CN101716270A

CN101716270A - Method for detecting quality of traditional Chinese herbal medicament compound preparation for invigorating blood and regulating menses

Info

Publication number: CN101716270A
Application number: CN200910186825A
Authority: CN
Inventors: 刘红英; 陈云辉; 刘晓鹏
Original assignee: JIANGXI XINGANJIANG PHARMACEUTICAL CO Ltd
Current assignee: Jiangxi New Ganjiang Pharmaceutical Co., Ltd.
Priority date: 2009-12-28
Filing date: 2009-12-28
Publication date: 2010-06-02
Anticipated expiration: 2029-12-28
Also published as: CN101716270B

Abstract

The invention discloses a method for detecting the quality of traditional Chinese herbal medicament compound preparation for invigorating blood and regulating menses. The traditional Chinese herbal medicament compound preparation consists of fifteen species of crude medicament, such as elecampane, ligusticum wallichii, rhizome corydalis processed with rice vinegar, angelica, prepared rehmannia root, the root of common peony, red flower, the root of three-nerved spicebush, bighead atractylodes rhizome, the root of red-rooted salvia, processed cyperus, the seed of Chinese dodder, fructus evodiae processed with decoction of radix glycyrrhizae, herba lycopi and suberect spatholobus stem. The method consists of thin-layer identification for the rhizome corydalis, the elecampane, the ligusticum wallichii, the angelica, the red flower and the root of red-rooted salvia and content determination for the root of common peony, thereby controlling the quality of the traditional Chinese herbal medicament compound preparation better.

Description

A kind of quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation

Technical field

The present invention relates to the detection method of compound Chinese medicinal preparation, particularly a kind of detection method of regulating menstruation and activating blood compound Chinese medicinal preparation.

Background technology

The regulating menstruation and activating blood sheet records in " Ministry of Health of the People's Republic of China's ministry standard " " WS3-B-0614-91 regulating menstruation and activating blood sheet " (1991:150), prescription consists of the Radix Aucklandiae, Rhizoma Chuanxiong, Rhizoma Corydalis (vinegar system), Radix Angelicae Sinensis, Radix Paeoniae Rubra etc., has regulating menstruation and activating blood, the effect of promoting the circulation of QI to relieve pain, be used for menoxenia, gynaecopathias such as menalgia, clinical efficacy is remarkable, instant effect, market reaction is good.In the existing quality standard, the discriminating item except microscopical identification, only has the thin layer chromatography of corydalis tuber medicinal material to differentiate more there is not assay down; For the compound preparation of being made up of ten Chinese medicine of the five flavours materials, such discrimination method obviously is difficult to control, reflects its real quality.Therefore, be necessary to set up the strong discrimination method of more specificities, seek suitable content assaying method in addition, thereby control the quality of preparation better, guarantee patient's drug safety.

Summary of the invention

The quality determining method that the purpose of this invention is to provide a kind of regulating menstruation and activating blood compound Chinese medicinal preparation.

In order to solve the problems of the technologies described above, the technical solution used in the present invention is:

The crude drug of regulating menstruation and activating blood compound Chinese medicinal preparation of the present invention is composed as follows:

Radix Aucklandiae 10-80 weight portion Rhizoma Chuanxiong 10-80 weight portion Rhizoma Corydalis (processed with vinegar) 10-80 weight portion

Radix Angelicae Sinensis 30-240 weight portion Radix Rehmanniae Preparata 20-160 weight portion Radix Paeoniae Rubra 20-160 weight portion

Flos Carthami 15-120 weight portion Radix Linderae 15-120 weight portion Rhizoma Atractylodis Macrocephalae 15-120 weight portion

Radix Salviae Miltiorrhizae 30-240 weight portion Rhizoma Cyperi (processed) 30-240 weight portion Caulis Spatholobi 30-240 weight portion

Liquorice beverage Fructus Evodiae (processed) 5-40 weight portion Herba Lycopi 30-240 weight portion Semen Cuscutae 40-320 weight portion

The crude drug of regulating menstruation and activating blood compound Chinese medicinal preparation of the present invention is formed preferred:

The Radix Aucklandiae 41.67 weight portion Rhizoma Chuanxiongs 41.67 weight portion Rhizoma Corydalis (processed with vinegar) 41.67 weight portions

Radix Angelicae Sinensis 125 weight portion Radix Rehmanniae Preparata 83.34 weight portion Radix Paeoniae Rubra 83.34 weight portions

The Flos Carthami 62.5 weight portion Radixs Linderae 62.5 weight portion Rhizoma Atractylodis Macrocephalaes 62.5 weight portions

Radix Salviae Miltiorrhizae 125 weight portion Rhizoma Cyperi (processed) 125 weight portion Caulis Spatholobis 125 weight portions

Liquorice beverage Fructus Evodiae (processed) 20.83 weight portion Herba Lycopi 125 weight portion Semen Cuscutae 166.67 weight portions

Get the above-mentioned raw materials medicine, technology adds conventional adjuvant and is prepared into any preparation of acceptable clinically, as tablet, capsule, granule, powder, pill, slow releasing preparation or controlled release preparation routinely.

The preparation technology of Chinese medicine compound capsule of the present invention is:

(1) Radix Angelicae Sinensis powder with the Radix Aucklandiae, Rhizoma Chuanxiong, Rhizoma Corydalis (processed with vinegar) and 2/3rds weight portions of total weight part is broken into fine powder.

(2) will remain the Radix Angelicae Sinensis of 1/3rd weight portions and Radix Rehmanniae Preparata, Radix Paeoniae Rubra, Flos Carthami, the Radix Linderae, the Rhizoma Atractylodis Macrocephalae, Radix Salviae Miltiorrhizae, Rhizoma Cyperi (processed), liquorice beverage Fructus Evodiae (processed), Herba Lycopi, Caulis Spatholobi, the Semen Cuscutae of total weight part decocts with water twice, 3 hours for the first time, 2 hours for the second time, merge decocting liquid, filter, filtrate is condensed into thick paste; Wherein preferred each 10 times of water gagings decoctions, thick paste that it is 1.32-1.35 that filtrate decompression is condensed into 60 ℃ of relative densities of adding.

(3) thick paste that step 2 is made adds the fine powder that step 1 makes, and mixing is made granule, and drying incapsulates, every dress 0.38g.

Described Chinese medicine compound capsule is oral, one time 4,3 times on the one, to be converted to the crude drug amount be 15.501g to day clothes dosage.

The day clothes dosage of the different preparations of Chinese medicine compound of the present invention determines, and the day clothes dosage of different preparations to be converted to the crude drug amount be identical.So detection method of the present invention is that unit takes by weighing the detection preparation with daily dosage.

The detection method of compound Chinese medicinal preparation of the present invention comprises the authentication method of following I, II, III, IV and V:

I, get described compound Chinese medicinal preparation daily dosage 5/6, porphyrize adds methanol 30ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes dissolving, add strong ammonia solution and transfer to alkalescence, extract 2 times, each 10ml with the ether jolting, merge ether solution, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent with volume ratio, launches, take out, dry, put in the iodine vapor and smoked about 3 minutes, take out, after waving the iodine that adsorbs on the most plate, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

II, get described compound Chinese medicinal preparation daily dosage 5/6, porphyrize adds ethyl acetate 20ml, reflux 30 minutes filters, filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Aucklandiae control medicinal material 0.3g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

III, get described compound Chinese medicinal preparation daily dosage 5/12, porphyrize adds 30-60 ℃ of petroleum ether 10ml, supersound process 20 minutes filters, filtrate is as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material, each 0.5g of Radix Angelicae Sinensis control medicinal material, makes control medicinal material solution respectively with method; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that normal hexane-ethyl acetate of 9: 1 is developing solvent with volume ratio, launches, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

IV, get described compound Chinese medicinal preparation daily dosage 1/4, porphyrize adds n-butyl alcohol 20ml, supersound process 30 minutes filters, filtrate is as need testing solution; Other gets Flos Carthami control medicinal material 0.3g, decocts with water 30 minutes, filters, and filtrate is concentrated into 20ml, adds water saturated n-butyl alcohol 20ml, and jolting is extracted, and gets n-butanol layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launch, take out, dry, spray is 1: 1 20% sodium nitrite solution-30% sodium molybdate solution mixed liquor with volume ratio, it is clear to be heated to speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

V, get described compound Chinese medicinal preparation daily dosage 5/6, add water 20ml, ground 10 minutes, centrifugal, get supernatant and add hydrochloric acid accent pH value to 2, add the ethyl acetate jolting and extract 2 times, each 20ml, combined ethyl acetate liquid, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets the protocatechualdehyde reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that chloroform-acetone-formic acid of 60: 5: 2 is developing solvent with volume ratio, launches, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The quality determining method of compound Chinese medicinal preparation of the present invention also comprises the assay of following high performance liquid chromatography:

Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler, with volume ratio is that methanol-0.05mol/L potassium dihydrogen phosphate of 22: 78 is a mobile phase, the detection wavelength is 230nm, theoretical cam curve is calculated by the peoniflorin peak should be not less than 2000, and the separating degree of peoniflorin peak and adjacent peak is greater than 1.5;

The preparation of reference substance solution: it is an amount of that precision takes by weighing the peoniflorin reference substance, adds 50% methanol solution and make the solution that every 1ml contains 25 μ g, promptly;

The preparation of need testing solution: get 5/3 of the daily dosage of described compound Chinese medicinal preparation, porphyrize takes by weighing 5/18 of the daily approximately dosage of fine powder, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol solution 50ml that adds, close plug claims to decide weight, supersound process 40 minutes, put coldly, claim once more to decide weight, supply the weight that subtracts mistake with 50% methanol solution, shake up, filter, get subsequent filtrate, promptly;

Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, calculate, promptly;

1/12 of the daily dosage of described compound Chinese medicinal preparation contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 0.30mg.

The quality determining method of capsule of the present invention comprises the authentication method of following I, II, III, IV and V

I, get described capsule content 3.8g, porphyrize adds methanol 30ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes dissolving, add strong ammonia solution and transfer to alkalescence, extract 2 times, each 10ml with the ether jolting, merge ether solution, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent with volume ratio, launches, take out, dry, put in the iodine vapor and smoked about 3 minutes, take out, after waving the iodine that adsorbs on the most plate, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

II, get described capsule content 3.8g, porphyrize adds ethyl acetate 20ml, and reflux 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Aucklandiae control medicinal material 0.3g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

III, get described capsule content 2g, porphyrize adds 30-60 ℃ of petroleum ether 10ml, and supersound process 20 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material, each 0.5g of Radix Angelicae Sinensis control medicinal material, makes control medicinal material solution respectively with method; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that normal hexane-ethyl acetate of 9: 1 is developing solvent with volume ratio, launches, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

IV, get described capsule content 1.2g, porphyrize adds n-butyl alcohol 20ml, and supersound process 30 minutes filters, and filtrate is as need testing solution; Other gets Flos Carthami control medicinal material 0.3g, decocts with water 30 minutes, filters, and filtrate is concentrated into 20ml, adds water saturated n-butyl alcohol 20ml, and jolting is extracted, and gets n-butanol layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launch, take out, dry, spray is 1: 1 20% sodium nitrite solution-30% sodium molybdate solution mixed liquor with volume ratio, it is clear to be heated to speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

V, get described capsule content 3.8g, add water 20ml, ground 10 minutes, centrifugal, get supernatant and add hydrochloric acid accent pH value to 2, add the ethyl acetate jolting and extract 2 times, each 20ml, combined ethyl acetate liquid, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets the protocatechualdehyde reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that chloroform-acetone-formic acid of 60: 5: 2 is developing solvent with volume ratio, launches, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The quality determining method of capsule of the present invention also comprises the assay of following high performance liquid chromatography:

The preparation of need testing solution: get described capsule content 7.6g, porphyrize takes by weighing the about 1g of fine powder, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol solution 50ml that adds, close plug claims to decide weight, supersound process 40 minutes, put coldly, claim once more to decide weight, supply the weight that subtracts mistake with 50% methanol solution, shake up, filter, get subsequent filtrate, promptly;

Every of described capsule contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 0.30mg.

Regulating menstruation and activating blood tablet quality standard ((WS ₃-B-0614-91) in, tetrahydropalmatine serves as to extract solvent with the big benzene of toxicity in the tablet, the present invention extracts solvent, supersound process 30 minutes replaces the big benzene of toxicity with this, has reduced toxicity; Experimental result proves that this pre-treating method is feasible.In addition, thin layer chromatography adopts common silica gel g thin-layer plate in this standard, and the alkaline environment of diethylamine when keeping launching added in developing solvent, but this method is subjected to the temperature of environment, humidity effect big.The present invention adopts the silica gel g thin-layer plate with the preparation of 1% sodium hydroxide solution, with normal hexane-chloroform-methanol (10: 6: 1) is developing solvent, do not drip diethylamine, after the expansion, put in the iodine vapor smoked, put under the uviol lamp (365nm) and inspect the speckle rounding, good separating effect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color, the chromatograph feature is obvious; Thin-layer developing is affected by environment little, and experimental result is more stable.

Rhizoma Corydalis thin layer of the present invention is differentiated in (discrimination method I), the aqueous solution that obtains behind the methanol extract liquid evaporate to dryness, transfer to PH＞7 as long as add strong ammonia solution, can realize that tetrahydropalmatine is transferred to the ether phase of organic solvent from water, thereby in thin layer chromatography, inspect out the speckle of tetrahydropalmatine.

Thin layer of the present invention differentiates no matter II, III, V adopt commercially available prefabricated silica gel g thin-layer plate, still adopts the lamellae of manual shop system, can realize separating effect of the present invention; Thin layer is differentiated I, IV because binding agent is had specific (special) requirements, then adopts the lamellae of manual shop system.

With the existing standard ratio, the present invention has increased the discriminating of protocatechualdehyde in the Radix Aucklandiae, Rhizoma Chuanxiong, Radix Angelicae Sinensis, Flos Carthami control medicinal material and the Radix Salviae Miltiorrhizae, and content of paeoniflorin is measured, taken into account with two parts medical material that former powder is used as medicine and the water extracted immersing paste is used as medicine, thereby made this quality determining method more fully control the quality of described compound Chinese medicinal preparation.

Preferred below by the preparation process condition of the described capsule of description of test, and be the foundation that example illustrates quality determining method of the present invention with the capsule.

Experimental example 1 described capsule formulation technology preferred

Each decoction amount of water is not stipulated among the preparation technology of existing regulating menstruation and activating blood sheet, and the quality that how much directly influences preparation of amount of water.The present invention adopts regulating menstruation and activating blood tablet recipe list doubly to measure, and, as investigating index amount of water is investigated with dry extract yield and content of paeoniflorin.Experiment is divided into groups and be the results are shown in Table 1.

Table 1 amount of water investigation table

According to experimental result, dried cream yield and paeoniflorin content are all lower when decocting amount of water and being 8 times, and when 10 times of amount of water and 12 times, dried cream yield and paeoniflorin content are more or less the same, and according to the practical situation of saving energy and reduce the cost in producing, determine preferred 10 times of amount of water.

Because the Radix Aucklandiae, Radix Angelicae Sinensis, Rhizoma Chuanxiong contain more active volatile ingredient, and be used as medicine with former powder, so the medicinal liquid of water boiling and extraction is when concentrating, the relative density of clear paste can be thicker relatively, also be easy to wet granulation, so concentrate the preferred 1.32-1.35 of relative density (60 ℃) of thick paste.

During particle drying that wet granulation obtains, scatter and disappear in order to prevent the Radix Aucklandiae, Radix Angelicae Sinensis, the contained active volatile ingredient of Rhizoma Chuanxiong, baking temperature is controlled at 60 ℃.

The specificity test that experimental example 2 Rhizoma Corydalis thin layers are differentiated

Get the preparation method that other medical material of not containing Rhizoma Corydalis in the prescription puts down in writing to specifications and make negative sample, get test sample 3.8g, negative sample 3.4g, (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110726-200208) the described method of by specification is mixed with need testing solution, negative sample solution, reference substance solution to the tetrahydropalmatine reference substance.Draw need testing solution 10 μ l, reference substance solution 2 μ l put respectively on same silica gel g thin-layer plate with the preparation of 1% sodium hydroxide solution, with volume ratio is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent, launches, and takes out, dry, put in the iodine vapor smoked about 3 minutes, and took out, wave the iodine that adsorbs on the most plate after, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 1)

The specificity test that experimental example 3 Radix Aucklandiae thin layers are differentiated

Get the preparation method that other medical material of not containing the Radix Aucklandiae in the prescription puts down in writing to specifications and make negative sample, get test sample 3.8g, negative sample 3.4g, (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 120921-20405) the described method of 0.3g by specification is mixed with need testing solution, negative sample solution, control medicinal material solution to Radix Aucklandiae control medicinal material.Drawing each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 2)

The specificity test that experimental example 4 Radix Angelicae Sinensis and Rhizoma Chuanxiong thin layer are differentiated

Get the preparation method that other medical material of not containing Radix Angelicae Sinensis and Rhizoma Chuanxiong in the prescription puts down in writing to specifications and make negative sample, get test sample 2g, negative sample 1.5g, Radix Angelicae Sinensis control medicinal material (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 120927-200310), (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 120918-200406) the described method of each 0.5g by specification is mixed with need testing solution, negative sample solution, control medicinal material solution to the Rhizoma Chuanxiong control medicinal material.Draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that normal hexane-ethyl acetate of 9: 1 is developing solvent, launch, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 3)

The specificity test that experimental example 5 Flos Carthami thin layers are differentiated

Get the preparation method that other medical material of not containing Flos Carthami in the prescription puts down in writing to specifications and make negative sample, get test sample 1.2g, negative sample 1.2g, (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 120906-200308) the described method of 0.3g by specification is mixed with need testing solution, negative sample solution, control medicinal material solution to the Flos Carthami control medicinal material.Draw each 5 μ l of above-mentioned solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launches, and takes out, dry, spray is 1: 1 20% sodium nitrite solution-30% a sodium molybdate solution mixed liquor with volume ratio, and it is clear to be heated to the speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 4)

The specificity test that experimental example 6 Radix Salviae Miltiorrhizae thin layers are differentiated

Get the preparation method that other medical material of not containing Radix Salviae Miltiorrhizae in the prescription puts down in writing to specifications and make negative sample, get test sample 2g, negative sample 2g, (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110810-200205) the described method of by specification is mixed with need testing solution, negative sample solution, reference substance solution to the protocatechualdehyde reference substance.Draw each 5 μ l of above-mentioned solution, put respectively on same silica gel g thin-layer plate, with volume ratio is that chloroform-acetone-formic acid of 60: 5: 2 is developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color; In the negative sample chromatograph with the corresponding position of reference substance chromatograph on no corresponding speckle.Illustrate negative noiseless.(see figure 5)

Experimental example 7 assays

1) instrument and reagent

Tianjin, island LC-10AD _VPThe solvent delivery pump, SPD-10AD _VPDetector, CTO-10AD _VPColumn oven, PhenomenexC ₁₈Chromatographic column, Startorius BS electronic balance; Chromatographically pure methanol, other reagent is analytical pure; Peoniflorin reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110736-200220).

2) system suitability test

With octadecylsilane chemically bonded silica is filler, is that methanol-0.05mol/L potassium dihydrogen phosphate of 22: 78 is a mobile phase with volume ratio, flow velocity 0.5-1.0ml/min, and the detection wavelength is 230nm, column temperature 30-40 ℃.Analysis condition like this, other component in peoniflorin and the preparation all can reach baseline separation (seeing Fig. 6 and 7), and separating degree is greater than 1.5, and theoretical cam curve is calculated by the peoniflorin peak should be not less than 2000.

3) wavelength is selected

Get the peoniflorin reference substance solution, put the interscan of TU-1900UV Win4.0 type ultraviolet device, peoniflorin has an absworption peak (see figure 8) at the 230nm place as a result, so select 230nm for detecting wavelength.

4) selection of mobile phase

Be that methanol-0.05mol/L potassium dihydrogen phosphate of 40: 65 is a mobile phase with volume ratio in the prior art; Find with volume ratio to be that methanol-0.05mol/L potassium dihydrogen phosphate of 22: 78 is a mobile phase through overtesting, peoniflorin and other component in the preparation can be separated (seeing Fig. 6 and 7).

5) preparation of need testing solution

Under the situation of extracting solvent species and given volume, the leaching rate of supersound process time to peoniflorin in the test sample has material impact, so investigated the influence of different ultrasonic times to the test sample assay, test is divided into groups and be the results are shown in Table 2.

Determining of table 2 supersound process time

As seen from the above table, supersound extraction 40 minutes, the peoniflorin in the sample just can fully propose, so determine that ultrasonic time is 40 minutes.

6) blank assay

Get the preparation method that other medical material of not containing Radix Paeoniae Rubra in the prescription puts down in writing to specifications and make negative sample, the need testing solution preparation method of middle record is made negative sample solution to specifications again, according to the content assaying method test of description record, feminine gender is noiseless as a result.(see figure 9)

7) linear relationship is investigated

Precision takes by weighing peoniflorin reference substance 15.0mg, put in the 50ml volumetric flask, add 50% methanol solution and make dissolving, and be diluted to scale, shake up, accurate respectively the absorption in right amount is diluted to the reference substance solution of variable concentrations: 6.2 μ g/ml, 14.4 μ g/ml, 18.6 μ g/ml, 24.8 μ g/ml, 31.0 μ g/ml with 50% methanol solution.Accurate respectively each the 20 μ l of reference substance solution that go up variable concentrations that draw, Ji Zai chromatographiccondition is measured peak area to specifications, the results are shown in Table 3.

Table 3 linear relationship is investigated the result

With the peak area integrated value is vertical coordinate, and the amount of peoniflorin is an abscissa drawing standard curve (see figure 10), calculates regression equation:

Y＝2489267X-35225，R＝0.9996

Show that peoniflorin has the good linear relation in 0.124 μ g-0.620 μ g scope.

8) precision test

Accurate peoniflorin reference substance solution (18.6 μ g/ml) the 20 μ l that draw repeat sample introduction 5 times, measure peak area (seeing Table 4), and RSD=0.59% shows that this law precision is good.

Table 4 Precision test result

9) stability test

Get the about 1g of described capsule content, the accurate title, decide, Ji Zai method is made need testing solution to specifications, under the chromatographic condition of description record, accurate described need testing solution 20 μ l, the injection chromatograph of liquid drawn, measure once every 2 hours sample introductions, in 8 hours, peak area is consistent substantially, RSD=1.02%.Show that peoniflorin is stable in 8 hours in 50% methanol solution, the results are shown in Table 5.

Table 5 stability test result

10) replica test

Get respectively with 5 parts of a collection of described capsule contents, the accurate title, decide, and Ji Zai method is made need testing solution to specifications, under the chromatographic condition of description record, measure content of paeoniflorin, RSD=0.37%, show that this law repeatability is good, the results are shown in Table 6.

Table 6 replica test result

11) average recovery test

Get with 5 parts of the described capsule contents of a collection of known content, every part of about 0.5g accurate claims surely, and it is an amount of to add the peoniflorin reference substance respectively, Ji Zai method is made need testing solution to specifications, under the chromatographic condition of description record, measure content of paeoniflorin, calculate recovery rate, average recovery rate is 99.7%, RSD=1.57% shows that this law response rate is good, the results are shown in Table 7.

Table 7 recovery test result

Description of drawings

Fig. 1 Rhizoma Corydalis thin layer differentiates that wherein 1 is the tetrahydropalmatine reference substance, and 2,3,4 is three batch samples, 5 negative samples

Fig. 2 is that Radix Aucklandiae thin layer differentiates that wherein 1 is Radix Aucklandiae control medicinal material, and 2,3,4 is three batch samples, 5 negative samples

Fig. 3 is that Radix Angelicae Sinensis and Rhizoma Chuanxiong thin layer differentiate that wherein 1 is the Rhizoma Chuanxiong control medicinal material, and 2 is the Radix Angelicae Sinensis control medicinal material, and 3,4,5 is three batch samples, 6 negative samples

Fig. 4 is that the Flos Carthami thin layer differentiates that wherein 1 is the Flos Carthami control medicinal material, and 2,3,4 is three batch samples, 5 negative samples

Fig. 5 is that the Radix Salviae Miltiorrhizae thin layer differentiates that wherein 1 is the protocatechualdehyde reference substance, and 2,3,4 is three batch samples, 5 negative samples

Fig. 6 is a peoniflorin reference substance solution HPLC collection of illustrative plates

Fig. 7 is described capsule need testing solution HPLC collection of illustrative plates

Fig. 8 is peoniflorin reference substance solution UV scanning figure

Fig. 9 is a negative sample Solution H PLC collection of illustrative plates

Figure 10 is a peoniflorin standard solution canonical plotting

The specific embodiment

Following embodiment all can realize the effect of above-mentioned experimental example, but the present invention is not limited to described embodiment.

Embodiment 1 capsule of the present invention

Radix Aucklandiae 41.67g Rhizoma Chuanxiong 41.67g Rhizoma Corydalis (processed with vinegar) 41.67g

Radix Angelicae Sinensis 125g Radix Rehmanniae Preparata 83.34g Radix Paeoniae Rubra 83.34g

Flos Carthami 62.5g Radix Linderae 62.5g Rhizoma Atractylodis Macrocephalae 62.5g

Radix Salviae Miltiorrhizae 125g Rhizoma Cyperi (processed) 125g Caulis Spatholobi 125g

Liquorice beverage Fructus Evodiae (processed) 20.83g Herba Lycopi 125g Semen Cuscutae 166.67g

More than ten five tastes, the Radix Aucklandiae, Rhizoma Chuanxiong, Rhizoma Corydalis and 83.33g Radix Angelicae Sinensis powder are broken into fine powder, 41.67g Radix Angelicae Sinensis and Radix Rehmanniae Preparata etc. ten are added 9 times of water gagings simply decoct twice, 3 hours for the first time, 2 hours for the second time, collecting decoction, filter, filtrate decompression is condensed into the thick paste of 60 ℃ of relative density 1.32-1.35, adds above-mentioned fine powder mixing, makes granule, 60 ℃ of dryings, incapsulate, make 1000, the 0.38g/ grain.

Instructions of taking and day taking dose: oral, one time 4, one day three times.

Embodiment 2 tablets of the present invention

Radix Aucklandiae 20g Rhizoma Chuanxiong 20g Rhizoma Corydalis (processed with vinegar) 20g Radix Angelicae Sinensis 60g Radix Rehmanniae Preparata 40g

Radix Paeoniae Rubra 40g Flos Carthami 30g Radix Linderae 30g Rhizoma Atractylodis Macrocephalae 30g Radix Salviae Miltiorrhizae 60g

Rhizoma Cyperi (processed) 60g Caulis Spatholobi 60g liquorice beverage Fructus Evodiae (processed) 10g Herba Lycopi 60g Semen Cuscutae 80g

More than ten five tastes, the Radix Aucklandiae, Rhizoma Chuanxiong, Rhizoma Corydalis and 40g Radix Angelicae Sinensis powder are broken into fine powder, Radix Angelicae Sinensis 20g and Radix Rehmanniae Preparata etc. ten are added 9 times of water gagings simply decoct twice, 3 hours for the first time, 2 hours for the second time, collecting decoction, filter, filtrate is condensed into the thick paste of 60 ℃ of relative density 1.32-1.35, adds above-mentioned fine powder and adjuvant, mixing, make granule, drying is pressed into 600, the 0.3g/ sheet.

Instructions of taking and day taking dose: oral, one time 5, one day three times.

Embodiment 3 granules of the present invention

Radix Aucklandiae 15g Rhizoma Chuanxiong 30g Rhizoma Corydalis (processed with vinegar) 30g Radix Angelicae Sinensis 80g Radix Rehmanniae Preparata 60g

Radix Paeoniae Rubra 80g Flos Carthami 30g Radix Linderae 30g Rhizoma Atractylodis Macrocephalae 60g Radix Salviae Miltiorrhizae 70g

Rhizoma Cyperi (processed) 60g Caulis Spatholobi 80g liquorice beverage Fructus Evodiae (processed) 5g Herba Lycopi 60g Semen Cuscutae 120g

More than ten five tastes, according to common process, add appropriate amount of auxiliary materials, make granule, the 1.0g/ bag.

Instructions of taking and day taking dose: boiled water is taken after mixing it with water, one time one bag, one day three times.

Embodiment 4 water-honeyed pill of the present invention

Radix Aucklandiae 10g Rhizoma Chuanxiong 10g Rhizoma Corydalis (processed with vinegar) 15g Radix Angelicae Sinensis 40g Radix Rehmanniae Preparata 30g

Radix Paeoniae Rubra 25g Flos Carthami 20g Radix Linderae 15g Rhizoma Atractylodis Macrocephalae 20g Radix Salviae Miltiorrhizae 40g

Rhizoma Cyperi (processed) 40g Caulis Spatholobi 60g liquorice beverage Fructus Evodiae (processed) 5g Herba Lycopi 30g Semen Cuscutae 40g

More than ten five tastes, according to common process, add appropriate amount of auxiliary materials, make water-honeyed pill, the 6.0g/ bag.

Instructions of taking and day clothes dosage: one time one bag, one day three times.

Embodiment 5 controlled release agent of the present invention

Radix Aucklandiae 80g Rhizoma Chuanxiong 80g Rhizoma Corydalis (processed with vinegar) 80g Radix Angelicae Sinensis 200g

Radix Rehmanniae Preparata 100g Radix Paeoniae Rubra 80g Flos Carthami 100g Radix Linderae 95g

Rhizoma Atractylodis Macrocephalae 120g Radix Salviae Miltiorrhizae 240g Rhizoma Cyperi (processed) 200g Caulis Spatholobi 240g

Liquorice beverage Fructus Evodiae (processed) 20g Herba Lycopi 180g Semen Cuscutae 300g

More than ten five tastes, according to common process, add appropriate amount of auxiliary materials, make controlled release agent.

Instructions of taking: oral, one day one to secondary.

The capsule method of quality control of embodiment 6 embodiment 1 preparation

[discriminating]

I, get described capsule content 3.8g, porphyrize adds methanol 30ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes dissolving, add strong ammonia solution and transfer to alkalescence, extract 2 times, each 10ml with the ether jolting, merge ether solution, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent with volume ratio, launches, take out, dry, put in the iodine vapor and smoked about 3 minutes, take out, after waving the iodine that adsorbs on the most plate, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

II, get described capsule content 3.8g, porphyrize adds ethyl acetate 20ml, and reflux 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Aucklandiae control medicinal material 0.3g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

III, get described capsule content 2g, porphyrize adds 30-60 ℃ of petroleum ether 10ml, and supersound process 20 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material, each 0.5g of Radix Angelicae Sinensis control medicinal material, makes control medicinal material solution respectively with method; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that normal hexane-ethyl acetate of 9: 1 is developing solvent with volume ratio, launches, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

IV, get described capsule content 1.2g, porphyrize adds n-butyl alcohol 20ml, and supersound process 30 minutes filters, and filtrate is as need testing solution; Other gets Flos Carthami control medicinal material 0.3g, decocts with water 30 minutes, filters, and filtrate is concentrated into 20ml, adds water saturated n-butyl alcohol 20ml, and jolting is extracted, and gets n-butanol layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launch, take out, dry, spray is 1: 1 20% sodium nitrite solution-30% sodium molybdate solution mixed liquor with volume ratio, it is clear to be heated to speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D)

Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler, with volume ratio is that methanol-0.05mol/L potassium dihydrogen phosphate of 22: 78 is a mobile phase, the detection wavelength is 230nm, theoretical cam curve is calculated by the peoniflorin peak should be not less than 2000, and the separating degree of peoniflorin peak and adjacent peak is greater than 1.5.

The preparation of reference substance solution: it is an amount of that precision takes by weighing the peoniflorin reference substance, adds 50% methanol solution and make the solution that every 1ml contains 25 μ g, promptly.

The preparation of need testing solution: get described capsule content 7.6g, porphyrize takes by weighing the about 1g of fine powder, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol solution 50ml that adds, close plug claims to decide weight, supersound process 40 minutes, put coldly, claim once more to decide weight, supply the weight that subtracts mistake with 50% methanol solution, shake up, filter, get subsequent filtrate, promptly.

Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, calculate, promptly.

The method of quality control of the tablet of embodiment 7 embodiment 2 preparations

I, get described tablet 3.75g, porphyrize adds methanol 30ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes dissolving, add strong ammonia solution and transfer to alkalescence, extract 2 times, each 10ml with the ether jolting, merge ether solution, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent with volume ratio, launches, take out, dry, put in the iodine vapor and smoked about 3 minutes, take out, after waving the iodine that adsorbs on the most plate, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

II, get described tablet 3.75g, porphyrize adds ethyl acetate 20ml, and reflux 30 minutes filters, and filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Aucklandiae control medicinal material 0.3g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

III, get described tablet 1.9g, porphyrize adds 30-60 ℃ of petroleum ether 10ml, and supersound process 20 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material, each 0.5g of Radix Angelicae Sinensis control medicinal material, makes control medicinal material solution respectively with method; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that normal hexane-ethyl acetate of 9: 1 is developing solvent with volume ratio, launches, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

IV, get described tablet 1.1g, porphyrize adds n-butyl alcohol 20ml, and supersound process 30 minutes filters, and filtrate is as need testing solution; Other gets Flos Carthami control medicinal material 0.3g, decocts with water 30 minutes, filters, and filtrate is concentrated into 20ml, adds water saturated n-butyl alcohol 20ml, and jolting is extracted, and gets n-butanol layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launch, take out, dry, spray is 1: 1 20% sodium nitrite solution-30% sodium molybdate solution mixed liquor with volume ratio, it is clear to be heated to speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

V, get described tablet 3.75g, add water 20ml, ground 10 minutes, centrifugal, get supernatant and add hydrochloric acid and transfer pH value to 2, add the ethyl acetate jolting and extract 2 times, each 20ml, combined ethyl acetate liquid, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets the protocatechualdehyde reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that chloroform-acetone-formic acid of 60: 5: 2 is developing solvent with volume ratio, launches, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The preparation of need testing solution: get 10 in described tablet, porphyrize takes by weighing the about 1g of fine powder, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol solution 50ml that adds, close plug claims to decide weight, supersound process 40 minutes, put coldly, claim once more to decide weight, supply the weight that subtracts mistake with 50% methanol solution, shake up, filter, get subsequent filtrate, promptly.

Every in described tablet contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 0.23mg.

The method of quality control of the granule of embodiment 8 embodiment 3 preparations

[discriminating]

I, get described granule daily dosage 5/6, porphyrize adds methanol 30ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes dissolving, add strong ammonia solution and transfer to alkalescence, extract 2 times, each 10ml with the ether jolting, merge ether solution, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent with volume ratio, launches, take out, dry, put in the iodine vapor and smoked about 3 minutes, take out, after waving the iodine that adsorbs on the most plate, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

II, get described granule daily dosage 5/6, porphyrize adds ethyl acetate 20ml, reflux 30 minutes filters, filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Aucklandiae control medicinal material 0.3g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

III, get described granule daily dosage 5/12, porphyrize adds 30-60 ℃ of petroleum ether 10ml, supersound process 20 minutes filters, filtrate is as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material, each 0.5g of Radix Angelicae Sinensis control medicinal material, makes control medicinal material solution respectively with method; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that normal hexane-ethyl acetate of 9: 1 is developing solvent with volume ratio, launches, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

IV, get described granule daily dosage 1/4, porphyrize adds n-butyl alcohol 20ml, supersound process 30 minutes filters, filtrate is as need testing solution; Other gets Flos Carthami control medicinal material 0.3g, decocts with water 30 minutes, filters, and filtrate is concentrated into 20ml, adds water saturated n-butyl alcohol 20ml, and jolting is extracted, and gets n-butanol layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launch, take out, dry, spray is 1: 1 20% sodium nitrite solution-30% sodium molybdate solution mixed liquor with volume ratio, it is clear to be heated to speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

V, get described granule daily dosage 5/6, add water 20ml, ground 10 minutes, centrifugal, get supernatant and add hydrochloric acid accent pH value to 2, add the ethyl acetate jolting and extract 2 times, each 20ml, combined ethyl acetate liquid, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets the protocatechualdehyde reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that chloroform-acetone-formic acid of 60: 5: 2 is developing solvent with volume ratio, launches, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The preparation of need testing solution: get 5/3 of the daily metering of described granule, porphyrize takes by weighing 5/18 of the daily approximately dosage of fine powder, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol solution 50ml that adds, close plug claims to decide weight, supersound process 40 minutes, put coldly, claim once more to decide weight, supply the weight that subtracts mistake with 50% methanol solution, shake up, filter, get subsequent filtrate, promptly.

1/12 of the daily dosage of described granule contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 0.30mg.

The method of quality control of the water-honeyed pill of embodiment 9 embodiment 4 preparations

[discriminating]

I, get described water-honeyed pill daily dosage 5/6, porphyrize adds methanol 30ml, supersound process 30 minutes filters the filtrate evaporate to dryness, residue adds water 10ml makes dissolving, add strong ammonia solution and transfer to alkalescence, extract 2 times, each 10ml with the ether jolting, merge ether solution, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Other gets the tetrahydropalmatine reference substance, adds ethanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution 10 μ l, reference substance solution 2 μ l, putting respectively on same silica gel g thin-layer plate with 1% sodium hydroxide solution preparation, is that normal hexane-chloroform-methanol of 10: 6: 1 is developing solvent with volume ratio, launches, take out, dry, put in the iodine vapor and smoked about 3 minutes, take out, after waving the iodine that adsorbs on the most plate, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

II, get described water-honeyed pill daily dosage 5/6, porphyrize adds ethyl acetate 20ml, reflux 30 minutes filters, filtrate evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets Radix Aucklandiae control medicinal material 0.3g, shines medical material solution in pairs with legal system; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that toluene-methanol of 27: 1 is developing solvent with volume ratio, launches, take out, dry, spray is with 10% vanillin sulfuric acid solution, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

III, get described water-honeyed pill daily dosage 5/12, porphyrize adds 30-60 ℃ of petroleum ether 10ml, supersound process 20 minutes filters, filtrate is as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material, each 0.5g of Radix Angelicae Sinensis control medicinal material, makes control medicinal material solution respectively with method; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, putting respectively on same silica gel g thin-layer plate, is that normal hexane-ethyl acetate of 9: 1 is developing solvent with volume ratio, launches, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

IV, get described water-honeyed pill daily dosage 1/4, porphyrize adds n-butyl alcohol 20ml, supersound process 30 minutes filters, filtrate is as need testing solution; Other gets Flos Carthami control medicinal material 0.3g, decocts with water 30 minutes, filters, and filtrate is concentrated into 20ml, adds water saturated n-butyl alcohol 20ml, and jolting is extracted, and gets n-butanol layer, and evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with volume ratio is 7: 2: 3: ethyl acetate-formic acid of 0.4-water-methanol is developing solvent, launch, take out, dry, spray is 1: 1 20% sodium nitrite solution-30% sodium molybdate solution mixed liquor with volume ratio, it is clear to be heated to speckle colour developing at 105 ℃, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

V, get described water-honeyed pill daily dosage 5/6, add water 20ml, ground 10 minutes, centrifugal, get supernatant and add hydrochloric acid accent pH value to 2, add the ethyl acetate jolting and extract 2 times, each 20ml, combined ethyl acetate liquid, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets the protocatechualdehyde reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, putting respectively on same silica gel g thin-layer plate, is that chloroform-acetone-formic acid of 60: 5: 2 is developing solvent with volume ratio, launches, take out, dry, spray is with 5% ferric chloride alcoholic solution, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Claims

1. the quality determining method of crude drug regulating menstruation and activating blood compound Chinese medicinal preparation composed as follows,

Radix Aucklandiae 10-80 weight portion, Rhizoma Chuanxiong 10-80 weight portion, Rhizoma Corydalis (processed with vinegar) 10-80 weight portion, Radix Angelicae Sinensis 30-240 weight portion, Radix Rehmanniae Preparata 20-160 weight portion, Radix Paeoniae Rubra 20-160 weight portion, Flos Carthami 15-120 weight portion, Radix Linderae 15-120 weight portion, Rhizoma Atractylodis Macrocephalae 15-120 weight portion, Radix Salviae Miltiorrhizae 30-240 weight portion, Rhizoma Cyperi (processed) 30-240 weight portion, Caulis Spatholobi 30-240 weight portion, liquorice beverage Fructus Evodiae (processed) 5-40 weight portion, Herba Lycopi 30-240 weight portion, Semen Cuscutae 40-320 weight portion;

Described compound Chinese medicinal preparation is meant: get the above-mentioned raw materials medicine, technology adds conventional adjuvant and is prepared into any preparation of acceptable clinically routinely; It is characterized in that described quality determining method comprises the authentication method of following I, II, III, IV and V

2, the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 1 is characterized in that described detection method also comprises the assay of following high performance liquid chromatography:

3. the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 1 and 2 is characterized in that described compound Chinese medicinal preparation is the capsule for preparing by the following method:

(1) Radix Angelicae Sinensis powder with the Radix Aucklandiae, Rhizoma Chuanxiong, Rhizoma Corydalis (processed with vinegar) and 2/3rds weight portions of total weight part is broken into fine powder;

(2) will remain the Radix Angelicae Sinensis of 1/3rd weight portions and Radix Rehmanniae Preparata, Radix Paeoniae Rubra, Flos Carthami, the Radix Linderae, the Rhizoma Atractylodis Macrocephalae, Radix Salviae Miltiorrhizae, Rhizoma Cyperi (processed), liquorice beverage Fructus Evodiae (processed), Herba Lycopi, Caulis Spatholobi, the Semen Cuscutae of total weight part decocts with water twice, 3 hours for the first time, 2 hours for the second time, merge decocting liquid, filter, filtrate is condensed into thick paste;

4. the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 3 is characterized in that in the described capsule preparation process 2, adds 10 times of water gagings and decocts twice; The thick paste that it is 1.32-1.35 that described filtrate decompression is condensed into 60 ℃ of relative densities; Described particle drying temperature is 60 ℃.

5. the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 3, the detection method that it is characterized in that described capsule comprises the authentication method of following I, II, III, IV and V

6. the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 3, the detection method that it is characterized in that described capsule also comprises the assay of following photograph high performance liquid chromatography:

The preparation of need testing solution: get described capsule 's content 7.6g, porphyrize takes by weighing the about 1g of fine powder, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol solution 50ml that adds, close plug claims to decide weight, supersound process 40 minutes, put coldly, claim once more to decide weight, supply the weight that subtracts mistake with 50% methanol solution, shake up, filter, get subsequent filtrate, promptly;

7. the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 4, the detection method that it is characterized in that described capsule comprises the authentication method of following I, II, III, IV and V

8. the quality determining method of regulating menstruation and activating blood compound Chinese medicinal preparation according to claim 4, the detection method that it is characterized in that described capsule also comprises the assay of following photograph high performance liquid chromatography: