Herba epimedii medicinal material as well as processing method and application thereof

Abstract

The invention relates to the technical field of preparation of rare flavonoids of herba epimedii, and particularly provides a herba epimedii medicinal material as well as a processing method and application thereof, and the processing method comprises the following steps: selecting a cultivated variety of epimedium sagittatum with main components of epimedium sagittatum C and icariin, and collecting overground parts of fresh herba epimedii including leaves and stalks from July to August; the leaves are naturally dried in the shade, and the water content of the leaves dried in the shade is kept to be 60% or above. The semi-shade-dried leaves are subjected to color changing treatment for 24-48 h under the condition of 55-65 DEG C in a completely sealed state. The treated leaves are completely changed from green to brown yellow, the surfaces of the leaves are still glossy, and the leaf veins are clear and free of mildew spots. And taking out the treated leaves, and drying at 40-60 DEG C for 12 hours until the water content is within 10%. According to the method, the existing enzymolysis conversion process is preposed, the conversion process is directly completed in the raw materials, special equipment and reagents are not needed, the process is simple, the conversion cost is low, the conversion process is free of pollution and environmentally friendly, and the method is suitable for industrial production.

Description

Technical Field

[0001] The invention relates to the technical field of preparation of rare flavonoids of epimedium, in particular to an epimedium medicinal material and a processing method and application thereof. Background Art

[0002] The Chinese medicinal material Epimedii Folium is the dried leaves of plants of the genus Epimedium, including E. brevicornum Maxim., E. sagittatum (Sieb. et Zucc.) Maxim., E. pubescens Maxim. and E. koreanum Nakai.

[0003] Epimedium was first recorded in Shennong's Herbal Classic and was listed as a top-grade herb. It is pungent, sweet, warm in nature, and enters the liver and kidney meridians. It is mainly used to treat male impotence, genital strain, and pain in the penis. It can promote urination, increase strength, and keep people in a good mental state. The sweet and warm nature can nourish the kidney and strengthen yang, and can treat impotence, frequent urination, weakness in the waist and knees, and infertility caused by insufficient kidney yang; the pungent and warm nature can dispel wind and dampness, and has a good effect on rheumatic pain, limb numbness, etc. It is especially suitable for patients with wind-cold dampness and kidney yang deficiency. Modern evidence-based medical research shows that epimedium can also be used to prevent osteoporosis, liver cancer, depression, and treat Alzheimer's disease, type 2 diabetes, and rheumatoid arthritis.

[0004] The main active ingredients in Epimedium are a class of flavonoid secondary metabolites with isopentenylation at the C8 position of ring A, such as icariin, epimedin A, epimedin B, epimedin C, baohuoside I, icariside I, 2"-O-rhamnosyl icariside II and icaritin, among which icariin, epimedin A, epimedin B and epimedin C are common and ubiquitous flavonol glycosides in the original medicinal materials of Epimedium, and are also the quality markers of Epimedium medicinal materials. In addition to the above four substances, Baohuoside I, Icariin I, Rhamnosyl Icariin II and Icariin are rare flavonoid compounds. Their content in Epimedium medicinal materials is extremely low, but they have higher economic value. For example, the Class 1.2 innovative drug "Icariin Soft Capsules" that has been approved for the treatment of middle and late stage liver cancer has icariin as its single ingredient.

[0005] Baohuoside I (also known as icariside II) is the main product of icariin metabolism in animals (7-O-glucoside on icariin is hydrolyzed by enzymes in the body), while 2"-rhamnosyl icariside II is the main product of chondrin C metabolism in animals (7-O-glucoside on chondrin C is hydrolyzed by enzymes in the body). Compared with icariin and chondrin C, Baohuoside I and 2"-rhamnosyl icariside II have similar functions and stronger biological activities. For example, Baohuoside I has stronger alpha-glucosidase (a membrane-bound enzyme present in the epithelial cells of the small intestine, which promotes the absorption of glucose in the small intestine by catalyzing the hydrolysis and cleavage of oligosaccharides into absorbable oligosaccharides) inhibitory activity than icariin, which can slow down the increase in blood sugar after carbohydrate meals and effectively manage non-insulin-dependent diabetes mellitus (Shen Y, Wang M, Zhou J, Chen Y, Wu M, Yang Z, Yang C, Xia G, Tam JP, Zhou C, Yang H, Jia A):1361-1372.doi:10.1016 / j.ijbiomac.2020.10.018.). 2"-rhamnosyl icariin II has the effects of promoting muscle cell proliferation or differentiation, increasing the number of myotubes, promoting myotube formation, increasing myotube length, increasing myotube width, promoting myoblast differentiation or promoting muscle cell fusion (CN113368123A, application of rhamnosyl icariin II in muscle repair and regeneration). In addition, due to the hydrolysis of 7-O-glucoside, the polarity of baohuoside I and 2"-rhamnosyl icariin II is reduced, and they are easier to penetrate the small intestinal mucosa than the original compounds icariin and choholidine C before hydrolysis, which is more conducive to the body's absorption and improves bioavailability.

[0006] Existing studies have shown that rare flavonoids in epimedium are mainly obtained by chemical cleavage or enzymatic hydrolysis of crude epimedium extracts (Peng Lei, Xu Siyu, Wu Bo, et al. Enzymatic conversion of epimedium glycosides A, B and C and preparation of rare epimedium glycosides [J]. Food and Fermentation Industries, 2023, 49(21): 95-100.). These conversion methods or processes have the problems of complicated steps, time-consuming and labor-intensive, high cost, or poor specificity and many by-products, and the prospects for industrial application are poor. How to efficiently increase the content of rare flavonoids in epimedium glycosides has become one of the technical problems that need to be solved urgently. Summary of the invention

[0007] In view of this, the present invention proposes an epimedium medicinal material and a processing method and application thereof. The present invention uses the hydrolase possessed by the raw material itself to advance the conversion process, and provides a method for directly converting in the raw material without extraction, so that icariin and chamoidine C in the raw material are hydrolyzed and completely converted to obtain epimedium rare flavonoids bajuroside I and 2"-rhamnosyl icariin II.

[0008] The technical solution of the present invention is achieved as follows: The present invention provides a method for processing Epimedium medicinal material, comprising the following steps:

[0009] Material collection: Select fresh Epimedium from July to August, and collect the aboveground parts, including leaves and stems;

[0010] Semi-drying treatment: The above-ground part collected is naturally dried in the shade, so that the water content of the leaves after drying in the shade is not less than 60%;

[0011] Color change treatment: Heat the semi-dried above-ground part in a completely sealed state to change the color of the leaves from green to brown;

[0012] Leaf drying: The discolored leaves are dried until their moisture content does not exceed 10%.

[0013] In some embodiments, the epimedium is Epimedium sagittatum.

[0014] In some embodiments, the color change treatment heating temperature is 55-65° C., and the treatment time is 24-48 hours.

[0015] In some embodiments, the leaf drying process is carried out at a temperature of 40-60° C. and a drying time of 12 hours.

[0016] The second aspect of the present invention also provides an Epimedium medicinal material obtained by the above processing method.

[0017] In a third aspect, the present invention further provides an application of the above-mentioned Epimedium medicinal material, wherein the prepared Epimedium medicinal material is used to prepare Epimedium rare flavonoids, wherein the rare flavonoids include 2"-rhamnosyl epimedin II and baohuoside I.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The invention completely converts the original compounds icariin and icariin C in the raw material of epimedium into rare flavonoid target products bajuoside I and 2"-rhamnosylicariin II respectively without other by-products. The existing enzymatic hydrolysis conversion process is pre-placed to directly complete the conversion process in the raw material without special equipment and reagents, and the process is simple, the conversion cost is low, the conversion process is pollution-free and environmentally friendly, and the method is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 Schematic diagram of substance conversion in Epimedium in the processing method of the present invention, wherein glc represents glucose and rha represents rhamnosyl;

[0022] Figure 2 This is a picture of the leaves of Epimedium after being treated in Example 1 of the present invention;

[0023] Figure 3 This is a color comparison diagram of Example 1 of the present invention and Comparative Example 1, wherein green is the dried leaves after the treatment of Comparative Example 1, and brown is the dried leaves after the treatment of Example 1;

[0024] Figure 4 It is a comparison diagram of the flavonoid extracts after treatment in Example 1 of the present invention and in Comparative Example 1, the light color is the sample of the flavonoid extract in Comparative Example 1, and the dark color is the sample of the flavonoid extract in Example 1;

[0025] Figure 5 The HPLC detection spectra of flavonoids before and after treatment of Example 1 and Comparative Example 1 are shown. DETAILED DESCRIPTION

[0026] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the embodiments of the present invention belong. If the definitions set forth in this section are contrary to or otherwise inconsistent with the definitions set forth in the patents, patent applications, published patent applications, and other publications incorporated herein by reference, the definitions listed in this section take precedence over the definitions incorporated herein by reference.

[0028] Unless otherwise specified, the methods used in the following examples are all conventional methods. Unless otherwise specified, the materials, reagents and instruments used are all conventional materials, reagents and instruments in the art, and can be obtained by those skilled in the art through commercial channels.

[0029] When an amount, concentration or other value or parameter is expressed as a range, a preferred range or a range defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is disclosed separately. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted as including the range "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described herein, unless otherwise stated, the range is intended to include its end values ​​and all integers and fractions within the range. In the present specification and claims, range definitions can be combined and / or interchanged, and if not otherwise stated, these ranges include all subranges contained therein.

[0030] At present, rare flavonoids of epimedium are mainly obtained by chemical cleavage or enzymatic hydrolysis of crude epimedium extracts. These conversion methods or processes have the problems of complicated steps, time-consuming and labor-intensive, high cost, poor specificity, and many by-products, and the prospects for industrial application are poor. The present invention uses the glycoside hydrolase in the raw material itself to advance the conversion process, and provides a method for direct conversion in the raw material without extraction, enzymatically hydrolyzing and completely converting the icariin and chondrin C in the raw material to obtain rare flavonoids of epimedium, baohuoside I and 2"-rhamnosyl icariin II.

[0031] In order to achieve the above purpose, the technical solution adopted is:

[0032] The sagittaria epimedium, the main components of which are epimedin C and icariin, were selected, and the fresh above-ground parts of epimedium, including leaves and stems, were collected in July and August.

[0033] The leaves are then dried naturally in the shade, and the moisture content of the leaves after drying is maintained above 60%.

[0034] The semi-shade-dried above-ground part is placed in a completely sealed state and subjected to color change treatment at 55-65℃ for 24-48h. After the treatment, the green leaves completely turn into brown-yellow, the leaf surface is still shiny, the leaf veins are clean, and there are no mold spots.

[0035] The treated above-ground part is taken out from the sealing device, and dried at 40-60° C. for 12 hours to make the water content of the leaves not exceed 10%, thereby obtaining the Epimedium medicinal material.

[0036] Example 1

[0037] This example uses fresh materials of the above-ground parts of three-year-old cultivated Epimedium sagittatum harvested in July-August 2024, and the specific processing method is as follows:

[0038] Material collection: Select fresh Epimedium from July to August, and collect the aboveground parts, including leaves and stems;

[0039] Semi-drying treatment: The above-ground part collected is naturally dried in the shade, so that the water content of the leaves after drying in the shade is not less than 60%;

[0040] Discoloration treatment: Heat the semi-dried above-ground part to 55℃ in a completely sealed state for 24 hours, so that the leaves change from green to brown-yellow, the leaf surface is still shiny, the veins are clear, and there is no mold spot;

[0041] Leaf drying: Dry the discolored leaves at 40°C for 12 hours until their moisture content does not exceed 10%.

[0042] Example 2

[0043] The only difference between this embodiment and embodiment 1 is that the color change treatment temperature is 65° C., the time is 24 h, and the drying treatment temperature is 45° C., the time is 12 h.

[0044] Example 3

[0045] The only difference between this embodiment and embodiment 1 is that the color change treatment temperature is 55° C., the time is 48 hours, and the drying treatment temperature is 50° C., the time is 12 hours.

[0046] Example 4

[0047] The only difference between this embodiment and embodiment 1 is that the color change treatment temperature is 65° C., the time is 48 hours, and the drying treatment temperature is 60° C., the time is 12 hours.

[0048] Comparative Example 1

[0049] Material collection: Select fresh Epimedium from July to August, and collect the aboveground parts, including leaves and stems;

[0050] The collected above-ground parts are naturally dried in the shade so that the water content of the leaves after drying in the shade does not exceed 10%, thereby obtaining the Epimedium medicinal material.

[0051] The leaf parts of the Herba Epimedii prepared in the above different embodiments and Comparative Example 1 were respectively subjected to extraction and detection of flavonoid components, and the flavonoid extraction method was referred to the 2020 edition of the Pharmacopoeia method. The conversion rates of icariin C to 2"-rhamnosylicariin II and icariin to baohuoside I were calculated.

[0052] Wherein: the conversion rate of the single substance of icariin or icariin = (peak area before treatment - peak area after treatment) / peak area before treatment × 100%.

[0053] The calculation results are shown in the following table:

[0054]

[0055] The HPLC spectra of flavonoids in Example 1 and Comparative Example 1 are as follows Figure 5 As shown, Figure A is a standard reference spectrum, Figure B is a spectrum of Example 1, and Figure C is a spectrum of Comparative Example 1. In the figure: Compound 1: Chaohuoding A; Compound 2: Chaohuoding B; Compound 3: Chaohuoding C; Compound 4: Icariin; Compound 5: Baohuoside II; Compound 6: 2"-rhamnosylicariin II; Compound 7: Baohuoside I.

[0056] It is not difficult to see from the comparison of the above data that the present invention provides a method for converting rare flavonoids of Epimedium, which utilizes the glycosyl hydrolase contained in the leaves of the original plant itself to hydrolyze the 7-O-glucoside on the icariin C and icariin compounds, and convert them into the corresponding rare flavonoids 2"-rhamnosylicariin II and baohuoside I, respectively. The method is simple and easy to implement, and the complete conversion of the original components can be achieved by controlling the processing temperature and time in a sealed state, without extracting the crude extract of Epimedium and adding additional heterologous hydrolases, and is suitable for the industrial production of rare flavonoids of Epimedium.

[0057] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for processing Epimedium medicinal material, characterized in that: The steps include: Material collection: Select fresh Epimedium from July to August, and collect the aboveground parts, including leaves and stems; Semi-drying treatment: The above-ground part collected is naturally dried in the shade, so that the water content of the leaves after drying in the shade is not less than 60%; Color change treatment: Heat the semi-dried above-ground part in a completely sealed state to change the color of the leaves from green to brown; Leaf drying: The discolored leaves are dried until their moisture content does not exceed 10%.

2. The method for processing the Herba Epimedii as claimed in claim 1, characterized in that: The epimedium is epimedium sagittatum.

3. The method for processing the Herba Epimedii as claimed in claim 1, characterized in that: The heating temperature for the color change treatment is 55-65°C and the treatment time is 24-48h.

4. The method for processing the Herba Epimedii as claimed in claim 1, characterized in that: The temperature of leaf drying treatment is 40-60℃ and the drying time is 12h.

5. A Herba Epimedii medicinal material, characterized in that: The product is obtained by processing using any one of the processing methods described in claims 1 to 4.

6. The use of the Epimedium medicinal material according to claim 5, characterized in that: The invention is used for preparing rare flavonoids of epimedium, wherein the rare flavonoids include 2"-rhamnosyl epimediside II and baohuoside I.

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