Dendrobium officinale extract as well as preparation method and application thereof

Abstract

The invention discloses a dendrobium officinale extract and a preparation method and application thereof, and relates to the technical field of traditional Chinese medicine, the preparation method comprises the following steps: crushing and sieving dendrobium officinale flowers to obtain a first material; performing oxygen-enriched microwave plasma treatment on the first material to obtain a second material; adding the second material into water for reflux extraction, and concentrating the obtained water extract to obtain an extract; and carrying out column-passing gradient elution on the extract, collecting an eluent, concentrating and drying to obtain the dendrobium officinale extract. According to the method, the in-vitro inhibition effect of the obtained dendrobium officinale extract on brain glioma U87 cells is remarkably improved by screening the medicinal parts and the extraction solvent of the dendrobium officinale and combining oxygen-enriched microwave plasma treatment and a gradient elution process, the application range of the dendrobium officinale extract is widened, and the application range of the dendrobium officinale extract is widened. And a novel clinical treatment means is provided for patients with brain glioma.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicines, and in particular to a Dendrobium officinale extract and a preparation method and application thereof. Background Art

[0002] Dendrobium officinale, as a precious medicinal material in the treasure house of traditional Chinese medicine, has played an important role in the treatment of various diseases with its unique efficacy since ancient times. With the advancement of modern science and technology and the deepening of Chinese medicine research, various bioactive components in Dendrobium officinale have been gradually revealed, and its potential in anti-oxidation, immunomodulation, anti-inflammatory and anti-tumor has received increasing attention. However, despite the wide range of pharmacological effects of Dendrobium officinale, there are no reports in the prior art on the use of Dendrobium officinale extracts for the treatment of specific diseases, especially tumors such as brain gliomas that are highly malignant and difficult to treat.

[0003] As the most common malignant tumor of the central nervous system, glioma has a complex pathogenesis, is difficult to treat, and is prone to recurrence, posing a serious threat to the life and health of patients. At present, although comprehensive treatments such as surgery, radiotherapy, and chemotherapy have prolonged the survival of patients to a certain extent, the prognosis of patients is still not optimistic. Therefore, finding new and effective adjuvant therapeutic drugs, especially those derived from nature, with low toxicity and high efficacy, has become an important direction of current research on the treatment of glioma.

[0004] In addition, the preparation method of traditional Dendrobium officinale extract has many limitations. On the one hand, the screening of medicinal parts is not detailed enough, and often only focuses on common parts such as stems and leaves, while ignoring parts such as flowers that may be rich in specific active ingredients. On the other hand, the selection of extraction solvents and the optimization of extraction conditions are insufficient, resulting in low content and purity of active ingredients in the extract, affecting its efficacy. In addition, traditional extraction methods may destroy some active ingredients during the processing process, further reducing the efficacy of the extract. Summary of the invention

[0005] In order to solve the above problems, the present invention provides a Dendrobium officinale extract and a preparation method and application thereof.

[0006] In a first aspect, the present invention provides a method for preparing a Dendrobium officinale extract, the preparation method comprising the following steps:

[0007] Grinding and screening the Dendrobium officinale flowers to obtain a first material;

[0008] The first material is subjected to oxygen-rich microwave plasma treatment to obtain a second material;

[0009] The second material is added into water for reflux extraction, and then the obtained water extract is concentrated to obtain an extract;

[0010] The extract is subjected to column gradient elution, and the eluate is collected and concentrated and dried to obtain the Dendrobium officinale extract.

[0011] Furthermore, the working condition parameters of the oxygen-rich microwave plasma treatment include: microwave power of 180 to 290 W, oxygen and argon with a volume ratio of 7:1 as the working medium, an inlet flow rate of the working medium of 65 to 75 mL / min, a working mode of glow discharge, a treatment time of 20 to 45 seconds, and oxygen is continued to be introduced for 5 to 10 minutes after the glow discharge treatment is completed.

[0012] Furthermore, the microwave power of the oxygen-enriched microwave plasma treatment is 225 W, and the time of the oxygen-enriched microwave plasma treatment is 30 seconds.

[0013] Furthermore, the particle size of the first material is 20-50 meshes.

[0014] Furthermore, the working condition parameters of the reflux extraction include: temperature of 100-110° C., time of 1-2 hours, number of times of 1-3 times, and a weight ratio of the second material to the water of (1-2):(8-12).

[0015] Furthermore, the extract is subjected to column gradient elution, the eluate is collected and concentrated and dried, and the step of obtaining the Dendrobium officinale extract comprises the following process:

[0016] The extract and D101 macroporous resin are uniformly mixed in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with petroleum ether having a volume of 5 times the column, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol, respectively, wherein the amount of each gradient eluent is 5 times the column volume, and the eluents having 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to obtain the Dendrobium officinale extract.

[0017] In a second aspect, the present invention provides a Dendrobium officinale extract, wherein the Dendrobium officinale extract is prepared by the preparation method of the Dendrobium officinale extract described in any one of the first aspects.

[0018] In a third aspect, the present invention provides use of the Dendrobium officinale extract described in any one of the second aspects in the preparation of a drug for treating brain glioma.

[0019] Furthermore, the dosage form of the drug for treating brain glioma includes tablets, capsules, powders, mixtures, pills or granules.

[0020] The above technical solution provided by the embodiment of the present invention has at least the following advantages compared with the prior art:

[0021] The embodiment of the present invention provides a Dendrobium officinale extract and a preparation method and application thereof. The present invention significantly improves the in vitro inhibitory effect of the obtained Dendrobium officinale extract on brain glioma U87 cells by screening the medicinal parts of Dendrobium officinale and the extraction solvent, and combining oxygen-rich microwave plasma treatment and gradient elution process, which not only broadens the application scope of the Dendrobium officinale extract, but also provides a new clinical treatment method for patients with brain glioma. Specifically:

[0022] 1. Selection of medicinal parts of Dendrobium candidum:

[0023] As a traditional Chinese medicinal material, the types and contents of active ingredients contained in different parts of Dendrobium candidum (such as stems, leaves, flowers, etc.) are different. The present invention selects Dendrobium candidum flowers as the extraction raw material, based on the research that the Dendrobium candidum flowers are rich in active ingredients such as polysaccharides, flavonoids, alkaloids, etc., and the in vitro inhibitory effect on brain glioma U87 cells is significantly enhanced compared with the effect of Dendrobium candidum stem extract.

[0024] 2. Oxygen-enriched microwave plasma treatment and extraction solvent selection:

[0025] Oxygen-rich microwave plasma technology can instantly generate a high-temperature, high-pressure plasma environment to modify the Dendrobium officinale flower powder. It can not only destroy the cell wall structure and promote the release of active ingredients in the cells, but also increase the hydrophilicity of its surface to make it easier to be extracted by water and enhance the biological activity of the active ingredients, thereby further improving the inhibitory effect of Dendrobium officinale extract on brain glioma U87 cells.

[0026] 3. Reflux extraction and column gradient elution:

[0027] The present invention performs column gradient elution on the extract obtained by reflux extraction, and gradually elutes with eluents of different polarities, so that the Dendrobium officinale extract can be further purified, and the target components with anti-tumor activity on brain glioma U87 cells are enriched, thereby improving the purity and efficacy of the Dendrobium officinale extract.

[0028] In summary, the present invention combines modern processing technology with traditional Chinese medicine extraction technology, screens the medicinal parts of Dendrobium officinale and extraction solvents, and combines oxygen-enriched microwave plasma treatment and gradient elution technology to achieve efficient preparation and activity enhancement of Dendrobium officinale extract, opening up a new way for the modernization of traditional Chinese medicine research and the clinical treatment of brain glioma. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0030] 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, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0031] Figure 1 A schematic flow chart of a method for preparing a Dendrobium officinale extract provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0032] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are 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.

[0033] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0034] In a first aspect, the present invention provides a method for preparing a Dendrobium officinale extract, such as Figure 1 As shown, the preparation method comprises the following steps:

[0035] Grinding and screening the Dendrobium officinale flowers to obtain a first material;

[0036] The first material is subjected to oxygen-rich microwave plasma treatment to obtain a second material;

[0037] The second material is added into water for reflux extraction, and then the obtained water extract is concentrated to obtain an extract;

[0038] The extract is subjected to column gradient elution, and the eluate is collected and concentrated and dried to obtain the Dendrobium officinale extract.

[0039] The embodiment of the present invention provides a method for preparing a Dendrobium officinale extract. The present invention significantly improves the in vitro inhibitory effect of the obtained Dendrobium officinale extract on brain glioma U87 cells through screening of medicinal parts of Dendrobium officinale and extraction solvents, and combining oxygen-rich microwave plasma treatment and gradient elution technology. This not only broadens the application scope of the Dendrobium officinale extract, but also provides a new clinical treatment method for patients with brain glioma.

[0040] In some specific embodiments, the working condition parameters of the oxygen-rich microwave plasma treatment include: microwave power of 180 to 290 W, oxygen and argon with a volume ratio of 7:1 as the working medium, an inlet flow rate of the working medium of 65 to 75 mL / min, a working mode of glow discharge, a treatment time of 20 to 45 seconds, and oxygen is continued to be introduced for 5 to 10 minutes after the glow discharge treatment is completed.

[0041] In some specific embodiments, the microwave power of the oxygen-enriched microwave plasma treatment is 225 W, and the time of the oxygen-enriched microwave plasma treatment is 30 seconds.

[0042] In some specific embodiments, the particle size of the first material is 20-50 mesh.

[0043] In some specific embodiments, the working condition parameters of the reflux extraction include: temperature of 100-110°C, time of 1-2 hours, number of times of 1-3 times, and a weight ratio of the second material to the water of (1-2):(8-12).

[0044] In some specific embodiments, the extract is subjected to column gradient elution, the eluate is collected and concentrated and dried, and the step of obtaining the Dendrobium officinale extract comprises the following process:

[0045] The extract and D101 macroporous resin are uniformly mixed in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with petroleum ether having a volume of 5 times the column, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol, respectively, wherein the amount of each gradient eluent is 5 times the column volume, and the eluents having 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to obtain the Dendrobium officinale extract.

[0046] In a second aspect, based on the same inventive concept, the present invention provides a Dendrobium officinale extract, wherein the Dendrobium officinale extract is prepared by the preparation method of the Dendrobium officinale extract described in any one of the first aspects.

[0047] The Dendrobium officinale extract provided in the embodiment of the present invention is realized based on the preparation method of the Dendrobium officinale extract described in any one of the first aspects, and therefore has at least the beneficial effects of the Dendrobium officinale extract described in any one of the first aspects, which will not be described one by one here.

[0048] In a third aspect, based on the same inventive concept, the present invention provides the use of the Dendrobium officinale extract described in any one of the second aspects in the preparation of a drug for treating brain glioma.

[0049] In some specific embodiments, the dosage form of the drug for treating brain glioma includes tablets, capsules, powders, mixtures, pills or granules.

[0050] It should be noted that the components and raw materials involved in the Dendrobium officinale extract and its preparation method and application provided by the embodiments of the present invention, unless otherwise specified or specified, can be directly commercially available products or homemade using existing public preparation methods; at the same time, the steps and parameters involved, unless otherwise specified or specified, can be carried out according to the existing processing technology of Dendrobium officinale extract or directly carried out using existing equipment, and the present invention document will not describe them one by one.

[0051] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually measured according to national standards. If there is no corresponding national standard, then the conditions recommended by the manufacturer are followed.

[0052] Example 1

[0053] This example provides a Dendrobium officinale extract, and the preparation method of the Dendrobium officinale extract comprises the following steps:

[0054] Step (1): crushing the Dendrobium officinale flower through a 30-mesh sieve to obtain a first material;

[0055] Step (2): placing the first material obtained in step (1) in a microwave plasma device, setting the microwave power to 225 W, the inlet gas flow rate to 70 mL / min, the working mode to glow discharge, introducing oxygen and argon with a volume ratio of 7:1 as the working medium to carry out oxygen-enriched microwave plasma treatment for 30 seconds, and continuing to introduce oxygen for 8 minutes after the glow discharge treatment to obtain the second material;

[0056] Step (3): adding the second material obtained in step (2) into water and stirring, wherein the weight ratio of the second material to the water is 1.5:11, and then performing reflux extraction at 105° C. for 1.5 hours, extracting twice, and then combining the obtained water extracts and concentrating to obtain an extract with a density of 1.2 g / cm3;

[0057] Step (4): the extract obtained in step (3) is mixed evenly with D101 macroporous resin in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with 5 times the column volume of petroleum ether, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol by volume fraction, wherein the amount of each gradient elution solution is 5 times the column volume, and the eluates of 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to constant weight to obtain the Dendrobium officinale extract.

[0058] Example 2

[0059] This example provides a Dendrobium officinale extract, and the preparation method of the Dendrobium officinale extract comprises the following steps:

[0060] Step (1): crushing the Dendrobium officinale flower through a 20-mesh sieve to obtain a first material;

[0061] Step (2): placing the first material obtained in step (1) in a microwave plasma device, setting the microwave power to 290 W, the inlet gas flow rate to 65 mL / min, the working mode to glow discharge, introducing oxygen and argon with a volume ratio of 7:1 as the working medium to perform oxygen-enriched microwave plasma treatment for 20 seconds, and continuing to introduce oxygen for 10 minutes after the glow discharge treatment to obtain a second material;

[0062] Step (3): adding the second material obtained in step (2) into water and stirring, wherein the weight ratio of the second material to the water is 1.5:11, and then performing reflux extraction at 105° C. for 1 hour, extracting twice, and then combining the obtained water extracts and concentrating to obtain an extract with a density of 1.2 g / cm3;

[0063] Step (4): the extract obtained in step (3) is mixed evenly with D101 macroporous resin in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with 5 times the column volume of petroleum ether, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol by volume fraction, wherein the amount of each gradient elution solution is 5 times the column volume, and the eluates of 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to constant weight to obtain the Dendrobium officinale extract.

[0064] Example 3

[0065] This example provides a Dendrobium officinale extract, and the preparation method of the Dendrobium officinale extract comprises the following steps:

[0066] Step (1): crushing the Dendrobium officinale flower through a 50-mesh sieve to obtain a first material;

[0067] Step (2): placing the first material obtained in step (1) in a microwave plasma device, setting the microwave power to 180 W, the inlet gas flow rate to 75 mL / min, the working mode to glow discharge, introducing oxygen and argon with a volume ratio of 7:1 as the working medium to carry out oxygen-enriched microwave plasma treatment for 45 seconds, and continuing to introduce oxygen for 5 minutes after the glow discharge treatment to obtain a second material;

[0068] Step (3): adding the second material obtained in step (2) into water and stirring, wherein the weight ratio of the second material to the water is 1.5:11, and then performing reflux extraction at 105° C. for 2 hours, extracting twice, and then combining the obtained water extracts and concentrating to obtain an extract with a density of 1.2 g / cm3;

[0069] Step (4): the extract obtained in step (3) is mixed evenly with D101 macroporous resin in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with 5 times the column volume of petroleum ether, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol by volume fraction, wherein the amount of each gradient elution solution is 5 times the column volume, and the eluates of 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to constant weight to obtain the Dendrobium officinale extract.

[0070] Comparative Example 1

[0071] This example provides a Dendrobium officinale extract and a preparation method thereof, which is different from Example 1 only in that the stem of Dendrobium officinale is used as the extraction raw material; the remaining steps and parameters are the same.

[0072] Comparative Example 2

[0073] This example provides a Dendrobium officinale extract and a preparation method thereof, which is different from Example 1 only in that step (2) oxygen-enriched microwave plasma treatment is not performed; the remaining steps and parameters are the same.

[0074] The preparation method of the Dendrobium officinale extract comprises the following steps:

[0075] Step (1): crushing the Dendrobium officinale flower through a 30-mesh sieve to obtain a first material;

[0076] Step (2): adding the first material obtained in step (1) into water and stirring, wherein the weight ratio of the first material to the water is 1.5:11, and then performing reflux extraction at 105° C. for 1.5 hours, extracting twice, and then combining the obtained water extracts and concentrating to obtain an extract with a density of 1.2 g / cm3;

[0077] Step (3): the extract obtained in step (2) is mixed evenly with D101 macroporous resin in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with 5 times the column volume of petroleum ether, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol by volume fraction, wherein the amount of each gradient elution solution is 5 times the column volume, and the eluates of 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to constant weight to obtain the Dendrobium officinale extract.

[0078] Comparative Example 3

[0079] This example provides a Dendrobium officinale extract and a preparation method thereof, which is different from Example 1 only in that the extraction solvent water in step (3) is replaced by ethyl acetate; the remaining steps and parameters are the same.

[0080] Comparative Example 4

[0081] This example provides a Dendrobium officinale extract and a preparation method thereof, which differs from Example 1 only in that the eluents collected in step (4) with 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are replaced by eluents collected with 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol and 80% ethanol; the remaining steps and parameters are the same.

[0082] Test Case

[0083] In this example, the in vitro inhibition test of brain glioma cells U87 was carried out on the Dendrobium officinale extracts obtained in the above Examples 1 to 3 and Comparative Examples 1 to 4 according to the in vitro cancer cell inhibition activity detection method disclosed in the prior art.

[0084] Test method: 1) Inoculate the brain glioma U87 cells in the logarithmic growth phase on a 96-well plate, treat the cells with the sample solution of Dendrobium officinale extract obtained in Examples 1 to 3 and Comparative Examples 1 to 4 for 48 hours, add 75 μL of 1% glacial acetic acid prepared sulforhodamine B staining solution (0.4%) to each well, and stain for 10 minutes at room temperature; discard the staining solution in each well, fully dissolve the sulforhodamine B staining solution, and use an enzyme reader to measure the absorbance OD value at a wavelength of 490 / 630nm. 2) According to the OD value measured by the enzyme reader, calculate the cell survival rate according to the following formula: survival rate (%) = treatment group OD / control group OD × 100%; inhibition rate (%) = 100% - survival rate (%), the test results are shown in Table 1.

[0085] Table 1

[0086] Test samples and concentrations Inhibition rate (%) Example 1, 20ug / mL 96.2 Example 2, 20ug / mL 89.7 Example 3, 20ug / mL 91.4 Comparative Example 1, 20ug / mL 53.9 Comparative Example 2, 20ug / mL 36.1 Comparative Example 3, 20ug / mL 42.5 Comparative Example 4, 20ug / mL 48.8

[0087] It can be seen from Table 1 that, compared with Comparative Examples 1 to 4, the Dendrobium officinale extract provided in the embodiment of the present invention significantly improves the in vitro inhibitory effect of the obtained Dendrobium officinale extract on U87 glioma cells by screening the medicinal parts of Dendrobium officinale and the extraction solvent, and combining oxygen-rich microwave plasma treatment and gradient elution process. The in vitro inhibition rate of U87 glioma cells can reach up to 96.2% at a low sample concentration of 20ug / mL, which broadens the application scope of the Dendrobium officinale extract and provides a new clinical treatment method for patients with glioma.

[0088] Various embodiments of the present invention may be presented in the form of a range; it should be understood that the description in the form of a range is only for convenience and brevity, and should not be understood as a rigid limitation on the scope of the present invention; therefore, the range description should be considered to have specifically disclosed all possible sub-ranges and single numerical values ​​within the range. For example, the range description from 1 to 6 should be considered to have specifically disclosed sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as single numbers within the range, such as 1, 2, 3, 4, 5 and 6, which apply regardless of the range. In addition, whenever a numerical range is indicated herein, it is meant to include any cited number (fractional or integer) within the indicated range.

[0089] The foregoing is merely a specific embodiment of the present invention, which enables those skilled in the art to understand or implement the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features claimed herein.

Claims

1. A method for preparing a Dendrobium officinale extract, characterized in that: The preparation method comprises the following steps: Grinding and screening the Dendrobium officinale flowers to obtain a first material; The first material is subjected to oxygen-rich microwave plasma treatment to obtain a second material; The second material is added into water for reflux extraction, and then the obtained water extract is concentrated to obtain an extract; The extract is subjected to column gradient elution, and the eluate is collected and concentrated and dried to obtain the Dendrobium officinale extract.

2. The method for preparing the Dendrobium officinale extract according to claim 1, characterized in that: The working condition parameters of the oxygen-enriched microwave plasma treatment include: microwave power of 180 to 290 W, oxygen and argon with a volume ratio of 7:1 as the working medium, an inlet flow rate of the working medium of 65 to 75 mL / min, a working mode of glow discharge, a treatment time of 20 to 45 seconds, and oxygen is continuously introduced for 5 to 10 minutes after the glow discharge treatment is completed.

3. The method for preparing the Dendrobium officinale extract according to claim 1, characterized in that: The microwave power of the oxygen-enriched microwave plasma treatment is 225 W, and the time of the oxygen-enriched microwave plasma treatment is 30 seconds.

4. The method for preparing the Dendrobium officinale extract according to claim 1, characterized in that: The particle size of the first material is 20-50 meshes.

5. The method for preparing the Dendrobium officinale extract according to claim 1, characterized in that: The working condition parameters of the reflux extraction include: temperature of 100-110° C., time of 1-2 hours, number of times of 1-3 times, and a weight ratio of the second material to the water of (1-2):(8-12).

6. The method for preparing the Dendrobium officinale extract according to claim 1, characterized in that: The extract is subjected to column gradient elution, and the eluate is collected and concentrated and dried to obtain the Dendrobium officinale extract, which comprises the following steps: The extract and D101 macroporous resin are uniformly mixed in a weight ratio of 1:1.2, and the mixture is loaded onto a column by a dry method. First, elution is performed with petroleum ether having a volume of 5 times the column, and then gradient elution is performed with 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 55% ethanol, 60% ethanol, 65% ethanol, 70% ethanol, 75% ethanol, 80% ethanol, 85% ethanol, 90% ethanol and 95% ethanol, respectively, wherein the amount of each gradient eluent is 5 times the column volume, and the eluents having 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol and 35% ethanol are collected, concentrated and dried to obtain the Dendrobium officinale extract.

7. A Dendrobium officinale extract, characterized in that: The Dendrobium officinale extract is prepared by the preparation method of the Dendrobium officinale extract according to any one of claims 1 to 6.

8. Use of the Dendrobium officinale extract according to claim 7 in the preparation of a drug for treating brain glioma.

9. The use according to claim 8, characterized in that: The dosage form of the drug for treating brain glioma includes tablets, capsules, powders, mixtures, pills or granules.

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