CN101293063A

CN101293063A - Composition for treating climacteric syndrome, preparation and quality control method thereof

Info

Publication number: CN101293063A
Application number: CNA2007100987719A
Authority: CN
Inventors: 付立家; 付建家
Original assignee: Beijing Asia East Bio Pharmaceutical Co Ltd
Current assignee: Beijing rich church Pharmaceutical Technology Co., Ltd.
Priority date: 2007-04-26
Filing date: 2007-04-26
Publication date: 2008-10-29
Anticipated expiration: 2027-04-26
Also published as: CN101293063B

Abstract

The invention discloses a pharmaceutical composition for treating women menopausal syndrome, a preparation method and a quality control method. The pharmaceutical composition of the invention is composed of pharmaceutical raw materials of glossy privet fruit, palmleaf raspberry fruit, south dodder seed, tuber fleeceflower root, cortex lycii radicis, ladybell root, dwarf lilyturf tuber, baical skullcap root, rehmannia root, white paeony root, red paeony, Chinese angelica, nacre and blackend swallowwort root; the preparation method is that: the pharmaceutical raw materials are selected, ground into fine powder, screened and evenly mixed; every 100g of powder is added by 42g of refining honey and appropriate amount of water, pill making and low-temperature drying are carried out, thus preparing the pharmaceutical composition. The preparation method adopts high performance liquid chromatography to carry out the content measurement on paeoniflorin. The pharmaceutical composition of the invention has good efficacy for treating women menopausal syndrome.

Description

The composition and method of making the same and the method for quality control of treatment climacteric syndrome

Technical field

The present invention relates to a kind of Chinese medicine composition and preparation method thereof and method of quality control, particularly relate to a kind of Chinese medicine composition for the treatment of climacteric syndrome and preparation method thereof and method of quality control.

Background technology

Climacteric syndrome is meant the women before and after menopause, because the ovarian function decline causes the endocrine disturbance, the a series of syndrome that the autonomic nervous dysfunction produced, as hectic fever, flushing is perspired, lather, blahs aypnia, mood disorders sometimes, being happy and angry uncertainly, for no reason cry and laugh at, have like the insane.The traditional Chinese medical science is thought the generation of primary disease, be since the women over seventy-seven, the kidney qi degradation, dash appoint lose empty, asthenia of essence and blood, or because of depressed emotion, the dark consumption of battalion, cause the imbalance of YIN and YANG of kidney, and then influence the heart, liver, all dysfunctions of spleen, thereby all signs of climacteric syndrome occur.

Doctor trained in Western medicine adopts complementing estrogen when this type of disease of treatment, because climacteric syndrome goes down or loses because of women's ovary merit palace and causes, so, if at this moment complementing estrogen replaces the estrogen of human body shortage to play a role, it can slow down or cure fully a series of symptoms that climacteric women causes because of estrogen deficiency.By can " recovering " function of ovary for women's complementarity hormone, thereby alleviate involutional various symptom.But some women can not use controversies in hormone replacement in the elderly as hysteromyoma etc. because of various reasons, and patients with uterine myoma uses estrogen might become cancer.The Western medicine that other alleviates menopause syndrome also all can have side effects to patient body, can not life-time service.

This type of disease of Chinese traditional treatment generally speaking is to be guiding principle with kidney yin, kidney yang dysequilibrium, and dialectical main points are that yin-yang attribute will be distinguished.Aspect the selection medicine, to have the medicine of enriching yin and nourishing kidney, benefiting QI and nourishing blood, promoting blood flow to regulate menstruation, the spleen invigorating effect of drying, very useful to involutional women, can promote the endocrine regulation of climacteric women, help health to set up new balance rapidly, recover normal physiological status.As enriching yin and nourishing kidney, can improve the unsettled situation of suspection that climacteric women occurs; Promoting blood flow to regulate menstruation, benefiting QI and nourishing blood are to pointed therapeutical effect of disease such as this women's of climacteric easy perspiration, hectic fever, insomnia and dreamful sleep, dizziness headaches.Chinese medicine is aspect the treatment climacteric syndrome, and not only curative effect is relatively good, and has avoided using the adverse consequences that estrogen brought.

Summary of the invention

The object of the invention is to provide a kind of Chinese medicine composition for the treatment of climacteric syndrome;

The object of the invention also is to provide a kind of Chinese medicine composition preparation method for the treatment of climacteric syndrome;

The object of the invention also is to provide a kind of method of quality control for the treatment of the Chinese medicine composition of climacteric syndrome.

The present invention seeks to be achieved through the following technical solutions:

The Chinese medicine composition of treatment climacteric syndrome of the present invention is to be made by the crude drug of following weight ratio:

Fructus Ligustri Lucidi (wine is processed) 10-60 weight portion Fructus Rubi 10-50 weight portion

Semen Cuscutae 10-50 weight portion Radix Polygoni Multiflori (black soya bean wine is processed) 10-50 weight portion

Cortex Lycii 10-60 weight portion Radix Adenophorae 10-60 weight portion

Radix Ophiopogonis 10-50 weight portion Radix Scutellariae 10-60 weight portion

Radix Rehmanniae 10-60 weight portion Radix Paeoniae Alba 20-100 weight portion

Radix Paeoniae Rubra 10-60 weight portion Radix Angelicae Sinensis 10-50 weight portion

Concha Margaritifera 20-100 weight portion Radix Cynanchi Atrati 10-60 weight portion;

The above-mentioned raw materials optimum ratio is:

Fructus Ligustri Lucidi (wine is processed) 40 weight portion Fructus Rubies 30 weight portions

Semen Cuscutae 30 weight portion Radix Polygoni Multiflori (black soya bean wine is processed) 30 weight portions

Cortex Lycii 30 weight portion Radix Adenophoraes 30 weight portions

Radix Ophiopogonis 20 weight portion Radix Scutellariaes, 30 weight portions

The Radix Rehmanniae 30 weight portion Radix Paeoniae Albas 60 weight portions

Radix Paeoniae Rubra 30 weight portion Radix Angelicae Sinensis 30 weight portions

Concha Margaritifera 60 weight portion Radix Cynanchi Atratis 40 weight portions;

The Chinese medicine composition of treatment climacteric syndrome of the present invention can be made by the crude drug of following weight ratio:

Cortex Lycii 10-60 weight portion Radix Adenophorae 10-60 weight portion

Radix Ophiopogonis 10-50 weight portion Radix Scutellariae 10-60 weight portion

Radix Rehmanniae 10-60 weight portion Radix Paeoniae Alba 20-100 weight portion

Concha Margaritifera 20-100 weight portion Radix Cynanchi Atrati 10-60 weight portion

Rhizoma Anemarrhenae 10-60 weight portion Carapax Et Plastrum Testudinis 10-60 weight portion

Caulis Spatholobi 20-100 weight portion Fructus Lycii 10-50 weight portion;

The above-mentioned raw materials optimum ratio is:

Cortex Lycii 30 weight portion Radix Adenophoraes 30 weight portions

Radix Ophiopogonis 20 weight portion Radix Scutellariaes, 30 weight portions

The Radix Rehmanniae 30 weight portion Radix Paeoniae Albas 60 weight portions

Concha Margaritifera 60 weight portion Radix Cynanchi Atratis 40 weight portions

The Rhizoma Anemarrhenae 30 weight portion Carapax Et Plastrum Testudiniss 15 weight portions

Caulis Spatholobi 50 weight portion Fructus Lycii 20 weight;

Cortex Lycii 10-60 weight portion Radix Adenophorae 10-60 weight portion

Radix Ophiopogonis 10-50 weight portion Radix Scutellariae 10-60 weight portion

Radix Rehmanniae 10-60 weight portion Radix Paeoniae Alba 20-100 weight portion

Rhizoma Anemarrhenae 10-60 weight portion Carapax Et Plastrum Testudinis 10-50 weight portion

Caulis Spatholobi 20-100 weight portion Fructus Lycii 10-50 weight portion

Herba Dendrobii 10-60 weight portion Flos Chrysanthemi 10-60 weight portion

Herba Ecliptae 20-100 weight portion Folium Mori 10-50 weight portion

Semen Ziziphi Spinosae (stir-fry) 10 weight portions;

Compositions of the present invention technology adding adjuvant is routinely made clinical acceptable forms such as tablet, capsule, oral liquid, drop pill, spray, granule; Described adjuvant comprises solvent, disintegrating agent, correctives, antiseptic, coloring agent, binding agent, lubricant, substrate etc.

The preparation method of Chinese medicinal composition preparation of the present invention is:

Choose described crude drug, partly or entirely be ground into fine powder, all the other flavour of a drug are through water extraction, and extracting solution concentrates, and drying is ground into fine powder, with the flavour of a drug fine powder mix homogeneously of above-mentioned direct pulverizing, sieve again, add appropriate amount of auxiliary materials, make clinical adaptable preparation.

The preparation method of Chinese medicine composition pill preparation of the present invention can for:

Method for making: choose described crude drug, be ground into fine powder, cross the 70-90 mesh sieve, mixing; 90-110 weight portion Mel is in the time of 116-118 ℃, and refining density is 1.37, promptly gets refined honey; Per 100 weight portion powder add the water of refined honey 35-55 weight portion and 5-20 weight portion, general ball, and 60-80 ℃ of drying, promptly.

Pharmaceutical composition method of quality control of the present invention comprises following discrimination method and/or assay:

Differentiate:

A. get the preparation of quite described composition material medicine 11-13g, the 20ml that adds diethyl ether, supersound process 10-25 minute, filter, filtrate is concentrated into 1ml, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and supersound process 10-25 minute, filter, filtrate is concentrated into 1ml, makes control medicinal material solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 8-12: 1-2 petroleum ether (60～90 ℃)-ethyl acetate is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical light blue white fluorescent speckle;

B. get the preparation of quite described composition material medicine 3.5-4.5g, the 35ml that adds diethyl ether, supersound extraction 20-40 minute, filter, filtrate volatilizes, and the 2ml that adds diethyl ether makes dissolving, as need testing solution; Get Radix Rehmanniae control medicinal material 1.0g again, add the 20ml ether, supersound extraction 20-40 minute, filter, filtrate volatilizes, and the control medicinal material solution that every 1ml contains 1.0g is made in the dissolving that adds diethyl ether; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 7-11: 1-3: lower floor's solution of 1-2 chloroform-methanol-ammonia is developing solvent, launches, and takes out, and dries; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

C. get the preparation of quite described composition material medicine 1.5-2.5g, porphyrize, 20ml adds diethyl ether, supersound process 15-25 minute, filter, discard ether, residue is put water-bath Back stroke ether to the greatest extent, adds ethyl acetate 30ml, reflux 0.5-2 hour, take out, put coldly, filter, medicinal residues volatilize ethyl acetate, add methanol 20ml, supersound process 10-25 minute, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, add 3 of strong ammonia solutions again, filter, filtrate adds 3 of hydrochloric acid, and is centrifugal, abandoning supernatant, precipitation adds methanol 2ml makes dissolving, filters, and filtrate is as test sample; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 4-7: 2-5: 1-2: 1-2 ethyl acetate-butanone-formic acid-water is developing solvent, launches, and takes out, and dries, and spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

According to high effective liquid chromatography for measuring: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 8-12: the 85-95 acetonitrile-water is a mobile phase; The detection wavelength is 230nm; The preparation of reference substance solution: precision takes by weighing peoniflorin reference substance 5mg, puts in the 100ml measuring bottle, adds dissolve with methanol and is diluted to scale, shakes up, promptly; The preparation of need testing solution: get the preparation of quite described composition material medicine 3-4.5g, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug claims to decide weight, supersound process 25-40 minute, take out, put coldly, claim to decide weight again, supply the weight that subtracts mistake with methanol, filter, precision is measured subsequent filtrate 10ml, puts in the 50ml volumetric flask and is diluted to scale with methanol, shake up, promptly; Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

Pharmaceutical composition method of quality control of the present invention is preferably as follows discrimination method and/or assay:

Differentiate:

A. get the preparation of quite described composition material medicine 11-13g, the 20ml that adds diethyl ether, supersound process 15 minutes filters, and filtrate is concentrated into 1ml, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and supersound process 15 minutes filters, and filtrate is concentrated into 1ml, makes control medicinal material solution; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 9: 1 petroleum ether (60～90 ℃)-ethyl acetates, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical light blue white fluorescent speckle;

B. get the preparation of quite described composition material medicine 3.5-4.5g, the 35ml that adds diethyl ether, supersound extraction 30 minutes filters, and filtrate volatilizes, and the 2ml that adds diethyl ether makes dissolving, as need testing solution; Get Radix Rehmanniae control medicinal material 1.0g again, add the 20ml ether, supersound extraction 30 minutes filters, and filtrate volatilizes, and the control medicinal material solution that every 1ml contains 1.0g is made in the dissolving that adds diethyl ether; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with lower floor's solution of 8: 2: 1 chloroform-methanol-ammonia, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

C. get the preparation of quite described composition material medicine 1.5-2.5g, porphyrize, 20ml adds diethyl ether, supersound process 20 minutes filters, and discards ether, residue is put water-bath Back stroke ether to the greatest extent, adds ethyl acetate 30ml, reflux 1 hour, take out, put coldly, filter, medicinal residues volatilize ethyl acetate, add methanol 20ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, add 3 of strong ammonia solutions again, filter, filtrate adds 3 of hydrochloric acid, and is centrifugal, abandoning supernatant, precipitation adds methanol 2ml makes dissolving, filters, and filtrate is as test sample; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 5: 3: 1: 1 ethyl acetate-butanone-formic acid-water is developing solvent, launches, and takes out, and dries, spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

According to high effective liquid chromatography for measuring: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 10: 90 acetonitrile-waters are mobile phase; The detection wavelength is 230nm; The preparation of reference substance solution: precision takes by weighing peoniflorin reference substance 5mg, puts in the 100ml measuring bottle, adds dissolve with methanol and is diluted to scale, shakes up, promptly; The preparation of need testing solution: get the preparation of quite described composition material medicine 3-4.5g, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug claims to decide weight, power 250W, frequency 40KHz supersound process 30 minutes is taken out, and puts cold, claim again to decide weight, supply the weight that subtracts mistake, filter with methanol, precision is measured subsequent filtrate 10ml, put in the 50ml volumetric flask and be diluted to scale, shake up, promptly with methanol; Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

The present composition has good drug effect, compares the easypro sheet of existing preparation climacteric and shows good drug effect.Be used for climacteric syndrome clinically, the menoxenia that the hepatic and renal YIN deficiency causes, the hectic fever hyperhidrosis, insomnia forgetfulness, susceptible to lose temper due to restlessness, dizziness and tinnitus, dry throat and mouth, aching and soreness in limb, disease curative effects such as arthralgia are clear and definite.

The method of quality control of Chinese medicine composition provided by the present invention, be by obtaining behind the creative experiment sieving of big measuring, pass through screening in the discrimination method to sample treatment, the selection of developing solvent, make and differentiate that specificity is fine, and method is economic and practical, the result is quick, and can both use different lamellaes.Pass through screening in the content assaying method to the test sample processing method, the selection of developing solvent, make content assaying method effectivelyly to carry out quality control, and will compare more stable that product that additive method measures shows on pharmacological effect with the product that this method is measured to product.

Following experimental example and embodiment are used to further specify but are not limited to the present invention.

Experimental example 1 test of pesticide effectiveness

Experiment medicine: medicine 1 (being prepared into pill) by embodiment 1 method

Fructus Ligustri Lucidi (wine is processed) 40g Fructus Rubi 30g Semen Cuscutae 30g

Radix Polygoni Multiflori (black soya bean wine is processed) 30g Cortex Lycii 30g Radix Adenophorae 30g

Radix Ophiopogonis 20g Radix Scutellariae 30g Radix Rehmanniae 30g

Radix Paeoniae Alba 60g Radix Paeoniae Rubra 30g Radix Angelicae Sinensis 30g

Concha Margaritifera 60g Radix Cynanchi Atrati 40g;

Medicine 2 (being prepared into pill) by embodiment 2 methods

Radix Ophiopogonis 20g Radix Scutellariae 30g Radix Rehmanniae 30g

Radix Paeoniae Alba 60g Radix Paeoniae Rubra 30g Radix Angelicae Sinensis 30g

Concha Margaritifera 60g Radix Cynanchi Atrati 40g Rhizoma Anemarrhenae 30g

Carapax Et Plastrum Testudinis 15g Caulis Spatholobi 50g Fructus Lycii 20g;

Medicine 3 (being prepared into pill) by embodiment 6 methods

Fructus Ligustri Lucidi (wine is processed) 30g Fructus Rubi 20g Semen Cuscutae 20g

Fructus Lycii 20g Radix Polygoni Multiflori (black soya bean wine is processed) 20g

Carapax Et Plastrum Testudinis 15g Cortex Lycii 30g Radix Adenophorae 30g

20g Semen Ziziphi Spinosae Radix Ophiopogonis (stir-fry) 10g Radix Scutellariae 30g

Radix Rehmanniae 30g Radix Paeoniae Alba 60g Radix Paeoniae Rubra 30g

Radix Angelicae Sinensis 20g Caulis Spatholobi 60g Concha Margaritifera 60g

Herba Dendrobii 30g Flos Chrysanthemi 30g Herba Ecliptae 40g

Folium Mori 20g Radix Cynanchi Atrati 30g Rhizoma Anemarrhenae 30g

The commercially available climacteric of the positive control medicine sheet that relaxes

(1) nourishing the liver and kidney effect:

Get the Wistar rat and be divided into totally 6 groups of normal group, hepatic and renal YIN deficiency group, 1,2,3 groups of hepatic and renal YIN deficiency Drug therapys of the present invention and positive controls at random.10 of normal group, 16 of hepatic and renal YIN deficiency groups, medicine group of the present invention and positive controls, 15 every group.Hepatic and renal YIN deficiency group, medicine group of the present invention and positive controls are in experiment the 1st day and later per 3 days subcutaneous injection of carbon tetrachloride Oleum Arachidis hypogaeae semen (50%) 0.3ml/100g body weight, and hepatic and renal YIN deficiency group gavages the hepatic and renal YIN deficiency in experiment and equal moulding medicine (reserpine 0.015mg the 1st day every day, thyroxine 1mg/100g body weight), normal group irritates stomach for the equivalent normal saline, 21 days finish, medicine group of the present invention and the experiment of positive drug matched group gavaged medicine of the present invention and positive control medicine 0.5g/100g body weight, totally 20 days in second day.Six groups are all carried out ecological observation, finish after 21 days, cut open the belly, and heart extracting blood, and get main internal organs and do histopathologic examination.

1, serum total cholesterol, serum triglycerides, serum high-density LP cholesteryl ester adopt homemade enzyme rapid test method, enzyme process, Sodium phosphotungstate magnesium micromethod respectively.The result is as follows:

Group	The example number	T-CHOL (mg/dl)	Triglyceride (mg/dl)	HDL-C (mg/dl)
Group	The example number	T-CHOL (mg/dl)	Triglyceride (mg/dl)	HDL-C (mg/dl)	Normal group	10	82.64±10.01	63.272±13.56	55.174±8.341
Hepatic and renal YIN deficiency group	16	100.19±20.57	78.90±18.66	51.833±16.578	Normal group	10	82.64±10.01	63.272±13.56	55.174±8.341
Hepatic and renal YIN deficiency group	16	100.19±20.57	78.90±18.66	51.833±16.578	Medicine 1 of the present invention	15	68.57±27.39△ △ ^**	53.89±10.34△△ ^* ^*	65.31±16.326△
Medicine 2 of the present invention	15	65.14±15.32△ △ ^**	51.56±15.82△△ ^* ^*	68.74±15.589△△ ^*	Medicine 1 of the present invention	15	68.57±27.39△ △ ^**	53.89±10.34△△ ^* ^*	65.31±16.326△
Medicine 2 of the present invention	15	65.14±15.32△ △ ^**	51.56±15.82△△ ^* ^*	68.74±15.589△△ ^*	Medicine 3 of the present invention	15	73.02±17.13△ △ ^*	56.73±13.35△△ ^*	63.16±14.836△
Positive control	15	79.26±12.35△ △	60.43±21.38△△	58.28±26.973	Medicine 3 of the present invention	15	73.02±17.13△ △ ^*	56.73±13.35△△ ^*	63.16±14.836△

Compare △ △ P＜0.01, △ P＜0.05 with hepatic and renal YIN deficiency group; Compare with the positive drug matched group ^*P＜0.01, ^*P＜0.05

The result shows: medicine of the present invention and positive controls medicine have remarkable reduction effect to T-CHOL, monoglyceride content in the serum of hepatic and renal YIN deficiency rat, and HDL-C content is had raising.Relatively there were significant differences to hypercholesterolemia reducing and glyceric acid fat content and high density lipoprotein increasing cholesterol level for medicine of the present invention and positive controls medicine, better effects if.And between the medicine of the present invention, the effect of medicine 2 is better than medicine 1, and the effect of medicine 1 is better than medicine 3, but respectively organizes there was no significant difference.

2, serum lipid peroxide employing method is gone into Mu Shi thiobarbituricacid algoscopy.The results are shown in following table:

Group	The example number	Lipid peroxide content
Group	The example number	Lipid peroxide content	Normal group	10	5.68±0.259
Hepatic and renal YIN deficiency group	16	6.521±1.12	Normal group	10	5.68±0.259
Hepatic and renal YIN deficiency group	16	6.521±1.12	Medicine 1	15	4.237±0.86△△ ^*
Medicine 2	15	4.125±0.47△△ ^*	Medicine 1	15	4.237±0.86△△ ^*
Medicine 2	15	4.125±0.47△△ ^*	Medicine 3	15	4.448±0.35△△ ^*
The positive control medicine	15	4.834±0.67△△	Medicine 3	15	4.448±0.35△△ ^*

Compare △ △ P＜0.01 with hepatic and renal YIN deficiency group; Compare with the positive drug matched group ^*P＜0.05

The result shows: medicine group of the present invention and positive drug matched group and hepatic and renal YIN deficiency group comparison serum peroxyester matter content have utmost point significance to reduce, medicine group of the present invention and positive controls relatively have the significance influence, there is not significant difference between of the present invention group of medicine, but the effect of medicine 2 is better than medicine 1, and the effect of medicine 1 is better than medicine 3.

3, liver cholesterol content adopts to cut open the belly and gets 2 livers, wear into homogenate after, mix with chloroform one methanol (1: 1) and to extract the enzymatic assays cholesterol.The results are shown in following table:

Group	The example number	Liver sterin content (mg/dl)
Group	The example number	Liver sterin content (mg/dl)	Normal group	10	86.48±30.40
Hepatic and renal YIN deficiency group	16	162.0±31.64	Normal group	10	86.48±30.40
Hepatic and renal YIN deficiency group	16	162.0±31.64	Medicine 1	15	98.7±24.83△△ ^*
Medicine 2	15	93.6±24.83△△ ^*	Medicine 1	15	98.7±24.83△△ ^*
Medicine 2	15	93.6±24.83△△ ^*	Medicine 3	15	105.04±24.83△

The positive control medicine

15

124.04±24.83△

Compare △ △ P＜0.01, △ P＜0.05 with hepatic and renal YIN deficiency group; Compare with the positive drug matched group ^*P＜0.05

The result shows: medicine group of the present invention and positive drug matched group and hepatic and renal YIN deficiency group comparison liver cholesterol content have significance to reduce, medicine group of the present invention and positive controls relatively have the significance influence, there is not significant difference between of the present invention group of medicine, but the effect of medicine 2 is better than medicine 1, and the effect of medicine 1 is better than medicine 3.

(2) tranquilizing effect:

Get 50 of Kunming mouses, body weight 18-22g, the male and female dual-purpose is divided into 5 groups at random, 10 every group.Normal saline matched group (negative control group): give normal saline 0.2ml/10g and irritate stomach, every day 1 time, totally 3 days.Medicine group of the present invention and positive drug matched group (medicine 1, medicine 2, medicine 3, positive control medicine): get the drug medication solution of the present invention and the positive control medicine solution 0.2ml/10g that prepare and irritate stomach, every day 1 time, totally 3 days.Each is organized last and irritates 1h behind the stomach, after mice is put into the spontaneous activity instrument and adapts to 3 minutes, begins to measure the spontaneous activity number of times and sleep surpasses 2 hours the length of one's sleep.The results are shown in following table:

Group	Dosage (g/kg)	The spontaneous activity number of times	The length of one's sleep (min)
Group	Dosage (g/kg)	The spontaneous activity number of times	The length of one's sleep (min)	Negative control group	-	169.7±63.1	40.0±15.5
Medicine 1	1.5	113.7±61.2△△ ^*	72.4±23.3△△	Negative control group	-	169.7±63.1	40.0±15.5
Medicine 1	1.5	113.7±61.2△△ ^*	72.4±23.3△△	Medicine 2	1.5	102.6±57.7△△ ^*	77.8±25.0△△ ^*
Medicine 3	1.5	121.8±68.5△△	68.5±23.9△△	Medicine 2	1.5	102.6±57.7△△ ^*	77.8±25.0△△ ^*
Medicine 3	1.5	121.8±68.5△△	68.5±23.9△△	The positive control medicine	1.5	134.5±64.3△	63.8±19.7△△

Compare △ △ P＜0.01, △ P＜0.05 with negative control group; Compare with the positive drug matched group ^*P＜0.05

The result shows: medicine group of the present invention and positive drug matched group and negative control group comparison tranquilizing effect have significance to improve, medicine group of the present invention and positive controls relatively have significant difference, tranquilizing effect is that the effect of medicine 2 is better than medicine 1 between of the present invention group of medicine, and the effect of medicine 1 is better than medicine 3.

(3) effect of nourishing blood:

Get 90 of body weight 18～21g mices, female, evenly be divided into 9 groups at random, wherein make the virtual model of losing blood for 8 groups.9 treated animals afterbody before experiment is got blood, survey Hb earlier, the RBC level, 8 groups of difference of modeling type afterbody blood-letting 0.5ml, animal is lost blood, in lose blood back 24h again afterbody get blood and survey Hb, RBC observes the degree that causes blood deficiency, gavage big then respectively, low dose of pharmaceutical preparation 5g/g of the present invention, 1.7g/kg, be equivalent to 30 times of clinical consumption respectively, 10 times), positive control medicine (2.5g/kg, the dosage that is equivalent to be grown up 30 times) and normal saline (20ml/kg), the blank group gavages the normal saline with volume, administration every day 1 time, successive administration 7 days is in last 1 administration 2h, afterbody is got blood and is surveyed Hb again, RBC the results are shown in following table:

^*Expression and model group is than P＜0.05, ^*Expression and model group be than P＜0.01, △ represent with the positive control medicine than P＜0.05

The result shows: medicine of the present invention and positive control medicine all can significantly improve Hb, RBC level.The large and small dosage of medicine of the present invention all has remarkable result, medicine of the present invention and positive control medicine comparative effectiveness have significance to improve, illustrate that the invention medicine has the good effect of nourishing blood, there is not significant difference between medicine of the present invention, but the effect of its Chinese medicine 2 is better than medicine 1, and the effect of medicine 1 is better than medicine 3.

(4) collateral dredging effect:

Get 36 of rabbit, be divided into 6 groups at random, the auris dextra depilation, first group is the blank group; Second group only gives the wax thermotherapy at every rat right hind leg; Third and fourth, five groups give medicine 1, medicine 2, medicine 3 respectively, add the wax thermotherapy; The 6th group gives the positive control medicine, adds the wax thermotherapy.Every day 1 time, continuous 7 days.After 7 days auris dextra was placed-10 ℃ of ethanol freezing 2 minutes, and under anatomical lens, measured fixed point arteriole diameter, freeze postfixed point arteriole diameter before calculating is frozen and change.The results are shown in following table:

Group	The swelling percentage rate
Group	The swelling percentage rate	The blank group	46.7±10.3
Wax thermotherapy group	33.1±16.0	The blank group	46.7±10.3
Wax thermotherapy group	33.1±16.0	Medicine 1	16.8±8.2*
Medicine 2	15.2±10.1*	Medicine 1	16.8±8.2*
Medicine 2	15.2±10.1*	Medicine 3	19.2±6.4*
Positive control medicine+wax thermotherapy group	23.2±5.8	Medicine 3	19.2±6.4*

Annotate: * and wax thermotherapy group are than P＜0.05

The result shows: medicine of the present invention adds wax thermotherapy group and wax thermotherapy group relatively to be prevented arteriole due to freezing to shrink to have significance to improve, and the collateral dredging effect of medicine of the present invention is higher than the positive control medicine.

Experimental example 2 is differentiated screening experiment

1, the thin layer of Radix Angelicae Sinensis is differentiated

1. the investigation of need testing solution extraction time

Get bolus of drug 15g of the present invention, grind, the 20ml that adds diethyl ether, supersound process filters, and filtrate is concentrated into 1ml, as need testing solution.Other gets Radix Angelicae Sinensis control medicinal material 1g, and the 10ml that adds diethyl ether shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-ethyl acetate (9: 1) is developing solvent, launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.Relatively extract different time need testing solution and the reference substance solution color developing effect on lamellae, the results are shown in following table:

Ultrasonic time	5min	10min	15min	20min
Ultrasonic time	5min	10min	15min	20min	Color developing effect	Test sample be can't see colour developing	Test sample is glimmering	Test sample and contrast	Test sample and contrast

The fluorescence speckle, the colour developing of reference substance fluorescence speckle is fuzzyyer

The colour developing of hot spot point is fuzzyyer

It is clear that product fluorescence speckle all develops the color

As can be seen from the above table, when extraction time be 15min when above need testing solution and reference substance solution on lamellae, develop the color clearly, reached requirement of experiment.So consider from saving experimental period, select supersound extraction 15min.

2. the selection of developing solvent

According to the preparation method of need testing solution described in 1. and reference substance solution, preparation need testing solution and reference substance solution.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, launch with different developing solvents, take out, dry, put under the ultra-violet lamp (365nm) and inspect.Relatively use different developing solvents and launch, need testing solution and the reference substance solution expansion effect on lamellae the results are shown in following table:

Developing solvent	Petroleum ether (60～90 ℃)-ethyl acetate (7: 3)	Petroleum ether (60～90 ℃)-ethyl acetate (8: 2)	Petroleum ether (60～90 ℃)-ethyl acetate (9: 1)	Normal hexane-ethyl acetate (7: 3)	Normal hexane-ethyl acetate (8: 2)	Normal hexane-ethyl acetate (9: 1)
Developing solvent	Petroleum ether (60～90 ℃)-ethyl acetate (7: 3)	Petroleum ether (60～90 ℃)-ethyl acetate (8: 2)	Petroleum ether (60～90 ℃)-ethyl acetate (9: 1)	Normal hexane-ethyl acetate (7: 3)	Normal hexane-ethyl acetate (8: 2)	Normal hexane-ethyl acetate (9: 1)	Launch effect	Separate unclearly, launch weak effect.	Separate unclearly, launch weak effect.	Separate clearly, launch effective.	Separate unclearly, launch weak effect, interference is arranged.	Separate unclearly, interference is arranged.	Separate unclearly, launch weak effect, interference is arranged.

From above experimental result as can be seen, need testing solution and reference substance solution were launched effectively on lamellae when developing solvent was a petroleum ether (60～90 ℃)-ethyl acetate (9: 1), occurred separating unclear, and it is bad to launch effect, interference etc. is arranged, meet requirement of experiment.

3. negative control test

Get the negative sample that lacks Rhizoma Chuanxiong, prepare negative sample solution, launch the back and on control medicinal material solution correspondence position, corresponding speckle do not occur, illustrate that selected identification experiment specificity is strong according to need testing solution preparation method in the above-mentioned discrimination method.

2, the thin layer of Radix Rehmanniae is differentiated

1. the examination of need testing solution extraction time

Get bolus of drug 5g of the present invention, porphyrize, the 35ml that adds diethyl ether, supersound extraction filters, and filtrate volatilizes, and the 2ml that adds diethyl ether makes dissolving, as need testing solution.Get Radix Rehmanniae control medicinal material 1.0g again, make the control medicinal material solution that every 1ml contains 1.0g with method.According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with lower floor's solution of chloroform-methanol-ammonia (8: 2: 1), launch, take out, dry.Relatively extract little with time need testing solution and reference substance solution the color developing effect on lamellae, the results are shown in following table:

Ultrasonic time	15min	25min	35min	50min
Ultrasonic time	15min	25min	35min	50min	Color developing effect	Test sample be can't see the speckle of colour developing, and the colour developing of reference substance fluorescence speckle is fuzzyyer	The colour developing of test sample speckle is fuzzyyer	It is clear that test sample and reference substance speckle all develop the color	It is clear that test sample and reference substance speckle all develop the color

As can be seen from the above table, when extraction time be 35min when above need testing solution and reference substance solution on lamellae, develop the color clearly, reached requirement of experiment.

2. the selection of developing solvent

Drawing need testing solution and each 5 μ l of reference substance solution of method for preparing, put respectively on same silica gel g thin-layer plate, is developing solvent with lower floor's solution of chloroform-methanol-water and the different proportionings of chloroform-methanol-ammonia, launches, and taking-up is dried.Relatively extract different time need testing solution and the reference substance solution expansion effect on lamellae, the results are shown in following table:

Developing solvent	Chloroform-methanol-ammonia (7: 3: 1)	Chloroform-methanol-ammonia (8: 2: 1)	Chloroform-methanol-water ammonia (9: 1: 1)	Chloroform-methanol-water (8: 2: 1)	Chloroform-methanol-water (13: 6: 1)	Chloroform-methanol-water (16: 6: 1)
Developing solvent	Chloroform-methanol-ammonia (7: 3: 1)	Chloroform-methanol-ammonia (8: 2: 1)	Chloroform-methanol-water ammonia (9: 1: 1)	Chloroform-methanol-water (8: 2: 1)	Chloroform-methanol-water (13: 6: 1)	Chloroform-methanol-water (16: 6: 1)	Launch effect	Launch weak effect, separate unclear	Separate clearly, launch effective	Launch weak effect, the tail of taking off is arranged	Interference is arranged after the expansion	Launch weak effect, interference is arranged	Launch weak effect, interference is arranged

As can be seen from the above table, be developing solvent with chloroform-methanol-ammonia (8: 2: 1), each the speckle developing solvent on lamellae of need testing solution and reference substance solution is effective, tail, inferior separating effect, interference etc. do not occur taking off.

3. negative control test

Get the negative sample that lacks Radix Rehmanniae, prepare negative sample solution, launch the back and on control medicinal material solution correspondence position, corresponding speckle do not occur, illustrate that selected identification experiment specificity is strong according to need testing solution preparation method in the above-mentioned discrimination method.

3, the thin layer of Radix Scutellariae is differentiated

1. the preparation method of need testing solution

Method one, get bolus of drug 2g of the present invention, porphyrize, 20ml adds diethyl ether, supersound process 20 minutes filters, and discards ether, residue is put water-bath Back stroke ether to the greatest extent, adds ethyl acetate 30ml, reflux 1 hour, take out, put coldly, filter, medicinal residues volatilize ethyl acetate, add methanol 20ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, add 3 of strong ammonia solutions again, filter, filtrate adds 3 of hydrochloric acid, and is centrifugal, supernatant discarded night, precipitation adds methanol 2ml makes dissolving, filters, and filtrate is as test sample.

Method two, get bolus of drug 2g of the present invention, porphyrize adds methanol 20ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 10ml makes dissolving, as need testing solution.

Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with ethyl acetate-butanone-formic acid-water (5: 3: 1: 1) be developing solvent, launch, take out, dry, spray is with 1% ferric chloride alcoholic solution.Compare Different Extraction Method the test sample expansion and the color developing effect at molten night, the results are shown in following table:

Preparation method	Method 1	Method 2
Preparation method	Method 1	Method 2	Launch and color developing effect	The need testing solution principal spot is analysed clearly, and is noiseless.	The need testing solution principal spot is unclear analyses, and disturbs big.

As can be seen from the above table, employing method one preparation need testing solution after the expansion, has not only been eliminated the interference of other impurity, and more clear the analysing of principal spot colour developing, and the experiment accuracy is higher.

In method 1, compare the supersound extraction that adds diethyl ether in the different step, added the ethyl acetate reflux, extract, and added time of methanol supersound extraction, in method 2, compared the time that adds the methanol supersound extraction, the result shows that the selected experimental period of above experiment is reasonable, science, meets requirement of experiment.

The selection of 3. developing solvent proportioning

Draw each 5 μ l of need testing solution and reference substance solution, put respectively on same silica gel g thin-layer plate, the developing solvent with the different proportionings of ethyl acetate-butanone-formic acid-water launches, and takes out, and dries, and spray is with 1% ferric chloride alcoholic solution.Under the more different proportioning developing solvents, need testing solution and the reference substance solution expansion effect on lamellae the results are shown in following table:

The developing solvent proportioning

3∶5∶1∶1

4∶4∶1∶1

5∶3∶1∶1

6∶2∶1∶1

Launch effect

Launch weak effect, the test sample principal spot separates unclear analysing.

Launch effectively, the test sample principal spot separates analyses clearly.

Launch weak effect, the test sample principal spot separates unclear analysing.

As can be seen from the above table, the proportioning of developing solvent ethyl acetate-butanone-formic acid-water is 5: 3: 1: 1 o'clock, it is all good that need testing solution and reference substance solution principal spot launch effect, and test sample principal spot do not occur and separates unclear analysing, phenomenons such as interference are arranged, meet requirement of experiment.

3. negative control test

Get the negative sample that lacks Radix Scutellariae, prepare negative sample solution, launch the back and on control medicinal material solution correspondence position, corresponding speckle do not occur, illustrate that selected identification experiment specificity is strong according to need testing solution preparation method in the above-mentioned discrimination method.

The part thin layer that more than is Radix Angelicae Sinensis, Radix Rehmanniae, Radix Scutellariae in the medicine of the present invention is differentiated screening test, and empirical tests can effectively be controlled the quality of drug combination preparation of the present invention from the qualitative detection aspect, and the quality of drug combination preparation of the present invention is improved.

Experimental example 3 assay screening experiment

Adopt the content of paeoniflorin in the high-efficient liquid phase color popularize law mensuration medicine of the present invention, to improve quality determining method of the present invention, part test the results are shown in down:

1. the preparation of need testing solution

Get bolus of drug 4g of the present invention, put in the tool plug conical flask, the accurate methanol 50ml that adds, close plug claims to decide weight, power 250W, frequency 40KHz supersound process, take out, put coldly, claim to decide weight again, supply the weight that subtracts mistake with methanol, filter, precision is measured subsequent filtrate 10ml, puts in the 50ml volumetric flask and is diluted to scale with methanol, shake up, promptly.Compared different ultrasonic times, content of paeoniflorin in the need testing solution, the result is as follows:

Ultrasonic time	10 minutes	20 minutes	30 minutes	50 minutes
Ultrasonic time	10 minutes	20 minutes	30 minutes	50 minutes	Paeoniflorin content (mg/g)	1.2307	1.9232	2.5187	2.5517

As can be seen from the above table, supersound extraction can be extracted the peoniflorin in the medicine of the present invention fully in 30 minutes, so the preparation of need testing solution selects supersound extraction to get final product in 30 minutes.

2. the selection of mobile phase

Get test sample molten night, respectively with the mobile phase of acetonitrile-water with methanol-different proportionings of 0.05mol/L potassium dihydrogen phosphate, the separating effect at each peak in the need testing solution chromatogram relatively the results are shown in following table:

Proportion of mobile phase	Acetonitrile-water (30: 70)	Acetonitrile-water (20: 80)	Acetonitrile-water (10: 90)	Methanol-0.05mol/L potassium dihydrogen phosphate (50: 55)	Methanol-0.05mol/L potassium dihydrogen phosphate (40: 65)	Methanol-0.05mol/L potassium dihydrogen phosphate (30: 75)
Proportion of mobile phase	Acetonitrile-water (30: 70)	Acetonitrile-water (20: 80)	Acetonitrile-water (10: 90)	Methanol-0.05mol/L potassium dihydrogen phosphate (50: 55)	Methanol-0.05mol/L potassium dihydrogen phosphate (40: 65)	Methanol-0.05mol/L potassium dihydrogen phosphate (30: 75)	The chromatographic peak separating effect	Inferior separating effect has interference	Inferior separating effect has interference	Good separating effect, noiseless	Inferior separating effect has interference	Inferior separating effect has interference	Inferior separating effect has interference

As can be seen from the above table, disturbing does not appear in the good separating effect at each peak in test sample chromatogram when mobile phase is acetonitrile-water (10: 90), main peak.

3. the methodological study of content detection

To the detection method of content that medicine of the present invention adopted, carried out related side's science of law from aspects such as linear relationship, stability, precision, repeatability, the response rate and investigated, concrete grammar and result are as follows:

Detecting instrument (room temperature detection): Agilent 1100 type High Performance Liquid Chromatography posts: (Zorbax C184.6 * 150mm, 5 μ m) producer: Agilent Techologies Anjelen Sci. ﹠ Tech. Inc (China)

Mobile phase of acetonitrile and water (10: 90) detects wavelength: 230nm

Flow velocity: 1.0ml/min column temperature: room temperature

The reference substance source: peoniflorin is purchased lot number: the 0736-9913 in Nat'l Pharmaceutical ﹠ Biological Products Control Institute

Assay method: accurate respectively each the 10 μ l of negative controls, reference substance liquid and need testing solution that draw, inject chromatograph of liquid, measure, promptly.

(1) stability test reference substance solution respectively at preparing the back 0,2,4,6,12,24 hour, is measured in accordance with the law, and the result shows that it is basicly stable in 24 hours, the results are shown in following table:

(2) linear relationship is investigated and to be got reference substance solution (48.5 μ g/ml) and shake up, accurate respectively 2,4,6,8,10, the 12 μ l of absorption inject high performance liquid chromatograph, measure peak area, the results are shown in following table, and drawing standard curve, show that peoniflorin is linear between 0.097 μ g-0.582 μ g, its regression equation is:

Area＝1356.2714*Amt+7.7626(r＝0.9998)

(3) the accurate need testing solution 10 μ l that draw of precision test repeat sample introduction 5 times, try to achieve relative standard deviation＜2%, the results are shown in following table:

(4) the text method is pressed in repeatability test, gets five parts of same bolus of drug of the present invention, and every part is measured, and tries to achieve relative standard deviation＜2%, the results are shown in following table:

(5) the recovery test precision take by weighing known content bolus of drug 2.5g same of the present invention more respectively precision take by weighing peoniflorin reference substance 50mg, be configured to the solution of 0.5mg/ml, precision is measured 10ml, preparation method operation by above-mentioned need testing solution, measure its content, and calculate its response rate, measurement result sees the following form:

Tested number	Sampling amount (g)	Peoniflorin amount mg in the sample)	Add peoniflorin amount (mg)	Measure peoniflorin amount (mg)	The response rate (%)	Average recovery rate (%)	RSD(％)
Tested number	Sampling amount (g)	Peoniflorin amount mg in the sample)	Add peoniflorin amount (mg)	Measure peoniflorin amount (mg)	The response rate (%)	Average recovery rate (%)	RSD(％)	1	2.5023	6.5579	5.00	11.3209	95.26
2	2.6321	6.5212	5.00	11.1807	93.19			1	2.5023	6.5579	5.00	11.3209	95.26
2	2.6321	6.5212	5.00	11.1807	93.19			3	2.4983	6.5475	5.00	11.3790	96.63	95.13	1.3732
4	2.5432	6.6651	5.00	11.3991	94.68			3	2.4983	6.5475	5.00	11.3790	96.63	95.13	1.3732
4	2.5432	6.6651	5.00	11.3991	94.68			5	2.5119	6.5831	5.00	11.3781	95.90

(6) blank assay

Ratio according to drug prescription taste of Chinese medicine of the present invention, press oral liquid formulations technology, preparation does not contain the negative sample of the Radix Paeoniae Alba, Radix Paeoniae Rubra, according to preparation of need testing solution preparation method and detection, negative sample solution is that the identical retention time of peoniflorin reference substance place does not have chromatographic peak as a result, so negative noiseless.

By above methodology examination result as can be seen, its linear relationship of the content assaying method that medicine of the present invention adopted, stability, precision, repeatability etc. are all good, can effectively control drug quality of the present invention.Following embodiment all can realize the described effect of above-mentioned experimental example

The specific embodiment

Embodiment 1: pill

Radix Ophiopogonis 20g Radix Scutellariae 30g Radix Rehmanniae 30g

Radix Paeoniae Alba 60g Radix Paeoniae Rubra 30g Radix Angelicae Sinensis 30g

Concha Margaritifera 60g Radix Cynanchi Atrati 40g;

More than 14 flavors, be ground into fine powder, sieve (be meant whole energy by 80 mesh sieves), mixing, drug powder.Mel is in the time of 116-118 ℃, and refining density is 1.37, promptly gets refined honey; Every 100g powder adds refined honey 42g and water 8g, general ball, and low temperature (60-80 ℃) drying, promptly.

Embodiment 2: pill

Radix Ophiopogonis 20g Radix Rehmanniae 30g Radix Scutellariae 30g

Radix Paeoniae Alba 60g Radix Paeoniae Rubra 30g Radix Angelicae Sinensis 30g

Concha Margaritifera 60g Radix Cynanchi Atrati 40g Rhizoma Anemarrhenae 30g

Carapax Et Plastrum Testudinis 15g Caulis Spatholobi 50g Fructus Lycii 20g;

More than 18 flavors, be ground into fine powder, sieve (be meant whole energy by 80 mesh sieves), mixing, drug powder.Mel is in the time of 116-118 ℃, and refining density is 1.37, promptly gets refined honey; Every 100g powder adds refined honey 42g and water 8.5g, general ball, and low temperature (60-80 ℃) drying, promptly.

Embodiment 3: effervescent

Fructus Ligustri Lucidi (wine is processed) 50g Fructus Rubi 10g Semen Cuscutae 50g

Fructus Lycii 10g Radix Polygoni Multiflori (black soya bean wine is processed) 50g

Carapax Et Plastrum Testudinis 10g Cortex Lycii 50g Radix Adenophorae 20g

40g Semen Ziziphi Spinosae Radix Ophiopogonis (stir-fry) 10g Radix Scutellariae 60g

Radix Rehmanniae 20g Radix Paeoniae Alba 80g Radix Paeoniae Rubra 20g

Radix Angelicae Sinensis 50g Caulis Spatholobi 50g Concha Margaritifera 80g

Herba Dendrobii 20g Flos Chrysanthemi 60g Herba Ecliptae 30g

Folium Mori 20g Radix Cynanchi Atrati 50g Rhizoma Anemarrhenae 30g

More than 23 flavors, except that Carapax Et Plastrum Testudinis, Semen Ziziphi Spinosae, Concha Margaritifera, all the other flavour of a drug add 8 times of water gagings and decoct 2 times, each 1.5 hours, filter, merging filtrate, being concentrated into relative density is 1.30 (50～55 ℃), vacuum drying is ground into fine powder, and is standby.Carapax Et Plastrum Testudinis, Semen Ziziphi Spinosae, Concha Margaritifera powder are broken to more than 100 orders, sieve,, be divided into two equal portions with above-mentioned fine powder, an amount of low-substituted hydroxypropyl cellulose, xanthan gum mix homogeneously.Add 16.1% sodium bicarbonate mixing in the portion, make alkali grain, drying; Another part adds 19.3% citric acid and 0.05% sweetener mixing, makes acid particles, and drying with two kinds of dried granule mixings, sprays into flavouring agent, and the sealing certain hour gets final product.

Embodiment 4: drop pill

Fructus Ligustri Lucidi (wine is processed) 60g Fructus Rubi 20g Semen Cuscutae 50g

Radix Polygoni Multiflori (black soya bean wine is processed) 20g Cortex Lycii 60g Radix Adenophorae 20g

Radix Ophiopogonis 10g Radix Rehmanniae 20g Radix Scutellariae 50g

Radix Paeoniae Alba 90g Radix Paeoniae Rubra 20g Radix Angelicae Sinensis 50g

Concha Margaritifera 50g Radix Cynanchi Atrati 70g Rhizoma Anemarrhenae 20g

Carapax Et Plastrum Testudinis 55g Caulis Spatholobi 30g Fructus Lycii 50g;

More than 18 the flavor, be ground into fine powder, sieve mixing, fine powder and an amount of fused Macrogol 4000 mix homogeneously, 70 ℃ of insulations, through the water dropper of a certain size caliber, constant speed splashes in the liquid paraffin liquid coolant, the pill that will solidify formation takes out, the flush away liquid coolant, and drying.

Embodiment 5: soft capsule

Fructus Ligustri Lucidi (wine is processed) 60g Fructus Rubi 20g Semen Cuscutae 60g

Radix Polygoni Multiflori (black soya bean wine is processed) 25g Cortex Lycii 60g Radix Adenophorae 20g

Radix Ophiopogonis 60g Radix Scutellariae 20g Radix Rehmanniae 30g

Radix Paeoniae Alba 80g Radix Paeoniae Rubra 20g Radix Angelicae Sinensis 30g

Concha Margaritifera 90g Radix Cynanchi Atrati 40g;

More than 14 flavors, except that Carapax Et Plastrum Testudinis, Semen Ziziphi Spinosae, Concha Margaritifera, all the other flavour of a drug add 8 times of water gagings and decoct 2 times, each 1.5 hours, filter, merging filtrate concentrates, drying is ground into fine powder (more than 100 orders), and is standby.Carapax Et Plastrum Testudinis, Semen Ziziphi Spinosae, Concha Margaritifera powder are broken to more than 100 orders, sieve,,, be pressed into soft capsule evenly mixed with an amount of vegetable oil and Cera Flava heating with above-mentioned fine powder mix homogeneously.Soft capsule shell is prepared from by a certain percentage by gelatin, glycerol, water.

Embodiment 6:

Carapax Et Plastrum Testudinis 15g Cortex Lycii 30g Radix Adenophorae 30g

Radix Rehmanniae 30g Radix Paeoniae Alba 60g Radix Paeoniae Rubra 30g

Radix Angelicae Sinensis 20g Caulis Spatholobi 60g Concha Margaritifera 60g

Herba Dendrobii 30g Flos Chrysanthemi 30g Herba Ecliptae 40g

Folium Mori 20g Radix Cynanchi Atrati 30g Rhizoma Anemarrhenae 30g

Method for making: above 23 flavors are ground into fine powder, sieve (being meant that whole energy are by 80 mesh sieves), mixing.Mel is in the time of 116-118 ℃, and refining density is 1.37, promptly gets refined honey; Every 100g powder adds refined honey 42g and water 8.4g, general ball, and (60-80 ℃) drying is made 8700;

Differentiate:

(1) get this product, put microscopically and observe: the parenchyma taupe brown is to dark brown, and the many shrinkages of cell include brown nuclear shape thing; Nonglandular hair mostly is 3 cells, and the middle part cell is longer, and obvious verruca is arranged, the anxious point of apical cell and lacking; In the fiber surface similar round cell, contain siliceous of tiny circle, be arranged in rows; Bordered pit vessel is big; The fibre bundle peripheral cell contains prism of calcium oxalate and forms crystalline cellulose; Plant skin palisade cells 2 row, the outer row of interior row are long; It is faint yellow to plant the skin stone cell, the wavy bending of wall, and cell contains brown thing; The pale brown color of endotesta cell, surface sight rectangle or class are square, and the anticlinal wall beaded thickens; Fiber is faint yellow, fusiformis, and wall thickness, the hole ditch is thin; Fiber often is connected with the ray cell; Parenchyma cell contains calcium oxalate sand crystal; Nonglandular hair is unicellular, wall thickness, and lignify, the back vestiges that come off are like the stone cell shape;

(2) get this product 1g, grind, add dilute hydrochloric acid 20ml, promptly produce a large amount of bubbles, filter, filtrate shows the various identifications of calcium salt;

(3) get this product 15g, grind, the 20ml that adds diethyl ether, supersound process 15 minutes filters, and filtrate is concentrated into 1ml, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and supersound process 15 minutes filters, and filtrate is concentrated into 1ml, makes control medicinal material solution; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with petroleum ether (60～90 ℃)-ethyl acetate (9: 1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical light blue white fluorescent speckle;

(4) get this product 5g, porphyrize, the 35ml that adds diethyl ether, supersound extraction 30 minutes filters, and filtrate volatilizes, and the 2ml that adds diethyl ether makes dissolving, as need testing solution; Get Radix Rehmanniae control medicinal material 1.0g again, Same methodMake the control medicinal material solution that every 1ml contains 1.0g; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with lower floor's solution of chloroform-methanol-ammonia (8: 2: 1), launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(5) get this product 2g, porphyrize, the 20ml that adds diethyl ether, supersound process 20 minutes filters, discard ether, residue is put water-bath Back stroke ether to the greatest extent, adds ethyl acetate 30ml, and reflux 1 hour is taken out, put coldly, filter, medicinal residues volatilize ethyl acetate, add methanol 20ml, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add water 10ml makes dissolving, add 3 of strong ammonia solutions again, filter, filtrate adds 3 of hydrochloric acid, and is centrifugal, abandoning supernatant, precipitation adds methanol 2ml makes dissolving, filters, and filtrate is as test sample; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, (5: 3: 1: 1) be developing solvent, launch that taking-up is dried, spray was with 1% ferric chloride alcoholic solution with ethyl acetate-butanone-formic acid-water; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

According to high effective liquid chromatography for measuring: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-water (10: 90) is a mobile phase; The detection wavelength is 230nm; The preparation of reference substance solution: precision takes by weighing peoniflorin reference substance 5mg, puts in the 100ml measuring bottle, adds dissolve with methanol and is diluted to scale, shakes up, and promptly gets (containing peoniflorin 50 μ g among every 1ml); The preparation of need testing solution: get the content under this product content uniformity item, porphyrize, 4g decided in accurate title, put in the tool plug conical flask accurate methanol 50ml, the close plug of adding, claim to decide weight, supersound process (power 250W, frequency 40KHz) 30 minutes, take out, put coldly, claim to decide weight again, supply the weight that subtracts mistake with methanol, filter, precision is measured subsequent filtrate 10ml, put in the 50ml volumetric flask and be diluted to scale, shake up, promptly with methanol; Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly; This product contains peoniflorin (C for every bag ₂₃H ₂₈O ₁₁), must not be less than 6.0mg.

Function cures mainly: nourishing the liver and kidney, tranquillizing and allaying excitement, blood nourishing acupuncture-stimulating.Be used for climacteric syndrome, the menoxenia that the hepatic and renal YIN deficiency causes, hectic fever hyperhidrosis, insomnia forgetfulness, susceptible to lose temper due to restlessness, dizziness and tinnitus, dry throat and mouth, aching and soreness in limb, diseases such as arthralgia.

Claims

1, a kind of pharmaceutical composition for the treatment of climacteric syndrome is characterized in that the crude drug of this pharmaceutical composition consists of:

Cortex Lycii 10-60 weight portion Radix Adenophorae 10-60 weight portion

Radix Ophiopogonis 10-50 weight portion Radix Scutellariae 10-60 weight portion

Radix Rehmanniae 10-60 weight portion Radix Paeoniae Alba 20-100 weight portion

Concha Margaritifera 20-100 weight portion Radix Cynanchi Atrati 10-60 weight portion.

2, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug in this pharmaceutical composition consists of:

Cortex Lycii 30 weight portion Radix Adenophoraes 30 weight portions

Radix Ophiopogonis 20 weight portion Radix Scutellariaes, 30 weight portions

The Radix Rehmanniae 30 weight portion Radix Paeoniae Albas 60 weight portions

Concha Margaritifera 60 weight portion Radix Cynanchi Atratis 40 weight portions.

3, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug of this pharmaceutical composition consists of:

Cortex Lycii 10-60 weight portion Radix Adenophorae 10-60 weight portion

Radix Ophiopogonis 10-50 weight portion Radix Scutellariae 10-60 weight portion

Radix Rehmanniae 10-60 weight portion Radix Paeoniae Alba 20-100 weight portion

Caulis Spatholobi 20-100 weight portion Fructus Lycii 10-50 weight portion.

4, pharmaceutical composition as claimed in claim 3 is characterized in that the crude drug of this pharmaceutical composition consists of:

Cortex Lycii 30 weight portion Radix Adenophoraes 30 weight portions

Radix Ophiopogonis 20 weight portion Radix Scutellariaes, 30 weight portions

The Radix Rehmanniae 30 weight portion Radix Paeoniae Albas 60 weight portions

Caulis Spatholobi 50 weight portion Fructus Lycii 20 weight portions

5, pharmaceutical composition as claimed in claim 3 is characterized in that the crude drug of this pharmaceutical composition consists of:

Cortex Lycii 10-60 weight portion Radix Adenophorae 10-60 weight portion

Radix Ophiopogonis 10-50 weight portion Radix Scutellariae 10-60 weight portion

Radix Rehmanniae 10-60 weight portion Radix Paeoniae Alba 20-100 weight portion

Caulis Spatholobi 20-100 weight portion Fructus Lycii 10-50 weight portion

Herba Dendrobii 10-60 weight portion Flos Chrysanthemi 10-60 weight portion

Herba Ecliptae 20-100 weight portion Folium Mori 10-50 weight portion

Semen Ziziphi Spinosae (stir-fry) 10 weight portions.

6,, it is characterized in that this preparation of drug combination method is as the described pharmaceutical composition of claim 1-5:

Choose described crude drug, be ground into fine powder, cross the 70-90 mesh sieve, mixing; 90-110 weight portion Mel is in the time of 116-118 ℃, and refining density is 1.37, promptly gets refined honey; Per 100 weight portion powder add the water of refined honey 35-55 weight portion and 5-20 weight portion, general ball, and 60-80 ℃ of drying, promptly.

7,, it is characterized in that this method of quality control comprises following discrimination method and/or content assaying method as the method for quality control of the described pharmaceutical composition of claim 1-5:

Differentiate:

A. get preparation and be equivalent to described composition material medicine 11-13g, the 20ml that adds diethyl ether, supersound process 10-25 minute, filter, filtrate is concentrated into 1ml, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and supersound process 10-25 minute, filter, filtrate is concentrated into 1ml, makes control medicinal material solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 8-12: 1-2 petroleum ether (60～90 ℃)-ethyl acetate is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical light blue white fluorescent speckle;

B. get preparation and be equivalent to described composition material medicine 3.5-4.5g, the 35ml that adds diethyl ether, supersound extraction 20-40 minute, filter, filtrate volatilizes, and the 2ml that adds diethyl ether makes dissolving, as need testing solution; Get Radix Rehmanniae control medicinal material 1.0g again, add the 20ml ether, supersound extraction 20-40 minute, filter, filtrate volatilizes, and the control medicinal material solution that every 1ml contains 1.0g is made in the dissolving that adds diethyl ether; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 7-11: 1-3: lower floor's solution of 1-2 chloroform-methanol-ammonia is developing solvent, launches, and takes out, and dries; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

C. get preparation and be equivalent to described composition material medicine 1.5-2.5g, porphyrize, 20ml adds diethyl ether, supersound process 15-25 minute, filter, discard ether, residue is put water-bath Back stroke ether to the greatest extent, adds ethyl acetate 30ml, reflux 0.5-2 hour, take out, put coldly, filter, medicinal residues volatilize ethyl acetate, add methanol 20ml, supersound process 10-25 minute, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, add 3 of strong ammonia solutions again, filter, filtrate adds 3 of hydrochloric acid, and is centrifugal, abandoning supernatant, precipitation adds methanol 2ml makes dissolving, filters, and filtrate is as test sample; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 4-7: 2-5: 1-2: 1-2 ethyl acetate-butanone-formic acid-water is developing solvent, launches, and takes out, and dries, and spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay:

8, the method for quality control of pharmaceutical composition as claimed in claim 7 is characterized in that this method of quality control comprises following discrimination method and/or content assaying method:

Differentiate:

A. get preparation and be equivalent to described composition material medicine 11-13g, the 20ml that adds diethyl ether, supersound process 15 minutes filters, and filtrate is concentrated into 1ml, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, the 10ml that adds diethyl ether, and supersound process 15 minutes filters, and filtrate is concentrated into 1ml, makes control medicinal material solution; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 9: 1 petroleum ether (60～90 ℃)-ethyl acetates, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical light blue white fluorescent speckle;

B. get the quite described composition material medicine of preparation 3.5-4.5g, the 35ml that adds diethyl ether, supersound extraction 30 minutes filters, and filtrate volatilizes, and the 2ml that adds diethyl ether makes dissolving, as need testing solution; Get Radix Rehmanniae control medicinal material 1.0g again, add the 20ml ether, supersound extraction 30 minutes filters, and filtrate volatilizes, and the control medicinal material solution that every 1ml contains 1.0g is made in the dissolving that adds diethyl ether; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with lower floor's solution of 8: 2: 1 chloroform-methanol-ammonia, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

C. get the quite described composition material medicine of preparation 1.5-2.5g, porphyrize, 20ml adds diethyl ether, supersound process 20 minutes filters, and discards ether, residue is put water-bath Back stroke ether to the greatest extent, adds ethyl acetate 30ml, reflux 1 hour, take out, put coldly, filter, medicinal residues volatilize ethyl acetate, add methanol 20ml, supersound process 20 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, add 3 of strong ammonia solutions again, filter, filtrate adds 3 of hydrochloric acid, and is centrifugal, abandoning supernatant, precipitation adds methanol 2ml makes dissolving, filters, and filtrate is as test sample; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 5: 3: 1: 1 ethyl acetate-butanone-formic acid-water is developing solvent, launches, and takes out, and dries, spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay: