A preparation method of high-purity erigeside II standard substance

Abstract

The present invention relates to the technical field of pharmaceutical chemistry, and discloses a preparation method of a high-purity erigeroside standard. It comprises the following steps: 1) Cut Erigeron breviscapus into small segments, then add ethanol and heat under reflux, filter, and concentrate to obtain an extract; 2) Add water to the extract obtained in step 1) for extraction, repeat the extraction 2-3 times, combine the aqueous phases, and concentrate to dryness; 3) Separate the product obtained in step 2) by macroporous resin, then add a mixed solvent for gradient elution, concentrate, and recrystallize; 4) Dispense into brown bottles, 10 mg per bottle, seal, and store at room temperature. The beneficial effects of the present invention are as follows: The preparation method of the present invention has high extraction efficiency, large output, low solvent consumption, basically no loss and no pollution of the sample, the purity of the erigeronoside product extracted is as high as over 99%, and this preparation method is applicable to the separation and purification of the crude Erigeron breviscapus prepared under various process conditions.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and particularly to a method for preparing a high-purity erigeside standard. Background Art

[0002] Erigeron breviscapus, also known as Erigeron breviscapus (Vant.) Hand-Mazz., is the dried whole herb of the Compositae plant Erigeron breviscapus. It has a pungent and slightly bitter taste and is warm in nature. It belongs to the heart and liver meridians and has the effects of promoting blood circulation to dredge collaterals and relieve pain, dispelling wind and cold. Erigeron breviscapus contains various chemical components such as flavonoids, caffeoyl derivatives, coumarins, lignans, and terpenoids. Among them, flavonoids and caffeoyl derivatives are the characteristic components of Erigeron breviscapus.

[0003] The traditional method for preparing erigeside is as follows: Erigeron breviscapus is decocted with water three times, each time for 1 hour. The decoctions are combined, filtered, and the filtrate is heated and concentrated to a relative density of 1.20. Appropriate excipients are added, mixed evenly, made into granules, and dried to obtain the product. However, the traditional water decoction process has the following defects: long preparation time, large amount of extracts obtained, unstable active ingredients due to long heating time, and insufficient extraction of active ingredients. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for preparing a high-purity erigeside standard to overcome the shortcomings of the prior art.

[0005] The purpose of the present invention is achieved through the following technical solutions: A method for preparing a high-purity erigeside standard, comprising the following steps:

[0006] 1) Cut Erigeron breviscapus into small pieces, then add ethanol and heat under reflux, filter, and concentrate to obtain an extract;

[0007] 2) Add water to the extract obtained in step 1) for extraction, repeat the extraction 2 - 3 times, combine the aqueous phases, and concentrate to dryness;

[0008] 3) Separate the product obtained in step 2) by macroporous resin, then add a mixed solvent for gradient elution, concentrate, and recrystallize;

[0009] 4) Dispense into brown bottles, 10 mg per bottle, seal, and store at room temperature.

[0010] Preferably, the weight ratio of Erigeron breviscapus to ethanol in step 1) is 1:1 - 1:5.

[0011] Preferably, the volume ratio of the extract to water in step 2) is 1:1 - 1:5.

[0012] Preferably, the mixed solvent in step 3) is a mixture of ethyl acetate and ethanol.

[0013] Preferably, the heating reflux time in step 1) is 1 hour.

[0014] Preferably, the volume ratio of the mixed solvent to the extract in step 3) is 1:10.

[0015] The present invention has the following advantages:

[0016] The preparation method of the present invention has high extraction efficiency, large output, low solvent consumption, basically no loss and no pollution of the sample, the purity of the scutellarin product extracted is as high as over 99%, and this preparation method is applicable to the separation and purification of the crude Erigeron breviscapus products prepared under various process conditions. Description of the Drawings

[0017] Figure 1 It is the chromatogram of Example 1;

[0018] Figure 2 It is the chromatogram of Comparative Example 1;

[0019] Figure 3 It is the chromatogram of Comparative Example 2. Detailed Description of the Invention

[0020] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are some but not all of the embodiments of the present invention. Usually, the components of the embodiments of the present invention described and shown in the drawings here can be arranged and designed in various different configurations.

[0021] Therefore, the following detailed description of the embodiments of the present invention provided in the drawings is not intended to limit the scope of the claimed present invention, but merely represents the selected embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0022] It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other.

[0023] It should be noted that: similar reference numerals and letters indicate similar items in the following drawings. Therefore, once an item is defined in one drawing, it does not need to be further defined and explained in subsequent drawings.

[0024] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings, or the orientation or positional relationship in which the inventive product is customarily placed during use, or the orientation or positional relationship commonly understood by those skilled in the art. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on the present invention. In addition, the terms "first", "second", etc. are only used for distinguishing descriptions and cannot be understood as indicating or implying relative importance.

[0025] In the description of the present invention, it should also be noted that unless otherwise clearly specified and limited, the terms "set", "install", "connect", and "couple" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the communication inside two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0026] Example 1:

[0027] A method for preparing a high-purity erigeroside standard product, comprising the following steps:

[0028] 1) Cut Erigeron breviscapus into small pieces, then add ethanol and heat under reflux for 1 hour, filter, and concentrate to obtain an extract. The weight ratio of Erigeron breviscapus to ethanol is 1:1 to 1:5;

[0029] 2) Add water to the extract obtained in step 1) for extraction, repeat the extraction 2 to 3 times, combine the aqueous phases, and concentrate to dryness. The volume ratio of the extract to water is 1:1 to 1:5;

[0030] 3) Separate the product obtained in step 2) by macroporous resin, then add a mixed solvent for gradient elution, concentrate, and recrystallize. The mixed solvent is a mixture of ethyl acetate and ethanol, and the volume ratio of the mixed solvent to the extract is 1:10. The purity of the recrystallized product is determined by HPLC peak normalization, and the purity reaches more than 99%;

[0031] 4) Package in brown bottles, 10 mg per bottle, seal, and store at room temperature.

[0032] Comparative Example 1:

[0033] A method for preparing a high-purity erigeroside standard product, comprising the following steps:

[0034] 1) Cut Erigeron breviscapus into small pieces, then add ethanol and heat under reflux for 1 hour, filter, and concentrate to obtain an extract. The weight ratio of Erigeron breviscapus to ethanol is 1:1 - 1:5;

[0035] 2) Add water to the extract obtained in step 1) for extraction, repeat the extraction 2 - 3 times, combine the aqueous phases, and concentrate to dryness. The volume ratio of the extract to water is 1:1 - 1:5;

[0036] 3) Separate the product obtained in step 2) by macroporous resin, then add ethyl acetate for gradient elution, concentrate, and recrystallize. The volume ratio of ethyl acetate to the extract is 1:10;

[0037] 4) Dispense into brown bottles, 10 mg per bottle, seal, and store at room temperature.

[0038] Comparative Example 2:

[0039] A method for preparing a high - purity scaposide standard, comprising the following steps:

[0040] 1) Cut Erigeron breviscapus into small pieces, then add ethanol and heat under reflux for 1 hour, filter, and concentrate to obtain an extract. The weight ratio of Erigeron breviscapus to ethanol is 1:1 - 1:5;

[0041] 2) Add water to the extract obtained in step 1) for extraction, repeat the extraction 2 - 3 times, combine the aqueous phases, and concentrate to dryness. The volume ratio of the extract to water is 1:1 - 1:5;

[0042] 3) Separate the product obtained in step 2) by macroporous resin, then add ethanol for gradient elution, concentrate, and recrystallize. The volume ratio of ethanol to the extract is 1:10;

[0043] 4) Dispense into brown bottles, 10 mg per bottle, seal, and store at room temperature.

[0044] The recrystallization products of the above examples were all detected by UPLC method, and the chromatographic conditions are as follows:

[0045] Stationary phase: C18

[0046] Mobile phase: Gradient elution with acetonitrile and 0.1 - 5% acid aqueous solution; 0 - 5 min, 10% - 16% acetonitrile; 5 - 18 min, 16% - 18% acetonitrile; 18 - 26 min, 18% - 24% acetonitrile. Finally, the chromatograms of each example are as shown in the accompanying drawings of the specification.

[0047] It can be seen from the chromatograms that only the chromatogram of Example 1 has good separation effect, small peak tailing, and is easy for the identification and accurate detection of more components.

[0048] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a high-purity erigeside standard, characterized in that, it comprises the following steps: 1) Cut Erigeron breviscapus into small pieces, then add ethanol and heat under reflux, filter, and concentrate to obtain an extract; 2) Add water to the extract obtained in step 1) for extraction, repeat the extraction 2 - 3 times, combine the aqueous phases, and concentrate to dryness; 3) Separate the product obtained in step 2) by macroporous resin, then add a mixed solvent for gradient elution, concentrate, and recrystallize. The mixed solvent is a mixture of ethyl acetate and ethanol, and the volume ratio of the mixed solvent to the extract is 1:10; 4) Dispense into brown bottles, 10 mg per bottle, seal, and store at room temperature.

2. The method for preparing a high-purity erigeside standard according to claim 1, characterized in that: the weight ratio of Erigeron breviscapus to ethanol in step 1) is 1:1 - 1:

5.

3. The method for preparing a high-purity erigeside standard according to claim 1, characterized in that: the volume ratio of the extract to water in step 2) is 1:1 - 1:

5.

4. The method for preparing a high-purity erigeside standard according to claim 1, characterized in that: the heating reflux time in step 1) is 1 hour.

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