CN102973783A - Lily rehmanniae decoction anti-depression effective part and preparation method and application thereof - Google Patents
Lily rehmanniae decoction anti-depression effective part and preparation method and application thereof Download PDFInfo
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- CN102973783A CN102973783A CN2012104520699A CN201210452069A CN102973783A CN 102973783 A CN102973783 A CN 102973783A CN 2012104520699 A CN2012104520699 A CN 2012104520699A CN 201210452069 A CN201210452069 A CN 201210452069A CN 102973783 A CN102973783 A CN 102973783A
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Abstract
The invention discloses a lily rehmanniae decoction anti-depression effective part and a preparation method and application thereof. The preparation method thereof comprises the following steps: (1) mixing medical materials of lily and rehmanniae and smashing the lily and the rehmanniae to obtain medical material powder, (2) adding water or alcohol liquid water with alcohol more than 0 and less than or equal to 95 percent, extracting for at least two times in a reflux mode, combining extracted solution, reducing pressure to concentrate the solution into concrete, adding alcohol sediment with the alcohol weight being 2 to 5 times of the concrete to the concrete, filtering to obtain supernate and concentrating the supernate into the concentrated solution, (3) separating the concentrated solution through macroporous absorption resin column chromatography, firstly using the water to elute the concentrated solution and remove impurities, then using the alcohol liquid water with the alcohol of 10 percent to 95 percent to elute the concentrated solution in a gradient mode, respectively combining the eluted solution of each gradient, concentrating, reducing pressure, drying and correspondingly obtaining the effective part of an eluted part with the alcohol liquid water of 10 percent to 95 percent. The method is simple to operate, environment-friendly, low in cost and suitable for industrialized production. The effective part prepared through the method has good anti-depression effects and can be used in preparation of anti-depression medicine.
Description
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to a kind of lilii and Rehmanniae Decoction antidepressant effective site and its preparation method and application.
Background technology
Depression (depression, DEP) is a kind of able-bodied affective disorder mental sickness that has a strong impact on.Day by day fierce with the quickening of rhythm of life and social competition, depression has become the commonly encountered diseases of modern society, high morbidity, its sickness rate is soaring rapidly, according to the WHO statistics, the sickness rate of global depression is about 11%, crowd more than 60 years old up to 20%.According to " World Health Organization's report in 2002 " that World Health Organization (WHO) delivers, depression has become the fourth-largest illness in the world at present, has 3%~5% lifelong prevalence at least, may become to the year two thousand twenty depression to be only second to cardiopathic second largest disease.Depression serious threat human physical and mental health is just becoming serious global problems.Therefore, strengthen depression mechanism and study on prevention and will have very important strategic importance.
The modern disease pharmacological research shows: depression mechanism is very complicated, and induced factor is more, often is difficult to obtain satisfactory effect for the medicine of certain single link.Synthetic antidepressants exist mostly that antidepressant spectrum is narrow, side effect large, the medicine valency is high and the easy defective such as recurrence.Therefore, both at home and abroad aspect exploitation, more and more pay attention to traditional drugs in the development of antidepressants, the Chinese herbal medicine of more than 2,000 year history is particularly arranged, by multipath, many target spots, play a role and reach the purpose of Cure of depression at many levels.
Recent domestic is more to the antidepressant activity research report of the extract of single medicinal plants, these plants mainly comprise Herba Hyperici Monogyni, Radix Morindae Officinalis, Semen Ginkgo, Valeriana Fauriei Briq., Ziziphora plant, Semen Arecae, hold hereby Subgenus, Inner Mongolia Astragalis, Tagetes and Abies pindrow etc., and separation has obtained many chemical compounds with antidepressant activity from these plants.The antidepressant activity plant component of having found mainly contains the type compounds such as benzo dianthrone class, flavone and oligosaccharides, alkaloids, phloroglucinol class, sesquiterpenoids, Diterpenes, triterpenes, saponin, organic acid.
Chinese medicine compound is the key component of Chinese medicine, and it is formed by the principle reasonable compatibility of multiple medical material by traditional Chinese medical science monarch, thereby has stable curative effect, characteristics that toxic and side effects is little, is the main mode that tcm clinical practice is used.
Chinese patent application CN201110081236.9 discloses a kind of Chinese medicine composition, pass through removing heat-phlegm, blood circulation promoting and blood stasis dispelling, liver-smoothing, qi-regulating, consciousness-restoring and orifice-opening, improve blood circulation, improve fast depressive symptom, effectively the depressed state of an illness of control reaches the purpose of Cure of depression, in order to reach this purpose, adopt following technical scheme: its contained active component is prepared from by the following raw materials in part by weight proportioning: Pericarpium Citri Reticulatae 6-12 part, Rhizoma Pinelliae Preparatum 4-8 part, Poria 3-6 part, Caulis Bambusae In Taenia 4-8 part, Fructus Aurantii Immaturus 4-8 part, Rhizoma Zingiberis Recens 2-4 part, Bulbus Lilii 4-8 part, Radix Rehmanniae 4-8 part, Rhizoma Anemarrhenae 4-8 part, Radix Glycyrrhizae Preparata 3-6 part, Fructus Jujubae 2-4 part, and having adopted corresponding preparation method, this Chinese medicine composition has the effect of Cure of depression.
Depression belongs to the traditional Chinese medical science " melancholia " category.Melancholia that Zhong Jing discusses sees in treatise on Febrile Diseases, " Medical Treasures of the Golden Chamber " all pieces of writing, and syndrome of powder SI NI, BANXIA HOUPU TANG card, GANMAI DAZAO TANG card, lilii and Rehmanniae Decoction card etc. are arranged, and can be used as the representative of melancholia.Wherein classics recipe lilii and Rehmanniae Decoction involving clinical study report is more, but very few to the research report on its active substance basis, the extraction separation method of lilii and Rehmanniae Decoction effective site that so far there are no and the report of antidepressant effect thereof.
Summary of the invention
The invention provides a kind of preparation method of lilii and Rehmanniae Decoction antidepressant effective site, the method can be separated the effective site that obtains having antidepressant effect.
The present invention also provides a kind of lilii and Rehmanniae Decoction antidepressant effective site, has significant antidepressant effect, can be applicable to the medicine for the preparation of Cure of depression.
A kind of preparation method of lilii and Rehmanniae Decoction antidepressant effective site comprises step:
(1) with after medical material Bulbus Lilii and the Radix Rehmanniae mixing, pulverizes, obtain the medical material powder;
(2) with the medical material powder with water or the alcohol amount of containing greater than 0 and less than or equal to 95% water alcohol liquid reflux, extract, at least 2 times, merge extractive liquid,, concentrating under reduced pressure becomes extractum, adds to account for the alcohol precipitation that extractum weight 2-5 doubly measures in extractum, crosses the leaching supernatant, is condensed into concentrated solution; When elected water alcohol liquid extracted, concentrating under reduced pressure can reclaim the alcohol in the extracting solution;
(3) concentrated solution is separated with macroporous adsorbent resin column chromatography, wash first the removal of impurity with water, obtain water elution liquid, again with containing the water alcohol liquid gradient elution that the alcohol amount is 10%-95%, the eluent that merges respectively each gradient, concentrated each eluent, the corresponding effective site that obtains 10%-95% water alcohol liquid eluting position of drying under reduced pressure.
In order to reach better invention effect, preferably:
In the step (1), the mass ratio of Bulbus Lilii and Radix Rehmanniae is 1:1.
In the step (2), the number of times of reflux, extract, is 2 times.
In the step (3), preferably: concentrated solution is separated with macroporous adsorbent resin column chromatography, wash first the removal of impurity with water, obtain water elution liquid, again with containing the water alcohol liquid gradient elution that the alcohol amount is 10%-95%, each eluting concentration is followed successively by 10% ethanol water, 30% ethanol water, 50% ethanol water, 70% ethanol water, 95% ethanol water, the eluent that merges respectively each Concentraton gradient, concentrated, drying under reduced pressure respectively different eluting position, be accordingly: the water elution position, 10% ethanol water eluting position, 30% ethanol water eluting position, 50% ethanol water eluting position, 70% ethanol water eluting position, 95% ethanol water eluting position.
Wherein, effective site is 50% water alcohol liquid eluting position, and antidepressant effect is the most remarkable.
In step (2) and the step (3), alcohol is ethanol.
A kind of lilii and Rehmanniae Decoction antidepressant effective site is the effective site of the preparation method preparation of adopting described lilii and Rehmanniae Decoction antidepressant effective site.The significant 50% water alcohol of preferred antidepressant effect liquid eluting position, namely with containing the water alcohol liquid gradient elution that the alcohol amount is 10%-95%, merge the eluent with 50% water alcohol liquid eluting, concentrate eluant, the corresponding effective site that obtains the pure liquid eluting of 50% water position of drying under reduced pressure.
Described lilii and Rehmanniae Decoction antidepressant effective site, especially lilii and Rehmanniae Decoction 50% water alcohol liquid eluting position can be applicable to the medicine for the preparation of Cure of depression.
% among the present invention if no special instructions, all refers to percent by volume.
Medical material Bulbus Lilii among the present invention and Radix Rehmanniae meet the description in the Pharmacopoeia of the People's Republic of China 2010 editions (an one), and be specific as follows:
Bulbus Lilii is the dry meat scale leaf of liliaceous plant tiger lily Lilium lancifolium Thunb., Bulbus Lilii Lilium brownii F.E.Brown var.viridulum Baker or Lilium tenuifolium Lilium pumilum DC..
Radix Rehmanniae is the fresh or dried root of scrophulariaceae rehmannia glutinosa plant Rehmannia glutinosa Libosch..
The present invention has following beneficial effect:
By to the Bulbus Lilii Radix Rehmanniae take water or the alcohol amount of containing for greater than 0 and extract less than or equal to 95% water alcohol liquid, concentrated solution resin column chromatography is carried out the antidepressant drug effect to each separating part and is screened, and has found preferably separation component of antidepressant effect.Prove that through zoopery the isolated component of the present invention can shorten the dead time of outstanding tail experiment small mouse and the dead time of forced swimming experiment small mouse.Show: the active component of the extraction separation method preparation that Bulbus Lilii ground ecliptic longitude is certain, have preferably antidepressant effect, can be applicable to preparation and exploitation antidepressant preparation.
In addition, prove through zoopery, lilii and Rehmanniae Decoction 50% water alcohol liquid eluting position component can significantly shorten the dead time of outstanding tail experiment small mouse and the dead time of forced swimming experiment small mouse, has significant antidepressant effect, its antidepressant effect can compare favourably with the good fluoxetine Hydrochloride of present antidepressant effect, because lilii and Rehmanniae Decoction 50% water alcohol liquid eluting position component is that the Chinese medicine extract side effect is little, has broad application prospects.
The inventive method is simple to operate, environmental protection, cost are low, is suitable for suitability for industrialized production.
The specific embodiment
(1) extraction separation method of lilii and Rehmanniae Decoction effective site
(1) medical material Bulbus Lilii and Radix Rehmanniae are pressed the 1:1 mass ratio and mix after, pulverize.
(2) medical material with water or the alcohol amount of containing greater than 0 and less than or equal to 95% water alcohol liquid reflux, extract, at least 2 times, merge extractive liquid,, concentrating under reduced pressure becomes extractum.The extractum adding accounts for the alcohol precipitation that extractum weight 2-5 doubly measures, and crosses the leaching supernatant, is condensed into concentrated solution, i.e. the lilii and Rehmanniae Decoction alcohol extract.
(3) concentrated solution separates with macroporous adsorbent resin column chromatography, wash first the removal of impurity with water, obtain water elution liquid, again with the water alcohol liquid gradient elution that contains alcohol amount 10%-95%, the eluent that merges respectively each gradient, concentrated each eluent, the corresponding effective site that obtains 10%-95% water alcohol liquid eluting position of drying under reduced pressure.
(2) select mice behavioral despair model (experiment of mice forced swimming and Tail suspension test), estimate the antidepressant activity of each separation component of lilii and Rehmanniae Decoction.Active component through experiment confirm can be applicable to preparation and exploitation antidepressant preparation.
In conjunction with specific embodiments, further set forth the present invention.
Embodiment 1: the preparation of lilii and Rehmanniae Decoction separation component
With Bulbus Lilii, each 2kg of Radix Rehmanniae medical material, after the mixing, pulverize, with 70% ethanol water reflux, extract, 2 times, merge extractive liquid,, decompression recycling ethanol becomes extractum.Extractum adds the ethanol precipitation that accounts for 4 times of amounts of extractum weight, crosses the leaching supernatant, is condensed into concentrated solution, i.e. the lilii and Rehmanniae Decoction alcohol extract.Concentrated solution separates with the D101 macroporous adsorbent resin column chromatography, wash with water first and remove impurity, again with the water alcohol liquid gradient elution that contains alcohol amount 10%-95%, each eluting concentration is followed successively by 10% ethanol water, 30% ethanol water, 50% ethanol water, 70% ethanol water, 95% ethanol water, the eluent that merges respectively each Concentraton gradient, concentrated, drying under reduced pressure respectively different eluting position, be accordingly: the water elution position, 10% ethanol water eluting position, 30% ethanol water eluting position, 50% ethanol water eluting position, 70% ethanol water eluting position, 95% ethanol water eluting position.
Embodiment 2: the zoopery of lilii and Rehmanniae Decoction separation component
Select mice behavioral despair model (Tail suspension test, the experiment of mice forced swimming) to estimate the antidepressant effect at lilii and Rehmanniae Decoction alcohol extract (lilii and Rehmanniae Decoction alcohol position) and 6 different eluting positions thereof.The result shows, the dead time of the outstanding tail of desperate model small mouse and forced swimming is all shortened at lilii and Rehmanniae Decoction alcohol extract and 6 different eluting positions thereof to some extent, wherein with 50% ethanol water eluting position significantly (P<0.01).The effective site that shows lilii and Rehmanniae Decoction mainly is distributed in 50% alcohol elution.
Table 1 lilii and Rehmanniae Decoction respectively extracts the impact of separated part on the mouse tail suspension dead time
Annotate: n is test number (TN), compares a:P<0.05, b:P<0.01 with the blank group.
Table 2 lilii and Rehmanniae Decoction respectively extracts separated part to the impact of mice forced swimming dead time (x ± s)
Annotate: compare a:P<0.05, b:P<0.01 with the blank group.
Experimental result shows: the isolated component of the present invention can shorten the dead time of outstanding tail experiment small mouse and the dead time of forced swimming experiment small mouse.Show: lilii and Rehmanniae Decoction has preferably antidepressant effect through the active component of certain extraction separation method preparation, can be applicable to preparation and exploitation antidepressant preparation.In addition, prove through zoopery, lilii and Rehmanniae Decoction 50% water alcohol liquid eluting position component can significantly shorten the dead time of outstanding tail experiment small mouse and the dead time of forced swimming experiment small mouse, has significant antidepressant effect, its antidepressant effect can compare favourably with the good fluoxetine Hydrochloride of present antidepressant effect, because lilii and Rehmanniae Decoction 50% water alcohol liquid eluting position component is that the Chinese medicine extract side effect is little, has broad application prospects.
Among the present invention, owing to adopt water or contain the alcohol amount greater than 0 and can guarantee that less than or equal to 95% water alcohol liquid reflux, extract, most of antidepressant effective ingredient is extracted at least for 2 times, the variation of parameter does not affect effective extraction and the separation of lilii and Rehmanniae Decoction antidepressant effective site in the preparation method of the present invention, therefore, the combination in any in preparation method parameter area of the present invention all can realize effective extraction and the separation of lilii and Rehmanniae Decoction antidepressant effective site.Do not repeat them here.
Claims (8)
1. the preparation method of a lilii and Rehmanniae Decoction antidepressant effective site is characterized in that, comprises step:
(1) with after medical material Bulbus Lilii and the Radix Rehmanniae mixing, pulverizes, obtain the medical material powder;
(2) with the medical material powder with water or the alcohol amount of containing greater than 0 and less than or equal to 95% water alcohol liquid reflux, extract, at least 2 times, merge extractive liquid,, concentrating under reduced pressure becomes extractum, adds to account for the alcohol precipitation that extractum weight 2-5 doubly measures in extractum, crosses the leaching supernatant, is condensed into concentrated solution;
(3) concentrated solution is separated with macroporous adsorbent resin column chromatography, wash first the removal of impurity with water, obtain water elution liquid, again with containing the water alcohol liquid gradient elution that the alcohol amount is 10%-95%, the eluent that merges respectively each gradient, concentrated each eluent, the corresponding effective site that obtains 10%-95% water alcohol liquid eluting position of drying under reduced pressure;
Described % is percent by volume.
2. the preparation method of lilii and Rehmanniae Decoction antidepressant effective site according to claim 1 is characterized in that, in the step (1), the mass ratio of Bulbus Lilii and Radix Rehmanniae is 1:1.
3. the preparation method of lilii and Rehmanniae Decoction antidepressant effective site according to claim 1 is characterized in that, in the step (2), the number of times of reflux, extract, is 2 times.
4. the preparation method of lilii and Rehmanniae Decoction antidepressant effective site according to claim 1 is characterized in that, in the step (3), effective site is 50% water alcohol liquid eluting position.
5. the preparation method of lilii and Rehmanniae Decoction antidepressant effective site according to claim 1 is characterized in that, in step (2) and the step (3), described alcohol is ethanol.
6. a lilii and Rehmanniae Decoction antidepressant effective site is characterized in that, the effective site that described effective site prepares for the preparation method that adopts each described lilii and Rehmanniae Decoction antidepressant effective site of claim 1-5.
7. lilii and Rehmanniae Decoction antidepressant effective site according to claim 6 is characterized in that, effective site is 50% water alcohol liquid eluting position.
8. according to claim 6 or the application of 7 described lilii and Rehmanniae Decoction antidepressant effective sites, it is characterized in that described lilii and Rehmanniae Decoction antidepressant effective site is for the preparation of the application in the medicine of Cure of depression.
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN104208383A (en) * | 2014-09-03 | 2014-12-17 | 河南中医学院 | Application of fresh lily bulb rehmannia powder in preparation of medicine for treating depressive disorder |
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Non-Patent Citations (3)
Title |
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《中国新药杂志》 20101231 张萍等 《百合地黄汤抗抑郁活性部位的筛选》 P1973-1975 2 第19卷, 第21期 * |
张萍等: "《百合地黄汤抗抑郁活性部位的筛选》", 《中国新药杂志》 * |
张萍等: "《药用植物化学与中药资源可持续发展学术研讨会论文集(上)》", 31 December 2009 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104208383A (en) * | 2014-09-03 | 2014-12-17 | 河南中医学院 | Application of fresh lily bulb rehmannia powder in preparation of medicine for treating depressive disorder |
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Application publication date: 20130320 |