CN105732653B - A kind of method that Oridonin is prepared from Isodon Japonica Hara - Google Patents

Abstract

The present invention relates to a method for preparing oridonin from tea leaves, which can effectively solve the problem of extracting oridonin from tea leaves with high efficiency, rapidity and high reproducibility. The technical solutions to be solved are: 1) preparation of loading solution; 2) resin pretreatment; 3) preparation of crude oridonin; 4) purification of oridonin; Tea vegetables were used as raw materials, and gel resin column chromatography was used for purification, and a method for preparing high-purity oridonin with simple process and good reproducibility was established. The method is simple, easy to operate, high in production efficiency, and reproducible. Good presentability, the obtained oridonin A has good quality and high purity, which is a great innovation in the preparation of oridonin A.

Description

A kind of method for preparing oridonin from the fragrant tea leaves

technical field

The invention relates to the field of medicines, in particular to a method for preparing oridonin from chrysanthemum tomentosa.

Background technique

Herbal tea is the dried whole herb of the genus Lamiaceae, and it is known as quadrangular pole, sage, and yanmingcao among the people. It has the effects of clearing away heat and detoxification, promoting blood circulation and reducing swelling. For a long time, it has been used as an antibacterial and anti-inflammatory herbal medicine in my country. The whole herb is used as medicine to treat hepatitis, pharyngitis, mastitis, amenorrhea, bruises, joint pain and snake bites. It is a plant of the same family and genus as Dongling grass, and it is distributed in Jiangsu, Henan, Shanxi, and Shaanxi in my country. The chemical composition suggested that the content of oridonin in this plant was much higher than that in Oridonin grass. However, how to quickly and efficiently prepare oridonin from Chacao basilicates has not been publicly reported so far.

Rubescensin A is the main active substance of Rubescens oriolis. In the 2015 edition of "Chinese Pharmacopoeia", the quality index ingredients of the herb of Donglingcao and the Chinese patent medicine Donglingcao Tablets, which use Rubescens oriolis as the main raw material, are all Oridscens A white. Oridonin has obvious anti-cancer, anti-inflammatory, anti-bacterial and central nervous system and other pharmacological effects. Clinical trials have proved that this compound has unique and obvious anti-cancer effects, especially for esophageal cancer and cardia cancer, and it is non-toxic Or weak toxicity, no obvious damage to important organs of the human body such as bone marrow, liver, kidney, etc., is currently undergoing clinical research as a class of drugs.

It has been reported in the literature that the method for preparing oridonin by adopting the macroporous adsorption resin method with Oridonium grass as raw material, although the macroporous adsorption resin is relatively cheap, its selectivity and directional enrichment ability are all poor, therefore, Macroporous adsorption resins are mostly used in the preparation of effective components. The preparation of monomeric compounds must be assisted by various methods such as column chromatography and organic solvent extraction. Therefore, the preparation process is cumbersome, the reproducibility is poor, and the purity and yield of the finished product are low. Therefore, it is imperative to develop a method for preparing oridonin.

Contents of the invention

In view of the above situation, in order to solve the defects of the prior art, the purpose of the present invention is to provide a kind of method for preparing oridonin from the tea leaves, which can effectively solve the problem of high efficiency, fast and high reproducibility. The problem of extracting oridonin from tea leaves.

The technical scheme solved by the present invention is that the molecular structural formula of this oridonin is:

Achieved by the following steps:

1) Preparation of the loading solution: dry the medicinal materials of the chrysanthemum chrysanthemum, crush them, pass through a 40-80 mesh sieve, add ethanol with a mass concentration of 50-95% of 10-12 times the weight of the chrysanthemum chrysanthemum each time, Heated and refluxed for 3 times, each time for 1-2 hours, combined the extracts for 3 times, recovered under reduced pressure until there was no alcohol smell, added water to disperse, and obtained a water dispersion, the concentration of the water dispersion was equivalent to 1-2g/mL crude drug, and the product was obtained. Sample solution (concentration is equivalent to 1-2g/ml of raw medicinal materials);

2) Resin pretreatment: the method is to soak the resin with methanol for 24 hours, then soak the resin with methanol with a volume concentration of 15-20% for 12 hours, replace the methanol every 2 hours, wet-pack the column, and then use a volume concentration of 15-20%. elute with 2 column volumes of methanol to obtain a pretreated resin;

3) Preparation of crude oridonin: Wet-pack the pretreated resin in step 2) to make the resin bed diameter-to-height ratio reach 1:10-15, and then pass the sample solution in step 1) through the resin The column is dynamically adsorbed at a natural flow rate. After the adsorption, the resin column is first eluted with 1-2 times the column volume of 15-20% methanol water, and then 2-4 times the column volume with 30% ethanol. , collecting the eluent whose volume concentration is 30% ethanol, and reclaiming ethanol under reduced pressure to obtain a thick extract with a relative density of 1.2-1.4;

4) Purification of Rubescensin A: Dissolve the thick extract with 15-25ml of solvent, let stand at -20°C-30°C for 24-72h, precipitate white powdery crystals, filter to obtain white crystalline powders, and The clear liquid was left to stand at -20°C-30°C for 24-72h, and white powdery crystals were precipitated, filtered to obtain a white crystalline powder, and two white crystalline powders were combined to obtain oridonin; The solvent is one of methanol, ethanol, propanol, isopropanol, acetonitrile or acetone.

In the present invention, the low-cost chrysanthemum tea leaves are used as raw materials, purified by gel resin column chromatography, and a method for preparing high-purity oridonin with simple process and good reproducibility is established, and the method is simple , easy to operate, high production efficiency, good reproducibility, and the prepared oridonin A has good quality and high purity, which is a great innovation in the preparation of oridonin A.

Description of drawings

Fig. 1 is a molecular structure diagram of oridonin of the present invention.

Figure 2 is a high performance liquid chromatogram of oridonin of the present invention.

detailed description

The specific implementation manners of the present invention will be described in further detail below in conjunction with the accompanying drawings and examples.

Example 1

1) Preparation of sample solution: crush 1 kg of dry medicinal materials of Amoy basilisk, pass through a 40-mesh pharmacopoeia sieve, add 10 L of ethanol with a mass concentration of 95% each time, extract 3 times, each time for 1.5 h, and combine the extracts 3 times , recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour out the supernatant, add 100ml of ethanol with a volume concentration of 15% to the insoluble matter Dissolve, centrifuge at 3600r/min for 30min, combine two supernatants to prepare sample solution (concentration is equivalent to 1g/ml of raw medicinal material);

2) Resin pretreatment: the method is, after soaking Sephadex LH-20 resin with methanol for 24 hours, pour off the methanol solution, soak it in methanol with a volume concentration of 15% for 12 hours, replace the solvent once every 2 hours, wet-pack the column, and use the volume concentration Elute 2 column volumes with 15% methanol to obtain pretreated Sephadex LH-20 resin;

3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:15, and then put the upper resin in step 1) The natural flow rate of 1000ml sample liquid is completely absorbed by the resin column;

4) Separation and purification: the resin column is first eluted with 15% methanol by volume concentration to elute 1500ml, then 4000ml with 30% methanol to be eluted, and the eluent whose volume concentration is 30% methanol is collected, and the solvent is reclaimed under reduced pressure to obtain the relative density It is a thick extract of 1.243;

5) Recrystallization: dissolve the above-mentioned thick extract with 20ml of methanol, let it stand at -20°C-35°C for 24-72h, remove the supernatant, precipitate white powdery crystals, filter to obtain white crystalline powder, supernatant The liquid continued to stand for 24-72h, and white powdery crystals were precipitated, filtered to obtain a white crystalline powder, and the white crystalline powder was combined twice to obtain 0.523g of oridonin, and its purity was calculated by the HPLC method area normalization method Greater than 96.81% (oridonin with a purity greater than 95% is also called oridonin monomer compound).

Example 2

1) Preparation of the sample solution: 1 kg of dried medicinal materials of C. basilisk, crushed, passed through a 60-mesh pharmacopoeia sieve, added 12 L of ethanol with a mass concentration of 95% each time, extracted 3 times, 2 hours each time, and combined the extracts 3 times , recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour out the supernatant, add 200ml of ethanol with a volume concentration of 20% to the insoluble matter Dissolve, centrifuge at 3600r/min for 30min, combine two supernatants to prepare sample solution (concentration is equivalent to raw medicinal material 2g/ml);

2) Resin pretreatment: The method is to soak Sephadex G resin with methanol for 24 hours, pour off the methanol solution, soak it with 20% methanol for 12 hours, replace the solvent once every 2 hours, wet-pack the column, and wash it with 20% methanol by volume concentration. Remove 2 column volumes to obtain pretreated Sephadex G resin;

3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:15, and then put the upper resin in step 1) The 500mL sample liquid passes through the resin column at a natural flow rate, and the adsorption is complete;

4) Separation and purification: the resin column is first eluted with 20% methanol by volume to 1000ml, then 3000ml with 30% methanol, and the eluate with volume concentration of 30% methanol is collected, and the solvent is recovered under reduced pressure to obtain the relative density It is a thick extract of 1.321;

5) Recrystallization: Dissolve 20ml of the above-mentioned thick extract in ethanol, let stand at -20°C-35°C for 24-72h, remove the supernatant, precipitate white powdery crystals, filter to obtain white crystalline powder, supernatant The solution was left to stand for 24-72h, and white powdery crystals were separated out, filtered to obtain a white crystalline powder, and the above-mentioned white crystalline powder was combined to obtain 0.5021g of oridonin, and its purity was calculated by the HPLC method area normalization method to be greater than 95.12%.

Example 3

1) Preparation of the sample solution: 1 kg of dried medicinal materials of C. basilisk, crushed, passed through an 80-mesh pharmacopoeia sieve, added 12 L of ethanol with a mass concentration of 75% each time, extracted 3 times, 2 hours each time, and combined the extracts 3 times , recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour out the supernatant, add 800ml of ethanol with a volume concentration of 20% to the insoluble matter Dissolve, centrifuge at 3600r/min for 30min, and combine two supernatants to prepare sample solution (concentration is equivalent to raw medicinal material 1.25g/ml);

2) Resin pretreatment: the method is, after the MCI resin is soaked in methanol for 24 hours, the methanol liquid is poured off, soaked in 20% methanol for 12 hours, the solvent is replaced every 2 hours, the column is wet-packed, and the volume concentration is 20% methanol washing Remove 2 column volumes to obtain pretreated MCI resin;

3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:15, and then put the upper resin in step 1) 800ml of the sample liquid passes through the resin column at a natural flow rate, and the adsorption is complete;

4) Separation and purification: the resin column is first eluted with 20% methanol by volume to 2000ml, then 3000ml with 30% methanol, and the eluate with volume concentration of 30% methanol is collected, and the solvent is recovered under reduced pressure to obtain the relative density 1.246 thick extract;

5) Recrystallization: Dissolve the above-mentioned thick extract with 20ml of propanol, let it stand at -20°C-35°C for 24-72h, remove the supernatant, precipitate white powdery crystals, filter to obtain white crystalline powders, and The clear liquid was left to stand for 24-72 hours, and white powdery crystals were precipitated, filtered to obtain a white crystalline powder, and the white crystalline powder was combined twice to obtain 0.489g of oridonin, which was calculated by the HPLC method area normalization method Its purity is greater than 96.87% (oridonin with a purity greater than 95% is also called oridonin monomer compound).

Example 4

1) Preparation of the sample solution: 1 kg of dried medicinal materials of C. basilisk, crushed, passed through an 80-mesh pharmacopoeia sieve, added 12 L of ethanol with a mass concentration of 50% each time, extracted 3 times, each time for 1.5 h, and combined the 3 extractions Liquid, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour out the supernatant, add 500ml of insoluble matter with a volume concentration of 20% Dissolve in ethanol, centrifuge at 3600r/min for 30min, and combine the two supernatants to prepare a sample solution (concentration equivalent to 1g/ml of raw medicinal materials);

2) Resin pretreatment: the method is, after soaking Sephadex LH-20 resin with methanol for 24 hours, pour off the methanol solution, soak it with 20% methanol for 12 hours, replace the solvent once every 2 hours, and wet-pack the column with a volume concentration of 20%. 2 column volumes of methanol elution to obtain pretreated Sephadex LH-20 resin;

3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:10, and then put the upper resin in step 1) The natural flow rate of 1L sample liquid passes through the resin column, and the adsorption is complete;

4) Separation and purification: the resin column was first eluted with 20% methanol by volume concentration to elute 1.9L, then 4000ml was eluted with 30% methanol, and the eluent with volume concentration of 30% methanol was collected, and the solvent was reclaimed under reduced pressure to obtain relative A thick extract with a density of 1.321;

5) Recrystallization: Dissolve 20ml of the above-mentioned thick extract in isopropanol, let stand at -20°C-35°C for 24-72h, white powdery crystals are precipitated, filter to obtain a white crystalline powder, and the supernatant continues to stand still Put it for 24-72h, precipitate white powdery crystals, filter to obtain white crystalline powders, and combine the above white crystalline powders to obtain 0.549g of oridonin, the purity of which is greater than 95.02% calculated by HPLC area normalization method .

The extract prepared above is determined by chromatographic analysis (see Figure 2), and its structural formula is as shown in Figure 1. It is determined to be Rubescensin A, white crystal (methanol), mp: 248-250 ° C, and can be dissolved in organic compounds such as chloroform. Solvent, insoluble in water, sprayed with anisaldehyde-concentrated sulfuric acid, heated at 110°C, blue, and dark spots observed under ultraviolet 254nm.

¹³ C-NMR (125MHz, CDCl ₃ ) gave a total of 20 carbon signals, including 6 carbon signals connected to oxygen (including 1 ketone carbonyl); according to the NMR spectrum δ _H 6.17 (1H, s) and 5.55 (lH, s) is the two proton signals on the terminal olefinic bond at C-17; δ _C 207.3, 151.3 and 120.9 are the signals at C-15, C-16 and C-17, respectively; δ _H 1.12(3H,s ) and 1.12(3H,s) are two geminal carbon-angle methyl signals, suggesting that the basic skeleton of the compound is ent-kaur-16-en-15-one.

According to the chemical shifts, splitting conditions and coupling constants of the enantio-kaurene diterpenes, 3.75(1H,dd,7.9,11.7) is the proton signal at the β position on C-1; δ _H 3.45(1H,d ,7.8) is the proton signal at the α position on C-6; δ _H 4.89(1H,s) is the proton signal at the α position at C-14; δ _H 4.29(1H,d,9.1) and 4.07(1H,d, 10.2) is a pair of AB-coupled proton signals at position C-20;

In the ¹³ C-NMR spectrum, δ _C 74.7, δ _C 73.8, δ _C 97.2, and δ _C 72.4 are the carbon signals of C-1, C-6, C-7 and C-14 respectively; The NMR data was compared with the literature value of oridonin, and the data were consistent. It was determined that the compound was oridonin, and its molecular formula was C ₂₀ H ₂₈ O ₆ . See Table 1 for details.

Table 1.NMR data for compound 15in CDCl ₃ (Jin Hz)

Because the quality and quantity of Oridonin A distributed in different medicinal plants are different, the key of the present invention is to combine the distribution characteristics of Oridonin A in Herba basilicates, and to avoid cumbersome operation and toxicity. Large, inefficient, and poorly reproducible purification methods. Massive and rapid preparation of high-purity oridonin has high technological innovation. Provide an economical medical raw material for the pharmaceutical industry, and provide a high-purity reference substance for the drug regulatory department. A cheap new material for the preparation of oridonin was provided by taking the chrysanthemum as raw material.

Oridonin A prepared by the present invention adopts a conventional method (i.e. existing ballochnine activity test method) experimental proof, and Rubescensin A prepared by the present invention has:

1. Rubescensine A has a specific and obvious inhibitory effect on transplanted sarcoma S180 solid tumors in mice, and has a certain inhibitory effect on brain tumors (B-22); it can significantly prolong the survival period of tumor-bearing mice, and is non-toxic side effect;

2. Rubescensine A has obvious inhibitory activity on tumor-bearing liver cancer ascites mice, and can significantly prolong the survival period of tumor-bearing mice.

3. The results of cytotoxicity experiments showed that Rubescensine A had significant cytotoxicity on HCT-116, HepG2, BGC-823, and A2780 cells.

Compared with the prior art, it has the following outstanding beneficial technical effects:

1. One of the innovations of the present invention is to find a new raw material for preparing oridonin instead of oridonin.

2. Combining the quality and quantity distribution characteristics of Rubescensin A in the tea leaves and the physical and chemical properties of Rubescensin A, a high-purity winter grass can be prepared by only one column chromatography. The method for the monomeric compound of iconidin has the advantages of simple process, good reproducibility, easy operation and industrialized production;

3. Provide a simple, directional, and reproducible technical guarantee and conditions for the preparation of reference substances for the quality control of Donglingcao medicinal materials and Chinese patent medicines using Donglingcao as raw materials;

4. A simple and reproducible process for preparing intermediates has been developed for the development of innovative new drugs using oridonin as a raw material.

Claims (4)

1. a method for preparing oridonin from the chrysanthemum tea dish, is characterized in that, comprises the following steps: 1) Preparation of sample solution: Grind 1 kg of dried medicinal materials of A. basilisk, pass through a 40-mesh pharmacopoeia sieve, add 10 L of ethanol with a mass concentration of 95% each time, extract 3 times, each time for 1.5 hours, and combine the extracts 3 times , recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, disperse with water, centrifuge at 3600 r/min for 30 min, pour out the supernatant, add 100ml of insoluble matter to make the volume concentration 15% Dissolve in ethanol, centrifuge at 3600 r/min for 30 min, and combine the two supernatants to prepare the sample solution; 2) Resin pretreatment: The method is, after soaking Sephadex LH-20 resin in methanol for 24 hours, pour off the methanol solution, soak in methanol with a volume concentration of 15% for 12 hours, replace the solvent every 2 hours, wet-pack the column, and use the volume concentration Elute 2 column volumes with 15% methanol to obtain pretreated Sephadex LH-20 resin; 3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:15, and then put the upper resin in step 1) The natural flow rate of 1000ml sample liquid is completely absorbed by the resin column; 4) Separation and purification: the resin column was first eluted with 15% methanol by volume to 1500ml, and then 4000ml by 30% methanol, and the eluate with volume concentration of 30% methanol was collected, and the solvent was recovered under reduced pressure to obtain the relative density It is a thick extract of 1.243; 5) Recrystallization: Dissolve the above thick extract in 20ml of methanol, let it stand at -20°C-35°C for 24-72h, remove the supernatant, precipitate white powdery crystals, filter to obtain white crystalline powder, supernatant The liquid continued to stand for 24-72h, and white powdery crystals were precipitated, filtered to obtain a white crystalline powder, and the white crystalline powder was combined twice to obtain 0.523g of oridonin, and its purity was calculated by the HPLC method area normalization method Greater than 96.81%. 2. A method for preparing oridonin from the chrysanthemum tea dish, is characterized in that, comprises the following steps: 1) Preparation of the sample solution: 1 kg of dried medicinal materials of C. basilisk, crushed, passed through a 60-mesh pharmacopoeia sieve, added 12 L of ethanol with a mass concentration of 95% each time, extracted 3 times, 2 hours each time, and combined the extracts 3 times , recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour the supernatant, add 200ml of ethanol with a volume concentration of 20% to the insoluble matter Dissolve, centrifuge at 3600r/min for 30min, combine the two supernatants to prepare the sample solution; 2) Resin pretreatment: The method is to soak Sephadex G resin in methanol for 24 hours, pour off the methanol solution, soak in 20% methanol for 12 hours, replace the solvent every 2 hours, wet-pack the column, and wash it with 20% methanol by volume. Remove 2 column volumes to obtain pretreated Sephadex G resin; 3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:15, and then put the upper resin in step 1) The 500mL sample liquid passes through the resin column at a natural flow rate, and the adsorption is complete; 4) Separation and purification: the resin column is first eluted with 20% methanol by volume to 1000ml, and then 3000ml by 30% methanol, and the eluate with volume concentration of 30% methanol is collected, and the solvent is recovered under reduced pressure to obtain the relative density It is a thick extract of 1.321; 5) Recrystallization: Dissolve 20 ml of the above thick extract in ethanol, let it stand at -20°C-35°C for 24-72h, remove the supernatant, precipitate white powdery crystals, filter to obtain white crystalline powders, and The clear liquid continued to stand for 24-72 hours, and white powdery crystals were precipitated, filtered to obtain white crystalline powders, and the above white crystalline powders were combined to obtain 0.5021g of oridonin, and the purity was calculated by HPLC area normalization method Greater than 95.12%. 3. A method for preparing oridonin from the fragrant tea leaves, characterized in that, comprising the following steps: 1) Preparation of the sample solution: 1 kg of dried medicinal materials of C. basilisk, crushed, passed through an 80-mesh pharmacopoeia sieve, added 12 L of ethanol with a mass concentration of 75% each time, extracted 3 times, 2 hours each time, and combined the extracts 3 times , recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour out the supernatant, add 800ml of ethanol with a volume concentration of 20% to the insoluble matter Dissolve, centrifuge at 3600r/min for 30min, combine the two supernatants to prepare the sample solution; 2) Resin pretreatment: The method is to soak the MCI resin in methanol for 24 hours, pour off the methanol liquid, soak it in 20% methanol for 12 hours, replace the solvent every 2 hours, wet-pack the column, and wash it with 20% methanol by volume. Remove 2 column volumes to obtain pretreated MCI resin; 3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5 cm, so that the resin bed diameter-height ratio reaches 1:15, and then put the upper resin in step 1) 800ml of the sample liquid passes through the resin column at a natural flow rate, and the adsorption is complete; 4) Separation and purification: the resin column was first eluted with 20% methanol by volume to 2000ml, then 3000ml by 30% methanol, collected the eluate with a volume concentration of 30% methanol, recovered the solvent under reduced pressure, and obtained the relative density 1.246 thick extract; 5) Recrystallization: Dissolve the above thick extract with 20ml of propanol, let it stand at -20°C-35°C for 24-72h, remove the supernatant, precipitate white powdery crystals, filter to obtain white crystalline powders, and The clear liquid was left to stand for 24-72 hours, and white powdery crystals were precipitated, filtered to obtain a white crystalline powder, and the white crystalline powder was combined twice to obtain 0.489g of oridonin, which was calculated by the HPLC method area normalization method Its purity is greater than 96.87%. 4. A method for preparing Oridonin from Herba Sinensis, characterized in that, comprising the following steps: 1) Preparation of the sample solution: 1 kg of dried medicinal materials of C. basilisk, crushed, passed through an 80-mesh pharmacopoeia sieve, added 12L of ethanol with a mass concentration of 50% each time, extracted 3 times, 1.5h each time, combined 3 extractions Liquid, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, recover the solvent under reduced pressure until there is no alcohol smell, add water to disperse, centrifuge at 3600r/min for 30min, pour out the supernatant, add 500ml of insoluble matter with a volume concentration of 20% Dissolve in ethanol, centrifuge at 3600r/min for 30min, combine the two supernatants to prepare the sample solution; 2) Resin pretreatment: The method is, after soaking Sephadex LH-20 resin in methanol for 24 hours, pour off the methanol solution, soak in 20% methanol for 12 hours, replace the solvent every 2 hours, and wet-pack the column with a volume concentration of 20% 2 column volumes of methanol elution to obtain pretreated Sephadex LH-20 resin; 3) Column loading and sample loading: Wet-pack the resin pretreated in step 2) into a chromatography column with an inner diameter of 5cm, so that the resin bed diameter-height ratio reaches 1:10, and then put the upper resin in step 1) The natural flow rate of 1L sample liquid passes through the resin column, and the adsorption is complete; 4) Separation and purification: the resin column was first eluted with 20% methanol by volume to 1.9L, and then 4000ml was eluted with 30% methanol, and the eluate with a volume concentration of 30% methanol was collected, and the solvent was recovered under reduced pressure to obtain a relative A thick extract with a density of 1.321; 5) Recrystallization: Dissolve 20ml of the above-mentioned thick extract in isopropanol, let stand at -20°C-35°C for 24-72h, white powdery crystals are precipitated, filter to obtain white crystalline powder, and the supernatant continues to stand still Set aside for 24-72h, precipitate white powdery crystals, filter to obtain white crystalline powders, and combine the above white crystalline powders to obtain 0.549g of oridonin, the purity of which is greater than 95.02% calculated by HPLC area normalization method .