CN105732653B - A kind of method that Oridonin is prepared from Isodon Japonica Hara - Google Patents
A kind of method that Oridonin is prepared from Isodon Japonica Hara Download PDFInfo
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- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
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Abstract
The present invention relates to a kind of method that Oridonin is prepared from Isodon Japonica Hara, can effectively solve the problems, such as that efficient, quick, reproduction degree is high and Oridonin is extracted from Isodon Japonica Hara, its technical scheme solved is 1）The preparation of load solution；2）Resin pre-processes；3）The preparation of thick Oridonin；4）The purifying of Oridonin；The present invention is using cheap Isodon Japonica Hara as raw material, purified using gel resin column chromatography, establish that a kind of technique is simple, the preparation method of the high-purity Oridonin of favorable reproducibility, method is simple, it is easy to operate, production efficiency is high, favorable reproducibility, and it is good that Oridonin quality is made, purity is high, is the big innovation on prepared by Oridonin.
Description
Technical field
The present invention relates to field of medicaments, particularly a kind of method that Oridonin is prepared from Isodon Japonica Hara.
Background technology
Mao Yexiang bitter edible plants dish is the drying herb of Labiatae Rabdosia plant Isodon Japonica Hara, among the people to be referred to as four rib bars, mountain
Perillaseed, prolong life grass.The effect of with clearing heat and detoxicating, activating blood circulation and reducing swelling.For a long time, China is among the people is used as anti-inflammation herbal medicine, entirely
Grass is used as medicine, and controls hepatitis, pharyngitis, mastitis, amenorrhoea, traumatic injury, arthralgia and snake bite and insect sting.Equal for Rabdosia rubescens belongs to together
Plant, it is distributed in China Jiangsu, Henan, Shanxi, Shaanxi.Chemical composition prompts the content of Oridonin in the plant remote
Higher than the content of Oridonin in Rabdosia rubescens.But how fast and efficiently to be prepared by Oridonin from Isodon Japonica Hara
Out, so far there are no open report.
Oridonin be Rabdosia rubescens main active substances, 2015 editions《Chinese Pharmacopoeia》Middle Rabdosia rubescens medicinal material and insulted with the winter
Grass is Oridonin for the quality index composition of the Chinese patent drug Dong ling cao tablet of primary raw material.Oridonin has obvious
The multiple pharmacological effects such as anticancer, anti-inflammatory, antibacterial and nervous centralis, it is unique and significantly anti-that clinical test proves that the compound has
Cancer effect, especially there are special efficacy, and nontoxic or weak poison to Esophageal carcinoma, to the human body important organ such as nothing such as marrow, liver, kidney
Obvious damage, clinical research is carried out currently as a kind of medicine.
There is document report, the method that Oridonin is prepared using Flavonoids by Macroporous Adsorption Resin as raw material using Rabdosia rubescens, macropore
Although polymeric adsorbent price is relatively relatively inexpensive, its selectivity and orienting enriching ability are poor, and therefore, macroporous absorbent resin is more
Preparation for active principle.Preparation for monomeric compound necessarily aids in more with column chromatography, organic solvent extractionprocess etc.
Kind method.So preparation technology is cumbersome, poor reproducibility, finished product purity and yield are relatively low.Therefore, develop and prepare Rabdosia rubescens
The method of A prime is imperative.
The content of the invention
For the above situation, to solve the defect of prior art, the purpose of the present invention is just to provide one kind from hair leaf scented tea
The method that Oridonin is prepared in dish, can effectively solve efficient, quick, the high extraction winter icepro from Isodon Japonica Hara of reproduction degree
The problem of careless A prime.
The technical scheme that the present invention solves is that the molecular structural formula of the Oridonin is：
Realized by following steps：
1) preparation of load solution：Isodon Japonica Hara is dried into medicinal material, crushed, crosses 40-80 mesh sieves, each Jia Maoye scented teas
10-12 times of mass concentration of dish weight is 50-95% ethanol, heating and refluxing extraction 3 times, each 1-2h, merges 3 extractions
Liquid, it is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, obtain aqueous dispersions, aqueous dispersions concentration produces equivalent to 1-2g/mL crude drugs
Sample liquid (concentration is equivalent to raw medicinal herbs 1-2g/ml)；
2) resin pre-processes：Method is, after resin is soaked into 24h with methanol, the methanol for being 15-20% with volumetric concentration soaks
12h is steeped, a methanol, wet method dress post, then the methanol for being 15-20% with volumetric concentration are replaced per 2h and elutes 2 column volumes, is obtained pre-
The resin handled well；
3) preparation of thick Oridonin：The resin wet method dress post that step 2) has been pre-processed, reaches resin bed blade diameter length ratio
1 ︰ 10-15, then the sample solution in step 1) is subjected to Dynamic Adsorption, after having adsorbed, resin column by resin column with natural flow velocity
First with 1-2 times of column volume 15-20% methanol water elution, then the ethanol elution for being 30% with the volumetric concentration of 2-4 times of column volume, receive
Collect the eluent that volumetric concentration is 30% ethanol, ethanol is recovered under reduced pressure, obtains relative density 1.2-1.4 thick medicinal extract；
4) purifying of Oridonin：Thick medicinal extract solubilizer 15-25ml is dissolved, 24- is stood at -20 DEG C -30 DEG C
72h, white powder crystallization is separated out, filtering obtains white crystals sprills, and supernatant continues to stand 24- at -20 DEG C -30 DEG C
72h, white powder crystallization is separated out, filtering, obtains white crystals sprills, merge white crystals sprills twice, the obtained winter insults
Careless A prime；Described solvent is one kind of methanol, ethanol, propyl alcohol, isopropanol, acetonitrile or acetone.
The present invention is purified using cheap Isodon Japonica Hara as raw material using gel resin column chromatography, is established
A kind of technique is simple, and the preparation method of the high-purity Oridonin of favorable reproducibility, method is simple, easy to operate, production efficiency
Height, favorable reproducibility, obtained Oridonin quality is good, and purity is high, is the big innovation on prepared by Oridonin.
Brief description of the drawings
Fig. 1 is the molecular structure of the Oridonin of the present invention.
Fig. 2 is the high-efficient liquid phase chromatogram of Oridonin of the present invention.
Embodiment
The embodiment of the present invention is described in further detail below in conjunction with drawings and examples.
Embodiment 1
1) preparation of sample solution：Isodon Japonica Hara is dried medicinal material 1kg and crushed, and crosses 40 mesh pharmacopeia sieve, adds mass concentration every time
For 95% ethanol 10L, extract 3 times, each 1.5h, merge No. 3 extract solutions, solvent is recovered under reduced pressure extremely without alcohol taste, adds moisture to dissipate,
Solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 100ml
The ethanol that volumetric concentration is 15% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution (concentration is made
Equivalent to raw medicinal herbs 1g/ml)；
2) resin pre-processes：Method is that after Sephadex LH-20 resins soak 24h with methanol, incline methanol solution, uses body
The methanol that product concentration is 15% soaks 12h, and a solvent is replaced per 2h, wet method dress post, is washed with the methanol that volumetric concentration is 15%
Take off 2 column volumes, the Sephadex LH-20 resins that must have been pre-processed；
3) post and loading are filled：The resin wet method dress post that step 2) has been pre-processed, in the chromatographic column that internal diameter is 5cm, make tree
Fat bed blade diameter length ratio is adsorbed completely up to 1 ︰ 15, then by the sample solution 1000ml natures flow velocity in step 1) by resin column；
4) isolate and purify：Resin column first elutes 1500ml with the methanol that volumetric concentration is 15%, then is washed with 30% methanol
De- 4000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, and obtain the thick leaching that relative density is 1.243
Cream；
5) recrystallize：Above-mentioned thick medicinal extract 20ml methanol is dissolved, 24-72h is stood at -20 DEG C -35 DEG C, removes supernatant
Liquid, white powder crystallization is separated out, filtering, obtains white crystals sprills, supernatant continues to place 24-72h, separates out white powder
Shape crystallizes, and filtering, obtains white crystals sprills, merges white crystals sprills twice, produces Oridonin 0.523g, uses
HPLC method area normalization methods calculate its purity, and more than 96.81%, (Oridonin of the purity more than 95% is also known as Rabdosia rubescens first
Plain monomeric compound).
Embodiment 2
1) preparation of sample solution：Isodon Japonica Hara dries medicinal material 1kg, crushes, and crosses 60 mesh pharmacopeia sieve, adds mass concentration every time
For 95% ethanol 12L, extract 3 times, each 2h, merge No. 3 extract solutions, solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, subtracts
Recycling design is pressed to add moisture to dissipate to without alcohol taste, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 200ml bodies
The ethanol that product concentration is 20% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution (concentration phase is made
When in raw medicinal herbs 2g/ml)；
2) resin pre-processes：Method is that Sephadex G resins soak 24h with methanol, and incline methanol solution, with 20% first
Alcohol soaks 12h, and a solvent is replaced per 2h, wet method dress post, 2 column volumes is eluted with the methanol that volumetric concentration is 20%, obtains pre-
The Sephadex G resins handled well；
3) post and loading are filled：The resin wet method dress post that step 2) has been pre-processed, in the chromatographic column that internal diameter is 5cm, make tree
Fat bed blade diameter length ratio is complete by resin column, absorption up to 1 ︰ 15, then by the sample solution 500mL natures flow velocity in step 1)；
4) isolate and purify：Resin column first elutes 1000ml with the methanol that volumetric concentration is 20%, then is washed with 30% methanol
De- 3000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, and obtain the thick leaching that relative density is 1.321
Cream；
5) recrystallize：Above-mentioned thick medicinal extract ethanol is dissolved into 20ml, 24-72h is stood at -20 DEG C -35 DEG C, removes supernatant
Liquid, white powder crystallization is separated out, filtering, obtains white crystals sprills, supernatant continues to place 24-72h, separates out white powder
Shape crystallizes, and filtering, obtains white crystals sprills, merges above-mentioned white crystals sprills, and Oridonin 0.5021g is made,
Its purity is calculated more than 95.12% using HPLC method area normalization methods.
Embodiment 3
1) preparation of sample solution：Isodon Japonica Hara dries medicinal material 1kg, crushes, and crosses 80 mesh pharmacopeia sieve, adds mass concentration every time
For 75% ethanol 12L, extract 3 times, each 2h, merge No. 3 extract solutions, solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, subtracts
Recycling design is pressed to add moisture to dissipate to without alcohol taste, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 800ml bodies
The ethanol that product concentration is 20% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution (concentration phase is made
When in raw medicinal herbs 1.25g/ml)；
2) resin pre-processes：Method is that after MCI resins soak 24h with methanol, incline methanol solution, is soaked with 20% methanol
12h is steeped, a solvent is replaced per 2h, wet method dress post, 2 column volumes is eluted with the methanol that volumetric concentration is 20%, must pre-process
Good MCI resins；
3) post and loading are filled：The resin wet method dress post that step 2) has been pre-processed, in the chromatographic column that internal diameter is 5cm, make tree
Fat bed blade diameter length ratio is complete by resin column, absorption up to 1 ︰ 15, then by the sample solution 800ml natures flow velocity in step 1)；
4) isolate and purify：Resin column first elutes 2000ml with the methanol that volumetric concentration is 20%, then is washed with 30% methanol
De- 3000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, obtains the thick medicinal extract of relative density 1.246；
5) recrystallize：Above-mentioned thick medicinal extract 20ml propyl alcohol is dissolved, 24-72h is stood at -20 DEG C -35 DEG C, removes supernatant
Liquid, white powder crystallization is separated out, filtering, obtains white crystals sprills, supernatant continues to place 24-72h, separates out white powder
Shape crystallizes, and filtering, obtains white crystals sprills, merges white crystals sprills twice, that is, Oridonin 0.489g is made, adopts
Calculating its purity with HPLC method area normalization methods, (Oridonin of the purity more than 95% is also known as Rabdosia rubescens more than 96.87%
A prime monomeric compound).
Embodiment 4
1) preparation of sample solution：Isodon Japonica Hara dries medicinal material 1kg, crushes, and crosses 80 mesh pharmacopeia sieve, adds mass concentration every time
For 50% ethanol 12L, extract 3 times, each 1.5h, merge No. 3 extract solutions, solvent is recovered under reduced pressure extremely without alcohol taste, adds moisture to dissipate,
Solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 500ml
The ethanol that volumetric concentration is 20% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution (concentration is made
Equivalent to raw medicinal herbs 1g/ml)；
2) resin pre-processes：Method is that after Sephadex LH-20 resins soak 24h with methanol, incline methanol solution, is used
20% methanol immersion 12h, a solvent is replaced per 2h, wet method dress post, 2 cylinders are eluted with the methanol that volumetric concentration is 20%
Product, the Sephadex LH-20 resins that must have been pre-processed；
3) post and loading are filled：The resin wet method dress post that step 2) has been pre-processed, in the chromatographic column that internal diameter is 5cm, make tree
Fat bed blade diameter length ratio is complete by resin column, absorption up to 1 ︰ 10, then by the sample solution 1L natures flow velocity in step 1)；
4) isolate and purify：Resin column first elutes 1.9L with the methanol that volumetric concentration is 20%, then is eluted with 30% methanol
4000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, and obtain the thick medicinal extract that relative density is 1.321；
5) recrystallize：Above-mentioned thick medicinal extract isopropanol is dissolved into 20ml, 24-72h is stood at -20 DEG C -35 DEG C, is separated out white
The powdered crystallization of color, filtering, white crystals sprills being obtained, supernatant continues to stand 24-72h, separates out white powder crystallization,
Filtering, obtains white crystals sprills, merges above-mentioned white crystals sprills, Oridonin 0.549g is produced, using HPLC
Method area normalization method calculates its purity more than 95.02%.
The extract of above-mentioned preparation determines (see Fig. 2) through chromatography, and its structural formula is shown in Fig. 1, is defined as Rabdosia rubescens
A prime, white crystals (methanol), mp：248-250 DEG C, the organic solvents such as chloroform can be dissolved in, not soluble in water, the anisaldehyde-concentrated sulfuric acid
Spraying, 110 DEG C of heating, shows blueness, and being observed under ultraviolet 254nm has blackening.
13C-NMR(125MHz,CDCl3) 20 carbon signals are provided altogether, wherein carbon signal (including 1 ketone carbonyl of 6 company's oxygen
Base)；According to H NMR spectroscopy δH6.17 (1H, s) and 5.55 (lH, s) are two proton signals on the terminal olefinic link of C-17 positions；δC207.3、
151.3 and 120.9 be the signal of C-15 positions, C-16 positions and C-17 positions respectively；δH1.12 (3H, s) and 1.12 (3H, s) be two
It is individual together with carbon angular methyl signal, prompt that compound basic framework is ent-kaur-16-en-15-one.
According to the chemical shift of ent kauranoid, split point situation and coupling constant rule, 3.75 (1H, dd,
7.9,11.7) be β positions on C-1 proton signal；δH3.45 (1H, d, 7.8) are the proton signals of α positions on C-6；δH4.89(1H,
S) be α positions at C-14 proton signal；δH4.29 (1H, d, 9.1) and 4.07 (1H, d, 10.2) are that a pair of AB of C-20 positions are coupled
Proton signal；
13δ in C-NMR spectrumsC74.7、δC73.8、δC 97.2、δC72.4 Fen Do are that C-1, C-6, C-7 and C-14 position are connected with hydroxyl
The carbon signal of base；The nuclear-magnetism number of compound 15 is contrasted with the literature value of Oridonin, data are consistent, are defined as compound and are
Oridonin (Oridonin), molecular formula C20H28O6, details are shown in Table 1.
Table 1.NMR data for compound 15in CDCl3(Jin Hz)
Because the quality and quantity that Oridonin is distributed in different medicinal plants is that discrepant, of the invention key is
With reference to Oridonin in Isodon Japonica Hara characteristic distributions, avoid that cumbersome, toxicity is big, efficiency is low, repeatable poor
Purification process.Largely, high-purity Oridonin is quickly prepared, there is higher technological innovation.One is provided for pharmaceuticals industry
The economic medical raw material of kind, the reference substance of high-purity is provided for Drug Administration department.Using Isodon Japonica Hara as raw material, there is provided
A kind of cheap new material for preparing Oridonin.
Oridonin prepared by the present invention use conventional method (i.e. existing pinostrobin activity test method) it is demonstrated experimentally that
Oridonin prepared by the present invention has：
1. Oridonin is to the specific obvious inhibitory action of mouse transplanted sarcoma S180 solid types tumour, to brain tumor
(B-22) there is certain inhibitory action；Tumor-bearing mice survival period can significantly be extended, and had no toxic side effect；
2. Oridonin has obvious inhibitory activity to lotus knurl liver ascites mouse, tumor-bearing mice can be obviously prolonged and deposited
Current.
3. Cyto toxic experiment showed shows, Oridonin has to HCT-116, HepG2, BGC-823, A2780 cell
Significant CDCC.
Compared with prior art, there are advantageous effects following prominent：
1st, one of novelty of the invention is to have found a kind of new raw material that Oridonin is prepared instead of Rabdosia rubescens.
2nd, Oridonin physics and chemistry of the characteristic distributions of quality and quantity and Oridonin in Isodon Japonica Hara can be combined
Matter and a kind of method that only can prepare high-purity Oridonin monomeric compound with 1 column chromatography developed, technique letter
Single, favorable reproducibility is easy to operate, is easy to industrialized production；
3rd, the quality control for Rabdosia rubescens medicinal material and using Rabdosia rubescens as the Chinese patent drug of raw material provide it is a kind of simple, can determine
The technology that reference substance is prepared to, favorable reproducibility ensures and condition；
4th, opened up for the development using Oridonin as the innovation new drug of raw material among a kind of simple, repeatable preparation
The technique of body.
Claims (4)
- A kind of 1. method that Oridonin is prepared from Isodon Japonica Hara, it is characterised in that comprise the following steps：1）The preparation of sample solution：Isodon Japonica Hara is dried medicinal material 1kg and crushed, and crosses 40 mesh pharmacopeia sieve, and every time plus mass concentration is 95% Ethanol 10L, extract 3 times, each 1.5h, merge No. 3 extract solutions, solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, depressurizes back Solvent is received to being dissipated without alcohol taste, moisture, 30 min are centrifuged under 3600 r/min, inclines and takes supernatant, insoluble matter adds 100ml volumes dense The ethanol dissolving for 15% is spent, 30 min are centrifuged under 3600 r/min, merges supernatant twice and sample solution is made；2）Resin pre-processes：Method is that after Sephadex LH-20 resins soak 24h with methanol, incline methanol solution, dense with volume Spend and soak 12h for 15% methanol, a solvent is replaced per 2h, wet method dress post, 2 posts are eluted with the methanol that volumetric concentration is 15% Volume, the Sephadex LH-20 resins that must have been pre-processed；3）Fill post and loading：By step 2）The resin wet method dress post pre-processed, in the chromatographic column that internal diameter is 5cm, make resin bed Blade diameter length ratio is up to 1 ︰ 15, then by step 1）In sample solution 1000ml natures flow velocity adsorbed by resin column it is complete；4）Isolate and purify：Resin column first elutes 1500ml with the methanol that volumetric concentration is 15%, then is eluted with 30% methanol 4000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, and obtain the thick medicinal extract that relative density is 1.243；5）Recrystallization：Above-mentioned thick medicinal extract 20ml methanol is dissolved, 24-72h is stood at -20 DEG C -35 DEG C, removes supernatant, White powder crystallization is separated out, filtering, obtains white crystals sprills, supernatant continues to place 24-72h, separates out white powder Crystallization, filtering, white crystals sprills are obtained, merge white crystals sprills twice, produce Oridonin 0.523g, used HPLC method area normalization methods calculate its purity more than 96.81%.
- A kind of 2. method that Oridonin is prepared from Isodon Japonica Hara, it is characterised in that comprise the following steps：1）The preparation of sample solution：Isodon Japonica Hara dries medicinal material 1kg, crushes, and crosses 60 mesh pharmacopeia sieve, and every time plus mass concentration is 95% ethanol 12L, extract 3 times, each 2h, merge No. 3 extract solutions, solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, depressurizes Recycling design adds moisture to dissipate, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 200ml volumes to without alcohol taste The ethanol that concentration is 20% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution is made；2）Resin pre-processes：Method is that Sephadex G resins soak 24h with methanol, and incline methanol solution, is soaked with 20% methanol 12h is steeped, a solvent is replaced per 2h, wet method dress post, 2 column volumes is eluted with the methanol that volumetric concentration is 20%, must pre-process Sephadex G resins；3）Fill post and loading：By step 2）The resin wet method dress post pre-processed, in the chromatographic column that internal diameter is 5cm, make resin bed Blade diameter length ratio is up to 1 ︰ 15, then by step 1）In sample solution 500mL natures flow velocity by resin column, absorption is complete；4）Isolate and purify：Resin column first elutes 1000ml with the methanol that volumetric concentration is 20%, then is eluted with 30% methanol 3000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, and obtain the thick medicinal extract that relative density is 1.321；5）Recrystallization：Above-mentioned thick medicinal extract ethanol is dissolved into 20 ml, 24-72h is stood at -20 DEG C -35 DEG C, removes supernatant, White powder crystallization is separated out, filtering, obtains white crystals sprills, supernatant continues to place 24-72h, separates out white powder Crystallization, filtering, obtains white crystals sprills, merges above-mentioned white crystals sprills, and Oridonin 0.5021g is made, adopts Its purity is calculated more than 95.12% with HPLC method area normalization methods.
- A kind of 3. method that Oridonin is prepared from Isodon Japonica Hara, it is characterised in that comprise the following steps：1）The preparation of sample solution：Isodon Japonica Hara dries medicinal material 1kg, crushes, and crosses 80 mesh pharmacopeia sieve, and every time plus mass concentration is 75% ethanol 12L, extract 3 times, each 2h, merge No. 3 extract solutions, solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, depressurizes Recycling design adds moisture to dissipate, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 800ml volumes to without alcohol taste The ethanol that concentration is 20% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution is made；2）Resin pre-processes：Method is that after MCI resins soak 24h with methanol, incline methanol solution, soaks 12h with 20% methanol, A solvent is replaced per 2h, wet method dress post, 2 column volumes, the MCI that must have been pre-processed are eluted with the methanol that volumetric concentration is 20% Resin；3）Fill post and loading：By step 2）The resin wet method dress post pre-processed, in the chromatographic column that internal diameter is 5cm, make resin bed Blade diameter length ratio is up to 1 ︰ 15, then by step 1）In sample solution 800ml natures flow velocity by resin column, absorption is complete；4）Isolate and purify：Resin column first elutes 2000ml with the methanol that volumetric concentration is 20%, then is eluted with 30% methanol 3000ml, collected volume concentration are the eluent of 30% methanol, and solvent is recovered under reduced pressure, obtains the thick medicinal extract of relative density 1.246；5）Recrystallization：Above-mentioned thick medicinal extract 20ml propyl alcohol is dissolved, 24-72h is stood at -20 DEG C -35 DEG C, removes supernatant, White powder crystallization is separated out, filtering, obtains white crystals sprills, supernatant continues to place 24-72h, separates out white powder Crystallization, filtering, white crystals sprills are obtained, merge white crystals sprills twice, that is, Oridonin 0.489g is made, used HPLC method area normalization methods calculate its purity more than 96.87%.
- A kind of 4. method that Oridonin is prepared from Isodon Japonica Hara, it is characterised in that comprise the following steps：1）The preparation of sample solution：Isodon Japonica Hara dries medicinal material 1kg, crushes, and crosses 80 mesh pharmacopeia sieve, and every time plus mass concentration is 50% ethanol 12L, extract 3 times, each 1.5h, merge No. 3 extract solutions, solvent is recovered under reduced pressure to without alcohol taste, adds moisture to dissipate, subtracts Recycling design is pressed to add moisture to dissipate to without alcohol taste, 30min is centrifuged under 3600r/min, incline and take supernatant, insoluble matter adds 500ml bodies The ethanol that product concentration is 20% is dissolved, and 30min is centrifuged under 3600r/min, merges supernatant twice and sample solution is made；2）Resin pre-processes：Method is that after Sephadex LH-20 resins soak 24h with methanol, incline methanol solution, with 20% Methanol soaks 12h, and a solvent is replaced per 2h, wet method dress post, 2 column volumes is eluted with the methanol that volumetric concentration is 20%, obtains pre- The Sephadex LH-20 resins handled well；3）Fill post and loading：By step 2）The resin wet method dress post pre-processed, in the chromatographic column that internal diameter is 5cm, make resin bed Blade diameter length ratio is up to 1 ︰ 10, then by step 1）In sample solution 1L natures flow velocity by resin column, absorption is complete；4）Isolate and purify：Resin column first elutes 1.9L with the methanol that volumetric concentration is 20%, then elutes 4000ml with 30% methanol, Collected volume concentration is the eluent of 30% methanol, and solvent is recovered under reduced pressure, and obtains the thick medicinal extract that relative density is 1.321；5）Recrystallization：Above-mentioned thick medicinal extract isopropanol is dissolved into 20ml, 24-72h is stood at -20 DEG C -35 DEG C, separates out white powder Last shape crystallization, filtering, white crystals sprills are obtained, supernatant continues to stand 24-72h, separates out white powder crystallization, mistake Filter, obtains white crystals sprills, merges above-mentioned white crystals sprills, produce Oridonin 0.549g, using HPLC methods Area normalization method calculates its purity more than 95.02%.
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| CN106631804B (en) * | 2016-12-21 | 2019-02-15 | 江苏省中医药研究院 | A kind of compound and the preparation method and application thereof isolated from Labiatae Rabdosia plant |
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