CN101961358A - Preparation method of hypericum perforatum extractive - Google Patents
Preparation method of hypericum perforatum extractive Download PDFInfo
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- CN101961358A CN101961358A CN 201010506239 CN201010506239A CN101961358A CN 101961358 A CN101961358 A CN 101961358A CN 201010506239 CN201010506239 CN 201010506239 CN 201010506239 A CN201010506239 A CN 201010506239A CN 101961358 A CN101961358 A CN 101961358A
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- herba hyperici
- hyperici perforati
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Abstract
The invention discloses a preparation method of hypericum perforatum extractive which is rich in hyperforin, which comprises the following steps of: taking the hypericum perforatum crude extractive which is sold in the market as the raw material; ultrasonically extracting in the ice water bath; preliminarily removing the impurity by means of two-phase extraction and the like to obtain 11-18% hyperforin-containing extractive; and separating and purifying by means of the silica column chromatography technology to obtain 37-57% hyperforin-containing extractive. The pharmacological test proves that the extractive has the functions of the depression resistant and the learning and memorizing promotion. The preparation method is simple and easy, is suitable for the small-scale preparation required by the pharmacological test and the like, and can save the extracting time and space.
Description
Technical field
The present invention relates to the extracting method field of effective ingredients in plant, relate in particular to the method that the Herba Hyperici perforati essence extract of hyperforine is rich in the commercially available Herba Hyperici perforati crude extract preparation of a kind of usefulness.
Background technology
Herba Hyperici perforati contains the number of chemical composition, mainly contains naphthalene a pair of horses going side by side dianthrone class, phloroglucinol class, flavonoid, volatile oil, amino acids, phenylpropyl alcohol alkanes and xanthone class etc.Wherein naphthalene a pair of horses going side by side dianthrone class, phloroglucinol class and flavonoid are considered to the main active of Herba Hyperici perforati.The phloroglucinol class mainly comprises hyperforine (hyperforin) and Adhyperforin. (adhyperforin), and hyperforine is a low-polarity component, and is extremely unstable to light, heat and oxygen.
Known hyperforine extracting method mainly contains extraction and co_2 supercritical fluid extraction method.Extraction is the method that utilization is the most extensive at present, cost is minimum, but its extraction time is longer, and extraction efficiency is low.The co_2 supercritical fluid extraction method is comparatively advanced at present extracting method, the product purity that obtains is also higher, with Herba Hyperici perforati with after the co_2 supercritical fluid extraction, reuse methanol and normal hexane extract repeatedly, the extract that obtains contains 70% hyperforine, but the instrument cost of CO 2 supercritical stream extraction is too high, and output is lower, does not also popularize at home.
The extracting method of traditional hyperforine generally is to pulverize dry crude drug, uses the organic solvent lixiviate.The Chinese invention patent that publication number is respectively CN1198097 and CN1251999 discloses the stable extract that contains hyperforine that German Dr. Willmar Schwabe Gmbh ﹠ Co. invents, the stabilizing agent that also adds q.s with aqueous methanol, aquiferous ethanol, normal heptane as the extraction solvent in the preparation, as organic mercaptide, ascorbic acid and derivant thereof, its Herba Hyperici Monogyni cellulose content 0.1%～2% or more than, can be steady in a long-term.Publication number is that the Chinese invention patent of CN1392130A discloses the method for extracting the purification hypericum perforatum component in the plants such as Herba Hyperici perforati of people such as Yang Depo invention, and purity can reach more than 95%.
Summary of the invention
The invention provides a kind of commercially available Herba Hyperici perforati crude extract that contains hyperforine 3～4% that utilizes, the method for the Herba Hyperici perforati essence extract of hyperforine is rich in the preparation of ultrasound wave auxiliary extraction at low temperatures.
A kind of preparation method of Herba Hyperici perforati essence extract may further comprise the steps:
A) get the Herba Hyperici perforati crude extract, mix, in ice-water bath, carry out ultrasonic extraction 30～60min, make extracting solution with ethyl acetate; The volumetric usage of described ethyl acetate (ml) is 4～6 times of Herba Hyperici perforati crude extract quality (g).
Described Herba Hyperici perforati crude extract refers to general commercially available Herba Hyperici perforati crude extract, and according to international quality control standard, wherein the content of hypericin is more than 0.3%.According to mensuration, wherein the Herba Hyperici Monogyni cellulose content is 3～4%.
B) the prepared extracting solution of step a) is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, after being 8～14 with the inorganic base adjust pH, sucking filtration, get filtrate, transfer pH to 0～2, with non-polar solven extraction 3～5 times with mineral acid, merge the non-polar solven phase, reclaim under reduced pressure gets extractum, 0～20 ℃ of vacuum drying, and the Herba Hyperici perforati extract that makes keeps in Dark Place at-20 ℃.Described inorganic base is KOH, NaOH or NaHCO
3, described mineral acid is HCl, H
2SO
4Or H
3PO
4Described non-polar solven is the low boiling petroleum ether, and the boiling point of described low boiling petroleum ether is at 30～60 ℃.
C) with b) step gained Herba Hyperici perforati extract is dissolved in and carries out silica gel column chromatography in the eluant, the wet method upper prop, to obtain the Herba Hyperici Monogyni cellulose content be 37～57% Herba Hyperici perforati essence extract through concentrating.Described eluant is one or more the mixture in petroleum ether, normal hexane, alcohol, ester and the organic acid.Described silica gel is that column chromatography silica gel H or thin layer are analysed silica gel H.
The ultrasonic extraction condition optimization is in the step a): with consumption is the ethyl acetate of 5ml/g Herba Hyperici perforati crude extract lucifuge, airtight extraction 60 minutes in ice-water bath.
As preferably, transfer pH=8 with NaOH earlier in the step b), reuse 20%HCl transfers pH=2.
As preferably, c) in each component of silica gel column chromatography eluant and volume ratio thereof be petroleum ether: ethyl acetate: acetic acid=80: 19: 1, more preferably petroleum ether: ethyl acetate=10: 1.
The ultimate principle of ultrasonic wave extraction is the judder that utilizes ultrasound wave to produce, high acceleration, intensive cavitation effect, stirring action etc., quicken ingredient and enter solvent, thereby raising extraction rate, shorten extraction time, compare with soxhlet extraction and can remove high temperature from extracting the influence of composition.Compare with conventional extraction, ultrasonic wave extraction have extraction time short, productive rate is high, need not advantages such as heating, in this field, have a good application prospect.
Detecting through high performance liquid chromatography, adopt step a), the b of the inventive method) the Herba Hyperici Monogyni cellulose content is 11～18% in the Herba Hyperici perforati extract for preparing, this extractum is stable at least between 1 month storage life, does not have obviously minimizing.Prove that through pharmacological evaluation this Herba Hyperici perforati extract not only has significant antidepressant effect, and can promote the animal learning memory ability.This Herba Hyperici perforati extract obtains being rich in the Herba Hyperici perforati essence extract of hyperforine again through silica gel column chromatography, and wherein the content of hyperforine can reach 57%.
The inventive method is applicable to the small lot preparation that pharmacological testing is required, and is simpler and easy, can save extraction time and space.
Description of drawings
Fig. 1 is the process chart of preparation method of the present invention.
The specific embodiment
The invention will be further described by the following examples.
Embodiment 1
Get Herba Hyperici perforati crude extract 50g, mix, in ice-water bath, carry out ultrasonic extraction 60min with the 250ml ethyl acetate.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, sucking filtration, get filtrate, transfer pH=2 with 20%HCl, use 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, 0～20 ℃ of vacuum drying 2 hours.The said extracted thing is dissolved in a small amount of eluant carries out silica gel column chromatography, eluant is a petroleum ether: ethyl acetate: acetic acid=80: 19: 1, an amount of pressurization, collect liquid through concentrating the Herba Hyperici perforati essence extract that obtains being rich in hyperforine, detect with HPLC, wherein the content of hyperforine is 57%.
Embodiment 2
Get Herba Hyperici perforati crude extract 50g, mix, in ice-water bath, carry out ultrasonic extraction 30min with the 250ml ethyl acetate.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, get filtrate after low-temperature and high-speed is centrifugal, transfer pH=2 with 20%HCl, with 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, 0～20 ℃ of vacuum drying 2 hours.The said extracted thing is dissolved in a small amount of eluant carries out silica gel column chromatography, eluant is a petroleum ether: ethyl acetate=10: 1, and an amount of pressurization is collected liquid through concentrating the essence extract that obtains being rich in hyperforine, after testing, wherein the content of hyperforine is 37%.
Embodiment 3
Get Herba Hyperici perforati crude extract 50g, with 250ml ethyl acetate lixiviate 24 hours.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, sucking filtration, get filtrate, transfer pH=2 with 20%HCl, use 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, 0～20 ℃ of vacuum drying 2 hours.The said extracted thing is dissolved in a small amount of eluant carries out silica gel column chromatography, eluant is a petroleum ether: ethyl acetate: acetic acid=80: 19: 1, and an amount of pressurization is collected liquid and is concentrated the essence extract that obtains being rich in hyperforine, after testing, wherein the content of hyperforine is 36%.
Embodiment 4
Get Herba Hyperici perforati crude extract 50g, mix, in ice-water bath, carry out ultrasonic extraction 45min with the 250ml ethyl acetate.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, transfer pH=2 with 20%HCl, with 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, and 0～20 ℃ of vacuum drying 2 hours obtains the Herba Hyperici Monogyni cellulose content and be 18% extract.
Embodiment 5
Get Herba Hyperici perforati crude extract 50g, add stabilizing agent, mix, in ice-water bath, carry out ultrasonic extraction 45min with the 250ml ethyl acetate.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, transfer pH=2 with 20%HCl, with 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, and 0～20 ℃ of vacuum drying 2 hours obtains the Herba Hyperici Monogyni cellulose content and be 18% extract.
Embodiment 6
Get Herba Hyperici perforati crude extract 50g, add stabilizing agent, mix, in ice-water bath, carry out ultrasonic extraction 45min with 250ml ethanol.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, transfer pH=2 with 20%HCl, with 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, and 0～20 ℃ of vacuum drying 2 hours obtains the Herba Hyperici Monogyni cellulose content and be 11% extract.
Embodiment 7
Get Herba Hyperici perforati crude extract 50g, add stabilizing agent, mix, in ice-water bath, carry out ultrasonic extraction 45min with 250ml acetone.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, transfer pH=2 with 20%HCl, with 150ml low boiling petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, and 0～20 ℃ of vacuum drying 2 hours obtains the Herba Hyperici Monogyni cellulose content and be 14% extract.
Embodiment 8
Get Herba Hyperici perforati crude extract 50g, mix, in ice-water bath, carry out ultrasonic extraction 45min with the 250ml ethyl acetate.Above-mentioned gained extracting solution is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, transfer pH=8 with NaOH, transfer pH=2 with 20%HCl, with 150ml common petroleum ether extraction 3～5 times, merge the petroleum ether phase, reclaim under reduced pressure gets extractum, 0～20 ℃ of vacuum drying 2 hours, obtain the Herba Hyperici Monogyni cellulose content and be 16% extract, petroleum ether has obviously residual.
Claims (10)
1. the preparation method of a Herba Hyperici perforati essence extract is characterized in that: may further comprise the steps:
A) get commercially available Herba Hyperici perforati crude extract, add ethyl acetate and mix, under ice-water bath, carried out ultrasonic extraction 30～60 minutes, make extracting solution; The consumption of described ethyl acetate is 4～6ml/g Herba Hyperici perforati crude extract;
B) extracting solution that step a) is made is condensed into the concentrated solution that contains Herba Hyperici perforati crude extract 0.5～5g/ml, after being 8～14 with the inorganic base adjust pH, sucking filtration, get filtrate, transferring pH with mineral acid is 0～2, with non-polar solven extraction 3～5 times, merge the non-polar solven phase, reclaim under reduced pressure gets extractum, 0～20 ℃ of vacuum drying, and the Herba Hyperici perforati extract that makes keeps in Dark Place at-20 ℃;
C) gained Herba Hyperici perforati extract in the step b) is dissolved in eluant and carries out silica gel column chromatography, the wet method upper prop concentrates the Herba Hyperici perforati essence extract that obtains being rich in hyperforine.
2. the preparation method of Herba Hyperici perforati essence extract according to claim 1, it is characterized in that: the ultrasonic extraction condition is in the step a): with ethyl acetate lucifuge, airtight extraction 30～60 minutes in ice-water bath, the consumption of described ethyl acetate is 4～6ml/g Herba Hyperici perforati crude extract.
3. the preparation method of Herba Hyperici perforati essence extract according to claim 1 is characterized in that: the inorganic base described in the step b) is KOH, NaOH or NaHCO
3
4. the preparation method of Herba Hyperici perforati essence extract according to claim 1 is characterized in that: the mineral acid described in the step b) is HCl, H
2SO
4Or H
3PO
4
5. the preparation method of Herba Hyperici perforati essence extract according to claim 1 is characterized in that: transferring pH with NaOH earlier in the step b) is 8～14, and it is 0～2 that reuse hydrochloric acid is transferred pH.
6. the preparation method of Herba Hyperici perforati essence extract according to claim 1 is characterized in that: the non-polar solven described in the step b) is the low boiling petroleum ether.
7. the preparation method of Herba Hyperici perforati essence extract according to claim 1 is characterized in that: the silica gel described in the step c) is that column chromatography silica gel H or thin layer are analysed silica gel H.
8. the preparation method of Herba Hyperici perforati essence extract according to claim 1 is characterized in that: the eluant described in the step c) is one or more the mixture in petroleum ether, normal hexane, alcohol, ester and the organic acid.
9. the preparation method of Herba Hyperici perforati essence extract according to claim 8 is characterized in that: each component and volume ratio thereof are petroleum ether in the eluant described in the step c): ethyl acetate: acetic acid=80: 19: 1.
10. the preparation method of Herba Hyperici perforati essence extract according to claim 8 is characterized in that: each component and volume ratio thereof are petroleum ether in the eluant described in the step c): ethyl acetate=10: 1.
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| CN101961358B CN101961358B (en) | 2012-07-04 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103288614A (en) * | 2013-06-27 | 2013-09-11 | 中国科学院昆明植物研究所 | Monocyclic phloroglucinol compounds with antineoplastic activity and pharmaceutical composition thereof |
| CN105693494A (en) * | 2016-02-27 | 2016-06-22 | 蚌埠医学院 | Process for extracting and purifying hypericin from hypericum perforatum |
| CN105777522A (en) * | 2014-12-18 | 2016-07-20 | 六安裕发农业科技有限公司 | Method for extracting hypericin from Hypericum perforatum |
| CN115872848A (en) * | 2022-12-27 | 2023-03-31 | 陕西嘉禾药业有限公司 | Method for preparing hypericin qualified in polycyclic aromatic hydrocarbon by using hypericum perforatum |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1215033C (en) * | 2002-07-18 | 2005-08-17 | 中山大学 | Process for preparing hypericum perforatum extract |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1215033C (en) * | 2002-07-18 | 2005-08-17 | 中山大学 | Process for preparing hypericum perforatum extract |
Non-Patent Citations (1)
| Title |
|---|
| 《中国优秀博硕士学位论文全文数据库（硕士）工程科技Ⅰ辑》 20050315 张筱 贯叶金丝桃素的分离纯化研究 B016-517 1-10 , 第1期 2 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103288614A (en) * | 2013-06-27 | 2013-09-11 | 中国科学院昆明植物研究所 | Monocyclic phloroglucinol compounds with antineoplastic activity and pharmaceutical composition thereof |
| CN105777522A (en) * | 2014-12-18 | 2016-07-20 | 六安裕发农业科技有限公司 | Method for extracting hypericin from Hypericum perforatum |
| CN105777522B (en) * | 2014-12-18 | 2017-12-26 | 安徽农达生物技术有限公司 | A kind of method that hypericin is extracted from hypericum perforatum |
| CN105693494A (en) * | 2016-02-27 | 2016-06-22 | 蚌埠医学院 | Process for extracting and purifying hypericin from hypericum perforatum |
| CN115872848A (en) * | 2022-12-27 | 2023-03-31 | 陕西嘉禾药业有限公司 | Method for preparing hypericin qualified in polycyclic aromatic hydrocarbon by using hypericum perforatum |
| CN115872848B (en) * | 2022-12-27 | 2024-03-26 | 陕西嘉禾药业有限公司 | Method for preparing hypericin qualified by polycyclic aromatic hydrocarbon by using Hypericum perforatum |
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| CN101961358B (en) | 2012-07-04 |
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