CN102389456A - Method for extracting isodon japonica var.galaucocalyx total diterpenoids or Glaucocalyxin A - Google Patents

Method for extracting isodon japonica var.galaucocalyx total diterpenoids or Glaucocalyxin A Download PDF

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CN102389456A
CN102389456A CN2011103871284A CN201110387128A CN102389456A CN 102389456 A CN102389456 A CN 102389456A CN 2011103871284 A CN2011103871284 A CN 2011103871284A CN 201110387128 A CN201110387128 A CN 201110387128A CN 102389456 A CN102389456 A CN 102389456A
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extracting
ethanol
japonica
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张健
姚士
沈晓丹
徐乃玉
褚纯隽
吴双庆
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Suzhou University
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Abstract

本发明属于中药提取领域，公开了一种制备蓝萼香茶菜总二萜或蓝萼甲素的方法：包括提取、洗脱、脱色步骤，将药材蓝萼香茶菜地上部分粉碎，在常压下，用乙醇水溶液回流提取1~3次，合并提取液，减压回收溶剂，得浓缩提取液；将所得浓缩提取液过已经处理好的大孔吸附树脂，50倍柱体积水洗脱色素，25倍柱体积30%乙醇洗脱杂质，30倍柱体积60%乙醇洗脱，收集此洗脱部分，回收溶剂，得到蓝萼香茶菜二萜提取物；将蓝萼香茶菜二萜提取物溶于乙醇，加入活性炭脱色，回收溶剂，干燥，得到蓝萼香茶菜总二萜，乙醇结晶，制备得到蓝萼甲素。采用本发明所述制备方法得到的蓝萼香茶菜总二萜含量高，蓝萼甲素纯度90%以上，得率7%以上。

Figure 201110387128

The invention belongs to the field of extraction of traditional Chinese medicines, and discloses a method for preparing total diterpenoids or indigosin: comprising steps of extraction, elution, and decolorization, and crushing the aboveground part of the medicinal material, Indigo indica, in normal Under pressure, reflux extraction with ethanol aqueous solution for 1 to 3 times, combine the extracts, and recover the solvent under reduced pressure to obtain a concentrated extract; pass the obtained concentrated extract through the treated macroporous adsorption resin, and elute the pigment with 50 times the column volume of water , 25 times of column volume 30% ethanol elutes impurities, 30 times of column volume 60% ethanol elutes, collects this elution part, reclaims solvent, obtains the diterpene extract of S. The extract is dissolved in ethanol, decolorized by adding activated carbon, the solvent is recovered, and dried to obtain the total diterpenes of Elephantia calyx, which is crystallized by ethanol to prepare Elephrine A. The total diterpene content of the orchid calyx that is obtained by adopting the preparation method of the invention is high, the purity of the orchid calyxin is over 90%, and the yield is over 7%.

Figure 201110387128

Description

A kind of method of extracting Rabdosia japonica total diterpene or glaucocalyxin
Technical field
The invention belongs to the Chinese medicine extraction field, relating to a kind of is the method for raw material preparing Rabdosia japonica total diterpene or glaucocalyxin with the medical material Rabdosia japonica.
Background technology
Rabdosia japonica ( Isodon japonicaVar glaucocalyx (Maxim) Hara) is Labiatae Rabdosia plant, extensively is distributed in the northeast of China, North China, areas such as East China; Have be good for the stomach, heat-clearing and toxic substances removing, invigorate blood circulation, the anti-inflammation isoreactivity, be used to treat gastritis, abdominal distention, hepatitis initial stage, cold, fever, mastitis, arthralgia etc.Rabdosia japonica mainly contains glaucocalyxin, Glaucocalyxin B, blue calyx third element, Glaucocalyxin D, blue calyx penta element, glaucocalyxin F, glaucocalyxin X; Deng mapping-Kaurane diterpine (referring to Zhao Bao Xiang; Et al. Two new diterpenoids from Rabdosia japonica var. glaucocalyx. Chinese Chemical Letters. 2008,19:852, Cao Luye etc.; The Rabdosia japonica progress; Yunnan Chinese medicine magazine, 2004,25 (3): 42.).Diterpenes composition in the Rabdosia japonica is its main active substances, and the rabbit platelet that glaucocalyxin can suppress calcimycin and stimulate generates platelet activating factor, and the rabbit platelet thromboxane A2 that can suppress arachidonic acid-induction generates; And the PGE2 that raises simultaneously, can significantly suppress ADP, AA; Inductive rabbit platelet such as PAF is assembled, and cAMP level in the remarkable increased platelets counts (referring to: Zhang B, Long K. Effects of glaucocalyxin A on aggtegation and cAMP levels of rabbit platelets in vitro. Acta Pharmacologica Sinica 1993; 14 (4): 347.) glaucocalyxin has certain inhibitory action to ehrlich carcinoma, in the body S180 solid tumor suppression ratio is reached 30.4% (10mg/kg, 7 days); (referring to: Zhang Shuxiang etc.; Glaucocalyxin external to DNA, the biosynthetic influence of RNA and protein, pharmacy Intelligence Newsletter; 1990,8 (3): 238.).Glaucocalyxin, Glaucocalyxin B, Glaucocalyxin D, blue calyx penta element, the own element of blue calyx have stronger cytotoxic activity to A549 (human lung carcinoma cell), LOVO (people's colon-cancer cell), 6T-CEM (human T cell leukemia cell), HL-60 (human leukemia cell); Can be used for preparing cancer therapy drug (referring to Chen Haisheng etc.; The application of diterpene-kind compound in the preparation cancer therapy drug in the Rabdosia japonica, the patent No. is 200910044882.0 Chinese invention patent); Glaucocalyxin B, blue calyx third element, Glaucocalyxin D, blue calyx penta element and the own element of blue calyx have carried out anti-hepatitis B surface antigen and HBcAg test; The result finds; They have significant anti-hepatitis B activity; Can be used for preparing the medicine (referring to Chen Haisheng etc., the application of diterpene-kind compound in the preparation anti-hepatitis virus medicament in the Rabdosia japonica, the patent No. is 200910044883.5 Chinese invention patent) of anti-hepatitis virus.
In the prior art, the method for from the medical material Rabdosia japonica, extracting glaucocalyxin is complex steps not only, and yield is on the low side; Generally be lower than 10%; And because employing has toxic solvent to organism, residual toxic solvent does not meet the pharmacopeia requirement, specifically may further comprise the steps:
Figure 2011103871284100002DEST_PATH_IMAGE002
gathers medical material Rabdosia japonica aerial parts and pulverizes; Under normal pressure; With percentage by volume 85% ~ 95% ethanol water reflux, extract, 2 ~ 3 times; Each reflux, extract, 2 hours; Merge extractive liquid,, decompression and solvent recovery, extremely every milliliter of solvent contains crude drug 3 ~ 10g;
Figure 2011103871284100002DEST_PATH_IMAGE004
is with isopyknic petroleum ether extraction three times; Petroleum ether layer discards; Water layer reuse equal-volume chloroform extraction three times; The combined chloroform layer also concentrates, and obtains chloroform extraction liquid;
Figure 2011103871284100002DEST_PATH_IMAGE006
adds silica gel and stirs in step
Figure 984982DEST_PATH_IMAGE004
gained chloroform extraction liquid; Drying under reduced pressure carries out silica gel column chromatography; With chloroform: methanol 200:1 or chloroform: methanol 100:1 eluting, with the thin layer chromatography contrast, collect the eluent that contains glaucocalyxin, eluent is reclaimed solvent;
Figure 2011103871284100002DEST_PATH_IMAGE008
is with dehydrated alcohol or methanol repeated crystallization; Get glaucocalyxin; Test purity is calculated yield.
The purity that adopts technique scheme gained glaucocalyxin is generally more than 90%; But yield is generally 7% ~ 9%, and has used deleterious solvent in the process, chloroform, methanol; And residual toxic solvent can influence the use of glaucocalyxin as medicine, does not meet the pharmacopeia requirement.
Publication number is the method for distilling that the Chinese invention patent application of CN 101914002 discloses a kind of glaucocalyxin, and extraction, silica gel column chromatography eluting pigment and impurity, purification step specifically may further comprise the steps:
Figure 528221DEST_PATH_IMAGE002
gathers crude drug Rabdosia japonica aerial parts and pulverizes; Under normal pressure; With percentage by volume 75 ~ 80% ethanol water reflux, extract, 2 ~ 3 times, each reflux, extract, 2 ~ 3 hours, merge extractive liquid; Partial solvent is removed in decompression, gets concentrated extracting solution;
Figure 370275DEST_PATH_IMAGE004
adds silica gel and stirs in step
Figure 827801DEST_PATH_IMAGE002
gained concentrated extracting solution; Drying under reduced pressure; Carry out silica gel column chromatography: with the silicagel column volume is 1 parts by volume; At first with the eluent A eluting fat-soluble pigment of 2 ~ 3 parts of parts by volume; Again with the eluent B eluting impurity of 4 ~ 6 parts of parts by volume, at last with the eluent C eluting of 10 ~ 15 parts of parts by volume; Collect eluent, concentrate the back and place, separate out crystal;
Wherein, said eluent A is the mixture of petroleum ether and ethyl acetate, and by volume, Shi You Mi ︰ ethyl acetate=8 ︰ 1; Said eluent B is the mixture of petroleum ether and ethyl acetate, and by volume, Shi You Mi ︰ ethyl acetate=4 ︰ 1; Said eluent C is the mixture of petroleum ether and ethyl acetate, and by volume, Shi You Mi ︰ ethyl acetate=2 ︰ 1;
Figure 474464DEST_PATH_IMAGE006
dehydrated alcohol recrystallization gets the glaucocalyxin crystal.
The purity of the glaucocalyxin that obtains through said method is 95 ~ 100%, and purity is high, is suitable for preparing various dosage forms; Yield is more than 10%, and yield is high, and the repeatability of method is strong; Measure according to organic solvent residual assay method in 2010 editions pharmacopeia appendix; Organic solvent-free is residual, and the organic solvent avirulence that is suitable for, and meets 2010 editions pharmacopeia requirements.
Macroporous adsorbent resin be one type not with the porous cross linked polymer of ion-exchange group, have the polymeric sorbent of macroporous structure.Physicochemical properties are stable, and specific surface area is big, and adsorption capacity is big, and selectivity is good; Adsorption rate is fast, and desorption condition is gentle, and is good to the Organic substance selectivity, the influence that not existed by inorganic salts and other strong ions, low molecule; Help absorption, wide in variety, the adsorbable multiple organic compound of different cultivars; Fluid resistance is less, and decoloring ability is high, and Regeneration Treatment is convenient, life cycle is long, be suitable for formation closed cycle, cost saving; Also can reduce the moisture absorption of product simultaneously, effectively remove heavy metal, solve the difficult problem of Chinese medicine heavy metals exceeding standard.Be mainly used in the separation and purification of Chinese herbal medicine at present, as to the medical material The pretreatment, to the separation and purification of single medicinal material extract and the separation and purification of centering recurrence due to taking drug formula extraction, the realization suitability for industrialized production that has has vast potential for future development.(referring to: Liu Shuaiying etc., the purification with macroreticular resin of peoniflorin research in the Radix Paeoniae Rubra, Chinese herbal medicine, 2010,41 (9): 1480.)
Silica gel is not re-usable column packing, Chemical Decomposition purification glaucocalyxin, and the silica gel consumption is big, and cost is higher, and macroporous resin is reusable column chromatography filler, and can suitability for industrialized production, can reduce cost greatly.
Glaucocalyxin and diterpene have significant pharmacologically active in the Rabdosia japonica, but do not see that the macroporous resin of bibliographical information glaucocalyxin and total diterpene method for preparing is arranged, and are necessary to carry out preparation technology's investigation.
Summary of the invention
Goal of the invention of the present invention provides a kind of method of extracting the Rabdosia japonica total diterpene.
For reaching the foregoing invention purpose, the technical scheme that the present invention adopts is: a kind of method of extracting the Rabdosia japonica total diterpene, comprise extraction, eluting, decolouring step, wherein,
Said extraction step is: medical material Rabdosia japonica aerial parts is pulverized, used the ethanol water reflux, extract,, obtain extracting solution, decompression and solvent recovery gets concentrated extracting solution, contains crude drug 0.5-1g in said every milliliter of concentrated extracting solution;
Said elution step is: the gained concentrated extracting solution is crossed macroporous adsorbent resin; With 50 times of column volume water elution pigments; 25 times of column volume 30% ethanol water eluting impurity, 30 times of column volume 60% ethanol water eluting are collected this eluting part; Reclaim solvent, obtain the Rabdosia japonica diterpene extract.
Said decolouring step is: the Rabdosia japonica diterpene extract is dissolved in the ethanol, adds activated carbon decolorizing then, reclaim solvent, drying obtains the Rabdosia japonica total diterpene.
In the technique scheme, the decolouring step is in order to remove pigment such as chlorophyll, to improve yield.
In the technique scheme, use the method for ethanol water reflux, extract, to be: under normal pressure, with percentage by volume 55-70% ethanol water reflux, extract, 1 ~ 3 time, reflux, extract, is 1 ~ 3 hour at every turn, merge extractive liquid.
In the technique scheme; Said macroporous adsorbent resin is the macroporous adsorbent resin of nonpolar or low pole; Comprise AB-8 type, D101 type, HPD-100, HPD-200A, HPD-200B, HPD-300, HPD-700, HPD-722 type, handle by prior art before use.
In the technique scheme, in the elution step, the macroporous resin quality: quality of medicinal material is 1: 1.0-2.7.
In the technique scheme, in the elution step, the macroporous resin column diameter: the post height is 1: 4-9.
In the technique scheme, in the elution step, the macroporous resin quality: column volume is 1mg: 1mL.
In the technique scheme, in the decolouring step, the quality of Rabdosia japonica diterpene extract and alcoholic acid volume ratio are 1: 250-300.
In the technique scheme, in the decolouring step, the quality of active carbon is 1～5% of a Rabdosia japonica diterpene extract quality.
Further in the technical scheme, get above-mentioned Rabdosia japonica total diterpene, alcohol crystal prepares glaucocalyxin, and purity is more than 90%, and yield is more than 7%.
Therefore, the present invention requires a kind of method of extracting glaucocalyxin of protection simultaneously again.
The glaucocalyxin and the total diterpene that adopt method for preparing according to the invention to prepare can be used to process dosage forms such as tablet, capsule, pill, are applied to compound recipe or single preparations of ephedrine.
Because the technique scheme utilization, the present invention compared with prior art has advantage:
The Rabdosia japonica total diterpene content that adopts method for preparing according to the invention to obtain is high, and glaucocalyxin purity is more than 90%, and yield is more than 7%, and this technological operation is simple, and is pollution-free, but suitability for industrialized production.
Description of drawings
Fig. 1 is the purity analysis chromatogram of glaucocalyxin among the embodiment one;
Fig. 2 is the purity analysis chromatogram of glaucocalyxin among the embodiment two;
Fig. 3 is the purity analysis chromatogram of glaucocalyxin among the embodiment three;
Fig. 4 is the hydrogen analysis of spectrum figure of glaucocalyxin among the embodiment five;
Fig. 5 is the carbon analysis of spectrum figure of glaucocalyxin among the embodiment five.
The specific embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is further described:
Embodiment one:
Get Rabdosia japonica aerial parts 100 gram, through pulverizing, in the atmospheric pressure reflux extraction element, with volume fraction 55% ethanol water reflux, extract, 2 times, reflux, extract, is 2 hours at every turn, merge extractive liquid,, and decompression and solvent recovery is to 200 milliliters; Through 80 gram HPD-100 type macroporous adsorbent resins (macroporous resin column is high 13 centimetres for 80 milliliters of column volumes, 2.8 centimetres of macroporous resin column diameters); With 50 times of water elution pigments; 25 times of volume fraction 30% ethanol water eluting impurity, 30 times of 60% volume fraction ethanol water eluting collected this eluting part; Reclaim solvent, obtain Rabdosia japonica diterpene extract 1.65 grams.The Rabdosia japonica diterpene extract is dissolved in 430 milliliters of ethanol, adds 0.08 gram activated carbon decolorizing element, drying to this solution; Obtain Rabdosia japonica total diterpene 1.08 grams, 41.3%, 2 milliliter of alcohol crystal of content; Get glaucocalyxin 0.02 gram, its purity is 95.6%, yield 8%.
In the technique scheme, the glaucocalyxin method for detecting purity is: 2 milligrams of glaucocalyxin, and dissolve with methanol standardize solution to 10 milliliter, chromatographic column is Sweden Kromasil C18 (4.6 mm * 250 mm, 5 μ m); Mobile phase: methanol: 0.1% phosphoric acid (70:30); Flow velocity: 1 mL/min; Detect wavelength: 231 nm; Sample size: 20 μ L; Column temperature: 30 ℃, detect gained figure shown in liquid chromatograph Fig. 1, data see the following form:
Peak (Peak) Retention time Area Highly Area % Tailing factor
1 2.726 114372 7462 2.098 1.165
2 3.613 105597 4615 1.937 0.959
3 5.127 5209313 423492 95.573 1.048
4 8.247 21349 1323 0.392 0.735
Summation ? 5450631 436892 100.000 ?
Embodiment two:
Get Rabdosia japonica aerial parts 1000 gram, through pulverizing, in the atmospheric pressure reflux extraction element, with 60% alcohol reflux 2 times; Each reflux, extract, 3 hours, merge extractive liquid,, decompression and solvent recovery is to 1500 milliliters; Through 600 gram D101 type macroporous adsorbent resins (macroporous resin column is high 26 centimetres for 600 milliliters of column volumes, 5.5 centimetres of macroporous resin column diameters); With 50 times of water elution pigments, 25 times of 30% ethanol elution impurity, 30 times of 60% ethanol elution; Collect this eluting part, reclaim solvent, obtain Rabdosia japonica diterpene extract 16.72 grams.The Rabdosia japonica diterpene extract is dissolved in 4190 milliliters of ethanol, adds 0.60 gram activated carbon decolorizing element, drying to this solution; Obtain Rabdosia japonica total diterpene 10.16 grams, 42.0%, 35 milliliter of alcohol crystal of content; Get glaucocalyxin 0.25 gram, its purity is 95.0%, yield 9%.
In the technique scheme, the glaucocalyxin method for detecting purity is: 2 milligrams of glaucocalyxin, and dissolve with methanol standardize solution to 10 milliliter, chromatographic column is Sweden Kromasil C18 (4.6 mm * 250 mm, 5 μ m); Mobile phase: methanol: 0.1% phosphoric acid (70:30); Flow velocity: 1 mL/min; Detect wavelength: 231 nm; Sample size: 20 μ L; Column temperature: 30 ℃, detect gained figure shown in liquid chromatograph Fig. 2, data see the following form:
Peak (Peak) Retention time Area Highly Area % Tailing factor
1 2.688 137605 9202 3.026 1.425
2 3.526 76711 4157 1.687 1.028
3 4.988 4320503 389744 95.018 1.128
4 6.616 12219 1312 0.269 1.062
Summation ? 4547037 404415 100.000 ?
Embodiment three:
Get Rabdosia japonica aerial parts 10000 gram, through pulverizing, in the atmospheric pressure reflux extraction element, with 70% alcohol reflux 2 times; Each reflux, extract, 2 hours, merge extractive liquid,, decompression and solvent recovery is to 16000 milliliters; Through 6000 gram AB-8 type macroporous adsorbent resins (macroporous resin column is high 77 centimetres for 6000 milliliters of column volumes, 10 centimetres of macroporous resin column diameters); With 50 times of water elution pigments, 25 times of 30% ethanol elution impurity, 30 times of 60% ethanol elution; Collect this eluting part, reclaim solvent, obtain Rabdosia japonica diterpene extract 170.20 grams.The Rabdosia japonica diterpene extract is dissolved in 4190 milliliters of ethanol, adds 5.98 gram activated carbon decolorizings elements, drying to this solution; Obtain Rabdosia japonica total diterpene 121.83 grams, 43.2%, 50 milliliter of alcohol crystal of content; Get glaucocalyxin 2.48 grams, its purity is 96.0%, yield 8%.
In the technique scheme, the glaucocalyxin method for detecting purity is: 2 milligrams of glaucocalyxin, and dissolve with methanol standardize solution to 10 milliliter, chromatographic column is Sweden Kromasil C18 (4.6 mm * 250 mm, 5 μ m); Mobile phase: methanol: 0.1% phosphoric acid (70:30); Flow velocity: 1 mL/min; Detect wavelength: 231 nm; Sample size: 20 μ L; Column temperature: 30 ℃, detect gained figure shown in liquid chromatograph Fig. 3, data see the following form:
Peak (Peak) Retention time Area Highly Area % Tailing factor
1 2.648 92322 5999 1.810 1.159
2 3.440 97539 4940 1.913 1.022
3 4.833 4897540 415590 96.035 1.040
4 6.404 12360 1286 0.242 1.018
Summation ? 5099761 427815 100.000 ?
Embodiment four: diterpene content in the Rabdosia japonica
Get purity and be 6 parts of 96.6% glaucocalyxin, dissolve with methanol, standardize solution are to 25ml, and 230nm surveys absorbance as absorbing wavelength.With the glaucocalyxin quality is abscissa, and absorbance is a vertical coordinate, tries to achieve regression equation Y=0.001X+1.9856, R 2=0.9997 explanation is linear good, and glaucocalyxin is good in 4.376-21.996 mg scope internal linear relation.
Get Rabdosia japonica total diterpene 0.25 gram respectively, dissolve with methanol, standardize solution to 250 milliliter under the 230nm wavelength, is surveyed absorbance, does parallel sample three times, calculates the average content of Rabdosia japonica total diterpene.(list of references: the assay of total diterpene in the Rabdosia rubescens, but treasure etc., the time precious traditional Chinese medical science traditional Chinese medicines, 2010,21 (4): 828-829.)
Sequence number Rabdosia japonica total diterpene quality (gram) Rabdosia japonica total diterpene average content %
1 1.08 41.3
2 10.16 42.0
3 121.83 43.2
Embodiment five:
The glaucocalyxin structure is identified: colourless block crystallization (ethanol), mp 220.0-222.0 ℃; Be soluble in organic solvents such as chloroform, ethyl acetate, acetone, methanol.
Simultaneously embodiment gained glaucocalyxin is carried out hydrogen spectrum and carbon analysis of spectrum, the result is referring to Fig. 4,5, and concrete data are as follows:
1H?NMR?(400MHz,?DMSO- d 6)?δ：6.00?and?5.46?(each?1H,?brs,?17-H a/b)，4.75?(1H,brs,?H-14α)，4.09?(1H,?m,?7β-H)，3.00?(1H,?brs,?13α-H)，1.08?(3H,?s)，1.04(3H,?s)，1.02?(3H,?s)。
13C?NMR?(100MHz,?DMSO- d 6)?δ：37.9?(1-C)，33.9?(2-C)，216.5?(3-C)，46.5?(4-C)，50.6?(5-C)，30.9?(6-C)，73.0?(7-C)，60.7?(8-C)，52.8?(9-C)，38.6?(10-C)，18.1?(11-C)，30.3?(12-C)，46.1?(13-C)，74.6?(14-C)，207.0?(15-C)，148.8?(16-C)，116.9?(17-C)，27.4?(18-C)，21.1?(19-C)，18.1?(20-C)。POP data and document (Kim Yong Il etc., Rabdosia japonica root chemical constitution study, CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2000,25 (11): 38-39.) unanimity is accredited as glaucocalyxin.

Claims (10)

1.一种提取蓝萼香茶菜总二萜的方法，包括提取、洗脱、脱色步骤，其特征在于： 1. A method for extracting the total diterpenes of S. japonica, comprising steps of extraction, elution, and decolorization, characterized in that: 所述提取步骤为：将药材蓝萼香茶菜地上部分粉碎，用乙醇水溶液回流提取，得到提取液，减压回收溶剂，得浓缩提取液，每毫升浓缩提取液中含原药材0.5-1g； The extraction step is as follows: pulverize the above-ground part of the medicinal material S. japonica, reflux and extract with an aqueous ethanol solution to obtain an extract, recover the solvent under reduced pressure to obtain a concentrated extract, and each milliliter of the concentrated extract contains 0.5-1g of the original medicinal material; 所述洗脱步骤为：将所得浓缩提取液过大孔吸附树脂，以50倍柱体积水洗脱色素，25倍柱体积的浓度为体积百分数30%乙醇水溶液洗脱杂质，30倍柱体积的浓度为体积百分数60%乙醇水溶液洗脱，收集此洗脱部分，回收溶剂，得到蓝萼香茶菜二萜提取物； The elution step is as follows: pass the obtained concentrated extract through a macroporous adsorption resin, elute the pigment with 50 times the column volume of water, 25 times the column volume concentration is 30% ethanol aqueous solution to elute impurities, and 30 times the column volume The concentration is 60% by volume of ethanol aqueous solution for elution, the eluted part is collected, and the solvent is recovered to obtain the diterpene extract of S. japonica; 所述脱色步骤为：将蓝萼香茶菜二萜提取物溶于乙醇中，然后加入活性炭脱色，回收溶剂，干燥，得到蓝萼香茶菜总二萜。 The decolorization step is as follows: dissolving the diterpene extract of S. japonica in ethanol, adding active carbon for decolorization, recovering the solvent, and drying to obtain the total diterpenes of S. japonica. 2.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，用乙醇水溶液回流提取的方法为：在常压下，用体积百分数55-70%乙醇水溶液回流提取1~3次，每次回流提取1~3小时，合并提取液。 2. according to claim 1, extract the method for the total diterpenoids of Indigo saponica, it is characterized in that, the method for reflux extraction with ethanol aqueous solution is: under normal pressure, with volume percentage 55-70% ethanol aqueous solution reflux extraction 1 ~3 times, reflux extraction for 1~3 hours each time, and combine the extracts. 3.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，所述大孔吸附树脂为非极性或弱极性的大孔吸附树脂。 3. the method for extracting the total diterpenes of Indigo sativa according to claim 1, characterized in that, the macroporous adsorption resin is a non-polar or weakly polar macroporous adsorption resin. 4.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，所述大孔吸附树脂包括AB-8型、D101型、HPD-100、HPD-200A、HPD-200B、HPD-300、HPD-700、HPD-722型。 4. according to the method for extracting the total diterpenoids of Indigo sativa according to claim 1, it is characterized in that, described macroporous adsorption resin comprises AB-8 type, D101 type, HPD-100, HPD-200A, HPD-200B , HPD-300, HPD-700, HPD-722. 5.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，洗脱步骤中，大孔树脂质量∶药材质量为1∶1.0-2.7。 5. The method for extracting the total diterpenes of S. japonica according to claim 1, characterized in that, in the elution step, the mass of the macroporous resin: the mass of medicinal materials is 1: 1.0-2.7. 6.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，洗脱步骤中，大孔树脂柱直径∶柱高为1∶4-9。 6. The method for extracting the total diterpenes of Indigo sativa according to claim 1, characterized in that, in the elution step, the macroporous resin column diameter:column height is 1:4-9. 7.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，洗脱步骤中，大孔树脂质量∶柱体积为1mg∶1mL。 7. The method for extracting the total diterpenes of Indigo sativa according to claim 1, characterized in that, in the eluting step, the mass of the macroporous resin: column volume is 1 mg: 1 mL. 8.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，脱色步骤中，蓝萼香茶菜二萜提取物的质量和乙醇的体积比为1∶250-300。 8. The method for extracting the total diterpenes of S. japonica according to claim 1, characterized in that, in the decolorization step, the volume ratio of the quality of the diterpene extract of S. japonica to ethanol is 1: 250-300 . 9.根据权利要求1所述提取蓝萼香茶菜总二萜的方法，其特征在于，脱色步骤中，活性炭的质量为蓝萼香茶菜二萜提取物质量的1～5%。 9. The method for extracting the total diterpenes of Indigo japonica according to claim 1, characterized in that, in the decolorization step, the quality of the activated carbon is 1% to 5% of the mass of the diterpene extract of Indigo japonica. 10.一种提取蓝萼甲素的方法，其特征在于，采用权利要求1所述提取蓝萼香茶菜总二萜的方法制备得到蓝萼香茶菜总二萜，乙醇结晶，制备得到蓝萼甲素。 10. A method for extracting indigosin, characterized in that, the method for extracting the total diterpenes of indigo-scented tea according to claim 1 is used to prepare the total diterpenes in indigo-scented tea, crystallized with ethanol, and prepare indigo calyxin.
CN2011103871284A 2011-11-29 2011-11-29 Method for extracting isodon japonica var.galaucocalyx total diterpenoids or Glaucocalyxin A Pending CN102389456A (en)

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