Method for extracting and preparing linarin from Jinsha river buddleia

Abstract

The invention relates to a method for extracting and preparing linarin from Jinsha River buddleia, which comprises the following steps: S1, drying and crushing Jinsha River buddleia branches and leaves, adding an ethanol water solution, and extracting by a reflux extraction method to obtain an extract; s2, the extract obtained in the step S1 is subjected to column chromatography, resin adopted for column chromatography is styrene-divinylbenzene copolymer resin, and ethanol water is added for gradient elution; and S3, collecting the eluent obtained in the step S2, concentrating, and separating out linarin. Compared with the prior art, the method has the advantages that Jinsha river buddleia is used as a raw material, the styrene-divinylbenzene copolymer resin is used as a separation material for column chromatography, only ethanol and water are used as solvents, the purity of the obtained linarin reaches 85% or above, and the requirements of the fields of medicine, health care products and the like for high-purity linarin are met.

Description

Technical Field

[0001] This invention relates to the field of natural product separation technology, and in particular to a method for extracting and preparing buddleja glycosides from Buddleja officinalis in the Jinsha River. Background Technology

[0002] Buddleja officinalis is a flavonoid glycoside compound with various pharmacological activities, such as antibacterial, hypoglycemic, and hepatoprotective effects, and has broad application prospects in the fields of medicine and health products. Currently, the main source of buddleja officinalis is extraction from natural plants, such as wild chrysanthemum (Chrysanthemumindicum L.), Buddleja officinalis Maxim., and Cirsium spicatum Matsum.

[0003] The extraction and separation methods for buddleja glycosides mainly involve reflux extraction and alkaline extraction with alcohol precipitation, while purification methods include extraction, column chromatography, and recrystallization. For example, patent 202211304406.X uses an evaporation reflux device to treat the solution prepared from wild chrysanthemum powder by evaporation reflux treatment, and the buddleja glycoside content in the buddleja glycoside extract obtained by this method is above 60%; patent 202111390714.4 uses wild chrysanthemum stems as raw material, and uses ethanol extraction, water precipitation to separate the initial product, and stepwise crystallization to purify buddleja glycosides; patent 202010244509.6 uses citric acid, cellulase, and pectinase to soak buddleja glycosides for enzymatic hydrolysis, followed by pulping, fermentation with mixed strains, and then extraction of buddleja glycosides, which has a high time cost; patent 201810527675.X uses dried buddleja glycosides as raw material, and uses pulverization, enzymatic hydrolysis, concentration, macroporous adsorption resin column chromatography, reagent recovery, and vacuum drying to obtain buddleja glycosides.

[0004] However, the methods for extracting strychnine from these plants all have certain drawbacks, such as low purity, cumbersome operation, long time consumption, and high cost. They cannot effectively meet the requirements of high purity and high yield in the industrial production of strychnine, thus limiting the large-scale production and application of strychnine.

[0005] Therefore, there is an urgent need to find new plant sources and develop more efficient extraction and preparation methods. Summary of the Invention

[0006] The purpose of this invention is to provide a method for extracting and preparing buddleja glycosides from Buddleja officinalis in order to meet the requirements of high purity and high yield for industrial production of buddleja glycosides.

[0007] The objective of this invention can be achieved through the following technical solutions:

[0008] The technical solution of the present invention is to provide a method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River, comprising the following steps:

[0009] S1. Dry and pulverize the branches and leaves of *Buddleja officinalis* from Jinsha River, add an ethanol-water solution, and extract by reflux extraction to obtain the extract;

[0010] S2. The extract obtained in step S1 is subjected to column chromatography. The resin used for column chromatography is styrene-divinylbenzene copolymer resin, and ethanol and water are added for gradient elution.

[0011] S3. Collect the eluent obtained in step S2, concentrate and precipitate to obtain buddleja glycoside.

[0012] In some specific embodiments, in step S1, the volume percentage of ethanol in the aqueous ethanol solution is 95%.

[0013] In some specific implementations, the extraction is performed 1 to 3 times in step S1.

[0014] In some specific implementations, in step S1, the extraction time for each extraction is (2-4) hours.

[0015] As a better option, the extraction time is 3 hours each time.

[0016] In some specific embodiments, in step S2, the extract is mixed with styrene-divinylbenzene copolymer resin before column chromatography.

[0017] In some specific embodiments, the mass ratio of the extract to the styrene-divinylbenzene copolymer resin is 1:(1.5 to 2.5).

[0018] As a more preferred embodiment, the mass ratio of the extract to the styrene-divinylbenzene copolymer resin is 1:2.

[0019] In some specific embodiments, in step S2, the volume percentage of ethanol in the gradient elution ethanol water is successively 20% to 100%.

[0020] In some specific embodiments, in step S2, the ethanol-water used for gradient elution is selected with ethanol contents of 20%, 40%, 60%, 80%, and 100%, respectively.

[0021] In some specific implementations, five column volumes are collected for each gradient.

[0022] In some specific embodiments, in step S3, the component obtained by elution with 60% ethanol-water is concentrated and precipitated to obtain buddleja glycoside.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] (1) This invention uses Buddleja nivea Duthie. as raw material to extract buddleja glycoside. Buddleja nivea Duthie. is a shrub of the Buddleja genus in the Loganiaceae family. It is 1 to 3 meters tall and grows in sparse forests in mountainous areas or in shrubbery on mountain slopes and valleys. It is rich in resources and provides a new plant source for the extraction of buddleja glycoside.

[0025] (2) The present invention uses styrene-divinylbenzene copolymer resin as the separation material for column chromatography. Compared with other separation materials (such as octadecyl bonded phase silica gel), it has the advantages of low price, wide applicability and high purification efficiency.

[0026] (3) In the extraction process of buddleja glycosides in this invention, only ethanol and water are used as solvents. Compared with organic solvents such as acetone and methanol, which are highly toxic, ethanol can be recycled and has low toxicity, reducing the waste of organic solvents and pollution to the environment.

[0027] (4) This invention uses Buddleja officinalis from Jinsha River as raw material, uses styrene-divinylbenzene copolymer resin as the separation material for column chromatography, and uses only ethanol and water as solvents. The purity of the obtained Buddleja officinalis glycosides reaches more than 85%, which meets the demand for high-purity Buddleja officinalis glycosides in the fields of medicine and health products. Attached Figure Description

[0028] Figure 1 This is the carbon NMR spectrum of strychnoside in this invention.

[0029] Figure 2 This is the high-resolution mass spectrum of strychnoside in this invention.

[0030] Figure 3 This is the high-performance liquid chromatogram of strychnoside in this invention. Detailed Implementation

[0031] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0032] In the following embodiments, unless otherwise specified, the raw materials or processing techniques are conventional commercially available raw materials or conventional processing techniques in the art.

[0033] Example 1

[0034] This embodiment provides a method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River, including the following steps:

[0035] (1) Take 100g of the branches and leaves of Buddleja officinalis from Jinsha River, air dry and crush them, then extract them by reflux with 95% ethanol water three times for three hours each time. After collecting the extract, concentrate it under reduced pressure to obtain a dark brown extract.

[0036] (2) The concentrated extract was mixed with styrene-divinylbenzene copolymer resin at a mass ratio of 1:2 to ensure thorough mixing, and the mixture was used as a sample for later use.

[0037] (3) Take a glass chromatography column, add styrene-divinylbenzene copolymer resin, sample, and buffer cotton respectively, and compact them to complete the sample loading.

[0038] (4) Elute sequentially with 20% ethanol-water, 40% ethanol-water, 60% ethanol-water, 80% ethanol-water, and 100% ethanol-water gradients. Elute 5 column volumes for each gradient.

[0039] (5) Collect the 60% ethanol-water eluent, concentrate it using a rotary evaporator, remove the ethanol, and precipitate. Dry the precipitate, weigh it, and obtain 2.23g of white solid, which is budesonin.

[0040] Its structure was determined by carbon NMR spectroscopy and high-resolution mass spectrometry:

[0041] NMR data of budesonide: 13 C NMR (100MHz, DMSO) δ182.0,164.0,163.0,162.4,161.2,157.0,128.5,122.7,114.7,105.5,103 .8,100.6,100.0,99.7,94.8,76.3,75.7,73.1,72.1,70.8,70.4,69.6,68.4,66.1,55.6,17.8.

[0042] High-resolution mass spectrometry (HR-ESI-MS) in negative ion mode shows [MH]. - The value is 591.1716 (Calc m / z 591.1719), confirming the molecular formula of the compound as C. 28 H 32 O 14 .

[0043] The purity of the isolated scutellarin was analyzed by high performance liquid chromatography (HPLC). The HPLC experimental conditions are shown in Table 1; the specific mobile phase method used in the experiment is shown in Table 2. The results showed that at 254 nm, the retention time of scutellarin was 16.118 min, and the peak area was 85.96%, indicating that the purity of the prepared scutellarin was 85.96%.

[0044] Table 1 Experimental conditions for high performance liquid chromatography

[0045]

[0046] Table 2 Mobile Phase Method for High Performance Liquid Chromatography

[0047]

[0048] The above description of the embodiments is provided to enable those skilled in the art to understand and use the invention. It will be apparent to those skilled in the art that various modifications can be made to these embodiments, and the general principles described herein can be applied to other embodiments without inventive effort. Therefore, the present invention is not limited to the above embodiments, and any improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the invention should be within the protection scope of the present invention.

Claims

1. A method for extracting and preparing buddleja glycosides from Buddleja officinalis in the Jinsha River, characterized in that, Includes the following steps: S1. Dry and pulverize the branches and leaves of *Buddleja officinalis* from Jinsha River, add an ethanol-water solution, and extract by reflux extraction to obtain the extract; S2. The extract obtained in step S1 is subjected to column chromatography. The resin used for column chromatography is styrene-divinylbenzene copolymer resin, and ethanol and water are added for gradient elution. S3. Collect the eluent obtained in step S2, concentrate and precipitate to obtain buddleja glycoside.

2. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 1, characterized in that, In step S1, the volume percentage of ethanol in the ethanol-water solution is 95%.

3. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 1, characterized in that, In step S1, the extraction is performed 1 to 3 times.

4. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 3, characterized in that, In step S1, the extraction time is 2 to 4 hours each time.

5. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 1, characterized in that, In step S2, the extract is mixed with styrene-divinylbenzene copolymer resin before column chromatography.

6. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 5, characterized in that, The mass ratio of the extract to styrene-divinylbenzene copolymer resin is 1:(1.5-2.5).

7. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 1, characterized in that, In step S2, the volume percentage of ethanol in the gradient elution ethanol water is 20% to 100% sequentially.

8. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 7, characterized in that, In step S2, the ethanol-water used for gradient elution is selected with ethanol contents of 20%, 40%, 60%, 80%, and 100%, respectively.

9. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 8, characterized in that, Five column volumes are collected for each gradient.

10. The method for extracting and preparing buddleja glycosides from Buddleja officinalis of the Jinsha River according to claim 1, characterized in that, In step S3, the fraction obtained by elution with 60% ethanol-water is concentrated and precipitated to obtain buddleja glycoside.

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