiltration at 55 ℃ until the medicinal materials are soaked, respectively adding 160g, 180g and 200g of 75% ethanol for percolation extraction for 2 times, merging filtrates, and inspecting the content of icariin. See table 4.

Table 4: 75% ethanol and 75% ethanol content survey table

As can be seen from the above table, it is preferable to add 75% ethanol icariin in an amount of 160g, 180g, or 200g, and particularly, to add 65% ethanol icariin in an amount of 180g or 200g, but from the viewpoint of cost saving, 180g of 75% ethanol is preferable for extraction.

The third method comprises the following steps: taking 10g of epimedium herb, crushing into coarse powder, adding water for low-temperature infiltration at 55 ℃ until the medicinal materials are soaked, respectively adding 160g, 180g and 200g of 80% ethanol for percolation extraction for 2 times, merging filtrates, and inspecting the content of icariin. See table 5.

Table 5: examination table of extraction solvent 80% ethanol and 80% ethanol

As can be seen from the above table, the content of 80% ethanol icariin added in 160g, 180g, 200g was reduced relative to 175g, and therefore, the optimal amount of solvent was determined as follows: 75% ethanol 180 g.

3. Extraction time review

The method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C, percolating with 180g 75% ethanol for 2 times (18 hr for two times), and observing icariin content. See table 6.

The second method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C, percolating with 180g 75% ethanol for 2 times, extracting for 20 hr, and observing icariin content. See table 6.

The third method comprises the following steps: pulverizing herba Epimedii into coarse powder, soaking in water at 55 deg.C, percolating with 180g 75% ethanol for 2 times (23 hr for two times), and observing icariin content. See table 6.

Table 6: extracting a time look-up table

As can be seen from the above table, the icariin content was high at 20h and 23h of extraction time, and 20h was preferable from the viewpoint of time cost.

Third, investigation of drying process

The method comprises the following steps: pulverizing herba Epimedii, soaking, percolating for 2 times, mixing filtrates, concentrating, purifying, concentrating, evaporating the concentrated solution, drying to obtain the final product, and observing the required drying time and icariin content. See table 7.

The second method comprises the following steps: pulverizing herba Epimedii, soaking, percolating for 2 times, mixing filtrates, concentrating, purifying, concentrating, spray drying the concentrated solution to obtain the final product, and observing the required drying time and icariin content. See table 7.

The third method comprises the following steps: pulverizing herba Epimedii, soaking, percolating for 2 times, mixing filtrates, concentrating, purifying, concentrating, vacuum freeze drying to obtain the final product, and observing drying time and icariin content. See table 7.

Table 7: drying process survey table

As can be seen from the above table, the vacuum freeze-drying method requires the shortest drying time and the least loss of icariin, compared to the evaporation drying and the spray drying, and thus, the drying method of the epimedium extract was determined to be the vacuum freeze-drying method.

Fourth, comparative test

The icariin contents of comparative examples 1 to 3 of the present invention and example 1 of the present invention were compared. See table 8.

Comparative example 1: the Chinese invention is CN 200910307558.3A yang invigorating capsule for treating kidney deficiency and yang deficiency and a preparation method thereof, the preparation method is disclosed as follows: weighing herba epimedii, crushing into coarse powder, soaking for 24 hours by using 70% ethanol as a solvent, slowly percolating, collecting percolate, recovering ethanol at 60 ℃ under reduced pressure, concentrating to obtain clear paste with the relative density of 1.09-1.11, drying in vacuum at 70 ℃, and crushing into fine powder to obtain the herba epimedii extract.

Comparative example 2: the Chinese invention CN 201711368332.5A method for extracting icariin from epimedium herb, which discloses the following extraction method: cleaning herba Epimedii, pulverizing, adding water, decocting at 40-60 deg.C for 30min for 2-4 times at low temperature for 2-4 times, adding acetic acid, and evaporating acetic acid by high temperature evaporation for 2-4 min; adding ethanol into the wetted epimedium, and soaking for 12-24 h; putting the soaked herba Epimedii into a percolation pipe, and continuously pouring ethanol from the top of the percolation pipe for percolation; and (3) filling ethanol into the infiltration tube for elution, and putting the filtrate and the eluent into a vacuum evaporation tube for evaporation and purification to obtain the epimedium extract.

Comparative example 3: the invention relates to a epimedium extract CN202011137960.4 and a preparation process thereof, and the preparation process is disclosed as follows: pulverizing dried herba Epimedii leaf, adding warm water, heating and concentrating to obtain concentrated solution, wherein the heating temperature is 55-70 deg.C, the heating time is 25-35min, and the concentration time is 3-5 times; adding 60-70% ethanol water solution by volume fraction into the concentrated solution, wherein the mass ratio of the epimedium leaves to the added ethanol solvent is 1: 8-11, preserving at constant temperature for 5-8h, wherein the preserving temperature is 35-45 ℃, then heating and concentrating to obtain concentrated solution, the heating temperature is kept between 55 ℃ and 70 ℃, the heating time is 25-35min, and the concentrating frequency is 3-5 times; filtering the concentrated solution by using a micro-filtration membrane to obtain filtrate; eluting the filtrate with macroporous adsorbent resin to obtain eluate; and evaporating and drying the eluent by using a heating pipe to obtain solid particles.

Table 8: comparative test icariin content meter

As can be seen from the above table, the icariin content in example 1 is superior to that in comparative examples 1, 2 and 3.

While the invention has been described in detail in the foregoing by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that certain changes and modifications may be made therein based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.