CN102928524A

CN102928524A - Quality detection method of milk-increasing paste and correlated preparation thereof

Info

Publication number: CN102928524A
Application number: CN2012102850196A
Authority: CN
Inventors: 潘杰; 黄进明; 洪绯; 于娟; 吴丽璇
Original assignee: Zhangzhou Pientzehuang Pharmaceutical Co Ltd
Current assignee: Zhangzhou Pientzehuang Pharmaceutical Co Ltd
Priority date: 2012-08-10
Filing date: 2012-08-10
Publication date: 2013-02-13

Abstract

The invention discloses a quality detection method of a milk-increasing paste and a correlated preparation thereof. The method comprises a content determination method and/or identification method. According to the content determination method, determination is carried out with a high-performance liquid chromatography method. 10-20:90-80 of acetonitrile-0.1% phosphoric acid is adopted as a mobile phase. A sample to be tested is subjected to a methanol ultrasonic extraction method. The method is simple, and peoniflorin can be completely extracted. Compared with current standards, a peoniflorin determination result is more accurate. As a result of methodological test, precision and reproducibility are good, and the method is feasible. Also, operation steps are simplified, such that practical application is facilitated. The milk-increasing paste and the correlated preparation thereof are prepared from the raw materials of, by weight: 100-160 parts of cowherb seed, 100-160 parts of rice paper plant pith, 130-190 parts of prepared rehmannia root, 50-90 parts of ligusticum, 90-130 parts of white peony root, 40-69 parts of Szechwan lovage rhizome, 100-160 parts of motherwort, and 100-160 parts of snakegourd root.

Description

A kind of quality determining method that increases emulsifiable paste and related preparations thereof

Technical field

The present invention relates to a kind of quality determining method of Chinese medicine preparation, be specifically related to a kind of quality determining method that increases emulsifiable paste and related preparations thereof, belong to medical technical field.

Background technology

It is the pure Chinese medicinal preparation of being produced by Zhongzhou Pianziguang Pharmaceutical Industry Co., Ltd. that the Pien Tze Huang board increases emulsifiable paste, and having enriches blood invigorates blood circulation, and the function of vein relaxing lactagogue is used for hypogalactia after delivery.Increasing emulsifiable paste records in national drug standards WS-5163 (B-0163)-2002, in the primary standard under the assay item with the front extracting method in the content of high effective liquid chromatography for measuring Paeoniflorin be: " sample thief 8g; accurately weighed adds water 20ml, fully stirs and makes dissolving; by D101 macroporous absorbent resin (8g wet method dress post; internal diameter 15mm), water 200ml wash-out discards water liquid; use ethanol 70ml wash-out again; collect ethanol, evaporate to dryness, residue add methyl alcohol 15ml, ultrasonic processing 15 minutes, filter, filtrate is transferred in the 25ml measuring bottle, adds methyl alcohol to scale, shake up, and get final product ".Because Paeoniflorin dissolves in the Diluted Alcohol, may cause the loss of Paeoniflorin and make the measurement result of Paeoniflorin not too accurate therefore discard eluent after in the method, washing with water, and the method is also too loaded down with trivial details in operation, is not easy to practical application.

Summary of the invention

The object of the invention is to provide a kind of quality determining method that increases emulsifiable paste and related preparations thereof.

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

Quality determining method of the present invention comprises following content assaying method, and concrete steps are:

Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 D);

Chromatographic condition and system suitability; Be filling agent with octadecylsilane chemically bonded silica; Take acetonitrile-0.1% phosphoric acid of 10～20:90～80 as mobile phase; The detection wavelength is 200～250nm; Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 3000;

The preparation of reference substance solution: get the Paeoniflorin reference substance an amount of, accurately weighed, add methyl alcohol and make per 1 parts by volume and contain the solution of Paeoniflorin 0.00001～0.001 weight portion, and get final product;

The preparation of need testing solution: get increase emulsifiable paste and related preparations day taking dose 1/30, accurately weighed, add methyl alcohol 20 parts by volume in 25 parts by volume volumetric flasks, ultrasonic processing 30～60 minutes (power 300W, frequency 50KHz), let cool, add methyl alcohol to scale, shake up, filter, get subsequent filtrate, and get final product;

Determination method: precision is drawn reference substance solution and each 0.001-0.01 parts by volume of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product;

Increase emulsifiable paste and related preparations thereof in day taking dose contain the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 31.5 mg.

Preferably, quality determining method of the present invention comprises following content assaying method:

Chromatographic condition and system suitability; Be filling agent with octadecylsilane chemically bonded silica; Acetonitrile take ratio as 14:86-0.1% phosphoric acid is mobile phase; The detection wavelength is 230nm; Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 3000;

The preparation of reference substance solution: get the Paeoniflorin reference substance an amount of, accurately weighed, add methyl alcohol and make per 1 parts by volume and contain the solution of Paeoniflorin 0.0001 weight portion, and get final product;

The preparation of need testing solution: get increase emulsifiable paste and related preparations day taking dose 1/30, accurately weighed, add methyl alcohol 20 parts by volume in the 25ml volumetric flask, 30 minutes (power 300W of ultrasonic processing, frequency 50KHz), lets cool, add methyl alcohol to scale, shake up, filter, get subsequent filtrate, and get final product;

Determination method: precision is drawn reference substance solution and each 0.005 parts by volume of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product;

The quality determining method that increases emulsifiable paste and related preparations thereof of the present invention also comprises one or more in the following discrimination method:

A. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20 parts by volume merge normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20 parts by volume, with saturated nacl aqueous solution washing 3 times, each 20 parts by volume are got normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15 parts by volume, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets seed of cowherb control medicinal material 5 weight portions, add boiling range 60-90 ℃ sherwood oil 20 parts by volume, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25 parts by volume, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution; Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate of 10-20:30-50:18-24:8-12-methanol-water, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color;

B. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, 30 parts by volume that add diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7 parts by volume, ether 3 parts by volume, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds the dissolving of ethanol 1 parts by volume, as need testing solution; Other gets prepared rhizome of rehmannia control medicinal material 5 weight portions, and section adds ethanol 20 parts by volume, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the boiling range 60-90 ℃ of petroleum ether-ethyl acetate of 1-3:1 as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting;

C. get lower need testing solution 15 parts by volume of [assay] item, evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that per 1 parts by volume contains 0.002 weight portion, in contrast product solution; Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 0.004 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take chloroform-ethyl acetate of 30-50:3-8:5-15:0.1-0.2-methyl alcohol-formic acid as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

D. get need testing solution under [discriminating] B item, as need testing solution; Other gets the citrulline reference substance, adds Diluted Alcohol and makes the solution that per 1 parts by volume contains 0.001 weight portion, in contrast product solution; Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw need testing solution 0.01 parts by volume, reference substance solution 0.001 parts by volume, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol-absolute ethyl alcohol of 5-11:1-3:1-3:2-4-glacial acetic acid-water as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with ninhydrin solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

Preferably, the quality determining method that increases emulsifiable paste and related preparations thereof of the present invention also comprises one or more in the following discrimination method:

A. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20 parts by volume merge normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20 parts by volume, with saturated nacl aqueous solution washing 3 times, each 20 parts by volume are got normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15 parts by volume, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets seed of cowherb control medicinal material 5 weight portions, add boiling range 60-90 ℃ sherwood oil 20 parts by volume, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25 parts by volume, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution; Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate of 15:40:22:10-methanol-water, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color;

B. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, 30 parts by volume that add diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7 parts by volume, ether 3 parts by volume, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds the dissolving of ethanol 1 parts by volume, as need testing solution; Other gets prepared rhizome of rehmannia control medicinal material 5 weight portions, and section adds ethanol 20 parts by volume, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the 60-90 ℃ of petroleum ether-ethyl acetate of 2:1 as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting;

C. get lower need testing solution 15 parts by volume of [assay] item, evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that per 1 parts by volume contains 0.002 weight portion, in contrast product solution; Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 0.004 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take chloroform-ethyl acetate of 40:5:10:0.2-methyl alcohol-formic acid as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

D. get need testing solution under [discriminating] B item, as need testing solution; Other gets the citrulline reference substance, adds Diluted Alcohol and makes the solution that per 1 parts by volume contains 0.001 weight portion, in contrast product solution; Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw need testing solution 0.01 parts by volume, reference substance solution 0.001 parts by volume, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol-absolute ethyl alcohol of 8:2:2:3-glacial acetic acid-water as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with ninhydrin solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

The unit corresponding relation of described weight portion and parts by volume is grams per milliliter.

The described reference substance of getting is decided by the routine sampling amount of Chinese Pharmacopoeia regulation in right amount.

Describedly increase emulsifiable paste and related preparations is to be made by following bulk drug and technique: seed of cowherb 100-160 weight portion, stem pith of the rice-paper plant 100-160 weight portion, prepared rhizome of rehmannia 130-190 weight portion, Radix Angelicae Sinensis 50-90 weight portion, root of herbaceous peony 90-130 weight portion, Ligusticum wallichii 40-69 weight portion, motherwort 100-160 weight portion, root of Chinese trichosanthes 100-160 weight portion; Above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, dipping spent the night rear forced circulation 2-4 hour, filtered, and merged two kinds of alcohol extracts, Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use; The stem pith of the rice-paper plant was soaked 1-3 hour, decocted 1-3 hour, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1-2 hour, filtered filtrate for later use; Root of Chinese trichosanthes soaked 1-3 hour with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃); Merge alcohol extracting concentrate and water extracting liquid, add conventional auxiliary material, make paste, tablet, capsule, granule or pill according to common process.

Describedly increase emulsifiable paste and related preparations is preferably made by following bulk drug and technique: the seed of cowherb 133 weight portions, the stem pith of the rice-paper plant 133 weight portions, prepared rhizome of rehmannia 167 weight portions, Radix Angelicae Sinensis 67 weight portions, the root of herbaceous peony 111 weight portions, Ligusticum wallichii 50 weight portions, motherwort 133 weight portions, root of Chinese trichosanthes 133 weight portions; Above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, flood the rear forced circulation 3 hours of spending the night, and filtration merges two kinds of alcohol extracts, and Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use; The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃); Merge alcohol extracting concentrate and water extracting liquid, add conventional auxiliary material, make paste, tablet, capsule, granule or pill according to common process.

The day dose dosage that the present invention increases emulsifiable paste is 90g, a 30g, 3 times on the one.In the situation that it is identical to guarantee day to take the crude drug amount, the day taking dose that increases the emulsifiable paste related preparations can calculate according to increasing emulsifiable paste day taking dose.

Quality determining method of the present invention is compared with act.std, test sample adopts the method for the ultrasonic extraction of methyl alcohol under the assay item, the method both extraction easy and Paeoniflorin is complete, make the measurement result of Paeoniflorin more accurate, methodology test shows that precision and reappearance are good, method is feasible, has simplified simultaneously operation steps, is convenient to practical application.

Description of drawings

Fig. 1 Paeoniflorin reference substance chromatogram

Fig. 2 increases emulsifiable paste test liquid chromatogram

Fig. 3 increases emulsifiable paste negative control test solution chromatogram

Fig. 4 Paeoniflorin canonical plotting

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

The experimental example high effective liquid chromatography for measuring increases the methodological study of paeoniflorin content in the emulsifiable paste (pressing embodiment 1 preparation)

1, instrument and reagent

The Agilent-1200 high performance liquid chromatograph comprises: vacuum Off mechanism of qi, quaternary gradient pump, automatic sampler, UV-detector, Agilent workstation; Superpure water machine (Sartorius); Ultrasonic generator (Elma 820/H).

(Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, lot number: 110736-200731), methyl alcohol is chromatographically pure to the Paeoniflorin reference substance, and acetonitrile is chromatographically pure, and water is ultrapure water, and phosphoric acid is pure for analyzing.

2, method and result

2.1 chromatographic condition

Chromatographic column: Ultimate ^TMThe XB-C18 post, (5 μ m, 4.6 * 250mm Welch Materials, Inc.); Mobile phase: acetonitrile-0.1% phosphoric acid (14:86); Detect wavelength 230nm; Column temperature: 20 ℃; Flow velocity: 0.8ml/min; Sample size: 10 μ l are under this chromatographic condition, and the effective constituent Paeoniflorin in the Chinese herbaceous peony can separate with other impurity peaks better, and the chromatogram of reference substance solution, need testing solution and negative control solution is seen Fig. 1-3.

2.2 system tries out the property test:

Press preceding method, under following test condition, to the theory plate number that collapses at sample and contrast peak, the parameters such as degree of separation are tested, and see Table 1.

Table 1 system tries out the property measurement result

2.3 the selection of extracting method:

2.3.1 extract the selection of solvent:

It is that 0807005 sample extracts to lot number that experiment is selected respectively with the solvent of opposed polarity.Be respectively＜1 select the water elution sample to cross 101 macroreticular resins, again with behind the ethanol elution, collect ethanol eluate, evaporate to dryness extracts with the methyl alcohol dissolving again;＜2〉select the ultrasonic extraction of Diluted Alcohol;＜3〉select the ultrasonic extraction of ethanol;＜4〉select the ultrasonic extraction of methyl alcohol.The result shows with methyl alcohol extraction sample more complete, sees Table 2.

Table 2 different solvents extracts the result of paeoniflorin content

Figure 2012102850196100002DEST_PATH_IMAGE002

2.3.2 the selection of extraction time:

Experiment adopts methyl alcohol to use respectively the ultrasonic extraction of different time, is respectively 30 minutes, and 45 minutes, 60 minutes.The result shows with methyl alcohol ultrasonic extraction and just Paeoniflorin can be extracted fully in 30 minutes, and method is easy, easily to operate, sees Table 3.

The measurement result of table 3 different extraction times

Figure 2012102850196100002DEST_PATH_IMAGE003

2.4 the preparation of reference substance solution, need testing solution and Chinese herbaceous peony negative control solution:

The preparation of reference substance solution: with embodiment 1.

The preparation of need testing solution: with embodiment 1.

The preparation of negative control solution: by a prescription inferior segment ingredients (except Chinese herbaceous peony), by making blank sample under the method for making item, and prepare negative control solution by the text method.

2.5 blank test:

Draw respectively Paeoniflorin reference substance solution, need testing solution and Chinese herbaceous peony negative control solution, injecting high performance liquid chromatograph measures, the result shows that the negative control chromatogram is occurring without chromatographic peak with the corresponding retention time of reference substance chromatogram place, and other ingredientss are not all disturbed the mensuration of Paeoniflorin in the side.

2.6 linear relationship is investigated:

It is an amount of that precision takes by weighing the Paeoniflorin reference substance respectively, adds methyl alcohol dissolving, makes respectively the reference substance solution (II) that contrast solution (I) that every ml contains 0.09839mg and every ml contain 0.4669mg.Accurate contrast solution (I) 1 μ l, 2 μ l, 4 μ l, 6 μ l, 8 μ l, 10 μ l and reference substance solution (II) the 10 μ l of drawing measure with above-mentioned chromatographic condition, the results are shown in Table 4.Take the reference substance sample size as horizontal ordinate, it is ordinate that front cover amasss signal value (mAU), the drawing standard curve, the results are shown in Figure 4, the regression equation y=1306.8x+6.5428 of Paeoniflorin, correlation coefficient r=0.9999 shows that Paeoniflorin is good linear relationship in 0.09839 μ g～4.669 μ g scopes.

Table 4 Paeoniflorin linear relationship investigation table

Figure 2012102850196100002DEST_PATH_IMAGE004

2.7 precision test

The every 1ml of accurate absorption contains the contrast solution 6 μ l of 98.39 μ g, repeats continuously the injection liquid chromatography 9 times, measures peak area by above-mentioned chromatographic condition, ask relative standard deviation, take Paeoniflorin peak area RSD as 1.52%, show that instrument precision is good, the results are shown in Table 5.

Table 5 Paeoniflorin Precision Experiment result

2.8 stability test

Get the need testing solution of lot number (0807005), at rear 0 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 16 hours sample introductions of preparation, carry out analysis result by above-mentioned chromatographic condition and see Table 6 respectively.The result shows that the RSD of Paeoniflorin peak area in 0 ~ 16 hour is 0.79%, shows that sample places in 16 hours basicly stable.

Table 6 Paeoniflorin stability experiment result

2.9 repeated experiment

Get the need testing solution of same lot number (0807005), carry out 5 replicate determinations by aforementioned content assaying method, the results are shown in Table 7, the RSD of paeoniflorin content is 0.35%, shows that the method repeatability is good.

Table 7 Paeoniflorin repeatability measurement result

2.10 recovery test

Getting the known content lot number is (0807005) sample, adopts the application of sample recovery test, and precision adds a certain amount of Paeoniflorin reference substance, press preceding method and measure, and calculate recovery rate, RSD is 0.02%.The results are shown in Table 8, show that the method is feasible.

Table 8 Paeoniflorin recovery test

Figure 2012102850196100002DEST_PATH_IMAGE008

2.11 sample size is measured

Respectively six batch samples are measured by the method for aforementioned paeoniflorin content, be the results are shown in Table 9.

Table 9 increases emulsifiable paste six batch sample assay results

By the pharmacopeia regulation, the content of Paeoniflorin must not be less than 1.6% in the white Peony Root, and the every gram of this preparation contains the Paeoniflorin theoretical value must not be less than 1.776mg.The every gram of regulation this product contains Chinese herbaceous peony with Paeoniflorin (C in the primary standard ₂₃H ₂₈O ₁₁) meter, must not be less than 0.25mg.

In conjunction with six batch sample test findings, this standard code is: the every gram of this product contains Chinese herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 0.35mg.

Embodiment

Following embodiment all can realize the described effect of above-mentioned experimental example.

Embodiment 1 increases the quality determining method of emulsifiable paste

[prescription] seed of cowherb 133g stem pith of the rice-paper plant 133g prepared rhizome of rehmannia 167g Radix Angelicae Sinensis 67g

Root of herbaceous peony 111g Ligusticum wallichii 50g motherwort 133g root of Chinese trichosanthes 133g

[method for making] above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, spend the night rear forced circulation 3 hours of dipping filters, and merges two kinds of alcohol extracts, Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use.The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃).Merge alcohol extracting concentrate and water extracting liquid, sucrose and Sodium Benzoate that adding was refined are an amount of, stir, and make 1000g, packing, and get final product.

[specification] be bottled (1) 150g(2 whenever) 250g.

[usage and consumption] is oral.A 30g, 3 times on the one.

[assay] measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 D).

Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Acetonitrile-0.1% phosphoric acid (14:86) is mobile phase; The detection wavelength is 230nm.Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 3000.

It is an amount of that the Paeoniflorin reference substance is got in the preparation of reference substance solution, accurately weighed, adds methyl alcohol and make the solution that every 1ml contains Paeoniflorin 0.1mg, and get final product.

This product 3g is got in the preparation of need testing solution, and is accurately weighed, adds methyl alcohol 20ml in the 25ml volumetric flask, and ultrasonic processing 30 minutes (power 300W, frequency 50KHz) lets cool, and adds methyl alcohol to scale, shakes up, and filters, and gets subsequent filtrate, and get final product.

Determination method is accurate reference substance solution and each 5 μ l of need testing solution of drawing respectively, and the injection liquid chromatography is measured, and be get final product.

The every 1g of this product contains the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 0.35mg.

Embodiment 2 increases the quality determining method of emulsifiable paste

Prescription and method for making are with embodiment 1.

This product 60g is got in [discriminating] (1), adds absolute ethyl alcohol 70ml, and the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20ml make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20ml merges normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20ml, with saturated nacl aqueous solution washing 3 times, each 20ml gets normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15ml, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Other gets seed of cowherb control medicinal material 5g, add sherwood oil (60-90 ℃) 20ml, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25ml, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20ml make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate-methanol-water (15:40:22:10), launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

(2) get this product 60g, add absolute ethyl alcohol 70ml, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, the 30ml that adds diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7ml, ether 3ml, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds ethanol 1ml dissolving, as need testing solution.Other gets prepared rhizome of rehmannia control medicinal material 5g, and section adds ethanol 20ml, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, in contrast medicinal material solution.According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take sherwood oil (60-90 ℃)-ethyl acetate (2:1) as developping agent, launch, take out, dry.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting.

(3) get the lower need testing solution 15ml of [assay] item, evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution.Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that every 1ml contains 2mg, in contrast product solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take chloroform-ethyl acetate-methyl alcohol-formic acid (40:5:10:0.2) as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

(4) get the lower need testing solution of [discriminating] (2) item, as need testing solution.Other gets the citrulline reference substance, adds Diluted Alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw need testing solution 10 μ l, reference substance solution 1 μ l, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol-absolute ethyl alcohol-glacial acetic acid-water (8:2:2:3) as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with ninhydrin solution.In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

Embodiment 3 increases the quality determining method of newborn tablet

[method for making] above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, spend the night rear forced circulation 3 hours of dipping filters, and merges two kinds of alcohol extracts, Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use.The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃); Merge alcohol extracting concentrate and water extracting liquid, routinely technique, add conventional auxiliary material, be prepared into 100, every 0.5 gram.

[usage and consumption] is oral.One

time

3,3 times on the one.

This product 0.15g is got in the preparation of need testing solution, and is accurately weighed, adds methyl alcohol 20ml in the 25ml volumetric flask, and ultrasonic processing 30 minutes (power 300W, frequency 50KHz) lets cool, and adds methyl alcohol to scale, shakes up, and filters, and gets subsequent filtrate, and get final product.

Every of this product contains the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 3.5mg.

Embodiment 4 increases the quality determining method of latex wafer

[method for making] above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, spend the night rear forced circulation 3 hours of dipping filters, and merges two kinds of alcohol extracts, Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use.The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃); Merge alcohol extracting concentrate and water extracting liquid, routinely technique, add conventional auxiliary material, be prepared into 100 capsules.

[usage and consumption] is oral.One

time

3,3 times on the one.

6 of this product are got in [discriminating] (1), add absolute ethyl alcohol 70ml, and the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20ml make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20ml merges normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20ml, with saturated nacl aqueous solution washing 3 times, each 20ml gets normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15ml, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Other gets seed of cowherb control medicinal material 5g, add sherwood oil (60-90 ℃) 20ml, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25ml, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20ml make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate-methanol-water (15:40:22:10), launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

(2) get 6 of this product, add absolute ethyl alcohol 70ml, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, the 30ml that adds diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7ml, ether 3ml, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds ethanol 1ml dissolving, as need testing solution.Other gets prepared rhizome of rehmannia control medicinal material 5g, and section adds ethanol 20ml, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, in contrast medicinal material solution.According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take sherwood oil (60-90 ℃)-ethyl acetate (2:1) as developping agent, launch, take out, dry.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting.

3/10 of 1 capsules is got in the preparation of need testing solution, and is accurately weighed, adds methyl alcohol 20ml in the 25ml volumetric flask, and ultrasonic processing 30 minutes (power 300W, frequency 50KHz) lets cool, and adds methyl alcohol to scale, shakes up, and filters, and gets subsequent filtrate, and get final product.

The every capsules of this product contains the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 3.5mg.

Embodiment 5 increases the quality determining method of newborn granule

[method for making] above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, spend the night rear forced circulation 3 hours of dipping filters, and merges two kinds of alcohol extracts, Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use.The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃); Merge alcohol extracting concentrate and water extracting liquid, routinely technique, add conventional auxiliary material, be prepared into 30 bags of granules.

[usage and consumption] is oral.One time 3 bags, 3 times on the one.

1/10 of 1 bag of particle is got in the preparation of need testing solution, and is accurately weighed, adds methyl alcohol 20ml in the 25ml volumetric flask, and ultrasonic processing 30 minutes (power 300W, frequency 50KHz) lets cool, and adds methyl alcohol to scale, shakes up, and filters, and gets subsequent filtrate, and get final product.

Every bag of this product contains the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 10.5mg.

Embodiment 6 increases the quality determining method of newborn pill

[method for making] above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, spend the night rear forced circulation 3 hours of dipping filters, and merges two kinds of alcohol extracts, Recycled ethanol is concentrated into relative density and is 1.15 ~ 1.30(90 ~ 100 ℃), for subsequent use.The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into relative density is 1.08 ~ 1.23(90-100 ℃); Merge alcohol extracting concentrate and water extracting liquid, routinely technique, add conventional auxiliary material, be prepared into 300 balls.Per 8 balls weigh 2 grams.

[usage and consumption] is oral.8 balls, 3 times on the one.

This product 16 balls are got in [discriminating] (1), add absolute ethyl alcohol 70ml, and the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20ml make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20ml merges normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20ml, with saturated nacl aqueous solution washing 3 times, each 20ml gets normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15ml, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1ml makes dissolving, as need testing solution.Other gets seed of cowherb control medicinal material 5g, add sherwood oil (60-90 ℃) 20ml, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25ml, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20ml make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution.Test according to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate-methanol-water (15:40:22:10), launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color.

(2) get this product 16 balls, add absolute ethyl alcohol 70ml, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, the 30ml that adds diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7ml, ether 3ml, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds ethanol 1ml dissolving, as need testing solution.Other gets prepared rhizome of rehmannia control medicinal material 5g, and section adds ethanol 20ml, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, in contrast medicinal material solution.According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take sherwood oil (60-90 ℃)-ethyl acetate (2:1) as developping agent, launch, take out, dry.In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting.

This product 0.3g is got in the preparation of need testing solution, and is accurately weighed, adds methyl alcohol 20ml in the 25ml volumetric flask, and ultrasonic processing 30 minutes (power 300W, frequency 50KHz) lets cool, and adds methyl alcohol to scale, shakes up, and filters, and gets subsequent filtrate, and get final product.

Per 8 balls of this product contain the root of herbaceous peony with Paeoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 10.5mg.

Claims

1. quality determining method that increases emulsifiable paste and related preparations thereof is characterized in that the method comprises following content assaying method:

According to high effective liquid chromatography for measuring; Chromatographic condition and system suitability are: be filling agent with octadecylsilane chemically bonded silica; Take acetonitrile-0.1% phosphoric acid of 10～20:90～80 as mobile phase; The detection wavelength is 200～250nm; Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 3000; The preparation of reference substance solution: get the Paeoniflorin reference substance an amount of, accurately weighed, add methyl alcohol and make per 1 parts by volume and contain the solution of Paeoniflorin 0.00001～0.001 weight portion, and get final product;

The preparation of need testing solution: get increase emulsifiable paste and related preparations day taking dose 1/30, accurately weighed, add methyl alcohol 20 parts by volume in 25 parts by volume volumetric flasks, ultrasonic processing 30～60 minutes, power 300W, frequency 50KHz lets cool, and adds methyl alcohol to scale, shake up, filter, get subsequent filtrate, and get final product;

Increase emulsifiable paste and related preparations thereof in day taking dose contain the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 31.5 mg;

Describedly increase emulsifiable paste and related preparations is to be made by following bulk drug and technique: seed of cowherb 100-160 weight portion, stem pith of the rice-paper plant 100-160 weight portion, prepared rhizome of rehmannia 130-190 weight portion, Radix Angelicae Sinensis 50-90 weight portion, root of herbaceous peony 90-130 weight portion, Ligusticum wallichii 40-69 weight portion, motherwort 100-160 weight portion, root of Chinese trichosanthes 100-160 weight portion; Above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, flood and spend the night rear forced circulation 2-4 hour, and filtration merges two kinds of alcohol extracts, and Recycled ethanol is concentrated into 90 ~ 100 ℃ of relative densities and is 1.15 ~ 1.30, and is for subsequent use; The stem pith of the rice-paper plant was soaked 1-3 hour, decocted 1-3 hour, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1-2 hour, filtered filtrate for later use; Root of Chinese trichosanthes soaked 1-3 hour with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into 90-100 ℃ of relative density is 1.08 ~ 1.23; Merge alcohol extracting concentrate and water extracting liquid, add conventional auxiliary material, make paste, tablet, capsule, granule or pill according to common process.

2. quality determining method as claimed in claim 1 is characterized in that the method is:

According to high effective liquid chromatography for measuring; Chromatographic condition and system suitability are: be filling agent with octadecylsilane chemically bonded silica; Acetonitrile take ratio as 14:86-0.1% phosphoric acid is mobile phase; The detection wavelength is 230nm; Number of theoretical plate calculates by the Paeoniflorin peak should be not less than 3000;

The preparation of need testing solution: get increase emulsifiable paste and related preparations day taking dose 1/30, accurately weighed, add methyl alcohol 20 parts by volume in the 25ml volumetric flask, ultrasonic processing 30 minutes, power 300W, frequency 50KHz lets cool, and adds methyl alcohol to scale, shake up, filter, get subsequent filtrate, and get final product;

Determination method: precision is drawn reference substance solution and each 0.005 weight portion of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product;

Increase emulsifiable paste and related preparations thereof in day taking dose contain the root of herbaceous peony with Paeoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 31.5 mg.

3. quality determining method as claimed in claim 1 or 2 is characterized in that increasing emulsifiable paste described in the method and related preparations is made by following bulk drug and technique:

The seed of cowherb 133 weight portions, the stem pith of the rice-paper plant 133 weight portions, prepared rhizome of rehmannia 167 weight portions, Radix Angelicae Sinensis 67 weight portions, the root of herbaceous peony 111 weight portions, Ligusticum wallichii 50 weight portions, motherwort 133 weight portions, root of Chinese trichosanthes 133 weight portions; Above eight flavors are got prepared rhizome of rehmannia and are made solvent with 80% ethanol, get Radix Angelicae Sinensis, Ligusticum wallichii is made solvent with 70% ethanol, flood the rear forced circulation 3 hours of spending the night, and filtration merges two kinds of alcohol extracts, and Recycled ethanol is concentrated into 90 ~ 100 ℃ of relative densities and is 1.15 ~ 1.30, and is for subsequent use; The stem pith of the rice-paper plant was soaked 2 hours, decocted 2 hours, filtered, and the filtrate adding seed of cowherb, the root of herbaceous peony, motherwort decocted 1.5 hours, filtered filtrate for later use; Root of Chinese trichosanthes soaked 2 hours with 80 ℃ of hot water temperature, filtered, and filtrate and above-mentioned water extraction filtrate merge, and being concentrated into 90-100 ℃ of relative density is 1.08 ~ 1.23; Merge alcohol extracting concentrate and water extracting liquid, add conventional auxiliary material, make paste, tablet, capsule, granule or pill according to common process.

4. quality determining method as claimed in claim 1 or 2 is characterized in that the method also comprises one or more in the following discriminating:

A. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20 parts by volume merge normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20 parts by volume, with saturated nacl aqueous solution washing 3 times, each 20 parts by volume are got normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15 parts by volume, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets seed of cowherb control medicinal material 5 weight portions, add boiling range 60-90 ℃ sherwood oil 20 parts by volume, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25 parts by volume, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution; Test according to thin-layered chromatography, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate of 10-20:30-50:18-24:8-12-methanol-water, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color;

B. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, 30 parts by volume that add diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7 parts by volume, ether 3 parts by volume, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds the dissolving of ethanol 1 parts by volume, as need testing solution; Other gets prepared rhizome of rehmannia control medicinal material 5 weight portions, and section adds ethanol 20 parts by volume, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the boiling range 60-90 ℃ of petroleum ether-ethyl acetate of 1-3:1 as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting;

C. get lower need testing solution 15 parts by volume of [assay] item, evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that per 1 parts by volume contains 0.002 weight portion, in contrast product solution; Test according to thin-layered chromatography, draw each 0.004 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take chloroform-ethyl acetate of 30-50:3-8:5-15:0.1-0.2-methyl alcohol-formic acid as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

D. get need testing solution under [discriminating] B item, as need testing solution; Other gets the citrulline reference substance, adds Diluted Alcohol and makes the solution that per 1 parts by volume contains 0.001 weight portion, in contrast product solution; Test according to thin-layered chromatography, draw need testing solution 0.01 parts by volume, reference substance solution 0.001 parts by volume, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol-absolute ethyl alcohol of 5-11:1-3:1-3:2-4-glacial acetic acid-water as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with ninhydrin solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

5. quality determining method as claimed in claim 3 is characterized in that the method also comprises one or more in the following discriminating:

6. quality determining method as claimed in claim 4 is characterized in that differentiating being following one or more:

A. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20 parts by volume merge normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20 parts by volume, with saturated nacl aqueous solution washing 3 times, each 20 parts by volume are got normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15 parts by volume, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets seed of cowherb control medicinal material 5 weight portions, add sherwood oil (60-90 ℃) 20 parts by volume, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25 parts by volume, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution; Test according to thin-layered chromatography, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate of 15:40:22:10-methanol-water, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color;

B. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, 30 parts by volume that add diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7 parts by volume, ether 3 parts by volume, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds the dissolving of ethanol 1 parts by volume, as need testing solution; Other gets prepared rhizome of rehmannia control medicinal material 5 weight portions, and section adds ethanol 20 parts by volume, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the sherwood oil (60-90 ℃) of 2:1-ethyl acetate as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting;

C. get lower need testing solution 15 parts by volume of [assay] item, evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that per 1 parts by volume contains 2 weight portions, in contrast product solution; Test according to thin-layered chromatography, draw each 0.004 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take chloroform-ethyl acetate of 40:5:10:0.2-methyl alcohol-formic acid as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

D. get need testing solution under [discriminating] B item, as need testing solution; Other gets the citrulline reference substance, adds Diluted Alcohol and makes the solution that per 1 parts by volume contains 1 weight portion, in contrast product solution; Test according to thin-layered chromatography, draw need testing solution 0.01 parts by volume, reference substance solution 0.001 parts by volume, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol-absolute ethyl alcohol of 8:2:2:3-glacial acetic acid-water as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with ninhydrin solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.

7. quality determining method as claimed in claim 5 is characterized in that differentiating being following one or more:

A. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, leave standstill, incline and get upper strata liquid evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, extract 3 times with water-saturated n-butanol, each 20 parts by volume merge normal butyl alcohol liquid, with 5% ammonia spirit washing 2 times, each 20 parts by volume, with saturated nacl aqueous solution washing 3 times, each 20 parts by volume are got normal butyl alcohol liquid evaporate to dryness again, residue adds methyl alcohol 15 parts by volume, ultrasonic processing 15 minutes filters the filtrate evaporate to dryness, residue adds methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets seed of cowherb control medicinal material 5 weight portions, add boiling range 60-90 ℃ sherwood oil 20 parts by volume, added hot reflux 1 hour, filter, filter residue volatilizes, add ethanol 25 parts by volume, added hot reflux 30 minutes, let cool, filter, filtrate evaporate to dryness, residue add saturated nacl aqueous solution 20 parts by volume make the dissolving and be transferred in the separating funnel, be made in the same way of control medicinal material solution; Test according to thin-layered chromatography, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, place lower floor's solution of 12 hours below 5 ~ 10 ℃ as developping agent take chloroform-ethyl acetate of 15:40:22:10-methanol-water, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with 10% ethanol solution of sulfuric acid; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, the spot of aobvious same color;

B. get increase emulsifiable paste and related preparations day taking dose 2/3, add absolute ethyl alcohol 70 parts by volume, the limit edged stirs, stirred again 10 minutes, and left standstill, incline and get upper strata liquid, 30 parts by volume that add diethyl ether, standing over night is inclined and is got upper strata liquid, steam near and do, add absolute ethyl alcohol 7 parts by volume, ether 3 parts by volume, ultrasonic processing 10 minutes is got upper strata liquid, evaporate to dryness, residue adds the dissolving of ethanol 1 parts by volume, as need testing solution; Other gets prepared rhizome of rehmannia control medicinal material 5 weight portions, and section adds ethanol 20 parts by volume, adds hot reflux 0.5 hour, lets cool, and filters, and filtrate evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, in contrast medicinal material solution; According to thin-layered chromatography test, draw each 0.005 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take the boiling range 60-90 ℃ of petroleum ether-ethyl acetate of 2:1 as developping agent, launch, take out, dry; In the test sample chromatogram, with the corresponding position of control medicinal material chromatogram on, aobvious identical yellow spotting;

C. get lower need testing solution 15 parts by volume of [assay] item, evaporate to dryness, residue add methyl alcohol 1 parts by volume makes dissolving, as need testing solution; Other gets the Paeoniflorin reference substance, adds methyl alcohol and makes the solution that per 1 parts by volume contains 0.002 weight portion, in contrast product solution; Test according to thin-layered chromatography, draw each 0.004 parts by volume of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, take chloroform-ethyl acetate of 40:5:10:0.2-methyl alcohol-formic acid as developping agent, launch, take out, dry, spray is with 5% vanillic aldehyde sulfuric acid solution, and it is clear that hot blast blows to the spot colour developing; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color;

D. get need testing solution under [discriminating] B item, as need testing solution; Other gets the citrulline reference substance, adds Diluted Alcohol and makes the solution that per 1 parts by volume contains 0.001 weight portion, in contrast product solution; Test according to thin-layered chromatography, draw need testing solution 0.01 parts by volume, reference substance solution 0.001 parts by volume, put respectively on same silica gel g thin-layer plate, take normal butyl alcohol-absolute ethyl alcohol of 8:2:2:3-glacial acetic acid-water as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with ninhydrin solution; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, the spot of aobvious same color.