CN1600364A

CN1600364A - Method for controlling quality of oral liquid of medlar-chrysanthemum-rehmannia root

Info

Publication number: CN1600364A
Application number: CN 200410060977
Authority: CN
Inventors: 卢建中; 王伟兰; 李诒光; 徐昌瑞
Original assignee: Jiangzhong Pharmaceutical Co Ltd
Current assignee: Jiangzhong Pharmaceutical Co Ltd
Priority date: 2004-10-12
Filing date: 2004-10-12
Publication date: 2005-03-30
Anticipated expiration: 2024-10-12
Also published as: CN100500190C

Abstract

A quality control method for the Chinese medicine 'Qijudihuang oral liquid' which is prepared from 8 Chinese-medicinal materials including wolfberry fruit, chrysanthemum flower, rehmannia root, tree peony bark, etc features that the efficient liquid-phase chromatography is used to measure the content of patoniflorin in said oral liquid.

Description

The method of quality control of QIJU DIHUANG KOUFUYE

Technical field

The present invention relates to a kind of method of quality control of the QIJU DIHUANG KOUFUYE of making by Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, particularly relate to content of paeoniflorin in this medicine of high effective liquid chromatography for measuring and differentiate the QIJU DIHUANG KOUFUYE method of quality control of Chinese crude drug Flos Chrysanthemi, Rhizoma Alismatis with thin layer chromatography.

Background technology

The ten (WS of " Drug Standard of Ministry of Public Health of the Peoples Republic of China " (Chinese traditional patent formulation preparation) ₃-B-2146-96) in the treatment hepatic and renal YIN deficiency, vertigo and tinnitus, the photophobia photophobia, blurring of vision disease medicament QIJU DIHUANG KOUFUYE, this pharmaceutical formulation is made up of Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, and above-mentioned raw materials is made 1000ml altogether; And issued its quality control standard, but in the existing QIJU DIHUANG KOUFUYE method of quality control that Ministry of Public Health is issued, pH value, relative density, loading amount and limit test of microbe are arranged under the detection method item, and assay, but there is not the medical material qualitative identification, specificity is not strong, and limitation is bigger, is difficult to control the quality of QIJU DIHUANG KOUFUYE; " the QIJU DIHUANG WAN method of quality control that records of 2000 editions one of Chinese pharmacopoeia adopts the thin layer of medical material Cortex Moutan, Fructus Corni to differentiate, and a microscopical identification is arranged, and does not have to contain the survey index, is difficult to control the quality of QIJU DIHUANG WAN; The present invention overcomes the deficiencies in the prior art, improve the quality control standard of QIJU DIHUANG KOUFUYE, improve assay detection method in the preparation, improve the bottom line of paeoniflorin content, increase the thin layer chromatography qualitative identification method of Flos Chrysanthemi, Rhizoma Alismatis two flavor medical materials, make sample treatment easier, guaranteed this compound preparation higher quality standard level.

Summary of the invention

The new method of quality control that the purpose of this invention is to provide QIJU DIHUANG KOUFUYE, the accuracy of the examination criteria of ensuring the quality of products and advance.

The ten (WS of " Drug Standard of Ministry of Public Health of the Peoples Republic of China " (Chinese traditional patent formulation preparation) ₃-B-2146-96) middle QIJU DIHUANG KOUFUYE prescription is made up of Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, main flavour of a drug in the Cortex Moutan side of being, dry root bark for ranunculaceae peony Paeonia suffruticosa Andr., has clearing away heat and cooling blood, the effect of promoting blood circulation to remove blood stasis mainly contains chemical constituents such as paeoniflorin, paeonol, the former glycoside of paeonol; Flos Chrysanthemi is the head inflorescence of feverfew chrysanthemum Chrysanthemum morifolium morifolium Ramat., has wind and heat dispersing, detoxifcation, the effect that makes eye bright; Rhizoma Alismatis is Alismataceae plant Rhizoma Alismatis Alisma orientale (Sam.) Juzep. tuber, and sweet in the mouth is light, cold, have promoting diuresis to eliminate damp pathogen, the effect that expels the heat-evil mainly contains chemical constituents such as Alisol A, B, C and Alisol A acetas, alisol B acetas, alisol C acetas; The technology of QIJU DIHUANG KOUFUYE is that water is carried, so microscopical identification is inapplicable, the chemical constituent ursolic acid in the Fructus Corni is a liposoluble constituent, is not suitable for as what set up this preparation to contain the survey index, in addition, the present invention's thin-layer qualitative of having increased QIJU DIHUANG KOUFUYE ZHONGJUHUA, Rhizoma Alismatis is differentiated.And further improved the content of paeoniflorin assay method, QIJU DIHUANG KOUFUYE is too big by the test sample concentration of primary standard preparation, the mobile phase separating effect is not good enough yet, impurity disturbs bigger, makes measurement result inaccurate, after deliberation, primary standard has been carried out following improvement: (1) is because this product is denseer, dilute and strengthened the volume of extractant water-saturated n-butanol during extraction earlier, help two phase stratification, reduce the loss; (2) last constant volume is decided to be 25ml, makes need testing solution main peak peak area and reference substance suitable, has reduced the interference of impurity peaks, has improved accuracy; (3) improved the bottom line of peoniflorin.Result of study shows that the standard after the raising can reflect and control the inherent quality of preparation better.

Finishing the present invention is achieved by following technical proposals: one is with content of paeoniflorin in this medicine of high effective liquid chromatography for measuring; Its dual-purpose thin layer chromatography is differentiated this medicine Chinese crude drug Flos Chrysanthemi and Rhizoma Alismatis.

The quality control detection method that concrete QIJU DIHUANG KOUFUYE is new, can pass through following steps in advance to realize:

Test apparatus, reagent and reagent

High performance liquid chromatograph: Waters600 type pump, Waters600 controller, Waters486 type UV-detector; Shen, river chromatograph working software (JS-3030 version, big Shen, Liaanjiang county separation science technology company); Ultrasonic washing instrument (SK5200H type, Shanghai High Kudos Science Instrument Co., Ltd.); METTLER electronic analytical balance (AE240 type, Mettler-Toledo Instrument (Shanghai) Co., Ltd.).

The peoniflorin reference substance is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute (lot number 0736-200116, assay is used).

Acetonitrile is a chromatographically pure, and water is distilled water, and other reagent is analytical pure.

Measuring content of paeoniflorin with high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000) comprises the following steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler, and acetonitrile-water is a mobile phase, column temperature 25-30 ℃, detects wavelength 230 ± 2nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 1500;

B. the preparation of reference substance solution: the peoniflorin reference substance that takes by weighing drying is an amount of, adds mobile phase or methanol is made solution;

C. the preparation of need testing solution: get this product and put in the separatory funnel, thin up adds water saturated n-butyl alcohol jolting and extracts repeatedly, merges n-butyl alcohol liquid, and water bath method, residue add mobile phase or methanol makes dissolving in right amount, add mobile phase or

Methanol dilution and standardize solution;

D. algoscopy: accurate respectively reference substance and the need testing solution drawn, inject chromatograph of liquid, measure;

The numerical range value of above-mentioned chromatographic condition mobile phase can be chosen acetonitrile-water (10-19: 81-90), preferred acetonitrile-water (12: 88); It is an amount of that the best precision of the preparation of reference substance solution takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase or methanol and make the reference substance solution that every 1ml contains 10-60 μ g; The test sample preparation adds the dilution of 5ml water, adds 10ml water-saturated n-butanol extraction conditions the best, and last constant volume can be at 25-50ml, and preferably standardize solution is 25ml; Accurate respectively reference substance and each 5-10 μ l of need testing solution of drawing injects chromatograph of liquid, measures, promptly.Every ml QIJU DIHUANG KOUFUYE contains Cortex Moutan in peoniflorin, must not be less than 0.07mg.

Chinese crude drug Flos Chrysanthemi thin layer chromatography of the present invention (an appendix VI of 2000 editions Chinese Pharmacopoeias B) is differentiated and is followed these steps to carry out:

A. the preparation of control medicinal material solution: get the Flos Chrysanthemi control medicinal material, add alcohol reflux, filter, the filtrate evaporate to dryness,

Residue is dissolved in water, and last polyamide column successively with water, ethanol elution, is collected ethanol elution, and evaporate to dryness, residue add ethyl acetate dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product, last polyamide column successively with water, ethanol elution, is collected ethanol elution, and evaporate to dryness, residue add the ethyl acetate dissolving, as need testing solution.

C. according to thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, be developing solvent with chloroform-ethyl acetate-methanol-formic acid, launch, take out, dry, spray is with the ferric chloride alcoholic solution;

Above-mentioned Flos Chrysanthemi thin layer differentiates that sample treatment can adopt the method for polyamide column, the preferred amide post is (3g, the 30-60 order, diameter 1cm), developing solvent is chloroform-ethyl acetate-methanol-formic acid, (5: 5: 2: be best 0.5), developer was ferric chloride alcoholic solution the best to the ratio of chloroform-ethyl acetate-methanol-formic acid.

Chinese crude drug Rhizoma Alismatis thin layer chromatography (appendix VIB of 2000 editions Chinese Pharmacopoeias) is differentiated

A. the preparation of control medicinal material solution: get the Rhizoma Alismatis control medicinal material, add ethanol, reflux, extract, filters, the filtrate evaporate to dryness, residue is dissolved in water, and adds the ethyl acetate jolting and extracts repeatedly, merges ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate dissolving, medical material solution in contrast;

B. the preparation of test sample: get this product, the jolting that adds diethyl ether is extracted repeatedly, merges ether solution, extracts repeatedly with the aqueous slkali jolting, discards alkaline solution, and ether solution adds the water jolting and extracts, and water liquid discards, and ether solution evaporate to dryness, residue add the ethyl acetate dissolving, as need testing solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether-ethyl acetate-formic acid is developing solvent, launches, and takes out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol, and it is clear that the room temperature placement developed the color to speckle in several minutes.

Above-mentioned Rhizoma Alismatis thin layer differentiates that sample treatment can adopt extracted with diethyl ether, and aqueous slkali remove impurity, developing solvent are petroleum ether-ethyl acetate-formic acid, and developer is acetic anhydride-sulphuric acid-dehydrated alcohol; Above-mentioned aqueous slkali can be used 5% sodium bicarbonate solution; Developing solvent is petroleum ether (30-60 ℃)-ethyl acetate-formic acid, and the ratio of developing solvent (6: 4: 0.1) is best; The developer ratio is acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5).

The thin layer chromatography (appendix VIB of 2000 editions Chinese Pharmacopoeias) of preferred Chinese crude drug Flos Chrysanthemi is differentiated and is followed these steps to carry out:

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 1-3g, add ethanol 50-150ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 20-60ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1-3ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 20-60ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 0.5-3ml dissolving, as need testing solution.

C. test according to thin layer chromatography (appendix VIB of 2000 editions Chinese Pharmacopoeias), draw each 5-20 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5) be developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle (2-3) of same color.

The thin layer chromatography (appendix VIB of 2000 editions Chinese Pharmacopoeias) of preferred Chinese crude drug Rhizoma Alismatis is differentiated and is followed these steps to carry out:

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 21081-200302) 4-10g, add 70% ethanol 50-150ml, reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add water 20ml dissolving, adding the ethyl acetate jolting extracts twice, each 10-30ml merges ethyl acetate liquid, evaporate to dryness, residue adds ethyl acetate 0.5-3ml dissolving, medical material solution in contrast.

C. the preparation of test sample: get this product 60-180ml, the jolting that adds diethyl ether is extracted three times, and each 30-90ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 15-30ml discards alkaline solution, and ether solution adds water 15-30ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 0.5-3ml dissolving, as need testing solution.

C. test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 5-20ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

More preferably Chinese crude drug Flos Chrysanthemi thin layer chromatography (appendix VIB of 2000 editions Chinese Pharmacopoeias) discrimination method step:

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 1g, add ethanol 50ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 20ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 20ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution.

C. test according to thin layer chromatography, draw each 10 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5) be developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution.

More preferably the thin layer chromatography of Chinese crude drug Rhizoma Alismatis is differentiated and is followed these steps to carry out:

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 21081-200302) 4g, add 70% ethanol 50ml, reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add water 20ml dissolving, adding the ethyl acetate jolting extracts twice, each 10ml merges ethyl acetate liquid, evaporate to dryness, residue adds ethyl acetate 0.5ml dissolving, medical material solution in contrast;

B. the preparation of test sample: get this product 60ml, the jolting that adds diethyl ether is extracted three times, and each 30ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 15ml discards alkaline solution, and ether solution adds water 15ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 0.5ml dissolving, as need testing solution;

C. test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30 ~ 60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

The treatment hepatic and renal YIN deficiency of the present invention, vertigo and tinnitus, the photophobia photophobia, the preparation method of blurring of vision disease QIJU DIHUANG KOUFUYE is by the ten (WS of " Drug Standard of Ministry of Public Health of the Peoples Republic of China " (Chinese traditional patent formulation preparation) ₃-preparation method in B-2146-96) is made, and every bag of capacity is 10ml, and is oral, a 10ml, 2 times on the one.

The TCL that the invention provides Flos Chrysanthemi, Rhizoma Alismatis differentiates, the quality standard of former assay is improved, and comprises the bottom line that improves content assaying method in the preparation and improve paeoniflorin content.Through repeated trials repeatedly, confirmation method is easy, the result is accurate, can be used as the QIJU DIHUANG KOUFUYE quality control and investigates the index of the stability of technology.

The methodological study of assay method

(1) linear relationship: precision is measured peoniflorin reference substance mother solution (0.2408mg/ml) 0.25,0.5,1,1.5, and 2ml puts in the 10ml measuring bottle, adds mobile phase and is diluted to scale, shakes up.By method of the present invention, inject chromatograph of liquid, measure the peoniflorin peak area.With sample size (m) is abscissa, and peak area (A) is a vertical coordinate, and the drawing standard curve calculates regression equation: A=11862479m-20262.4, r=0.9998, straight-line pass initial point, the range of linearity 0.0301～0.2408 μ g.

(2) precision test: getting concentration is the peoniflorin reference substance solution of 14.08 μ g/ml, and the accurate 5 μ l that draw inject chromatograph of liquid, measure the peoniflorin peak area, and METHOD FOR CONTINUOUS DETERMINATION 6 times the results are shown in Table 2.

Table 2, Precision test result

Sequence number peak area meansigma methods RSD% (n=6)

1 1359899

2 1422759

3 1406292

1389283 1.74

4 1366895

5 1398489

6 1381366

Above result shows that this method precision is good.

(3) stability test: get this product need testing solution (lot number 0301001), at regular intervals, inject chromatograph of liquid, measure the peoniflorin peak area, the results are shown in Table 3.

Table 3, stability test result

Sequence number blanking time (hour) peak area RSD% (n=5)

1 0 979969

2 1 981891

3 2 988169 0.81

4 3 988544

5 4 969000

Above result shows that sample is stable in 4 hours, and this method has good stability.

(4) repeatability test: get 5 parts of this product (lot number 0301001), measure, the results are shown in Table 4 by the inventive method.

Table 4, reproducible test results

Sequence number content mg/ml average content mg/ml RSD% (n=5)

1 0.117

2 0.111

3 0.114 0.114 1.96

4 0.113

5 0.115

Above result shows that this method repeatability is good.

(5) recovery test: precision is measured this product 2ml, and the accurate reference substance mother solution that adds is an amount of, measures paeoniflorin content by the inventive method, is calculated as follows the response rate, the results are shown in Table 5.

Table 5, recovery test result

The known quantity addition amount of recording response rate meansigma methods RSD%

Sequence number

mg mg mg ％％ (n＝5)

1 0.2280 0.2408 0.4752 102.66

2 0.2280 0.2408 0.4727 101.62

3 0.2280 0.2408 0.4786 100.75 102.04 1.02

4 0.2280 0.2408 0.4771 103.45

5 0.2280 0.2408 0.4729 101.70

Above result shows that this method response rate is good.

(6) negative interference test: get and do not contain the negative control product that the Cortex Moutan medical material is made, measure the peoniflorin peak area by the inventive method, the result shows noiseless.

(7) sample determination: get 8 batches of this product,, measure paeoniflorin content, the results are shown in Table 6 by the inventive method.

Table 6,8 batch sample measurement results

Sequence number lot number content mg/ml meansigma methods mg/ml

0.114

1 0301001 0.114

0.113

0.120

2 0301002 0.120

0.120

0.117

3 0301003 0.116

0.114

4 0302001 0.114

0.115

0.124

5 0302002 0.126

0.128

0.120

6 0304001 0.121

0.122

0.090

7 0304002 0.090

0.089

0.095

8 0304003 0.095

0.094

According to above result and primary standard, the every 1ml of regulation this product contains Cortex Moutan in peoniflorin (C23H28O11), must not be less than 0.07mg.

The specific embodiment

Embodiment 1, get Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, above-mentioned raw materials Flos Chrysanthemi, Cortex Moutan were with 20 times of medical material water yield vapor distillations 4 hours, distillate carries out redistillation again, and it is an amount of to collect re-distilled liquid; Medicinal residues add 10 times of medical material amount of water and decocted 1.5 hours, filter, and medicinal liquid is standby.Fructus Lycii, Radix Rehmanniae Preparata, Fructus Corni, Rhizoma Dioscoreae, Rhizoma Alismatis add 8 times of water gagings of medical material and decoct twice, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered.Filtrate and Flos Chrysanthemi, Cortex Moutan decocting liquid merge, and are concentrated into about 880ml, put coldly, add ethanol and make and contain the alcohol amount and reach 70%, stir evenly, and leave standstill 48 hours, filter recovery ethanol.Poria adds 20 times of water gagings and decocts secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and filtrate is concentrated in right amount.Distillate and above-mentioned each medicinal liquid are merged, add simple syrup 250ml, sodium benzoate 2.5g, add water to 1000ml, stir evenly, filter, embedding, every bag of capacity is 10ml.

1. measure content of paeoniflorin with high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000):

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (12: 88) is a mobile phase, and 28 ℃ of column temperatures detect wavelength 230nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 1500;

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase and make the solution that every 1ml contains 20 μ g;

C. the preparation of need testing solution: precision is measured this product 5ml, puts in the separatory funnel, adds water 5ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly;

D. algoscopy: accurate respectively reference substance and each 5 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

Every ml QIJU DIHUANG KOUFUYE contains Cortex Moutan in peoniflorin, must not be less than 0.07mg.

2. differentiate this medicine Chinese crude drug Flos Chrysanthemi with thin layer chromatography

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 1g, add ethanol 50ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 20ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1ml dissolving, medical material solution in contrast;

D. the preparation of test sample: get this product 20ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution;

C. test according to thin layer chromatography (appendix VIB of 2000 editions Chinese Pharmacopoeias), draw each 10 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5) be developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle (2-3) of same color.

3. differentiate this medicine Chinese crude drug Rhizoma Alismatis with thin layer chromatography

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 21081-200302) 4g, add 70% ethanol 50ml, reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add water 20ml dissolving, adding the ethyl acetate jolting extracts twice, each 10ml merges ethyl acetate liquid, evaporate to dryness, residue adds ethyl acetate 0.5ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 60ml, the jolting that adds diethyl ether is extracted three times, and each 30ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 15ml discards alkaline solution, and ether solution adds water 15ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 0.5ml dissolving, as need testing solution.

C. test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

With the index of said method as the stability of this drug quality control and investigation technology.

Embodiment 2

Get Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, above-mentioned raw materials Flos Chrysanthemi, Cortex Moutan were with 20 times of medical material water yield vapor distillations 4 hours, distillate carries out redistillation again, and it is an amount of to collect re-distilled liquid; Medicinal residues add 10 times of medical material amount of water and decocted 1.5 hours, filter, and medicinal liquid is standby.Fructus Lycii, Radix Rehmanniae Preparata, Fructus Corni, Rhizoma Dioscoreae, Rhizoma Alismatis add 8 times of water gagings of medical material and decoct twice, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered.Filtrate and Flos Chrysanthemi, Cortex Moutan decocting liquid merge, and are concentrated into about 880ml, put coldly, add ethanol and make and contain the alcohol amount and reach 70%, stir evenly, and leave standstill 48 hours, filter recovery ethanol.Poria adds 20 times of water gagings and decocts secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and filtrate is concentrated in right amount.Distillate and above-mentioned each medicinal liquid are merged, add simple syrup 250ml, sodium benzoate 2.5g, add water to 1000ml, stir evenly, filter, embedding, every bag of capacity is 10ml.1. measure content of paeoniflorin with high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000):

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase and make the solution that every 1ml contains 20 μ g.

C. the preparation of need testing solution: precision is measured this product 5ml, puts in the separatory funnel, adds water 5ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly.

D. algoscopy: accurate respectively reference substance and each 5 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.Every ml QIJU DIHUANG KOUFUYE contains Cortex Moutan in peoniflorin, must not be less than 0.07mg.

2. differentiate this medicine Chinese crude drug Flos Chrysanthemi with thin layer chromatography:

C. test according to thin layer chromatography (an appendix VI of 2000 editions Chinese Pharmacopoeias B), draw each 10 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5) be developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle (2-3) of same color.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30 ~ 60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

4. differentiate this medicine Chinese crude drug Cortex Moutan with thin layer chromatography:

A. the preparation of reference substance: it is an amount of to get the paeonol reference substance, adds dissolve with methanol and becomes every 1ml to contain the solution of 0.1mg, product solution in contrast.

B. the preparation of test sample: get this product 20ml, add the ethyl acetate jolting and extract twice, each 20ml merges, and low temperature evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution.

C. test according to thin layer chromatography (an appendix VI of 2000 editions Chinese Pharmacopoeias B), draw each 10 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with cyclohexane extraction-ethyl acetate (3: 1) 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 speckle of same color.

Embodiment 3:

Get Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, above-mentioned raw materials Flos Chrysanthemi, Cortex Moutan were with 20 times of medical material water yield vapor distillations 4 hours, distillate carries out redistillation again, and it is an amount of to collect re-distilled liquid; Medicinal residues add 10 times of medical material amount of water and decocted 1.5 hours, filter, and medicinal liquid is standby.Fructus Lycii, Radix Rehmanniae Preparata, Fructus Corni, Rhizoma Dioscoreae, Rhizoma Alismatis add 8 times of water gagings of medical material and decoct twice, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered.Filtrate and Flos Chrysanthemi, Cortex Moutan decocting liquid merge, and are concentrated into about 880ml, put coldly, add ethanol and make and contain the alcohol amount and reach 70%, stir evenly, and leave standstill 48 hours, filter recovery ethanol.Poria adds 20 times of water gagings and decocts secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and filtrate is concentrated in right amount.Distillate and above-mentioned each medicinal liquid are merged, add simple syrup 250ml, sodium benzoate 2.5g, add water to 1000ml, stir evenly, filter, embedding, every bag of capacity is 10ml.The quality control detection method that QIJU DIHUANG KOUFUYE is new, this method can be undertaken by following determination step:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (10: 90) is a mobile phase, and 25 ℃ of column temperatures detect wavelength 230nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 1500.

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase and make the solution that every 1ml contains 10 μ g.

C. the preparation of need testing solution: precision is measured this product 5ml, puts in the separatory funnel, adds water 5ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase makes dissolving in right amount, in the dislocation 50ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly.

D. algoscopy: accurate respectively reference substance and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.Every ml QIJU DIHUANG KOUFUYE contains Cortex Moutan in peoniflorin, must not be less than 0.07mg.

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 3g, add ethanol 150ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 60ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1-3ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 60ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 3ml dissolving, as need testing solution.

3. this medicine Chinese crude drug Rhizoma Alismatis is differentiated with thin layer chromatography

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material 10g, add 70% ethanol 150ml, reflux, extract, 1 hour filters, the filtrate evaporate to dryness, residue adds water 20ml dissolving, adds the ethyl acetate jolting and extracts twice, each 30ml, merge ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 3ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 180ml, the jolting that adds diethyl ether is extracted three times, and each 90ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 30ml discards alkaline solution, and ether solution adds water 15-30ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 3ml dissolving, as need testing solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 15ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Embodiment 4:

Get Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g, above-mentioned raw materials Flos Chrysanthemi, Cortex Moutan were with 20 times of medical material water yield vapor distillations 4 hours, distillate carries out redistillation again, and it is an amount of to collect re-distilled liquid; Medicinal residues add 10 times of medical material amount of water and decocted 1.5 hours, filter, and medicinal liquid is standby.Fructus Lycii, Radix Rehmanniae Preparata, Fructus Corni, Rhizoma Dioscoreae, Rhizoma Alismatis add 8 times of water gagings of medical material and decoct twice, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered.Filtrate and Flos Chrysanthemi, Cortex Moutan decocting liquid merge, and are concentrated into about 880ml, put coldly, add ethanol and make and contain the alcohol amount and reach 70%, stir evenly, and leave standstill 48 hours, filter recovery ethanol.Poria adds 20 times of water gagings and decocts secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and filtrate is concentrated in right amount.Distillate and above-mentioned each medicinal liquid are merged, add simple syrup 250ml, sodium benzoate 2.5g, add water to 1000ml, stir evenly, filter, embedding, every bag of capacity is 10ml.

The quality control detection method that QIJU DIHUANG KOUFUYE is new, this method can be undertaken by following determination step:

1. measuring content of paeoniflorin with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D) measures and standard:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (19: 81) is a mobile phase, and 27 ℃ of column temperatures detect wavelength 230nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 1500.

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds methanol and make the solution that every 1ml contains 30 μ g.

C. the preparation of need testing solution: precision is measured this product 6ml, puts in the separatory funnel, adds water 5ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds methanol makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds methanol and is diluted to scale, shakes up, promptly.

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 2g, add ethanol 50-150ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 40ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1-3ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 40ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 2ml dissolving, as need testing solution.

C. test according to thin layer chromatography (an appendix VI of 2000 editions Chinese Pharmacopoeias B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5) be developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle (2-3) of same color.

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 21081-200302) 6g, add 70% ethanol 100ml, reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add water 20ml dissolving, adding the ethyl acetate jolting extracts twice, each 30ml merges ethyl acetate liquid, evaporate to dryness, residue adds ethyl acetate 1ml dissolving, medical material solution in contrast.

D. the preparation of test sample: get this product 100ml, the jolting that adds diethyl ether is extracted three times, and each 60ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 20ml discards alkaline solution, and ether solution adds water 20ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Embodiment 5

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (15: 85) is a mobile phase, and 26 ℃ of column temperatures detect wavelength 230nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 1500.

E. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase and make the solution that every 1ml contains 40 μ g.

F. the preparation of need testing solution: precision is measured this product 10ml, puts in the separatory funnel, adds water 10ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly.

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 2g, add ethanol 100ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 60ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 40ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution.

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material 8g, add 70% ethanol 100ml, reflux, extract, 1 hour filters, the filtrate evaporate to dryness, residue adds water 20ml dissolving, adds the ethyl acetate jolting and extracts twice, each 30ml, merge ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 2ml dissolving, medical material solution in contrast.

B. the preparation of test sample: get this product 100ml, the jolting that adds diethyl ether is extracted three times, and each 60ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 20ml discards alkaline solution, and ether solution adds water 15-30ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 20ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launch, take out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5), and it is clear that the room temperature placement developed the color to speckle in several minutes.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Embodiment 6

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (14: 86) is a mobile phase, and 29 ℃ of column temperatures detect wavelength 230nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 1500.

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase and make the solution that every 1ml contains 30 μ g.

C. the preparation of need testing solution: precision is measured this product 8ml, puts in the separatory funnel, adds water 10ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds mobile phase and is diluted to scale, shakes up, promptly.

B. the preparation of test sample: get this product 20ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 0.5ml dissolving, as need testing solution.

Embodiment 7

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds methanol and make the solution that every 1ml contains 20 μ g.

C. the preparation of need testing solution: precision is measured this product 5ml, puts in the separatory funnel, adds water 5ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds methanol makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds methanol and is diluted to scale, shakes up, promptly.

Claims

1. the method for quality control of a QIJU DIHUANG KOUFUYE of being made by Chinese crude drug Fructus Lycii 33g, Flos Chrysanthemi 33g, Radix Rehmanniae Preparata 130g, Fructus Corni (system) 65g, Cortex Moutan 50g, Rhizoma Dioscoreae 65g, Poria 50g, Rhizoma Alismatis 50g is characterized in that this method comprises the following steps:

(1), with content of paeoniflorin in this medicine of high effective liquid chromatography for measuring;

(2), differentiate this medicine Chinese crude drug Flos Chrysanthemi with thin layer chromatography;

(3), differentiate this medicine Chinese crude drug Rhizoma Alismatis with thin layer chromatography.

2. according to the method for quality control of the QIJU DIHUANG KOUFUYE of claim 1, it is characterized in that this method comprises the following steps:

(1), with content of paeoniflorin in this medicine of high effective liquid chromatography for measuring:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler, and acetonitrile-water is a mobile phase, column temperature 25-30 ℃, detects wavelength 230 ± 2nm;

C. the preparation of need testing solution: get this product and put in the separatory funnel, thin up adds water saturated n-butyl alcohol jolting and extracts repeatedly, merges n-butyl alcohol liquid, and water bath method, residue add mobile phase or methanol makes dissolving in right amount, adds dilution of mobile phase or methanol and standardize solution;

(2). differentiate this medicine Chinese crude drug Flos Chrysanthemi with thin layer chromatography:

A. the preparation of control medicinal material solution: get the Flos Chrysanthemi control medicinal material, add alcohol reflux, filter the filtrate evaporate to dryness, residue is dissolved in water, and last polyamide column successively with water, ethanol elution, is collected ethanol elution, evaporate to dryness, residue add ethyl acetate dissolving, medical material solution in contrast;

B. the preparation of test sample: get this product, last polyamide column successively with water, ethanol elution, is collected ethanol elution, and evaporate to dryness, residue add the ethyl acetate dissolving, as need testing solution;

(3). the medicine Chinese crude drug is differentiated with thin layer chromatography by Rhizoma Alismatis

C. according to the thin layer chromatography test, drawing above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, is developing solvent with petroleum ether-ethyl acetate-formic acid, launches, and takes out, and dries, and spray is with acetic anhydride-sulphuric acid-dehydrated alcohol.

3. according to the method for quality control of the QIJU DIHUANG KOUFUYE of claim 2, it is characterized in that this method comprises the following steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler, acetonitrile-water (10-19: 81-90) be mobile phase, column temperature 25-30 ℃, detect wavelength 230 ± 2nm;

B. the preparation of reference substance solution: it is an amount of that precision takes by weighing dry 48 hours peoniflorin reference substance of phosphorus pentoxide, adds mobile phase or methanol and make the solution that every 1ml contains 10-60 μ g;

C. the preparation of need testing solution: precision is measured this product 5-10ml, puts in the separatory funnel, adds water 5-10ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase or methanol makes dissolving in right amount, in the dislocation 25-50ml measuring bottle, adds mobile phase or methanol is diluted to scale, shakes up;

D. algoscopy: accurate respectively reference substance and each 5-10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure.

A. the preparation of control medicinal material solution: get Flos Chrysanthemi control medicinal material 1-3g, add ethanol 50-150ml reflux, extract, 1 hour, filter, the filtrate evaporate to dryness, residue adds water 20-60ml dissolving, polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution, evaporate to dryness with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, residue adds ethyl acetate 1-3ml dissolving, medical material solution in contrast;

B. the preparation of test sample: get this product 20-60ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 0.5-3ml dissolving, as need testing solution;

C. test according to thin layer chromatography, draw each 5-20 μ l of above-mentioned two kinds of solution, put respectively same be on the silica gel g thin-layer plate of binding agent with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5) be developing solvent, launch, take out, dry, spray is with 5% ferric chloride alcoholic solution.

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material 4-10g, add 70% ethanol 50-150ml, reflux, extract, 1 hour filters, the filtrate evaporate to dryness, residue adds water 20ml dissolving, adds the ethyl acetate jolting and extracts twice, each 10-30ml, merge ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 0.5-3ml dissolving, medical material solution in contrast;

B. the preparation of test sample: get this product 60-180ml, the jolting that adds diethyl ether is extracted three times, and each 30-90ml merges ether solution, extract twice with 5% sodium bicarbonate solution jolting, each 15-30ml discards alkaline solution, and ether solution adds water 15-30ml jolting and extracts, water liquid discards, ether solution evaporate to dryness, residue add ethyl acetate 0.5-3ml dissolving, as need testing solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 5-20ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launches, and takes out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5).

4. the method for quality control of QIJU DIHUANG KOUFUYE according to claim 3 is characterized in that this method comprises the following steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Acetonitrile-water (12: 88) is a mobile phase, and 28 ℃ of column temperatures detect wavelength 230nm;

C. the preparation of need testing solution: precision is measured this product 5ml, puts in the separatory funnel, adds water 5ml dilution, adds water saturated n-butyl alcohol jolting and extracts four times, and each 10ml merges n-butyl alcohol liquid, water bath method.Residue adds mobile phase makes dissolving in right amount, in the dislocation 25ml measuring bottle, adds mobile phase and is diluted to scale, shakes up;

D. content of paeoniflorin assay method: accurate respectively reference substance and each 5 μ l of need testing solution of drawing, inject chromatograph of liquid, measure content of paeoniflorin.

(2) medicine Chinese crude drug Flos Chrysanthemi is differentiated with thin layer chromatography

B. the preparation of test sample: get this product 20ml, last polyamide column (3g, 30-60 order, diameter 1cm), successively, collect 95% ethanol elution with water, 10% ethanol, 30% ethanol, each 50ml eluting of 95% ethanol, evaporate to dryness, residue add ethyl acetate 1ml dissolving, as need testing solution;

(3) medicine Chinese crude drug Rhizoma Alismatis is differentiated with thin layer chromatography

A. the preparation of control medicinal material solution: get Rhizoma Alismatis control medicinal material 4g, add 70% ethanol 50ml, reflux, extract, 1 hour filters, the filtrate evaporate to dryness, residue adds water 20ml dissolving, adds the ethyl acetate jolting and extracts twice, each 10ml, merge ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 0.5ml dissolving, medical material solution in contrast;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 10ul of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1) is developing solvent, launches, and takes out, dry, spray is with acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5).

5. according to the method for quality control of the arbitrary described QIJU DIHUANG KOUFUYE of claim 2-4, it is characterized in that: in the content assaying method

A. the test sample thin up adds the water-saturated n-butanol extraction;

B. last constant volume is decided to be 25-50ml;

C. the bottom line of peoniflorin is 0.07mg/ml.

6. the method for quality control of QIJU DIHUANG KOUFUYE according to claim 5 is characterized in that:

A. test sample adds the dilution of 5ml water, adds the extraction of 10ml water-saturated n-butanol;

B. last constant volume is decided to be 25ml;

C. the bottom line of peoniflorin is 0.07mg/ml.

7. according to the method for quality control of the arbitrary described QIJU DIHUANG KOUFUYE of claim 2-4, it is characterized in that: in the Chinese crude drug Flos Chrysanthemi thin layer chromatography discrimination method on the sample polyamide column, developing solvent be chloroform-ethyl acetate-methanol-formic acid, developer ferric chloride alcoholic solution.

8. according to the method for quality control of claim 7 QIJU DIHUANG KOUFUYE, it is characterized in that: polyamide column (3g, 30-60 order, diameter 1cm), developing solvent chloroform-ethyl acetate-methanol-formic acid (5: 5: 2: 0.5).

9. according to the method for quality control of the arbitrary described QIJU DIHUANG KOUFUYE of claim 2-4, it is characterized in that: the preparation of test sample employing extracted with diethyl ether, aqueous slkali remove impurity, developing solvent are petroleum ether-ethyl acetate-formic acid in the Chinese crude drug Rhizoma Alismatis thin layer chromatography discrimination method, and developer is acetic anhydride-sulphuric acid-dehydrated alcohol.

10. the method for quality control of QIJU DIHUANG KOUFUYE according to claim 9, it is characterized in that: adopting extracted with diethyl ether, 5% sodium bicarbonate solution remove impurity, developing solvent is petroleum ether (30-60 ℃)-ethyl acetate-formic acid (6: 4: 0.1), and developer is acetic anhydride-sulphuric acid-dehydrated alcohol (1: 1: 5).