CN1457702A - Wild jujube kernel soup active part and its preparing process and new use - Google Patents

Abstract

The active parts of wild jujube kernel include two parts B and E. The preparation process includes the following steps: soaking, multiple distillation, multiple filtering, collecting filtrate, collecting distillate, multiple alcohol deposition, collecting precipitate as the polysaccharide part B; depression concentration of mother liquid, extraction with ethyl ether to obtain extracted liquid and water solution, concentrating water solution, water and alcohol elution, merging the same parts, hydrochloric acid and magnesium powder inspection and depression drying to obtain flavone part E. The active parts of wild jujube kernel and wild jujube kernel decoction has the function of resisting dysphoria.

Description

Suanzaoren decoction active site and preparation technology thereof, new purposes

Invention field

The present invention relates to a kind of Chinese patent drug active site, particularly relate to suanzaoren decoction active site and preparation technology thereof, new purposes.

Background technology

(Suanzaoren Tang SZRT) comes from " Synopsis Golden Chamber " to suanzaoren decoction, is made up of spina date seed, Radix Glycyrrhizae, the wind-weed, Poria cocos, Ligusticum wallichii, and having enriches blood transfers liver, mental-tranquilization, clearing heat and relieving fidgetness effect.

Modern face on the basis of card experience inheriting forefathers, further enlarged our range of application, intractable aypnia is one of disease of being most widely used in spiritual nervous system of suanzaoren decoction; In addition, the hyperfunction disease of heart beta receptor, congenital nonhemolytic jaundice, the cutaneous diseases, hemospermia syndrome, recurrent oral ulceration, duodenal ulcer, uterine bleeding, spontaneous perspiration, the night sweat that cause because of reason such as nervous, excited, as to drink, have a liking for sour disease, virus B hepatitis, Meniere disease, illness such as dizzy is all used to some extent.

Summary of the invention

One object of the present invention is to provide the suanzaoren decoction active site; The present invention also aims to provide the preparation technology of suanzaoren decoction active site; The present invention also aims to provide the new purposes of suanzaoren decoction and active site thereof.

A kind of suanzaoren decoction active site comprises B polysaccharide 260-300 weight portion, E flavones 4-7 weight portion.This active site prepares by the following method: the suanzaoren decoction medicinal material, doubly measure (multiple of weight) water logging bubble 20-40min with 8-11 after, steam distillation 0.8-1.5 hour, collect distillate, filter, the dregs of a decoction were with 7-9 times of water gaging steam distillation 0.3-0.5 hour, collect distillate, filter; Merging filtrate and distillate get filtrate I and distillate I; When filtrate I continues to be distilled to the 60-85% of cumulative volume, get distillate II and filtrate I remainder-filtrate II; Distillate II redistillation stops distillation during to the 40-60% of original volume; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, adding 95% ethanol, to transfer to alcoholic degree be 55-65%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol, and to transfer to alcoholic degree be 75-85%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get B polysaccharide part; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; The aqueous solution be concentrated into crude drug heavy with volume ratio be 10: 1, last sample, polyamide column chromatography; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution; Inspection is known, and merges same area; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets E flavones part.

The active site of suanzaoren decoction of the present invention also can be made up of A, B, C, D, six parts of E, F; These six positions are prepared by following method: the suanzaoren decoction medicinal material, doubly measure (multiple of weight) water logging bubble 20-40min with 8-11 after, steam distillation 0.8-1.5 hour, collect distillate, filter, the dregs of a decoction were with 7-9 times of water gaging steam distillation 0.3-0.5 hour, collect distillate, filter; Merging filtrate and distillate get filtrate I and distillate I; When filtrate I continues to be distilled to the 60-85% of cumulative volume, get distillate II and remainder-filtrate II; Distillate II redistillation stops distillation during to the 40-60% of original volume; Heavily steam the liquid boiling point and be 30 ℃～60 ℃ petroleum ether extraction, reclaim benzinum, A volatile oil part (A is a volatile oil, because of amount very little, fill a prescription being used for so be dissolved in 500ml benzinum constant volume); When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, adding 95% ethanol, to transfer to alcoholic degree be 55-65%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol, and to transfer to alcoholic degree be 75-85%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get 260-300 weight portion B polysaccharide part; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, do not have the alcohol flavor, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; Ether extraction liquid recovered under reduced pressure ether gets 3-4.5 weight portion C extracted with diethyl ether part part; The aqueous solution be evaporated to crude drug heavy with volume ratio be 10: 1, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution; Inspection is known, and merges same area; Water elution partly, reaction negative is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection and aceticanhydride-strong sulfuric acid response is positive partly, 50% ethanol elution partly merges, and drying under reduced pressure gets 300-340 weight portion D saponin(e and aglycon part; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets 4-7 weight portion E flavones part; 95% ethanol eluate, recovered under reduced pressure gets 1-3 weight portion F (95% ethanol elution part).

It is to adopt following method: A, the anisaldehyde-strong sulfuric acid response positive that the above inspection is known; B, the Molish reacting positive; C, Molish reaction negative, the aceticanhydride-strong sulfuric acid response positive, hydrochloric acid-magnesium powder reacting positive; D, Molish reacting positive, the aceticanhydride-strong sulfuric acid response positive, hydrochloric acid-magnesium powder reaction negative; E, hydrochloric acid-magnesium powder reacting positive; F, more than reaction all is negative.Prompting contains volatile oil, polysaccharide, aglycon and flavones ingredient, saponin(e and aglycon constituents, flavonoids and mixtae composition respectively.

Press practice of pharmacy, the active site of suanzaoren decoction of the present invention can be prepared into the various clinical pharmaceutical dosage form, comprise the formulation of oral formulations or parenterai administration.Said oral formulations is selected from a kind of in the middle of the tablet, capsule, pill, granule, supensoid agent, dripping pill, oral liquid; Said parenterai administration formulation is selected from a kind of in the middle of injection, aerosol, suppository or the subcutaneous administration formulation.Medicine of the present invention also can add conventional drug excipient, as solvent, disintegrant, flavouring, anticorrisive agent, colouring agent etc.

The active site of suanzaoren decoction of the present invention and suanzaoren decoction have effect antianxity.

In overhead cross labyrinth (elevated plus-maze, EPM) in the anxiety model, the suanzaoren decoction component SZRT that effectively fills a prescription ₆(B+E) can significantly raise rat (7.5g/kg) and mouse (20g/kg) enter the open arms number of times than (0E%) and the open arms time of staying than (OT%) (p＜0.05), and rat head-dips number in open arms significantly raise (p＜0.05); But the number of times (total arm entries) that enters that rat, mouse open arms and closure arm are total is compared no significant difference (p＞0.05) with the whole composition control groups of decocting liquid.These parameters, SZRT ₆With the SZRT that can represent the whole compositions of SZRT decocting liquid substantially ₁No significant difference (p＞0.05).The result shows: under this dosage, and SZRT ₆Have definite angst resistance effect, and irrelevant with sedative-hypnotic effect.Its effect and the SZRT that can represent the whole compositions of SZRT decocting liquid substantially ₁Quite.Conclusion: the active site of suanzaoren decoction of the present invention is suitable with the effect of suanzaoren decoction decocting liquid, and efficacy component is more clear and definite.Antianxity good effect arranged on.

Following experimental example further specifies the present invention.Experimental example

For inquiring in SZRT (suanzaoren decoction) water extract relation about component and angst resistance effect, initial gross separation at the SZRT water extract goes out on the component basis, adopt international anxiety animal model elevated plus-maze (EPM, overhead cross labyrinth), successively investigate the influence of the component of SZRT and different compatibility thereof respectively to rat and mouse anxiety model.

1. material

1.1 animal

Kunming mouse, male, body weight 28 ± 3g, Institute of Experimental Animals, Chinese Academy of Medical Sciences provides, the animal quality certification number: SCXK11-00-0066;

The Wistar rat, male, body weight 250 ± 20g, Chinese pharmaceutical biological product identify that institute's Experimental Animal Center provides, the animal quality certification number: SCXK11-00-0010.

All animals are carried the last fortnight and buy, this chamber animal housing natural feeding.21 ± 2 ℃ of room temperatures.

1.2 medicine and reagent

1.2.1 DZP (diazepam, diazepam, stable) suspension: raw material DZP tablet (2.5mg * 20 slice), the accurate word (1996) of medicine is defended No. 154011 in the capital, product batch number: 0204090, Beijing Yimin Pharmaceutical Factory produces.The DZP slice lapping, 2%CMC-Na is an amount of in adding, is made into 0.2mg/ml (mouse is used) and two kinds of suspensions of 0.1mg/ml (rat is used) with distilled water.4 ℃ of preservations are standby, shake up during use.1.2.2 the component prescription of suanzaoren decoction: SZRT extracts isolated 6 parts (A, B, C, D, E, F) and is made by research one method.

1.3 instrument

1.3.1 the overhead cross of mouse labyrinth (EPM)

Make: overhead cross labyrinth is by two relative open arms (open arm, length * wide * height be respectively 30 * 5cm) with two relative closure arm (enclosed arm, close arm, length * wide * height is respectively 30 * 5 * 15cm) and forms, a central platform (centralplatform who connects four arms, 5cm * 5cm) (that is: open arms-central platform-open arms or closure arm-central platform-closure arm, this two orthogonal becoming " ten " is shape (plus)).This labyrinth makes (0.5cm is thick) by homemade lucite, and except that the base plate and the central platform of four arms is the black, all the other positions are water white transparency.Be provided with the high edge of 0.25cm around the open arms.Lucite part overall fixed is on (being made up of " ten " the font yokes of length and width such as black and stainless steel base) support, and the labyrinth base plate is apart from 60cm place, ground, laboratory.

1.3.2 the overhead cross of rat labyrinth (EPM)

Make: the overhead cross of rat labyrinth is by two relative open arms (open arm, long * the wide 50cm * 10cm) of being respectively, two relative closure arm (enclosed arm, close arm, length * wide * height is respectively 50cm * 10cm * 40cm), a central platform (centralplatform who connects four arms, 10cm * 10cm) (that is: open arms-central platform-open arms or closure arm-central platform-closure arm, this two orthogonal becoming " ten " is shape (plus)) and the high short baffle plate of 1cm that is centered around the open arms edge are formed (purpose is to prevent that animal from sliding the labyrinth accidentally in the process of probing into).This labyrinth makes (0.5cm is thick) by homemade lucite, and except that the base plate and the central platform of four arms is the black, all the other positions are water white transparency.Lucite part overall fixed makes the labyrinth base plate be adjusted to 50cm place, ground, distance laboratory on (being made up of " ten " yoke of length and width such as black and stainless steel liftable base) support.

2. method

2.1 component prescription

Press L ₈(2 ⁷) orthogonal table ^[5]Carry out component prescription (seeing attached list): SZRT respectively ₁(A+B+C+D+E+F), SZRT ₂(A+B+C), SZRT ₃(A+D+E), SZRT ₄(A+F), SZRT ₅(B+D+F), SZRT ₆(B+E), SZRT ₇(C+D), SZRT ₈(C+E+F) totally 8 prescriptions, the component compatibility consumption in each prescription are to calculate according to the suitable medicinal material amount of this component and former side's proportion compatibility get, prepare with distilled water or suspension (adding Tween-80), and 4 ℃ of preservations are standby, shake up during use.

SZRT component prescription L ₈(2 ⁷) orthogonal table

SZRT A B C D E F

Component fills a prescription 1234567

1 1 1 1 1 1 1 1

2 1 1 1 2 2 2 2

3 1 2 2 1 1 2 2

4 1 2 2 2 2 1 1

5 2 1 2 1 2 1 2

6 2 1 2 2 1 2 1

7 2 2 1 1 2 2 1

8 2 2 1 2 1 1 2

In the table there be the representative of 1 level, the representative of 2 levels does not have.

2.2 animal grouping and disposal

10 in the every cage of mouse, 7～8 in the every cage of rat is not limit drinking water.(animal housing keeps quite illumination rhythm and pace of moving things 12L: 12D for 7:00～19:00), room temperature (21 ± 2) ℃.

Mouse is divided into 10 groups at random, i.e. control group, DZP group, SZRT ₁Group, SZRT ₂Group, SZRT ₃Group, SZRT ₄Group, SZRT ₅Group, SZRT ₆Group, SZRT ₇Group, SZRT ₈Group, totally 10 groups, 20 every group.SZRT wherein ₁Group～SZRT ₈Group gives SZRT by being equivalent to full side's crude drug 20g/kg body weight respectively ₁～SZRT ₈Irritate stomach, the DZP group by every day 2.0mg/kg give the DZP suspension oral gavage, normal group gives isometric(al) distilled water and irritates stomach, irritates at every turn that body of stomach is long-pending to be the 0.1ml/10g body weight.

Rat is divided into 4 groups at random, i.e. control group, DZP, SZRT ₁Group, SZRT ₆Group, 14 every group; SZRT wherein ₁Group, SZRT ₆Group is respectively by giving SZRT by being equivalent to full side's crude drug 7.5g/kg body weight ₁, SZRT ₆Irritate stomach, DZP organizes by 1.0mg/kg filling every day stomach, and normal group gives isometric distilled water and irritates stomach, and irritate body of stomach long-pending is the 1ml/100g body weight at every turn.

Duration of test, the next day 8:00～9:30AM weigh, put back to again behind the animals administer in the former raising box.1.5h after mouse, rat successive administration 10 days, Chinese drug-treated group animal were irritated stomach on 10th, DZP group and control animals 0.5h behind lumbar injection do the behaviouristics test.All animals entered the laboratory in advance in 2 hours, and after performance testing finished, the animal broken end was got blood, takes out brain tissue fast.

2.3 index test

2.3.1 test environment

Rather dark in the laboratory (minimum brightness that can distinguish the trickle activity of mouse with the 1.5m distance is as the criterion) also keeps permanent bright, and room temperature remains on (21 ± 2) ℃, peace and quiet.Mouse, rat EPM place one jiao in laboratory, and cloth is with the dull background of the high black of 2m on every side.Have only one to participate in daily treatment people administration and carrying animal in the experimentation.By 2 personnel that carried out the experimental record training in advance but be unfamiliar with grouping situation activity of observed and recorded animal respectively at distance test case 1.5m place.The test of mouse behaviouristics is all carried out between 8:00～15:30, and rat behavior is learned test and all carried out between 8:00～12:30.Every animal testing 5min, next animal testing is carried out in middle earlier wet cloth wiping labyrinth again after cleaning with dried cloth.

2.3.2 mouse behaviouristics observation index

Every mouse places the central platform place of EPM, makes its head over against one of them open arms, is following index in the opening entry 5min after the release:

(1) enter the open arms number of times (open arm entry, OE): enter into the number of times of arbitrary open arms, all enter in the arm with four claws of mouse and be as the criterion, a claw withdraws from then fully from this arm and finishes for this time activity of entering midway; (2) the open arms time (open arm time, OT): enter the time of open arms, unit: second; (3) enter the closure arm number of times (close arm entry, CE): enter into the number of times of arbitrary closure arm, all enter in the arm with four claws of mouse and be as the criterion; (4) the closure arm time (close armtime, CT): enter the time of closure arm, unit: second; (5) probe into number of times (head-dips) downwards: when mouse is placed oneself in the midst of open arms, with fore paw hold labyrinth edge and on one side head and shoulder stretched out behavior number of times that the edge of open arms to labyrinth below probe on one side.

Calculate respectively by (1)～(4): 1. total number of times (total armentries) that enters of open arms and closure arm, that is: OE+CE, the motion vigor (locomotor activity) of expression mouse; Open arms enters number of times ratio (OE%), and promptly open arms enters number of times/(open arms enters number of times+closure arm and enters number of times) * 100%; Open arms time of staying ratio (OT%), i.e. open arms time of staying/(open arms entry time+closure arm entry time) * 100%.

2.3.3 rat behavior is learned observation index

Observation index, computational methods and experiment are handled the same.

2.4 data are handled

Each is organized data and represents that with x ± S q check [6] is adopted in the significance test between many groups.All data all use software SPSS 10.0 for windows (one-way ANOVA) to handle.SZRT soup different component prescription is to the influence of OE% and OT%, employing has L8 (27) the orthogonal experiment analysis of repetition with showing, with the influence of each factor of F test evaluation to OE% and OT%, all data all use software SPSS 10.0 forwindows (General Linear Model) to handle.

3. result

3.1SZRT the different component prescription is to the influence of mouse EPM model behavior

3.1.1SZRT the different component compatibility is to the influence of mouse OE% and OT%

The results are shown in Table 1.As can be seen from Table 1, compare with control group, DZP group OE%, OT% all are significantly higher than control group (P＜0.01).SZRT ₁Group, SZRT ₃Group, SZRT ₅Group, SZRT6 ₆Group mouse OE% all is significantly higher than control group (P＜0.05); SZRT ₁Group, SZRT ₆Group OT% all is significantly higher than control group (P＜0.05).OE%, OT% all do not have significant difference (P＞0.05) between each group of SZRT component prescription.Table one: SZRT component prescription SZRT ₁～SZRT ₈To EPM anxiety model mice

Open arms enter number of times than (OE%) and the open arms time of staying than the influence of (OT%) (x ± S)

Group Dose(/kg) N OE％ OT％

Vehicle 0 19 0.101±0.118 0.061±0.102

DZP 2mg 20 0.326±0.159** 0.358±0.203**

SZRT ₁ 15g 18 0.203±0.129* 0.225±0.168*

SZRT ₂ 15g 20 0.151±0.099 0.079±0.072

SZRT ₃ 15g 18 0.220±0.206* 0.200±0.258

SZRT ₄ 15g 19 0.193±0.136 0.163±0.146

SZRT ₅ 15g 20 0.214±0.175* 0.139±0.168

SZRT ₆ 15g 20 0.222±0.159* 0.241±0.202*

SZRT ₇ 15g 19 0.168±0.142 0.151±0.186

SZRT ₈ 15g 18 0.130±0.133 0.152±0.178

*P＜0.05，**P＜0.01，compared?with?vehicle-treated?group.

3.1.2SZRT the different component compatibility is to the influence of mouse Head-dips behavior

The results are shown in Table 2.As can be seen from Table 2, compare with control group, head-dips number of DZP group significantly increases (P＜0.01).SZRT respectively organizes mouse does not have remarkable increase at head-dips number average of open arms, but SZRT ₁Group, SZRT ₆Organize head-dips number rising trend is arranged.3.1.3SZRT the different component compatibility to mouse open arms and closure arm total enter number of times (total armentries)

The results are shown in Table 2.Can find out that from table 2 DZP group mouse open arms and the total number of times that enters of closure arm do not have significant difference (P＞0.05) with the control group ratio.SZRT component prescription respectively organizes the mouse open arms and the total number of times that enters of closure arm does not have significant difference (P＞0.05) with the control group ratio.Table two: SZRT component prescription SZRT ₁～SZRT ₈To the EPM anxiety model mice open arms influence of exploring number of times (Head-dips) and locomitivity (Total arm the entries) (x ± S) that bows

Group Dose(/kg) N Head-dips Total?arm?entries

Vehicle 0 19 3.63±8.27 16.00±2.77

DZP 2mg 20 21.40±13.91** 20.85±6.10

SZRT ₁ 15g 18 20.16±19.35 15.15±3.86

SZRT ₂ 15g 20 3.45±3.76 15.05±6.51

SZRT ₃ 15g 18 3.94±5.96 12.94±3.56

SZRT ₄ 15g 19 6.90±8.56 15.70±3.95

SZRT ₅ 15g 20 5.65±9.22 15.05±5.21

SZRT ₆ 15g 20 19.60±21.00 14.05±3.46

SZRT ₇ 15g 19 1.95±2.95 14.65±5.08

SZRT ₈ 15g 18 5.58±10.61 14.11±4.47

**P＜0.01，compared?with?vehicle-treated?group.

The SZRT 3.2SZRT component is effectively filled a prescription ₁, SZRT ₆Influence to rat EPM model behavior

SZRT1, SZRT6 the influence 3.2.1SZRT component is effectively filled a prescription to rat OE% and OT%

The results are shown in Table 3.As can be seen from Table 3, compare with control group, DZP group rat OE% and OT% value obviously raise, and utmost point significant difference (P＜0.01) is arranged; SZRT ₁, SZRT ₆Group rat OE% and OT% value obviously raise, and significant difference (P＜0.05) is arranged.SZRT ₁, SZRT ₆Group OE% and OT% value there was no significant difference (P＞0.05).Table three: the SZRT component SZRT that effectively fills a prescription ₁And SZRT ₆To EPM anxiety rat model

Open arms enter number of times than (OE%) and the open arms time of staying than the influence of (OT%) (x ± S)

Group Dose(/kg) N OE％ OT％

Vehicle 0 12 0.378±0.082 0.451±0.083

DZP 1mg 13 0.547±0.128** 0.622±0.134**

SZRT ₁ 7.5g 12 0.480±0.074* 0.553±0.118*

SZRT ₆ 7.5g 13 0.455±0.043* 0.537±0.074*

*P＜0.05，**P＜0.01，compared?with?vehicle-treated?group.

The SZRT 3.2.2SZRT component is effectively filled a prescription ₁, SZRT ₆Influence to rat head-dips behavior

The results are shown in Table 4.As can be seen from Table 4, compare with control group, head-dips number of DZP group increases, and utmost point significant difference (P＜0.01) is arranged; SZRT ₁, SZRT ₆Organizing head-dips number increases, and significant difference (P＜0.05) is arranged.But SZRT ₁Group, SZRT ₆Head-dips the no significant difference of number (P＞0.05) between group.Table four: the SZRT component SZRT that effectively fills a prescription ₁And SZRT ₆To the EPM anxiety model mice open arms influence of exploring number of times (Head-dips) and locomitivity (Total arm the entries) (x ± S) that bows

Group Dose(/kg) N Head-dips Total?arm?entries

Vehicle 0 12 16.92±4.19 21.00±3.93

DZP 1mg 13 30.31±9.51** 21.77±5.39

SZRT ₁ 7.5g 12 23.00±7.08* 22.83±3.79

SZRT ₆ 7.5g 13 24.23±5.70* 20.92±5.35

*P＜0.05，**P＜0.01，compared?with?vehicle-treated?group.

3.2.3SZRT the different component compatibility to rat open arms and closure arm total enter number of times (totalarm entries)

The results are shown in Table 4.Can find out that from table 4 DZP organizes and SZRT respectively organizes the rat open arms and the total number of times that enters of closure arm does not have significant difference (P＞0.05) with the control group ratio.

3.1.4 SZRT soup different component is to the influence of mouse OE% and OT%

3.1.4.1SZRT the soup different component is to the quadrature analysis of the influence of OE%

The results are shown in Table 5.As can be seen from Table 5, each component of SZRT only C has appreciable impact (P＜0.05) to mouse OE%, and all the other each components all do not have appreciable impact.When using C, its OE% is 0.163 ± 0.017, and time spent OE% is not 0.212 ± 0.017.Table five: the SZRT different component enters the influence of number of times than (OE%) to EPM anxiety model mice open arms

Source Sum?of?squares Df MS F Sig.

A 0.0026 1 0.0026 0.114 0.736

B 0.0145 1 0.0145 0.645 0.423

C 0.0920 1 0.0920 4.083 0.045*

D 0.0280 1 0.0280 1.242 0.267

E 0.0057 1 0.0057 0.253 0.615

F 0.00086 1 0.00086 0.038 0.845

Error 3.221 143 0.0225

* compare with other each groups P＜0.05.

3.1.4.2SZRT the soup different component is to the quadrature analysis of OT% influence

The results are shown in Table 6.As can be seen from Table 6, each component of SZRT only E has appreciable impact (P＜0.05) to mouse OT%, and all the other each components all do not have appreciable impact.When using E, its OT% is 0.205 ± 0.21, and time spent OT% is not 0.133 ± 0.20.Table six: the SZRT different component is to the influence of the EPM anxiety model mice open arms time of staying than (OT%)

Source Sum?of Df MS F Sig.

squares

A 0.00095 1 0.00095 0.030 0.863

B 0.00059 1 0.00059 0.018 0.892

C 0.04296 1 0.04296 1.346 0.248

D 0.01414 1 0.01414 0.443 0.507

E 0.195 1 0.195 6.111 0.015*

F 0.00028 1 0.00028 0.009 0.926

Compare with other each groups Error 4.565 143 0.03193*P＜0.05.

4. conclusion

This experiment is observed, in 8 prescriptions of each component compatibility, and SZRT ₁With SZRT ₆Rat, the main behavioral indicator of mouse (OE%, OT%) all there are appreciable impact, and do not influence the locomitivity of rat, mouse, show that these two prescriptions have significant angst resistance effect.In mouse EPM, SZRT ₁With SZRT ₆There is the rising mouse to explore the trend of number of times (head-dips) bowing of open arms (non-protection area); In rat EPM, SZRT ₁With SZRT ₆The rat that can raise is explored number of times (head-dips) bowing of open arms (non-protection area), supports its angst resistance effect to a certain extent.Previous studies show that SZRT decocting liquid mouse 20g/kg, rat 7.5g/kg have significant angst resistance effect, and this is also observed, quite dosage, include the whole components prescription of former side SZRT ₁Also have same function, prompting is in the extraction separation process of SZRT decocting liquid, and the live part of angst resistance effect is not obviously lost among the former side, and the extraction separation scheme of drafting in the research one about suanzaoren decoction is rational.

Experiment is observed, the SZRT in each prescription of SZRT component ₁, SZRT ₃, SZRT ₅And SZRT ₆Angst resistance effect is in various degree all arranged, SZRT ₂, SZRT ₄, SZRT ₇And SZRT ₈Then almost there is not angst resistance effect.At four groups of effective SZRT ₁, SZRT ₃, SZRT ₅And SZRT ₆In the group, the effect index of each group is different, as SZRT ₁, and SZRT ₆Effective to OE%, OT%; And SZRT ₃, and SZRT ₅Then only effective to OE%.The relevance of the contained component of SZRT and different formulations and antianxiety effectiveness is different in the prompting experiment, wherein SZRT ₆The closest with the angst resistance effect relation.

Conclusion: the prescription by B, E two component compatibilities has significant angst resistance effect, and the full side of its action intensity and suanzaoren decoction quite.

The following example all can be realized the effect of above-mentioned experimental example. Embodiment 1The extraction of active site B, E

Suanzaoren decoction 2484g, behind 10 times of water gagings immersion 30min, steam distillation 1 hour is collected distillate, filters, and the dregs of a decoction are collected distillate with 8 times of water gaging steam distillation 0.4h, filter; Merging filtrate and distillate get filtrate I and distillate I; Filtrate I continues to be distilled to 4/5 o'clock of cumulative volume, distillate II and remainder-filtrate II; Distillate II redistillation stopped distillation to 1/2 o'clock of original volume; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, add 95% ethanol and transfer to alcoholic degree and be 60%, 4 ℃ and place 24h, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol and transfers to alcoholic degree and be 80%, 4 ℃ and place 24h, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get 285.40g B polysaccharide part; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; The aqueous solution be concentrated into crude drug heavy with volume ratio be 10: 1, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution, (column internal diameter 10cm, the high 50cm of post); Inspection is known, and merges same area; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets 5.41g E flavones part. Embodiment 2The extraction of active site A, B, C, D, E, F

Suanzaoren decoction 2484g, behind 10 times of water gagings immersion 30min, steam distillation 1h hour, collect distillate, filter, the dregs of a decoction are collected distillate with 8 times of water gaging steam distillation 0.4h, filter; Merging filtrate and distillate get filtrate I and distillate I; Filtrate I continues to be distilled to 4/5 o'clock of cumulative volume, distillate II and filtrate I remainder-filtrate II; Distillate II redistillation stopped distillation to 1/2 o'clock of original volume; Heavily steam the liquid boiling point and be 30 ℃ ~ 60 ℃ petroleum ether extraction, reclaim benzinum, A volatile oil part; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, add 95% ethanol and transfer to alcoholic degree and be 60%, 4 ℃ and place 24h, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol and transfers to alcoholic degree and be 80%, 4 ℃ and place 24h, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get B (polysaccharide, 285.40g) part; Mother liquor I and mother liquor II merge, and are evaporated to proper volume, do not have the alcohol flavor, and extracted with diethyl ether gets the ether extraction liquid and the aqueous solution; Ether extraction liquid recovered under reduced pressure ether gets C (extracted with diethyl ether part, 3.71g) part; The aqueous solution is concentrated into suitable volume, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution (column internal diameter 10cm, the high 50cm of post); Inspection is known, and merges same area; Reaction negative and the positive part of aceticanhydride-strong sulfuric acid response are known in water elution part, 30% ethanol eluate hydrochloric acid-magnesium powder inspection, 50% ethanol merges, drying under reduced pressure, D (saponin(e and aglycon part, 330.51g); The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets E (flavones, 5.41g) part; 95% ethanol eluate, recovered under reduced pressure gets F (95% ethanol elution part, 1.62g) part. Experimental example 3The preparation of capsule

Active component B of the present invention (polysaccharide, 285.40g), (flavones, 5.41g), medical starch 1000g mixes E, the capsule of packing into No. 1, every 0.2g, each oral 2-3 grain, twice of every day. Experimental example 4The preparation of capsule

(A is a volatile oil to active component A of the present invention, because of measuring very little, so be dissolved in 500ml benzinum constant volume to be used for prescription), B285.40g, C3.71g, D330.51g, E5.41g, F1.62g, medical starch 1000g, mix the capsule of packing into No. 1, every 0.2g, each oral 2-3 grain, twice of every day. Embodiment 5The preparation of tablet

Active component B300g of the present invention, E6g, medical starch 200g, dextrin 50g mixes, and uses an amount of alcohol granulation, through the whole grain of pelletizing machine, compressing tablet, every 0.25g, oral, each 2, twice of every day.

Claims (10)

1, a kind of active component of suanzaoren decoction is characterized in that this active component comprises B polysaccharide 260-300 weight portion, E flavones 4-7 weight portion; This active site prepares by the following method: the suanzaoren decoction medicinal material, and behind the 8-11 times of water gaging immersion 20-40min with medicinal material weight, steam distillation 0.8-1.5 hour, collect distillate, filter, the dregs of a decoction were with 7-9 times of water gaging steam distillation 0.3-0.5 hour, collect distillate, filter; Merging filtrate and distillate get filtrate I and distillate I; When filtrate I continues to be distilled to the 60-85% of cumulative volume, get distillate II and filtrate I remainder-filtrate II; Distillate II redistillation stops distillation during to the 40-60% of original volume; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, adding 95% ethanol, to transfer to alcoholic degree be 55-65%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol, and to transfer to alcoholic degree be 75-85%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get B polysaccharide part; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; The aqueous solution be concentrated into crude drug heavy with volume ratio be 10: 1, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution; Inspection is known, and merges same area; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets E flavones part.

2, active component as claimed in claim 1 is characterized in that suanzaoren decoction, and behind 10 times of water gagings immersion 30min, steam distillation 1 hour is collected distillate, filters, and the dregs of a decoction are collected distillate with 8 times of water gaging steam distillation 0.4h, filter; Merging filtrate and distillate get filtrate I and distillate I; Filtrate I continues to be distilled to 4/5 o'clock of cumulative volume, distillate II and filtrate I remainder-filtrate II; Distillate II redistillation stopped distillation to 1/2 o'clock of original volume; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, add 95% ethanol and transfer to alcoholic degree and be 60%, 4 ℃ and place 24h, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol and transfers to alcoholic degree and be 80%, 4 ℃ and place 24h, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get 285.40 weight portion B polysaccharide parts; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; The aqueous solution be concentrated into crude drug heavy with volume ratio be 10: 1, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution, column internal diameter 10cm, the high 50cm of post; Inspection is known, and merges same area; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets 5.41 weight portion E flavones parts.

3, active component as claimed in claim 1 is characterized in that this active component is made up of A, B, C, D, six parts of E, F; These six positions are prepared by following method: the suanzaoren decoction medicinal material, and behind the 8-11 times of water gaging immersion 20-40min with medicinal material weight, steam distillation 0.8-1.5 hour, collect distillate, filter, the dregs of a decoction were with 7-9 times of water gaging steam distillation 0.3-0.5 hour, collect distillate, filter; Merging filtrate and distillate get filtrate I and distillate I; When filtrate I continues to be distilled to the 60-85% of cumulative volume, get distillate II and remainder-filtrate II; Distillate II redistillation stops distillation during to the 40-60% of original volume; Heavily steam the liquid boiling point and be 30 ℃～60 ℃ petroleum ether extraction, reclaim benzinum, A volatile oil part; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, adding 95% ethanol, to transfer to alcoholic degree be 55-65%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol, and to transfer to alcoholic degree be 75-85%, placed 22-26 hour for 3-5 ℃, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get 260-300 weight portion B polysaccharide part; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, do not have the alcohol flavor, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; Ether extraction liquid recovered under reduced pressure ether gets 3-4.5 weight portion C extracted with diethyl ether part part; The aqueous solution be concentrated into crude drug heavy with volume ratio be 10: 1, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution; Inspection is known, and merges same area; Water elution part, 30% ethanol eluate hydrochloric acid-magnesium powder examine the knowledge reaction negative and aceticanhydride-strong sulfuric acid response is positive partly, 50% ethanol merges, and drying under reduced pressure gets 300-340 weight portion D saponin(e and aglycon part; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets 4-7 weight portion E flavones part; 95% ethanol eluate, recovered under reduced pressure gets 1-3 weight portion F part.

4, active component as claimed in claim 3, it is characterized in that suanzaoren decoction soaks 30min with 10 times of water gagings after, steam distillation 1h hour, collect distillate, filter, the dregs of a decoction are collected distillate with 8 times of water gaging steam distillation 0.4h, filtration; Merging filtrate and distillate get filtrate I and distillate I; Filtrate I continues to be distilled to 4/5 o'clock of cumulative volume, distillate II and filtrate I remainder-filtrate II; Distillate II redistillation stopped distillation to 1/2 o'clock of original volume; Heavily steam the liquid boiling point and be 30 ℃～60 ℃ petroleum ether extraction, reclaim benzinum, A volatile oil part; When filtrate II is evaporated to crude drug heavy with volume ratio be 1: 1, add 95% ethanol and transfer to alcoholic degree and be 60%, 4 ℃ and place 24h, suction filtration, mother liquor I and precipitation I; Mother liquor I continues to add ethanol and transfers to alcoholic degree and be 80%, 4 ℃ and place 24h, suction filtration, mother liquor II and precipitation II; Precipitation I and precipitation II merge, and get 285.40 weight portion B polysaccharide parts; Mother liquor I and mother liquor II merge, be evaporated to crude drug heavy with volume ratio be 4: 1, do not have the alcohol flavor, extracted with diethyl ether, the ether extraction liquid and the aqueous solution; Ether extraction liquid recovered under reduced pressure ether gets 3.71 weight portion C extracted with diethyl ether part parts; The aqueous solution be concentrated into crude drug heavy with volume ratio be 10: 1, last sample; Difference water, 30% ethanol, 50% ethanol, 95% ethanol elution, column internal diameter 10cm, the high 50cm of post; Inspection is known, and merges same area; Water elution part, 30% ethanol eluate hydrochloric acid-magnesium powder examine the knowledge reaction negative and aceticanhydride-strong sulfuric acid response is positive partly, 50% ethanol merges, and drying under reduced pressure gets 330.51 weight portion D saponin(es and aglycon part; The reacting positive part is known in 30% ethanol eluate hydrochloric acid-magnesium powder inspection, and drying under reduced pressure gets 5.41 weight portion E flavones parts; 95% ethanol eluate, recovered under reduced pressure gets 1.62 weight portion F parts.

5, as the described active component of claim 1-4, it is characterized in that it is to adopt anisaldehyde-strong sulfuric acid response that described inspection is known, the Molish reaction, aceticanhydride-strong sulfuric acid response, hydrochloric acid-magnesium powder react points out the composition that contains respectively.

6, active component as claimed in claim 5 is characterized in that also can adding pharmaceutically acceptable dressing or excipient.

7, active component as claimed in claim 5 is characterized in that can be made into clinical acceptable forms.

8, active component as claimed in claim 7 is characterized in that the formulation that can be made into is a kind of in the middle of tablet, capsule, pill, granule, supensoid agent, dripping pill, oral liquid, injection, aerosol, suppository or the subcutaneous administration formulation.

9, the application of active component as claimed in claim 5 in preparation treatment anxiety disorder medicine.

10, the application of suanzaoren decoction in preparation treatment anxiety disorder medicine.