Preparation method of epimedium sagittatum extract

Abstract

The invention discloses a preparation method of an epimedium sagittatum extract. The preparation method comprises the steps of blanching, back washing, enzyme treatment, ethanol extraction, duplex filtration, column purification and concentration drying. According to the method disclosed by the invention, food-grade sodium carbonate used during blanching is safe and non-toxic, and a waxy coat on the surface of epimedium sagittatum is dissolved by a method of combining blanching and enzyme treatment, so that the problem of blockage of a subsequent duplex filter is solved, and the extraction rate of icariin in the epimedium extract is also improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of extract preparation, and particularly relates to a preparation method of Epimedium sagittatum extract. BACKGROUND

[0002] Epimedium is the dry leaves of Epimedium brevicornu Maxim., Epimedium sagittatum (Sieb.et Zucc.) Maxim., Epimedium pubescens Maxim. or Epimedium koreanum Nakai, belonging to perennial herb, and the leaves are used for treating kidney yang deficiency, impotence and spermatorrhea, and softening of muscles and bones. At present, Epimedium brevicornu is mainly used to prepare Epimedium extract in China, but with the shortage of wild resources of Epimedium brevicornu, it is necessary to find a substitute for Epimedium. Since Epimedium sagittatum is abundant in resources, it becomes the first choice for the substitute of Epimedium brevicornu.

[0003] At present, there are reports on the preparation of Epimedium extract by using Epimedium sagittatum. However, the phenomenon of clogging the screen of the extraction liquid of Epimedium sagittatum is prone to occur in the extraction process, which is mainly due to the fact that there is a wax layer on the surface of the Epimedium sagittatum medicinal material, and the wax layer enters the extraction liquid in the extraction process, resulting in clogging of the double filter during filtration. Therefore, it is of great significance to find an extraction method for decomposing the wax layer on the surface of Epimedium sagittatum for efficient use of Epimedium. The present application uses the method of scalding combined with enzyme treatment to pretreat the Epimedium sagittatum medicinal material, which not only solves the problem of difficult filtration of the Epimedium sagittatum extraction liquid, but also improves the extraction rate of icariin in the Epimedium sagittatum extract. SUMMARY

[0004] The present application aims to provide a preparation method of Epimedium sagittatum extract, so that the obtained Epimedium sagittatum extraction liquid is easy to filter, and the extract after drying has the characteristics of high icariin extraction rate.

[0005] The technical scheme adopted by the present application to solve the technical problems is as follows:

[0006] A preparation method of Epimedium sagittatum extract, comprising the following steps:

[0007] Step one, scalding: put Epimedium sagittatum into an extraction tank, add a certain volume of hot water containing sodium carbonate to scald for a period of time, then empty the solution in the extraction tank, and reserve the Epimedium sagittatum medicinal material;

[0008] Step two, re-washing: add a certain volume of pure water to the extraction tank, rotate and stir for a period of time, then empty the liquid in the extraction tank, repeat 2-3 times, and drain the water on the surface of the Epimedium sagittatum leaves in the extraction tank;

[0009] Step three, enzyme treatment: add a certain volume of pure water containing complex enzymes to the extraction tank, heat the extraction tank to a certain temperature, stir for a period of time, and then empty the solution in the extraction tank;

[0010] Step four, ethanol extraction: add a certain volume of ethanol solution to the extraction tank, heat and stir for a period of time;

[0011] Step five, double filtration: the extraction liquid after extraction is filtered through a double filter to obtain an Epimedium sagittatum extract;

[0012] Step six, column purification: the Epimedium sagittatum extract obtained in step five is concentrated to 1 / 2 of the original volume, then loaded onto a D101 macroporous resin, eluted with an ethanol solution, and the eluate is collected;

[0013] Step seven, concentration and drying: the Epimedium sagittatum eluate is concentrated and dried to obtain an Epimedium sagittatum extract.

[0014] Preferably, in step one, the mass-volume ratio of Epimedium sagittatum to hot water is 1:10-18 g / mL, the temperature of the hot water is 60-90°C, the mass concentration of food-grade sodium carbonate in the hot water is 4-25 g / L, and the blanching time is 1-5 min.

[0015] Preferably, in step two, the mass-volume ratio of Epimedium sagittatum to pure water is 1:10-18 g / mL, the stirring rate is 30-50 rpm / min, and the stirring time is 10-30 min.

[0016] Preferably, in step three, the mass-volume ratio of Epimedium sagittatum to pure water is 1:10-18 g / mL, the complex enzyme is papain and bromelain, the mass fraction of the complex enzyme is 1%-3%, the mass ratio of papain to bromelain is 1:1-3, the enzymolysis temperature is 30-50°C, the stirring rate is 30-50 rpm / min, and the stirring time is 5-10 min.

[0017] Preferably, in step four, the mass-volume ratio of Epimedium sagittatum to ethanol solution is 1:10-18 g / mL, the concentration of the ethanol solution is 20-30%, the stirring rate is 30-50 rpm / min, the extraction time is 2-3 h, and the extraction temperature is 60-90°C.

[0018] Preferably, in step five, the mesh size of the double filter is 250-400.

[0019] Preferably, in the step six, the feeding rate of the Epimedium sagittatum Maxim. concentrated solution is 0.8-1.5 BV / h, the volume fraction of the ethanol eluent is 40-60%, the elution rate is 0.8-1.5 BV / h, and the eluent amount is 1.5-2.5 BV.

[0020] Preferably, in the step seven, the drying method is plate vacuum drying or belt drying.

[0021] The present application has the following beneficial effects:

[0022] 1. The present application dissolves the wax layer on the surface of Epimedium sagittatum Maxim. by the method of scouring and enzyme treatment, solves the problem of subsequent double filter blockage, improves the filtering efficiency, and reduces the labor cost.

[0023] 2. The production process of the present application is simple, and no toxic solvent is used in the whole production process, which is green and safe.

[0024] 3. Compared with the conventional ethanol extraction, the extraction rate of icariin in the Epimedium sagittatum Maxim. extract obtained by the present application is increased by more than 1.2 times. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present application will be described below in a clear and complete manner. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0026] Embodiment 1

[0027] The present embodiment provides a preparation method of Epimedium sagittatum Maxim. extract, and the method steps are specifically as follows:

[0028] (1) 1 kg of Epimedium sagittatum Maxim. is weighed into an extraction tank, 10 L of hot water with a temperature of 60 DEG C is added, 40 g of sodium carbonate is dissolved in the hot water, the Epimedium sagittatum Maxim. is scoured in the hot water for 1 min, then the solution in the extraction tank is emptied, and the Epimedium sagittatum Maxim. is reserved. 10 L of pure water is added to the extraction tank again, and the extraction tank is rotated and stirred at a speed of 30 rpm / min for 10 min, then the liquid in the extraction tank is emptied, and the process is repeated twice to drain the water on the surface of the Epimedium sagittatum Maxim. in the extraction tank. Then 10 L of pure water containing 5 g of papain and 5 g of bromelain is added to the extraction tank, the extraction tank is heated to 30 DEG C, and the solution in the extraction tank is emptied after stirring at a speed of 30 rpm / min for 5 min.

[0029] (2) Add 10L of 20% ethanol solution to the extraction tank, and heat to 60°C. Stir at a speed of 30 rpm / min for 2h. After the extraction is completed, the extraction liquid is discharged through a 250-mesh double filter to obtain 9.3L of Epimedium sagittatum extract. Then, the Epimedium sagittatum extract is concentrated to 4.65L, and loaded onto a D101 macroporous resin column at a loading rate of 0.8BV / h. After loading is completed, 1.5L of 40% ethanol solution is used to elute at an elution rate of 0.8BV / h, and the eluate is collected. The Epimedium sagittatum eluate is concentrated and dried by plate vacuum drying to obtain 46.7g of Epimedium sagittatum extract.

[0030] Example 2

[0031] The present embodiment discloses a preparation method of Epimedium sagittatum extract, and the method steps are specifically as follows:

[0032] (1) Take 1kg of Epimedium sagittatum into the extraction tank, and add 15L of hot water with a temperature of 75°C. Dissolve 120g of sodium carbonate in the hot water. After blanching the Epimedium sagittatum for 3min, the solution in the extraction tank is discharged, and the Epimedium sagittatum medicinal material is reserved. Then, 15L of pure water is added to the extraction tank, and stirred at a speed of 40 rpm / min for 20min. The liquid in the extraction tank is discharged repeatedly for 2 times, and the water on the surface of the Epimedium sagittatum in the extraction tank is drained. Then, 15L of pure water containing 8g of papain and 8g of bromelain is added to the extraction tank. The extraction tank is heated to 40°C, and stirred at a speed of 40 rpm / min for 7min. Then, the solution in the extraction tank is discharged.

[0033] (2) Add 15L of 25% ethanol solution to the extraction tank, and heat to 75°C. Stir at a speed of 40 rpm / min for 2.5h. After the extraction is completed, the extraction liquid is discharged through a 300-mesh double filter to obtain 14.4L of Epimedium sagittatum extract. Then, the Epimedium sagittatum extract is concentrated to 7.2L, and loaded onto a D101 macroporous resin column at a loading rate of 1.2BV / h. After loading is completed, 2L of 50% ethanol solution is used to elute at an elution rate of 1.2BV / h, and the eluate is collected. The Epimedium sagittatum eluate is concentrated and dried by plate vacuum drying to obtain 53.2g of Epimedium sagittatum extract.

[0034] Example 3

[0035] The present embodiment discloses a preparation method of Epimedium sagittatum extract, and the method steps are specifically as follows:

[0036] (1) Take 1 kg of Epimedium sagittatum (L.) Maxim. to an extraction tank, add 18 L of hot water with a temperature of 90 °C, and dissolve 450 g of sodium carbonate in the hot water. After blanching Epimedium sagittatum (L.) Maxim. for 5 min, empty the solution in the extraction tank, and reserve the Epimedium sagittatum (L.) Maxim. Again, add 18 L of pure water to the extraction tank, rotate and stir at a speed of 50 rpm / min for 30 min, then empty the liquid in the extraction tank, repeat for 3 times, and drain the water on the surface of Epimedium sagittatum (L.) Maxim. in the extraction tank. Then add 18 L of pure water containing 10 g of papain and 20 g of bromelain to the extraction tank, heat the extraction tank to 50 °C, stir at a speed of 50 rpm / min for 10 min, and then empty the solution in the extraction tank.

[0037] (2) Add 18 L of 30% ethanol solution to the extraction tank, and heat to 90 °C. Stir at a speed of 50 rpm / min for 3 h. After the extraction is completed, discharge the extraction liquid through a 400-mesh double filter to obtain 17.4 L of Epimedium sagittatum (L.) Maxim. extraction liquid. Then concentrate the Epimedium sagittatum (L.) Maxim. extraction liquid to 8.7 L, and load a D101 macroporous resin column at a loading rate of 1.5 BV / h. After loading is completed, elute with 2.5 L of 60% ethanol solution at an elution rate of 1.5 BV / h, and collect the eluate. Concentrate the Epimedium sagittatum (L.) Maxim. eluate, and dry in a plate vacuum drying manner to obtain 57.2 g of Epimedium sagittatum (L.) Maxim. extract.

[0038] Comparative Example 1

[0039] In this comparative example, step one is not used, and the remaining steps are the same as in Example 1.

[0040] Comparative Example 2

[0041] In this comparative example, step two is not used, and the remaining steps are the same as in Example 1.

[0042] Comparative Example 3

[0043] In this comparative example, step three is not used, and the remaining steps are the same as in Example 1.

[0044] Comparative Example 4

[0045] In this comparative example, steps one, two, and three are not used, and the remaining steps are the same as in Example 1. The specific steps are as follows:

[0046] Take 1 kg of Epimedium sagittatum in the extraction tank, add 10 L of 20% ethanol solution to the extraction tank, heat to 60°C, and stir at a speed of 30 rpm / min for 2 h. After the extraction is completed, the extract is discharged through a 250-mesh double filter to obtain 9.3 L of Epimedium sagittatum extract. Then the Epimedium sagittatum extract is concentrated to 4.65 L, and loaded onto a D101 macroporous resin column at a loading rate of 0.8 BV / h. After loading, the column is eluted with 1.5 L of 40% ethanol solution at an elution rate of 0.8 BV / h, and the eluate is collected. The Epimedium sagittatum eluate is concentrated and dried by plate vacuum drying to obtain 39.2 g of Epimedium sagittatum extract.

[0047] Test Example 1

[0048] The content of icariin in the extract is determined by liquid chromatography. The chromatographic conditions are as follows: Agilent Eclipse XDB-C18 chromatographic column (4.6 mm x 250 mm, 5 μm); detection wavelength 270 nm; flow rate 1.0 mL / min; column temperature 25°C; injection volume 10 μL; mobile phase: acetonitrile (A)-water (B); gradient elution conditions: 0-20 min, 24%-26% A; 20-30 min, 26%-30% A; 30-45 min, 30%-45% A; 45-50 min, 45%-60% A; 50-60 min, 60%-24% A.

[0049] The extraction rate of icariin is calculated by the following formula:

[0050]

[0051] In the formula, C is the concentration of icariin in the Epimedium sagittatum eluate, unit: g / L; V is the volume of the Epimedium sagittatum eluate in the example, unit: L.

[0052] The extraction rates of icariin in the examples and comparative examples are shown in Table 1 below.

[0053] Table 1 Extraction rates of icariin in different examples and comparative examples

[0054]

[0055]

[0056] Note: * indicates a significant difference compared with Comparative Example 4, P<0.05.

[0057] As can be seen from Table 1, the extraction rates of icariin in Examples 1-3 are significantly higher than those in the comparative examples, indicating that the present application can significantly improve the extraction rate of icariin compared with the traditional alcohol extraction method.

[0058] Test Example 2

[0059] Experiments were carried out using the method in each example and comparative example, and the clogging of the double filter was observed during the experiment, and the results are shown in Table 2.

[0060] Table 2: Filtration of the extract in different examples and comparative examples

[0061] Name Clogging of double filter Example 1 No Example 2 No Example 3 No Comparative Example 1 Yes Comparative Example 2 Yes Comparative Example 3 Yes Comparative Example 4 Yes

[0062] As can be seen from Table 2, examples 1-3 in the present application do not clog the double filter, while comparative examples 1-4 all clog the double filter, which shows that the absence of any one of steps 1-3 in the present application will cause the extract to clog the double filter, affecting product production. Steps 1, 2 and 3 in the present application have integrity.

[0063] Based on the above data analysis results, the method of the present application has the characteristics of green safety, can better dissolve the wax layer on the surface of Epimedium sagittatum, solve the problem of double filter clogging, and at the same time can improve the extraction rate of icariin in Epimedium sagittatum. In summary, the above examples only illustrate the preferred mode of the present application and do not include all the scope of the present application. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present application, therefore the protection scope of the present application is limited by the scope defined by the claims.

Claims

1. A method for preparing an extract of Epimedium sagittatum, characterized in that: Includes the following steps: Step 1, Blanching: Put the Epimedium sagittatum into the extraction tank, add a certain volume of hot water containing sodium carbonate and blanch for a period of time, then drain the solution in the extraction tank and retain the Epimedium sagittatum medicinal material. Step 2, Re-wash: Add a certain volume of purified water to the extraction tank again, rotate and stir for a period of time, then drain the liquid in the extraction tank. Repeat 2-3 times to drain the water from the surface of the Epimedium brevicornu in the extraction tank. Step 3, Enzyme Treatment: Add a certain volume of purified water containing the compound enzyme to the extraction tank, heat the extraction tank to a certain temperature, stir for a period of time, and then drain the solution from the extraction tank. Step 4: Ethanol extraction: Add a certain volume of ethanol solution to the extraction tank, heat and stir for a period of time. Step 5, Double Filtration: Pass the extracted liquid through a double filter to obtain Epimedium brevicornu extract. Step 6, Column purification: Concentrate the Epimedium extract obtained in Step 5 to 1 / 2 of its original volume, then load it onto a D101 macroporous resin, elute with ethanol solution, and collect the eluent. Step 7: Concentration and Drying: Concentrate and dry the eluent of Epimedium sagittatum to obtain Epimedium sagittatum extract.

2. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step one, the mass-to-volume ratio of Epimedium sagittatum to hot water is 1:10-18 g / mL, the hot water temperature is 60-90℃, the mass concentration of food-grade sodium carbonate in the hot water is 4-25 g / L, and the blanching time is 1-5 min.

3. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step two, the mass-to-volume ratio of Epimedium sagittatum to purified water is 1:10-18 g / mL, the stirring rate is 30-50 rpm / min, and the stirring time is 10-30 min.

4. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step three, the mass-to-volume ratio of Epimedium sagittatum to purified water is 1:10-18 g / mL, the complex enzyme is papain and bromelain, the mass fraction of the complex enzyme is 1%-3%, the mass ratio of papain to bromelain is 1:1-3, the enzymatic hydrolysis temperature is 30-50℃, the stirring rate is 30-50 rpm / min, and the stirring time is 5-10 min.

5. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step four, the mass-to-volume ratio of Epimedium sagittatum to ethanol solution is 1:10-18 g / mL, the concentration of ethanol solution is 20-30%, the stirring rate is 30-50 rpm / min, the extraction time is 2-3 h, and the extraction temperature is 60-90℃.

6. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step five, the mesh size of the dual filter screen is 250-400 mesh.

7. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step six, the loading rate of the concentrated Epimedium brevicornu solution is 0.8-1.5 BV / h, the ethanol elution liquid fraction is 40-60%, the elution rate is 0.8-1.5 BV / h, and the eluent volume is 1.5-2.5 BV.

8. The method for preparing an extract of Epimedium sagittatum according to claim 1, characterized in that: In step seven, the drying method is plate vacuum drying or belt drying.

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