CN101265282A - Method for separating catalpol from fresh rehmanniae root - Google Patents
Method for separating catalpol from fresh rehmanniae root Download PDFInfo
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- CN101265282A CN101265282A CNA2008100160058A CN200810016005A CN101265282A CN 101265282 A CN101265282 A CN 101265282A CN A2008100160058 A CNA2008100160058 A CN A2008100160058A CN 200810016005 A CN200810016005 A CN 200810016005A CN 101265282 A CN101265282 A CN 101265282A
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Abstract
The invention discloses a method for separating catalpol from fresh rehmannia root, which includes pulverizing and homogenizing fresh rehmannia root to obtain catalpol-containing sample solution; separating macroporous absorption resin; and performing chromatography with a silica gel column or re-crystallizing. The method maximally separates the catalpol from saccharide component in the rehmannia root to clear away obstacles for the preparation of high purity catalpol. The method has high production capability of the high purity catalpol of up to 100 kg catalpol/year, energy saving effect, environmental friendliness, and low production cost, and provides raw material guarantee for research, drug development, and health product development.
Description
Technical field
The present invention relates to a kind of method of separating Catalpol, relate in particular to a kind of by the method for separating Catalpol in the Rehmannia Root.
Background technology
Glutinous rehmannia is a kind of large Chinese medicinal materials commonly used, and about 20000 tons/year of demand exports about 1000 tons/year.Because the difference of drug effect, it can be used as different medicinal materials and is used as medicine, and is typically to concoct and medicinal material that the property of medicine changes through processing; It is divided into Rehmannia Root, Radix Rehmanniae and Radix Rehmanniae Preparata.The Rehmannia Root clearing heat and promoting fluid, cooling blood for hemostasis is used for consumption of YIN caused by febrile disease, the deep red polydipsia of tongue; The Radix Rehmanniae clearing heat and cooling blood, nourishing Yin and promoting production of body fluid is used for the deep red polydipsia of tongue, interior-heat caused by deficiency of YIN; The Radix Rehmanniae Preparata nourishing YIN and supplementing blood, beneficial essence is filled out marrow, is used for the hepatic and renal YIN deficiency, and the deficiency of blood is sallow.
Catalpol belongs to iridoid glucose glycoside; modern pharmacology experimental results show that; effects such as that Catalpol has is anticancer, neuroprotective, hypoglycemic, laxative, diuresis, anti-inflammatory, spasmolysis; be one of effective constituent of glutinous rehmannia; be 2005 editions " quality control indexs of Chinese pharmacopoeia Radix Rehmanniae; Catalpol content in Rehmannia Root the highest (about 1%), be processed into Radix Rehmanniae after the Catalpol amount decline to a great extent, concoct almost all degradeds of Catalpol behind the ripe glutinous rehmannia.
For the separation of Catalpol with produce, existing report focuses mostly on aspect stripping Catalpol degree from glutinous rehmannia, comprises solvent for use, extraction time, and factors such as extracting method, as: Li Gengsheng etc.
[1]With the glutinous rehmannia chopping, water reflux 1h uses n-butanol extraction again, supplies examination with dissolve with methanol behind the recovery propyl carbinol; Permanent green grass or young crops etc. all
[2]Glutinous rehmannia is cut into small pieces, extracts 1h with methanol eddy; Wang Hongjie etc.
[3]With grinding powder after the Rehmannia Root lyophilize (crossing 40 mesh sieves), with methyl alcohol supersound extraction 20min; Limit Po Lam etc.
[4]With Hao Wuchang etc.
[5]All, cross 80 mesh sieves, with methyl alcohol room temperature lixiviate 2h with the glutinous rehmannia crushed after being dried; Wang Cheng is far away etc.
[6]Glutinous rehmannia is cut into small pieces, repeatedly with water logging, supersound extraction; Luo Yanyan etc.
[7]Glutinous rehmannia is cut into small pieces, with 20% alcohol immersion 15min, homogenate, homogenate is jolting frequently, soaks 2h; Zhao Xinfeng etc.
[8]With scissors the glutinous rehmannia sample is shredded, with alcohol heating reflux 0.5h.
Luo Yanyan etc.
[7]Investigated the influence of extraction solvent and extraction time, respectively with methyl alcohol, water, 15% ethanol, 20% ethanol, 30% ethanol and 50% extraction using alcohol 1,2,4,6,8,26h, high effective liquid chromatography for measuring catalpol content.Determine with 20% ethanol lixiviate 2h to be the best.
Zhang Ling etc.
[9]Investigated the extraction process of glutinous rehmannia with the dual-wavelength lamellar scanning method, compared the catalpol content that water is carried 1h, 2h, 3h, 4h, 5h and 95% extraction using alcohol 2h, 4h, determined the highlyest with water extraction 1h-2h catalpol content.
Liu Ming etc.
[10]With the Catalpol is to detect index, adopt high performance liquid chromatography, investigated the influence of factors such as extraction solvent, extracting method, extraction time and solvent consumption with 4 factors, 2 horizontal quadrature test design, determined that optimum process condition is to extract 2 times with 3 times of amount methanol eddies, return time is 2h.
Li Jiping etc.
[11]Investigated water, methyl alcohol and 10%, 20%, 50%, 70% and 95% ethanol as solvent, found that catalpol content is more or less the same in methyl alcohol, 20% ethanol and 50% ethanol extract; Same glutinous rehmannia sample is handled with backflow 1h, ultrasonic 1h, the method for soaking 1h respectively, and catalpol content is obviously difference not; Same sample ultrasonic processing 10,20,30,40,60min, catalpol content does not have notable difference yet.
Song Zirong etc.
[12]With HPLC is content assaying method, adopts repetition-orthogonal experiment method, with alcohol concn, quantity of solvent, extraction time and 4 factors of extraction time, each factor is chosen 3 levels and is experimentized, preferably determine with 8 times of amounts of 60% ethanol, refluxing extraction 2 times, each 1.5h is the preferable extraction conditions of Catalpol
Zhang Hanzhong
[13]Deng adopting preferably flavol lixiviate taking technique of orthogonal test, serve as to investigate index with pure medicinal extract yield, Catalpol is qualitative for referencial use, determines optimum extraction process.Flavol lixiviate taking technique extracts 3 times with 4 times of amount 60% ethanol definitely, and each 6d is an optimum extraction process.
The method that prepare at present in a small amount (milligram level or gram grade) high purity (content is higher than 98%) mostly is laboratory method (as with conventional plant separation means---zwitterion resin etc.; technology such as low pressure chromatography); complex steps; preparation amount is little, or uses the preparative high performance liquid chromatography instrument, but the cost height; waste time and energy; preparation amount is little, all is not suitable for industrial scale production, as: Wang Chengyuan etc.
[14]Separate Catalpol in the Radix Rehmanniae with the nonpolar macroporous adsorption resin combined utilization with natural clarifying agent.It is glutinous rehmannia pulverizing medicinal materials, alcohol immersion that Wang Huisen etc. adopt repeatedly silica gel column chromatography to separate to obtain Catalpol, step, concentrate, D101 macroporous adsorbent resin, silica gel column chromatography etc.Li Gengsheng etc.
[15]Adopt the low pressure column chromatography method and separate and to obtain Catalpol, but need enrichment repeatedly and be milligram grade.By retrieval, still not having producer or unit at present provides feather weight purity to be higher than 98% Catalpol report.
Reference:
[1] Li Gengsheng, Liu Changhe, Wang Huisen, etc. catalpol content compares [J] in the glutinous rehmannia in the different places of production. herbal medicine, 2002,33 (2): 126.
[2] all permanent blue or green, Li Zhaoxi, Liu Gencheng, etc. the quality approach of glutinous rehmannia [J]. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1992,17 (6): 327.
[3] Wang Hongjie, the limit Po Lam, Yang Jian, etc. the discussion of Catalpol change condition [J] in the glutinous rehmannia. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1997,22 (7): 408.
[4] limit Po Lam, Yang Jian, Wang Hongjie, etc. different drying conditionss are to the influence [J] of catalpol content in the Rehmannia Root. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1996,21 (6): 346.
[5] Hao Wuchang, Zhu Yuhong, Zhu Zhifeng. concoct influence [J] to catalpol content in the glutinous rehmannia. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1997,22 (6): 345.
[6] Wang Chengyuan, Zhang Hao, Meng Li waits .HPLC to measure the content [J] of Catalpol in glutinous rehmannia and the preparation thereof. West China pharmaceutical journal, 2003,18 (2): 134.
[7] Luo Yanyan, Zhang Shaoqing, Suo Jianzheng, etc. the content of Catalpol [J] in the high effective liquid chromatography for measuring glutinous rehmannia. Chinese Pharmaceutical Journal, 1994,29 (1): 38.
[8] Zhao Xinfeng, Sun Yuqing. the capillary zone electrophoresis method is measured the content [J] of Catalpol in the glutinous rehmannia. pharmaceutical analysis magazine, 2002,22 (6): 471.
[9] Zhang Ling, Xu Xingang, time delay increases, etc. glutinous rehmannia Study on extraction [J]. herbal medicine, 1998,29 (5): 308
[10] Liu Ming, Li Gengsheng. Catalpol extraction process [J] in the Rehmannia Root. the time precious traditional Chinese medical science traditional Chinese medicines, 2000,11 (4): 301
[11] Li Jiping, Ma Hua, Wang Yuesheng, etc. the comparison [J] of Catalpol, carbohydrate content content in Rehmannia Root and the Radix Rehmanniae. Chinese Pharmaceutical Journal, 2001,36 (5): 300.
[12] Song Zirong, Tan Zijun, Chen Xisong, white hypo. the optimization of glutinous rehmannia extraction process, Chinese experimental pharmacology of traditional Chinese medical formulae magazine [J], 2006,12:8.
[13] Zhang Hanzhong, Zhang Hanzhen, Wu Yang. orthogonal test is flavol lixiviate taking technique [J] preferably. Chinese Hospitals pharmaceutical journal, 2001,21 (11): 678.
[14] Wang Chengyuan, Zhang Hao, Zhang Chengping, Meng Li. the purifies and separates technology [J] of iridoids in the Radix Rehmanniae. medical Leader, 2003,22 (10): 707.
[15] Li Gengsheng, Wang Huisen, all permanent blue or green, etc. liquid chromatography separates preparation glutinous rehmannia blood sugar reducing component Catalpol [J]. traditional Chinese medical science research, 1997,10 (3): 24.
Summary of the invention
At the deficiencies in the prior art, the problem to be solved in the present invention provides the method that a kind of a large amount of separation prepares Catalpol, and the inventive method process is simple and direct, and equipment is simple, and environmental protection and energy saving are fit to commercial scale production.
Separate the method for Catalpol in the Rehmannia Root of the present invention, realize by following steps:
(1) gets Rehmannia Root, discard mildew and rot position, clean, be cut into the section that thickness is 1mm～2cm;
(2) immediately the Rehmannia Root section is heated to 40 ℃～100 ℃, insulation 1min～120min; Or directly the water heat of steam is handled 1min～120min; Then with 3000rpm～8000rpm rotating speed homogenate 5min～20min;
Perhaps, immediately with Rehmannia Root section with its etc. 40 ℃～100 ℃ hot water of weight mix, jointly with 3000rpm～8000rpm rotating speed homogenate 5min～20min;
(3) conventional filtration homogenate is removed not broken Rehmannia Root tissue and insoluble impurities, must contain the sample liquid of Catalpol;
(4) get macroporous adsorbent resin wet method dress post, the sample liquid that contains Catalpol is carried out macroporous adsorbent resin separate, the weight of sample solution and macroporous adsorbent resin volume ratio are 10g/L～10kg/L; Earlier with water elution, check with molish reaction bonded TLC, to the molish reaction negative, till Catalpol occurred simultaneously, changing eluting solvent was ethanol or methyl alcohol, continued to be eluted to no Catalpol and only detected, each part elutriant of collecting is detected with TLC respectively, observe the position at Catalpol place, merge the elutriant that contains Catalpol, volatilization crystallization or revolve steam the Catalpol crude product;
(5) get 95% dissolve with ethanol of Catalpol crude product with 1～3 times of its volume, filtration under diminished pressure, remove insoluble impurity and get sample liquid, get column chromatography then and adorn post, sample on the wet method with the silica gel wet method, chloroform-methanol=9: 2 wash-outs, with the Catalpol standard control, TLC detects, and merges the component that contains Catalpol of collecting, volatilize solvent, getting purity is 98% above Catalpol;
Perhaps, get the Catalpol crude product and add 40 ℃～60 ℃ hot methanols, splash into chloroform after treating to dissolve fully and end, put more than 20 hours to crystal just having occurred with 1～2 times of its volume, decompress filter, 60 ℃ of dryings of product, repeat this step 2～5 times the pure product of Catalpol.
Separate in the method for Catalpol in the above-mentioned Rehmannia Root: the described Rehmannia Root of step (1) is preferably peeled before section, removes Rehmannia Root middle part white and light yellow chrysanthemum core part (see figure 1) in addition as far as possible.
Separate in the method for Catalpol in the above-mentioned Rehmannia Root: the described method of step (2) is preferably carried out under oxygen free condition, and wherein Heating temperature is preferred 70 ℃～100 ℃, insulation 30min～90min; Preferred 5000rpm～the 6000rpm of homogenate rotating speed.
Separate in the method for Catalpol in the above-mentioned Rehmannia Root: the described homogenate of step (3) can add the activated carbon adsorption homogenate of its weight 0.1%～10% if color is darker, and solution colour is shoaled.
Separate in the above-mentioned Rehmannia Root in the method for Catalpol: the described macroporous adsorbent resin of step (4) is one or several balanced mix in HPD-600, HPD-500, HPD-300, the HPD-100 model preferably; Series such as corresponding resin model such as D101 resin.
Separate in the above-mentioned Rehmannia Root in the method for Catalpol: the macroporous adsorbent resin after the described sample separation of step (4) is that organic solvent wash-out more than 20% is regenerated with concentration of volume percent preferably.
Wherein: described organic solvent is ethanol or methyl alcohol preferably.
Because macroporous adsorbent resin needs to use repeatedly, so macroporous adsorbent resin needs aftertreatment carrying out the separation of next batch, the regenerate solvent of required wash-out of macroporous adsorbent resin is a content greater than similar organic solvents such as 10% ethanol, methyl alcohol.This technology utilization apparatus,Soxhlet's principle can make solvent cycle use, and stream is washed resin repeatedly, has saved solvent, has alleviated workload.Concrete device is seen Fig. 2, and right figure is integral construction figure, and left side figure is local.
Separate in the above-mentioned Rehmannia Root in the method for Catalpol: described eluting solvent ethanol of step (4) or methyl alcohol preferably concentration of volume percent are ethanol or methyl alcohol more than 20%.
Separate in the method for Catalpol in the above-mentioned Rehmannia Root: the described TLC in step (4) or (5) detects and adopts tlc silica gel G, chloroform: methyl alcohol=launch at 1: 1, the colour developing of aubepine sulfuric acid.
Separate in the above-mentioned Rehmannia Root in the method for Catalpol: the described column chromatography of step (5) is with silica gel 60 orders～100 purpose silica gel preferably.
The present invention is at state natural sciences fund (fund number: 30270101) finish under subsidizing, because of having beta-glucosidase in the Rehmannia Root cell, the gordian technique of the inventive method is that thereby the enzyme that decomposes Catalpol is carried out deactivation guarantees that the Catalpol that itself contains in the Rehmannia Root is not destroyed into, and then carry out later separation, and from Radix Rehmanniae and Radix Rehmanniae Preparata, extract Catalpol, because of itself catalpol content is low, cause the preparation cost height.
The inventive method can (account for more than 13% of fresh weight with carbohydrate part in Catalpol and the glutinous rehmannia greatly, be more than ten times of catalpol content, thereby) separate and remove the obstacles for the preparation of high-purity Catalpol, the inventive method prepares high-purity (content is greater than 98%), and the Catalpol ability can reach 100kg/, is expected to the pilot scale amplification and carries out suitability for industrialized production, yearly capacity reaches 10 tons, the production process energy-conserving and environment-protective, production cost declines to a great extent, and can be scientific research, new drug development, healthcare products are researched and developed the assurance of supplying raw materials.
Description of drawings
Fig. 1: Rehmannia Root transverse section figure, show Rehmannia Root middle part white and light yellow chrysanthemum core.
Fig. 2: macroporous adsorbent resin organic solvent wash-out regenerating unit figure, wherein: right figure is integral construction figure, left side figure is local.
Embodiment
Embodiment 1:
1) the broken homogenate of Rehmannia Root
Get Rehmannia Root, discard mildew and rot position, clean, peel (removing Rehmannia Root middle part white and light yellow chrysanthemum core part in addition) is cut into the section that thickness is 1mm～2cm;
Under oxygen free condition, immediately the Rehmannia Root section is heated to 80 ℃～100 ℃, insulation 70min～80min; Or directly the water heat of steam is handled 60min～80min; Then with 6000rpm rotating speed homogenate 10min;
Perhaps, immediately with Rehmannia Root section with its etc. 100 ℃ hot water of weight mix, jointly with 5000rpm rotating speed homogenate 20min;
The conventional filtration homogenate is removed not broken Rehmannia Root tissue and insoluble impurities, must contain the sample liquid of Catalpol.
2) macroporous adsorbent resin separates
Get the about 5kg wet method dress of macroporous adsorbent resin HPD-100 post, the about 5000mL of column volume.Sample solution 1000ml, elder generation check with molish reaction bonded TLC that with water elution to the molish reaction negative, till Catalpol occurred simultaneously, changing eluting solvent was 20% ethanol, continued to be eluted to no Catalpol and only detected.
Each part elutriant of collecting is detected (tlc silica gel G, chloroform: methyl alcohol=launch at 1: 1, the colour developing of aubepine sulfuric acid) with TLC respectively, with the Catalpol standard substance is contrast, observe the position at Catalpol place, merge the elutriant that contains Catalpol, volatilization crystallization or revolve steam the Catalpol crude product;
The macroporous adsorbent resin that will finish to the sample liquid separation that contains Catalpol makes resin regeneration with 95% ethanol elution.
3) silica gel column chromatography
Get the about 20g of Catalpol crude product and dissolve with 95% ethanol 50ml, filtration under diminished pressure is removed insoluble impurity.Get the 100g column chromatography and mix sample with silica gel (60-100 order).Get column chromatography silica gel 2.0kg, sample on the wet method dress post wet method, chloroform-methanol=9: 2 wash-outs.With the Catalpol standard control, TLC detects, and merges the component that contains Catalpol, volatilizes solvent, and getting purity is 98% above Catalpol.
Perhaps 4) recrystallization
Get the Catalpol crude product and add 50 ℃～60 ℃ hot methanols, splash into chloroform after treating to dissolve fully and end, put more than 20 hours to crystal just having occurred with 1.5 times of its volumes, decompress filter, 60 ℃ of dryings of product, repeat this step 3 times the pure product of Catalpol.
Catalpol extraction substep yield sees the following form in the Rehmannia Root:
Embodiment 2:
1) the broken homogenate of Rehmannia Root
Get Rehmannia Root, discard mildew and rot position, clean, peel (removing Rehmannia Root middle part white and light yellow chrysanthemum core part in addition) is cut into the section that thickness is 1mm～2cm;
Immediately the Rehmannia Root section is heated to 40 ℃～50 ℃, insulation 100min～120min; Or directly the water heat of steam is handled 90min; Then with 5000rpm rotating speed homogenate 15min;
Perhaps, immediately with Rehmannia Root section with its etc. 60 ℃ hot water of weight mix, jointly with 5000rpm rotating speed homogenate 20min;
The conventional filtration homogenate is removed not broken Rehmannia Root tissue and insoluble impurities, must contain the sample liquid of Catalpol.
2) macroporous adsorbent resin separates
Get the about 5kg wet method dress of macroporous adsorbent resin HPD-100 post, the about 5000mL of column volume.Sample solution 1000ml, elder generation check with molish reaction bonded TLC that with water elution to the molish reaction negative, till Catalpol occurred simultaneously, changing eluting solvent was 30% ethanol, continued to be eluted to no Catalpol and only detected.
Each part elutriant of collecting is detected (tlc silica gel G, chloroform: methyl alcohol=launch at 1: 1, the colour developing of aubepine sulfuric acid) with TLC respectively, with the Catalpol standard substance is contrast, observe the position at Catalpol place, merge the elutriant that contains Catalpol, volatilization crystallization or revolve steam the Catalpol crude product;
The macroporous adsorbent resin that will finish to the sample liquid separation that contains Catalpol makes resin regeneration with 95% ethanol elution.
3) silica gel column chromatography
Get the about 20g of Catalpol crude product and dissolve with 95% ethanol 50ml, filtration under diminished pressure is removed insoluble impurity.Get the 100g column chromatography and mix sample with silica gel (60-100 order).Get column chromatography silica gel 2.0kg, sample on the wet method dress post wet method, chloroform-methanol=9: 2 wash-outs.With the Catalpol standard control, TLC detects, and merges the component that contains Catalpol, volatilizes solvent, and getting purity is 98% above Catalpol.
Perhaps 4) recrystallization
Get the Catalpol crude product and add 40 ℃ of hot methanols, splash into chloroform after treating to dissolve fully and end, put more than 20 hours to crystal just having occurred with 1 times of its volume, decompress filter, 60 ℃ of dryings of product, repeat this step 5 times the pure product of Catalpol.
Embodiment 3:
1) the broken homogenate of Rehmannia Root
Get Rehmannia Root, discard mildew and rot position, clean, peel (removing Rehmannia Root middle part white and light yellow chrysanthemum core part in addition) is cut into the section that thickness is 1mm～2cm;
Immediately the Rehmannia Root section is heated to 100 ℃, insulation 20min; Or directly the water heat of steam is handled 30min; Then with 8000rpm rotating speed homogenate 5min;
Perhaps, immediately with Rehmannia Root section with its etc. 80 ℃ hot water of weight mix, jointly with 6000rpm rotating speed homogenate 20min;
The conventional filtration homogenate is removed not broken Rehmannia Root tissue and insoluble impurities, must contain the sample liquid of Catalpol.
2) macroporous adsorbent resin separates
Get the about 5kg wet method dress of macroporous adsorbent resin HPD-100 post, the about 5000mL of column volume.Sample solution 1000ml, elder generation check with molish reaction bonded TLC that with water elution to the molish reaction negative, till Catalpol occurred simultaneously, changing eluting solvent was 30% ethanol, continued to be eluted to no Catalpol and only detected.
Each part elutriant of collecting is detected (tlc silica gel G, chloroform: methyl alcohol=launch at 1: 1, the colour developing of aubepine sulfuric acid) with TLC respectively, with the Catalpol standard substance is contrast, observe the position at Catalpol place, merge the elutriant that contains Catalpol, volatilization crystallization or revolve steam the Catalpol crude product;
The macroporous adsorbent resin that will finish to the sample liquid separation that contains Catalpol makes resin regeneration with 95% ethanol elution.
3) silica gel column chromatography
Get the about 20g of Catalpol crude product and dissolve with 95% ethanol 50ml, filtration under diminished pressure is removed insoluble impurity.Get the 100g column chromatography and mix sample with silica gel (60-100 order).Get column chromatography silica gel 2.0kg, sample on the wet method dress post wet method, chloroform-methanol=9: 2 wash-outs.With the Catalpol standard control, TLC detects, and merges the component that contains Catalpol, volatilizes solvent, and getting purity is 98% above Catalpol.
Perhaps 4) recrystallization
Get the Catalpol crude product and add 60 ℃ of hot methanols, splash into chloroform after treating to dissolve fully and end, put more than 20 hours to crystal just having occurred with 1 times of its volume, decompress filter, 60 ℃ of dryings of product, repeat this step 4 times the pure product of Catalpol.
Claims (10)
1. separate the method for Catalpol in the Rehmannia Root, realize by following steps:
(1) gets Rehmannia Root, discard mildew and rot position, clean, be cut into the section that thickness is 1mm～2cm;
(2) immediately the Rehmannia Root section is heated to 40 ℃～100 ℃, insulation 1min～120min; Or directly the water heat of steam is handled 1min～120min; Then with 3000rpm～8000rpm rotating speed homogenate 5min～20min;
Perhaps, immediately with Rehmannia Root section with its etc. 40 ℃～100 ℃ hot water of weight mix, jointly with 3000rpm～8000rpm rotating speed homogenate 5min～20min;
(3) conventional filtration homogenate is removed not broken Rehmannia Root tissue and insoluble impurities, must contain the sample liquid of Catalpol;
(4) get macroporous adsorbent resin wet method dress post, the sample liquid that contains Catalpol is carried out macroporous adsorbent resin separate, the weight of sample solution and macroporous adsorbent resin volume ratio are 10g/L～10kg/L; Earlier with water elution, check with molish reaction bonded TLC, to the molish reaction negative, till Catalpol occurred simultaneously, changing eluting solvent was ethanol or methyl alcohol, continued to be eluted to no Catalpol and only detected, each part elutriant of collecting is detected with TLC respectively, observe the position at Catalpol place, merge the elutriant that contains Catalpol, volatilization crystallization or revolve steam the Catalpol crude product;
(5) get 95% dissolve with ethanol of Catalpol crude product with 1～3 times of its volume, filtration under diminished pressure, remove insoluble impurity and get sample liquid, get column chromatography then and adorn post, sample on the wet method with the silica gel wet method, chloroform-methanol=9: 2 wash-outs, with the Catalpol standard control, TLC detects, and merges the component that contains Catalpol of collecting, volatilize solvent, getting purity is 98% above Catalpol;
Perhaps, get the Catalpol crude product and add 40 ℃～60 ℃ hot methanols, splash into chloroform after treating to dissolve fully and end, put more than 20 hours to crystal just having occurred with 1～2 times of its volume, decompress filter, 60 ℃ of dryings of product, repeat this step 2～5 times the pure product of Catalpol.
2. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the described Rehmannia Root of step (1) is peeled before section, removes Rehmannia Root middle part white and light yellow chrysanthemum core part in addition.
3. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the described method of step (2) is carried out under oxygen free condition.
4. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the described homogenate of step (3) adds the activated carbon adsorption homogenate of its weight 0.01%～10% if color is darker, and solution colour is shoaled.
5. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the described macroporous adsorbent resin of step (4) is one or several balanced mix in HPD-600, HPD-500, HPD-300, HPD-100 model or the corresponding model.
6. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the macroporous adsorbent resin concentration of volume percent after the described sample separation of step (4) is that the organic solvent wash-out more than 20% is regenerated.
7. as separating the method for Catalpol in the Rehmannia Root as described in the claim 6, it is characterized in that: described organic solvent is ethanol or methyl alcohol.
8. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: described eluting solvent ethanol of step (4) or methyl alcohol are that concentration of volume percent is ethanol or the methyl alcohol more than 20%.
9. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the described TLC in step (4) or (5) detects and adopts tlc silica gel G, chloroform: methyl alcohol=launch at 1: 1, the colour developing of aubepine sulfuric acid.
10. separate the method for Catalpol according to claim 1 in the Rehmannia Root, it is characterized in that: the described column chromatography silica gel of step (5) is 60 orders～300 purpose silica gel.
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| CN102023194A (en) * | 2010-10-13 | 2011-04-20 | 河北科星药业有限公司 | Method for judging quality of radix rehmanniae |
| CN102408461A (en) * | 2011-09-16 | 2012-04-11 | 河南省中医药研究院 | Method for extracting and purifying catalpol by use of active carbon column chromatography and crystallization technology |
| CN102863489A (en) * | 2011-07-04 | 2013-01-09 | 苏州玉森新药开发有限公司 | Method for extracting catalpol from rehmannia stem |
| CN103623118A (en) * | 2013-12-05 | 2014-03-12 | 云南昆船设计研究院 | Rehmannia glutinosa libosch processing method |
| CN106220695A (en) * | 2016-07-21 | 2016-12-14 | 西安岳达生物科技股份有限公司 | A kind of industrial production process of catalpol |
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Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100457765C (en) * | 2007-01-16 | 2009-02-04 | 广东太阳神集团有限公司 | Method for producing stabhyose, and method for producing stabhyose and catalpol by using rehmannia root |
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| CN102023194A (en) * | 2010-10-13 | 2011-04-20 | 河北科星药业有限公司 | Method for judging quality of radix rehmanniae |
| CN102863489A (en) * | 2011-07-04 | 2013-01-09 | 苏州玉森新药开发有限公司 | Method for extracting catalpol from rehmannia stem |
| CN102408461A (en) * | 2011-09-16 | 2012-04-11 | 河南省中医药研究院 | Method for extracting and purifying catalpol by use of active carbon column chromatography and crystallization technology |
| CN103623118A (en) * | 2013-12-05 | 2014-03-12 | 云南昆船设计研究院 | Rehmannia glutinosa libosch processing method |
| CN106220695A (en) * | 2016-07-21 | 2016-12-14 | 西安岳达生物科技股份有限公司 | A kind of industrial production process of catalpol |
| CN108034681A (en) * | 2017-12-08 | 2018-05-15 | 新乡医学院 | Utilize the method for the stem of Radix Codonopsis lanceolatae forming layer stem cell production Catalpol for the culture that suspends |
| CN108034681B (en) * | 2017-12-08 | 2021-08-10 | 新乡医学院 | Method for producing catalpol by using suspension culture rehmannia stem cambium stem cells |
| CN111892665A (en) * | 2020-08-11 | 2020-11-06 | 西安天美生物科技股份有限公司 | Method for separating high-content catalpol and high-content rehmannia polysaccharide from rehmannia |
| CN113480581A (en) * | 2021-07-21 | 2021-10-08 | 湖南朗林生物资源股份有限公司 | Method for extracting iridoid glycoside from rehmannia |
| CN113480581B (en) * | 2021-07-21 | 2022-05-24 | 湖南朗林生物资源股份有限公司 | Method for extracting iridoid glycoside from rehmannia |
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