CN100402058C

CN100402058C - Quality control method of compound polygonium oriental preparation

Info

Publication number: CN100402058C
Application number: CNB2005100033148A
Authority: CN
Inventors: 叶湘武; 王泽坤
Original assignee: Guizhou Yibai Pharmaceutical Co Ltd
Current assignee: Guizhou Yibai Pharmaceutical Co Ltd
Priority date: 2005-12-09
Filing date: 2005-12-09
Publication date: 2008-07-16
Anticipated expiration: 2025-12-09
Also published as: CN1785263A

Abstract

The present invention discloses a quality control method for a compound orientin preparation in order to effectively control the quality of the preparation and ensure curative effect with the continuously enlarging of the application of the compound Hongcao preparation. The method adopts a high-efficiency liquid-phase chromatography to measure the content of wild baicalin, isoorientin and orientin in the medicament, and has the advantages of simplicity, good repeatability and accurate result. The present invention adopts thin layer chromatography to carry out discrimination to the orientin and fleabane, the method is exclusive and feasible, the accuracy and the advancement of quality detection standard are ensured, and the quality of the compound orientin preparation can be controlled effectively. The present invention can be used as the quality control method of compound orientin freeze-dried powder injection, injection and oral taken formulations. The quality control method has the advantage of good precision, sensitivity and stability, and the quality of products can be guaranteed to be uniform, stable, effective and controllable.

Description

The method of quality control of compound oriental smartweed preparation

Technical field:

The present invention relates to a kind of method of quality control of Chinese medicine preparation, particularly the compound oriental smartweed preparation method of quality control.

Background technology:

The prescription of compound oriental smartweed preparation comes from folk remedy, crude drug is made up of Herba Polygoni Orientalis and Herba Erigerontis, early eighties, the shady chief physician of the cardiovascular diseases expert great-grandfather of Guizhou Province etc. has carried out a large amount of big pharmacology and clinical research work to this side, on the basis of application among the people and experimental study, now it is developed into freeze-dried powder, injection and oral formulations and is used for the clinical treatment angina pectoris.But discover that through us existing compound oriental smartweed preparation exists method of quality control to fall behind the uppity shortcoming of product quality.Herba Polygoni Orientalis and Herba Erigerontis are not differentiated in the existing method of quality control, or effective ingredient in Herba Polygoni Orientalis and the Herba Erigerontis is carried out assay.So existing method of quality control can not effectively be controlled the quality of compound oriental smartweed preparation, thereby will influence the production of product and ensure the quality of products.

Continuous expansion along with above-mentioned preparation practical application, for effectively controlling the quality of preparation, guarantee curative effect, we propose the method for quality control of compound oriental smartweed preparation, this method adopts scutellarin in this medicine of high effective liquid chromatography for measuring, isorientin, the content of orientin, its method is easy, favorable reproducibility, result are accurate; Adopt thin layer chromatography that Herba Polygoni Orientalis and Herba Erigerontis are differentiated, its method is exclusive, feasible, the accuracy and the advance of quality inspection standard have been guaranteed, can control the quality of compound oriental smartweed preparation effectively, can be used as the compound oriental smartweed freeze-dried powder, the method for quality control of injection and oral formulations.This method of quality control precision, sensitivity, stability are all good, guarantee " homogeneous, stable, effective, controlled " of product quality.

Summary of the invention:

The object of the present invention is to provide the method for quality control of compound oriental smartweed preparation.

The present invention be directed to compound oriental smartweed preparation and propose method of quality control, this compound oriental smartweed preparation comprises other Chinese medicine preparation such as oral liquid, syrup, granule, freeze-dried powder and injection.

Described compound oriental smartweed preparation of the present invention is made by following parts by weight of Chinese traditional medicine raw material:

Herba Polygoni Orientalis 5000-10000 part and Herba Erigerontis 1000-2000 part.

Preferably

1500 parts of 7500 parts of Herba Polygoni Orientaliss and Herba Erigerontiss.

In more than forming, every part of representative be weight portion, weight is calculated with crude drug, if be unit with the gram, more than form and can be made into 1000 doses of pharmaceutical preparatioies, described 1000 doses of fingers, the final drug preparation of making, as make 1000 of capsule preparations, 1000 in tablet, granule 1000g, liquid preparation 1000ml etc.

More than form being by weight as proportioning, can increasing or reduce according to corresponding proportion when producing, can be unit with the kilogram as large-scale production, or is unit with the ton.

The preparation method of compound oriental smartweed preparation of the present invention adopts following method:

By the raw material of Chinese medicine of above-mentioned prescription is processed through extraction or other modes, making pharmaceutically active substance, subsequently, is raw material with this material, add the medicine acceptable carrier when needing, make oral liquid, syrup, granule, freeze-dried powder and injection according to the routine techniques of galenic pharmacy.Described active substance can obtain by the method that is selected from following mode, as: by pulverize, squeeze, calcine, grind, sieve, percolation, extraction, water are carried, alcohol extraction, ester are carried, methods such as ketone is carried, chromatography obtain, these active substances can be the materials of extractum form, can be that dry extract also can be a fluid extract, can also be the high-purity extract, make different concentration according to the different needs decision of preparation.

Compound oriental smartweed preparation of the present invention, when making preparation, can add the medicine acceptable carrier as required, these carriers can be any carriers that is fit to make compound oriental smartweed preparation, as: benzoic acid or potassium salt, mannitol, sorbitol, sorbic acid or potassium salt, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, Nepal's methyl ester, Nepal's ethyl ester, Nepal's propyl ester, Nepal's butyl ester, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone, the EDTA disodium, EDTA calcium sodium, the alkali-metal carbonate of monovalence, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulphuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, propylene glycol, ethanol, soil temperature 60-80, Arlacel-80, Cera Flava, lanoline, liquid paraffin, hexadecanol, gallate ester, agar, triethanolamine, basic amino acid, carbamide, allantoin, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, phospholipid material etc.

The present invention is to provide method of quality control, be the quality of control compound oriental smartweed preparation at above compound oriental smartweed preparation.At its characteristics and prescription of the present invention, we provide following method of quality control:

Method of quality control of the present invention may further comprise the steps:

Character is observed, and official method checks content, to the scutellarin that contains, and isorientin, orientin carries out assay, and content Herba Polygoni Orientalis and Herba Erigerontis are differentiated.

Therefore, the main step of method of quality control of the present invention is:

A. measure scutellarin in the pharmaceutical preparation of the present invention, isorientin, the content of orientin;

B. Herba Polygoni Orientalis in the pharmaceutical preparation of the present invention and Herba Erigerontis are differentiated;

Wherein the concrete steps of A mensuration content are as follows:

Measure scutellarin in the pharmaceutical preparation of the present invention, isorientin, the step of the content of orientin;

Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version-appendix VI D of portion in 2000),

Instrument, reagent and sample instrument: Tianjin, island HPLC system comprises the LC-10ATvp pump, LC-10Avp UV, visible light optical detector; Rheodyne7725 hand sampling valve (U.S. Rheodyne company); Weil-McLain dragon chromatographic work station (Guangxi Weil-McLain dragon scientific ﹠ technical corporation); TCQ-250 ultrasonic washing unit (Beijing's armarium two factories, 250W, 19-33Hz).It is double distilled water that acetonitrile, methanol are chromatographically pure (U.S. TEDIA company), 95% ethanol (analytical pure), phosphoric acid (analytical pure), water.Reference substance: isorientin and orientin (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, and for assay usefulness, lot number is 818-200001); Reference substance: scutellarin (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, and for assay usefulness, lot number is 880-200001);

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is second eyeball-0.1% phosphoric acid solution; Set flow velocity: 1ml/min, temperature and detection wavelength; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600.

B. it is an amount of that isorientin, orientin and scutellarin reference substance are got in the preparation of reference substance solution respectively, drying, and accurate the title, decide, add the first dissolving, make every 1ml and contain the solution that isorientin 0.025mg, every 1ml contain orientin 0.02mg and scutellarin 0.08mg, shake up, promptly.

C. the preparation precision of need testing solution is measured the content under the loading amount item, and mixing is got 50ml or 50mg, puts in the 100ml measuring bottle, and accurate the title decides, and adds water and makes dissolving in right amount and be diluted to scale, shakes up, and gets supernatant and filters, promptly.

D. accurate respectively reference substance solution and the need testing solution drawn of algoscopy injects chromatograph of liquid, measures;

The numerical range of above-mentioned chromatographic condition mobile phase can be chosen for second eyeball-0.1% phosphoric acid solution (14-25: 69-91), preferred proportion is second eyeball-0.1% phosphoric acid solution (18: 82), temperature: 37-42 ℃, is preferably 40 ℃, the detection wavelength is 350 ± 2nm, is preferably 350nm; The best precision of the preparation of reference substance solution takes by weighing and reaches 24 hours through the phosphorus pentoxide dried overnight, uses dissolve with methanol; The preparation of need testing solution is preferably got supernatant 0.45 μ m and is filtered; Accurate respectively reference substance solution and the need testing solution 5-10 μ l injecting chromatograph mensuration drawn.The every 1ml of this product contains Herba Polygoni Orientalis with isorientin (C ₂₁H ₂₀O ₁₁) and orientin (C ₂₁H ₂₀O ₁₁) the total amount meter, must not be less than 7.0mg, contain Herba Erigerontis with scutellarin (C ₂₁H ₁₈O ₂) meter, must not be less than 14.0mg.

Wherein B concrete steps that the Herba Polygoni Orientalis in the pharmaceutical preparation of the present invention and Herba Erigerontis are differentiated are as follows:

Herba Polygoni Orientalis is differentiated: thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get this product, add methanol, supersound process, centrifugal, get supernatant as need testing solution.

B. the preparation of reference substance solution: get isorientin, orientin reference substance, add methanol and make mixed solution, in contrast product solution.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with methanol-water-glacial acetic acid, launches, and takes out, and dries, and the spray developer is put under the ultra-violet lamp and inspected.

Above-mentioned Herba Polygoni Orientalis thin layer is differentiated the processing employing methanol extraction of sample, developing solvent is methanol-water-glacial acetic acid, methanol-water-glacial acetic acid ratio (4: 1: 1) is best, and developer 2% aluminum chloride ethanol liquid be the best, and inspects under ultra-violet lamp (365nm) and be the best.

Or follow these steps to carry out:

B. the preparation of reference substance solution: get protocatechuic acid, gallic acid reference substance, add methanol and make the mixed solution that contains protocatechuic acid and contain gallic acid, product solution in contrast.

C. according to thin layer chromatography test, draw need testing solution, reference substance solution is put respectively in on-the silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid, launches, and takes out, dry, and the spray developer, heating is put daylight lamp and is inspected.

Ether and ethyl acetate extraction are adopted in the processing of above-mentioned Herba Polygoni Orientalis thin layer discriminating sample successively; Developing solvent is chloroform-ethyl acetate-formic acid, and chloroform-ethyl acetate-formic acid ratio (5: 3: 1) is best, and the ferric chloride alcoholic solution of developer 2% is best, and it is best heating 105 ℃.

Herba Erigerontis is differentiated: thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

B. the preparation of reference substance solution: get the scutellarin reference substance, add methanol and make solution, in contrast product solution.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with glacial acetic acid-ethanol, launches, and takes out, and dries, and the spray developer is put under the daylight lamp and inspected.

Above-mentioned Herba Erigerontis thin layer is differentiated the processing employing dehydrated alcohol extraction of sample, and developing solvent is glacial acetic acid-ethanol, and glacial acetic acid-proportion of ethanol (4: 1) is best, and developer 2% ferric chloride ethanol liquid is best.

The present invention has also carried out preferably the condition of the inventive method, below is preferred version:

Preferred Herba Polygoni Orientalis is differentiated: thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

A. the preparation of need testing solution: get this product 10mg or 10ml, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution.

B. the preparation of reference substance solution: get isorientin, orientin reference substance, add methanol and make the mixed solution that every 1ml contains 0.5mg, in contrast product solution.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, with methanol-water-glacial acetic acid (2.4-7.5: 0.5-2.5: 0.5-2.5) be developing solvent, launch, take out, dry, the spray developer is put under the ultra-violet lamp and is inspected.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

Or follow these steps to carry out:

B. the preparation of reference substance solution: get protocatechuic acid, gallic acid reference substance, add methanol and make the mixed solution that every 1ml contains protocatechuic acid 0.5mg and contains gallic acid 0.2mg, product solution in contrast.

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 2-3ul, put respectively on same silica gel g thin-layer plate, with chloroform-ethyl acetate-formic acid (2-8: 1.8--5: 0.5-2.5) be developing solvent, launch, take out, dry, spray ferric chloride alcoholic solution with 2%.It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Preferred Herba Erigerontis is differentiated: thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

B. the preparation of reference substance solution: get isorientin and orientin, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 3ul of solution, ribbon is put on same polyamide membrane respectively, with glacial acetic acid-ethanol (2.4-7.5: 0.5-2.5) be developing solvent, launch, take out, dry, spray ferric chloride alcoholic solution, put under the daylight lamp and inspect with 2%.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Preferred Herba Polygoni Orientalis thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, with methanol-water-glacial acetic acid (4: 1: 1) is developing solvent, launches, and takes out, dry, spray aluminum chloride alcoholic solution, put under the ultra-violet lamp (365mn) and inspect with 2%.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

Or follow these steps to carry out:

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 3ul puts respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid (5: 3: 1), launches, and takes out, and dries, and sprays the ferric chloride alcoholic solution with 2%.It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Preferred Herba Erigerontis thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) is differentiated and is followed these steps to carry out:

B. the preparation of reference substance solution: get the scutellarin reference substance, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with glacial acetic acid-ethanol (4: 1), launches, and takes out, and dries, and sprays the ferric chloride alcoholic solution with 2%, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

This product described in the above method is meant that final drug of the present invention is as oral liquid, syrup, granule, freeze-dried powder or injection.

The TCL that the invention provides Herba Polygoni Orientalis and Herba Erigerontis differentiates.Increased the quality determining method to isorientin, orientin and scutellarin content in the preparation, repeatedly repeated trials confirms that this method of quality control is comparatively stable, easy, the result is accurate, repeatability is good.Can be used as the compound oriental smartweed freeze-dried powder, the index of the stability of injection and oral formulations quality control and investigation technology.

The used preparation of experimental example test sample of the present invention adopts the preparation described in each embodiment.

The content assaying method research of experimental example one isorientin of the present invention, orientin and scutellarin

(1) instrument, reagent and sample instrument: Tianjin, island HPLC system comprises the LC-10ATvp pump, LC-10Avp UV, visible light optical detector; Rheodyne7725 hand sampling valve (U.S. Rheodyne company); Weil-McLain dragon chromatographic work station (Guangxi Weil-McLain dragon scientific ﹠ technical corporation); TCQ-250 ultrasonic washing unit (Beijing's armarium two factories, 250W, 19-33Hz).

It is double distilled water that acetonitrile, methanol are chromatographically pure (U.S. TEDIA company), 95% ethanol (analytical pure), phosphoric acid (analytical pure), water.Reference substance: isorientin and orientin (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, and for assay usefulness, lot number is 818-200001); Reference substance: scutellarin (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, and for assay usefulness, lot number is 880-200001); Chromatographic column: Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite); KromasilC ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite); Diamonrsil C ₁₈(5um, 4.6mm * 250mm i.d. Di Ma company); Guard column is Security Guard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company).

(2) test condition is selected 1. chromatographic condition chromatographic columns: Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite); Kromasil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite) and InertsilODS-2 (5um, 5mm * 500mmi.d. Japanese firm); Guard column is SecurityGuard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company); Mobile phase is with methanol-water, methanol-0.1% phosphoric acid solution, and acetonitrile-0.1% phosphoric acid solution is a mobile phase, and the result is with the InertsilODS-2 chromatographic column, and acetonitrile-0.1% phosphoric acid solution (18: 82) separates best to isorientin in the Herba Polygoni Orientalis with orientin.

2. detect the HPLC-PDA ultraviolet spectrogram result of the selection of wavelength according to isorientin, orientin and scutellarin reference substance solution, isorientin and orientin have absorption maximum at the 350nm place, scutellarin has absorption maximum at the 335nm place, under above-mentioned two wavelength conditions, respectively same need testing solution is carried out assay determination, the results are shown in Table 1.

The different wavelength that detect of table 1 detect same need testing solution result relatively

Result of the test shows not have significant difference at 350nm and 335nm place mensuration isorientin, orientin and scutellarin content, and method selects 350nm for detecting wavelength to simplify the analysis.

3, the separating degree of isorientin, orientin and scutellarin and other component peaks should be greater than 1.5, theoretical cam curve is respectively 7600,8100 and 10000 in isorientin, orientin and scutellarin peak, so the theorem opinion number of plates is not less than 7600 in the scutellarin peak.

4, linear relationship is investigated

4.1 the preparation of standard solution

Precision takes by weighing isorientin 13.7mg, with dissolve with methanol and be settled to 50ml, orientin 12.3mg, in the 100ml measuring bottle, add dissolve with methanol and be diluted to scale, scutellarin 21.3mg is in the 25ml volumetric flask, with dissolve with methanol and standardize solution, make every 1ml respectively and contain isorientin 0.274mg, orientin 0.123mg, the reference substance liquid of scutellarin 0.819mg.

4.2 the drafting of standard curve

Above-mentioned each reference substance solution 1 of accurate respectively absorption, 2,4,6,8 μ l are in the 25ml volumetric flask, add methanol to scale, shake up, get every 1ml and contain isorientin 0.01096,0.02192,0.04384,0.06576,0.08768mg, orientin 0.00492,0.00984,0.01968,0.02952,0.03936mg, scutellarin 0.03276,0.06552,0.13104,0.19656,0.26208mg reference substance hybrid working liquid, the above-mentioned working solution 10ul of accurate respectively absorption injects liquid chromatograph, the record chromatograph, with peak area Y sample size X being carried out linear regression calculates, get regression equation, see Table 2,3,4.After getting need testing solution mensuration, with substitution equation of linear regression and the fit equation calculating respectively of isorientin, orientin and scutellarin peak area, the relative deviation of isorientin is 1.18% as a result, and the relative deviation of orientin is 0.21%, and the relative deviation of scutellarin is 0.43%.Visual thus equation of linear regression intercept is 0, so this paper adopts the appearance one point method to calculate content.

Table 2 isorientin linear relationship is investigated

Table 3 orientin linear relationship is investigated

Table 4 scutellarin linear relationship is investigated

Presentation of results, isorientin sample size are at 0.11 μ g～0.88 μ g, and the orientin sample size is at 0.05 μ g～0.39 μ g, and the scutellarin sample size is the good linear relation in 0.33 μ g～2.6 μ g scopes.

(3) precision test

Precision is measured the content under the loading amount item, and mixing prepares test liquid by the preparation method of test liquid under the assay item in the quality standard of the present invention, repeats sample introduction 5 times, measures peak area (table 5).

Table 5 preparation precision is investigated (peak area)

The result as seen, this method precision is good.

(4) stability test

Precision is measured the content under the loading amount item, and mixing prepares test liquid by the preparation method of test liquid under the assay item in the quality standard of the present invention, respectively at measuring isorientin, orientin and scutellarin peak area (seeing Table 5) in 0,1,2,4,8 hour.

Table 5 preparation stability is investigated (peak area)

The result shows that isorientin, orientin and scutellarin are stable in 8 hours in the need testing solution.

(5) repeatability test

Precision is measured the content under the loading amount item, mixing, by 5 parts of test liquids of preparation method preparation of test liquid under the assay item in the quality standard of the present invention, sample introduction is measured peak area, result of calculation is listed table 6 in, the isorientin average content is 5.00%, and RSD is 2.01%, the orientin average content is 3.53%, and RSD is 2.13%, the scutellarin average content is 15.9%, and RSD is 1.52%.

Isorientin, orientin and scutellarin assay repeatability are investigated in table 6 preparation

The result as seen, the repeatability of isorientin, orientin and scutellarin content assaying method is good.

(6) recovery test

Adopt the application of sample absorption method, the accurate respectively 5 parts of samples of having measured content (content: isorientin is 5%, orientin is 3.53%, scutellarin 15.9%) of measuring, it is an amount of to add isorientin, orientin and scutellarin, measure by above-mentioned chromatographic condition sample introduction, the record chromatograph, calculate recovery rate the results are shown in Table 7,8,9, and isorientin average recovery rate 99.0%, RSD are 2.70%; Orientin average recovery rate 99.2%, RSD are 2.01%; Scutellarin average recovery rate 101.1%, RSD are 2.35%.

Isorientin recovery test in the table 7 preparation need testing solution

Orientin recovery test in the table 8 preparation need testing solution

Scutellarin recovery test in the table 9 preparation need testing solution

(7) sample determination

Precision is measured the content under the loading amount item, mixing, and according to preparing need testing solution and reference substance solution by quality standard of the present invention, sample introduction 10 μ l write down chromatograph respectively, measure peak area, calculate content by following formula:

In the formula: A _A: need testing solution isorientin peak area A _B: need testing solution orientin peak area

As _A: the peak area As of isorientin reference substance _B: the peak area of orientin reference substance

Cs _A: isorientin reference substance solution concentration (mg/ml) Cs _B: orientin reference substance solution concentration (mg/ml)

Wi: average loading amount (mg/ props up) W: test sample sample weighting amount (mg)

100: need testing solution constant volume (ml)

In the formula: A _i: need testing solution scutellarin peak area

As: the peak area of scutellarin reference substance

Cs: scutellarin reference substance solution concentration (mg/ml)

Wi: average loading amount (mg/ props up)

100: need testing solution constant volume (ml)

According to prepare 10 batch samples in strict accordance with production technology, measurement result sees Table 10.

The assay result of table 1010 batch sample

Assay data from 10 batches of test specimens, the meansigma methods that this compound oriental smartweed preparation unit dose contains isorientin and orientin total amount is 8.69mg, content range is that 7-10mg/ props up, 100mg or 100ml, the content meansigma methods of wild Radix Scutellariae is 15.78mg, content range is that 14-17mg/ props up, 100mg or 100ml

Experimental example two of the present invention

1, the research that the thin layer chromatography of Herba Polygoni Orientalis medical material in the preparation is differentiated

1.1 the preparation of need testing solution: get this product 10mg or 10ml, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution.

1.2 the preparation of reference substance solution: get isorientin, orientin reference substance, add methanol and make the mixed solution that every 1ml contains 0.5mg, in contrast product solution.

1.3 the preparation of negative control product solution: it is an amount of to get the Herba Erigerontis medical material, extracts by preparation technology, the preparation method by need testing solution makes negative control product solution again.

1.4 lamellae polyamide (Taizhou, Zhejiang Province city biochemical material factory), silica gel G (Qingdao Haiyang cyclisation factory), 0.3% sodium carboxymethyl cellulose with solution from making sheet, thickness 300um.

1.5 each point sample 2-3ul of point sample amount need testing solution and reference substance solution polyamide membrane, ribbon point sample, each point sample 10ul of silica gel G plate, spot application.

1.6 the expansion mode ascending development, polyamide membrane is opened up apart from 4-5cm.The silica gel plate exhibition is apart from 8-12cm.

1.7 developing solvent is developing solvent with methanol-water-glacial acetic acid (4: 1: 1), 36% acetic acid, ethyl acetate-formic acid-water (10: 1.5: 1), acetone-alcohol-water (2: 1: 2), launches, and takes out, and dries.

1.8 chromatograph identification spray is put under the ultra-violet lamp (365mn) and is inspected with 2% aluminum chloride alcoholic solution.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

1.9 the result through repetition test, is that adsorbent, methanol-water-glacial acetic acid (4: 1: 1) are developing solvent with the polyamide, launches, the speckle separating degree is good, and it is clear to develop the color, and negative control is noiseless, so list quality standard of the present invention in.

2, the research that the thin layer chromatography of Herba Erigerontis medical material in the preparation is differentiated

2.1 the preparation of need testing solution: get this product 10mg or 10ml, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution.

2.2 the preparation of reference substance solution: get the scutellarin reference substance, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.

2.3 the preparation of negative control product solution: it is an amount of to get the Herba Polygoni Orientalis medical material, extracts by preparation technology, the preparation method by need testing solution makes negative control product solution again.

2.4 lamellae polyamide (Taizhou, Zhejiang Province city biochemical material factory), silica gel G (Qingdao Haiyang cyclisation factory), 0.3% sodium carboxymethyl cellulose with solution from making sheet, thickness 300um.

2.5 each point sample 2-3ul of point sample amount need testing solution and reference substance solution polyamide membrane, ribbon point sample, each point sample 10ul of silica gel G plate, spot application.

2.6 the expansion mode ascending development, polyamide membrane is opened up apart from 4-5cm.The silica gel plate exhibition is apart from 8-12cm.

2.7 developing solvent is developing solvent with glacial acetic acid, glacial acetic acid-water (4: 1), glacial acetic acid-methanol-water (5: 1: 0.5), launches, and takes out, and dries.

2.8 chromatograph identification spray is put under the ultra-violet lamp (365mn) and is inspected with 2% ferric chloride alcoholic solution.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

2.9 the result through repetition test, is that adsorbent, glacial acetic acid-ethanol (4: 1) are developing solvent with the polyamide, launches, the speckle separating degree is good, and it is clear to develop the color, and negative control is noiseless, so list quality standard of the present invention in.

3, the research that the thin layer chromatography of Herba Polygoni Orientalis medical material in the preparation is differentiated

3.1 the preparation of need testing solution: get this product 10mg or 10ml, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution.

3.2 the preparation of reference substance solution: get protocatechuic acid, gallic acid reference substance, add methanol and make the mixed solution that every 1ml contains protocatechuic acid 0.5mg and contains gallic acid 0.2mg, product solution in contrast.

3.3 the preparation of negative control product solution: it is an amount of to get the Herba Erigerontis medical material, extracts by preparation technology, the preparation method by need testing solution makes negative control product solution again.

3.4 lamellae silica gel G (Qingdao Haiyang cyclisation factory), 0.3% sodium carboxymethyl cellulose with solution from making sheet, thickness 300um.

3.5 each point sample 10ul of point sample amount need testing solution and reference substance solution.

3.6 the expansion mode ascending development, silica gel plate is opened up apart from 8-12cm.

3.7 developing solvent with chloroform-ethyl acetate-formic acid (5: 3: 1), 36% acetic acid, chloroform-acetone-formic acid (8: 1: 1), be developing solvent, launch, take out, dry.

3.8 chromatograph identification spray is with 2% ferric chloride alcoholic solution, it is clear to be heated to the speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

3.9 the result through repetition test, is that adsorbent, chloroform-ethyl acetate-formic acid (5: 3: 1) are developing solvent with the polyamide, launches, the speckle separating degree is good, and it is clear to develop the color, and negative control is noiseless, so list quality standard of the present invention in.

The invention has the advantages that: method of quality control of the present invention has guaranteed that the quality inspection standard of preparation of the present invention can be than the qualitative character of effectively controlling preparation comprehensively, have accuracy and advance, can be used as the effective technology means of the stability of quality control and investigation technology.Be of great importance to improving the quality of products.

The specific embodiment

Further specify the present invention by the following examples, but not as limitation of the present invention.

The quality control of embodiment 1 compound oriental smartweed oral liquid

Get Herba Polygoni Orientalis 7500g and Herba Erigerontis 1500g, more than two the flavor, Herba Polygoni Orientalis adds 10 times of decoctings and boils 3 times, each 1 hour, filter merging filtrate, being evaporated to relative density is 1.05-1.07 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 65%, stirs, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.04-1.06 (50 ℃), transfer pH value to 7 with saturated sodium carbonate, fully stir, left standstill 3 hours, get supernatant and filter filtrate for later use; Herba Erigerontis adds 10 times of decoctings and boils 3 times, each 0.5 hour, filter, merging filtrate, being evaporated to relative density is 1.09-1.11 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 55%, constantly stir, left standstill sucking filtration, decompression filtrate recycling ethanol and to be concentrated into relative density 1.10-1.12 (50 ℃) standby 12 hours.Add simple syrup 500ml, sodium benzoate 2.5g,, add water to 1000ml, stir evenly, filter with above-mentioned two kinds of concentrated solution mixings, embedding, every bottle of capacity is 10ml, promptly.

[character] this product is the translucent liquid of light brown; Sweet and the little hardship of distinguishing the flavor of.

[inspection] should meet every regulation relevant under the oral liquid item (appendix IC of Chinese Pharmacopoeia version in 2000).

[assay]

1, measure the content of isorientin, orientin and scutellarin with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D):

A. chromatographic condition and system suitability test: use Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite) is filler; Security Guard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company) is guard column; Mobile phase is second eyeball-0.1% phosphoric acid solution (18: 82); Set flow velocity: 1ml/min, 40 ℃ of temperature detect wavelength: 350nm; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600.

B. it is an amount of that isorientin, orientin and scutellarin reference substance are got in the preparation of reference substance solution respectively, through phosphorus pentoxide dried overnight 24 hours, the accurate title, decide, add the first dissolving, make every 1ml and contain the solution that isorientin 0.025mg, every 1ml contain orientin 0.02mg and scutellarin 0.08mg, shake up, promptly.

C. the preparation precision of need testing solution is measured the content under the loading amount item, and mixing is got 50ml, puts in the 100ml measuring bottle, and accurate the title decides, and adds water and makes dissolving in right amount and be diluted to scale, shakes up, and gets supernatant 0.45 μ m and filters, promptly.

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

The every 10ml of this product contains Herba Polygoni Orientalis with isorientin (C ₂₁H ₂₀O ₁₁) and orientin (C ₂₁H ₂₀O ₁₁) the total amount meter, must not be less than 7.0mg, contain Herba Erigerontis with scutellarin (C ₂₁H ₁₈O ₂) meter, must not be less than 14.0mg.

[discriminating]

2, differentiate this medicine Chinese crude drug Herba Polygoni Orientalis with thin layer chromatography

A. the preparation of need testing solution: get this product 10ml, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution.

3, differentiate this medicine Chinese crude drug Herba Erigerontis with thin layer chromatography

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 3ul of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with glacial acetic acid-ethanol (4: 1), launches, and takes out, and dries, and sprays the ferric chloride alcoholic solution with 2%, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

4, differentiate this medicine Chinese crude drug Herba Polygoni Orientalis with thin layer chromatography

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 2-3ul, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid (5: 3: 1), launches, take out, dry, spray ferric chloride alcoholic solution with 2%.It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The syrupy quality control of embodiment 2 compound oriental smartweeds

Get Herba Polygoni Orientalis 7500g and Herba Erigerontis 1500g, more than two the flavor, Herba Polygoni Orientalis adds 10 times of decoctings and boils 3 times, each 1 hour, filter merging filtrate, being evaporated to relative density is 1.05-1.07 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 65%, stirs, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.04-1.06 (50 ℃), transfer pH value to 7 with saturated sodium carbonate, fully stir, left standstill 3 hours, get supernatant and filter filtrate for later use; Herba Erigerontis adds 10 times of decoctings and boils 3 times, each 0.5 hour, filter, merging filtrate, being evaporated to relative density is 1.09-1.11 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 55%, constantly stir, left standstill sucking filtration, decompression filtrate recycling ethanol and to be concentrated into relative density 1.10-1.12 (50 ℃) standby 12 hours.Other gets sucrose 264g, and it is an amount of to add water, boil dissolving after, filter, with above-mentioned two kinds of concentrated solution mixings, continuing to be concentrated into relative density is 1.10 (90 ℃), puts coldly, adds sodium benzoate 3g, thin up stirs evenly to 1000ml, every bottle of capacity is 10ml, promptly.Promptly.

[character] this product is the supernatant liquid of light brown; Sweet and the little hardship of distinguishing the flavor of.

[inspection] should meet every regulation relevant under the syrup item (an appendix I of Chinese Pharmacopoeia version in 2000 H).

[assay]

A. chromatographic condition and system suitability test: use Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite) is filler; Security Guard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company) is guard column; Mobile phase is second eyeball-0.1% phosphoric acid solution (14: 69); Set flow velocity: 1ml/min, 37 ℃ of temperature detect wavelength: 348nm; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600.

[discriminating]

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with methanol-water-glacial acetic acid (2.4: 0.5: 0.5), launches, and takes out, and dries, and the spray developer is put under the ultra-violet lamp and inspected.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 3ul of solution, ribbon is put on same polyamide membrane respectively, with glacial acetic acid-ethanol (2.4: 0.5) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution, put under the daylight lamp and inspect with 2%.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 2-3ul, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid (2: 1.8: 0.5), launches, take out, dry, spray ferric chloride alcoholic solution with 2%.It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The quality control of embodiment 3 compound oriental smartweed granules

Get Herba Polygoni Orientalis 7500g and Herba Erigerontis 1500g, more than two the flavor, Herba Polygoni Orientalis adds 10 times of decoctings and boils 3 times, each 1 hour, filter merging filtrate, being evaporated to relative density is 1.05-1.07 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 65%, stirs, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.04-1.06 (50 ℃), transfer pH value to 7 with saturated sodium carbonate, fully stir, left standstill 3 hours, get supernatant and filter filtrate for later use; Herba Erigerontis adds 10 times of decoctings and boils 3 times, each 0.5 hour, filter, merging filtrate, being evaporated to relative density is 1.09-1.11 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 55%, constantly stir, left standstill sucking filtration, decompression filtrate recycling ethanol and to be concentrated into relative density 1.10-1.12 (50 ℃) standby 12 hours.Other gets sucrose 500g and above-mentioned two kinds of extractum mixing granulations, drying, and granulate is with every bag of 10g packing, promptly.

[character] this product is brown granule; Sweet and the little hardship of distinguishing the flavor of.

[inspection] should meet every regulation relevant under the granule item (an appendix I of Chinese Pharmacopoeia version in 2000 H).

[assay]

1, measures the content of isorientin, orientin and scutellarin with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D)

A. chromatographic condition and system suitability test: use Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite) is filler; Security Guard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company) is guard column; Mobile phase is second eyeball-0.1% phosphoric acid solution (25: 91); Set flow velocity: 1ml/min, 42 ℃ of temperature detect wavelength: 352nm; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600.

The every 10g of this product contains Herba Polygoni Orientalis with isorientin (C ₂₁H ₂₀O ₁₁) and orientin (C ₂₁H ₂₀O ₁₁) the total amount meter, must not be less than 7.0mg, contain Herba Erigerontis with scutellarin (C ₂₁H ₁₈O ₂) meter, must not be less than 14.0mg.

[discriminating]

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with methanol-water-glacial acetic acid (7.5: 2.5: 2.5), launches, and takes out, and dries, and the spray developer is put under the ultra-violet lamp and inspected.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 3ul of solution, ribbon is put on same polyamide membrane respectively, with glacial acetic acid-ethanol (7.5: 2.5) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution, put under the daylight lamp and inspect with 2%.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 2-3ul, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid (8: 5: 2.5), launches, take out, dry, spray ferric chloride alcoholic solution with 2%.It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

The quality control of embodiment 4 compound oriental smartweed injection

Get Herba Polygoni Orientalis 7500g and Herba Erigerontis 1500g, more than two the flavor, Herba Polygoni Orientalis adds 10 times of decoctings and boils 3 times, each 1 hour, filter merging filtrate, being evaporated to relative density is 1.05-1.07 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 65%, stirs, left standstill 12 hours, sucking filtration, decompression filtrate recycling ethanol also is concentrated into relative density 1.04-1.06 (50 ℃), transfer pH value to 7 with saturated sodium carbonate, fully stir, left standstill 3 hours, getting supernatant filters, filtrate discards ethyl acetate liquid, combining water layer liquid with the ethyl acetate extraction of 1/2 times of amount 3 times, transfer PH3 with hydrochloric acid, with the saturated n-butanol extraction of 1/2 times of water gaging 4 times, merge n-butyl alcohol liquid, wash 2 times with 1/8 times of water gaging, the reclaim under reduced pressure n-butyl alcohol, residue adds 80% ethanol 2500ml dissolving, last polyamide column (500g, Ф 8cm, blade diameter length ratio 1: 6, absorption flow velocity: 1.5BV/h), collect stream and wear liquid and eluent, reclaim ethanol, the residue vacuum drying gets the Herba Polygoni Orientalis extract;

Herba Erigerontis adds 10 times of decoctings and boils 3 times, each 0.5 hour, filters, merging filtrate, being evaporated to relative density is 1.09-1.11 (50 ℃), adds ethanol and makes and contain alcohol amount and reach 55%, constantly stir, left standstill sucking filtration 12 hours, decompression filtrate recycling ethanol also is concentrated into relative density 1.10-1.12 (50 ℃), transfers PH2 with hydrochloric acid, and 55 ℃ are incubated 6 hours, the tipping supernatant, sucking filtration, precipitation washes with water to PH3-4, vacuum drying gets Herba Erigerontis extract.

Get above-mentioned two kinds of extracts, add 1800 milliliters of waters for injection, stirring makes molten, it is an amount of to add glycerol again, mixing is transferred PH6.8-7.8 with saturated sodium carbonate, adds water for injection to 10000 milliliter, mixing, filter filtrate sterile filling, every 10ml with 0.45um and 0.22um microporous filter membrane, sterilized 60 minutes down, promptly get 1000 injection for 105 ℃.

[assay]

A. chromatographic condition and system suitability test:: use Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite) is filler; Security Guard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company) is guard column; Mobile phase is second eyeball-0.1% phosphoric acid solution (18: 82); Set flow velocity: 1ml/min, 40 ℃ of temperature detect wavelength: 350nm; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600.

C. the preparation precision of need testing solution is measured the content under the loading amount item, and mixing is got 50ml or 50mg, puts in the 100ml measuring bottle, and accurate the title decides, and adds water and makes dissolving in right amount and be diluted to scale, shakes up, and gets supernatant 0.45 μ m and filters, promptly.

Every of this product contains Herba Polygoni Orientalis with isorientin (C ₂₁H ₂₀O ₁₁) and orientin (C ₂₁H ₂₀O ₁₁) the total amount meter, must not be less than 7.0mg, contain Herba Erigerontis with scutellarin (C ₂₁H ₁₈O ₂) meter, must not be less than 14.0mg.

[discriminating]

The quality control of embodiment 5 compound oriental smartweed freeze-dried powders

Remove above-mentioned two kinds of extracts, add 1800 milliliters of waters for injection, stir and to make moltenly, it is an amount of to add mannitol again, mixing is transferred PH6.8-7.8 with saturated sodium carbonate, adds water for injection to 2000 milliliter, mixing is sub-packed in the infusion bottle of having handled well, fills in butyl rubber bung and thin film, roll aluminium lid, boiling sterilization 30 minutes, cold preservation 24 hours, filter with 0.45um and 0.22um microporous filter membrane, filtrate is aseptic subpackaged, 2.5 milliliters every bottle, lyophilization, gland promptly gets 1000 freeze-dried powders.After the lyophilizing, every heavily about 100mg of dried solid.

[assay]

[discriminating]

A. the preparation of need testing solution: get this product 10mg, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution.

The quality control of embodiment 6 compound oriental smartweed oral liquids

Herba Polygoni Orientalis 7500g and Herba Erigerontis 1500g.

[method for making], [character], [inspection] are identical with embodiment 1.

Its method of quality control is:

A. chromatographic condition: use Hypersil C ₁₈(5um, 5.0mm * 200mm i.d. Dalian Yi Lite) is filler; SecurityGuard C ₁₈(5um, 4mm * 3mm i.d.PheodyMmenex company) is guard column; Mobile phase is second eyeball-0.1% phosphoric acid solution (21: 86); Set flow velocity: 1ml/min, 39 ℃ of temperature detect wavelength: 351nm; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600.

C. test according to thin layer chromatography, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, with methanol-water-glacial acetic acid (3.5: 1.2: 1.2) is developing solvent, launch, take out, dry, spray is put under the ultra-violet lamp (365mn) and is inspected with 2% aluminum chloride alcoholic solution.In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color.

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 3ul of solution, ribbon is put on same polyamide membrane respectively, with glacial acetic acid-ethanol (3.5: 1.2) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution, put under the daylight lamp and inspect with 2%.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 2-3ul, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid (6: 2.5: 2.5), launches, take out, dry, spray ferric chloride alcoholic solution with 2%.It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Claims

1. the method for quality control of a compound oriental smartweed preparation, it is characterized in that comprise character is observed, official method is checked content, to the scutellarin that contains, isorientin, orientin carries out assay, and content Herba Polygoni Orientalis and Herba Erigerontis are differentiated, wherein said scutellarin to containing, isorientin, orientin carries out assay, may further comprise the steps:

According to an appendix VI of Chinese Pharmacopoeia version in 2000 D high effective liquid chromatography for measuring,

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is acetonitrile-0.1% phosphoric acid solution; Set flow velocity: 1ml/min, temperature and detection wavelength; Number of theoretical plate calculates by the scutellarin peak should be not less than 7600;

B. it is an amount of that isorientin, orientin and scutellarin reference substance are got in the preparation of reference substance solution respectively, drying, and accurate the title, decide, add dissolve with methanol, make every 1ml and contain the solution that isorientin 0.025mg, every 1ml contain orientin 0.02mg and scutellarin 0.08mg, shake up, promptly;

C. the preparation precision of need testing solution is measured the content under the loading amount item, and mixing is got 50ml or 50mg, puts in the 100ml measuring bottle, and accurate the title decides, and adds water and makes dissolving in right amount and be diluted to scale, shakes up, and gets supernatant and filters, promptly;

The numerical range of above-mentioned chromatographic condition mobile phase is acetonitrile-0.1% phosphoric acid solution 14-25: 69-91, temperature: 37-42 ℃, the detection wavelength is 350 ± 2nm, the preparation of reference substance solution: precision takes by weighing and reaches 24 hours through the phosphorus pentoxide dried overnight, uses dissolve with methanol; The preparation of need testing solution: get supernatant 0.45 μ m and filter; Accurate respectively reference substance solution and the need testing solution 5-10 μ l injecting chromatograph mensuration drawn; The every 1ml of this product contains Herba Polygoni Orientalis with isorientin C ₂₁H ₂₀O ₁₁With orientin C ₂₁H ₂₀O ₁₁The total amount meter, must not be less than 7.0mg, contain Herba Erigerontis with scutellarin C ₂₁H ₁₈O ₂Meter must not be less than 14.0mg;

Wherein said content Herba Polygoni Orientalis and Herba Erigerontis are differentiated, be may further comprise the steps:

Herba Polygoni Orientalis is differentiated:

A. the preparation of need testing solution: get this product, add methanol, supersound process, centrifugal, get supernatant as need testing solution;

B. the preparation of reference substance solution: get isorientin, orientin reference substance, add methanol and make mixed solution, in contrast product solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with methanol-water-glacial acetic acid, launches, and takes out, and dries, and the spray developer is put under the ultra-violet lamp and inspected;

Above-mentioned Herba Polygoni Orientalis thin layer differentiates that the processing of sample adopts methanol extraction, and developing solvent is methanol-water-glacial acetic acid, methanol-water-glacial acetic acid ratio 4: 1: 1, and developer 2% aluminum chloride ethanol liquid, and under the 365nm ultra-violet lamp, examine;

Or follow these steps to carry out:

B. the preparation of reference substance solution: get protocatechuic acid, gallic acid reference substance, add methanol and make the mixed solution that contains protocatechuic acid and contain gallic acid, product solution in contrast;

C. according to the thin layer chromatography test, draw need testing solution, reference substance solution is put respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid, launches, and takes out, dry, and the spray developer, heating is put daylight lamp and is inspected;

Ether and ethyl acetate extraction are adopted in the processing of above-mentioned Herba Polygoni Orientalis thin layer discriminating sample successively; Developing solvent is chloroform-ethyl acetate-formic acid, chloroform-ethyl acetate-formic acid ratio 5: 3: 1, and the ferric chloride alcoholic solution of developer 2% heats 105 ℃;

Herba Erigerontis is differentiated:

B. the preparation of reference substance solution: get the scutellarin reference substance, add methanol and make solution, in contrast product solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of solution, ribbon is put on same polyamide membrane respectively, is developing solvent with glacial acetic acid-ethanol, launches, and takes out, and dries, and the spray developer is put under the daylight lamp and inspected;

Above-mentioned Herba Erigerontis thin layer is differentiated the processing employing dehydrated alcohol extraction of sample, and developing solvent is glacial acetic acid-ethanol, glacial acetic acid-proportion of ethanol 4: 1, developer 2% ferric chloride ethanol liquid.

2. the method for claim 1 is characterized in that, wherein said content Herba Polygoni Orientalis and Herba Erigerontis is differentiated, may further comprise the steps:

Herba Polygoni Orientalis is differentiated:

A. the preparation of need testing solution: get this product 10mg or 10ml, add methanol 2ml, ultrasonic 5 minutes, centrifugal, get supernatant as need testing solution;

B. the preparation of reference substance solution: get isorientin, orientin reference substance, add methanol and make the mixed solution that every 1ml contains 0.5mg, in contrast product solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, with methanol-water-glacial acetic acid 2.4-7.5: 0.5-2.5: 0.5-2.5 is developing solvent, launches, and takes out, dry, the spray developer is put under the ultra-violet lamp and is inspected; In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color;

Or follow these steps to carry out:

B. the preparation of reference substance solution: get protocatechuic acid, gallic acid reference substance, add methanol and make the mixed solution that every 1ml contains protocatechuic acid 0.5mg and contains gallic acid 0.2mg, product solution in contrast;

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 2-3ul, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-ethyl acetate-formic acid 2-8: 1.8--5: 0.5-2.5, launches, take out, dry, spray ferric chloride alcoholic solution with 2%; It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Herba Erigerontis is differentiated:

B. the preparation of reference substance solution: get scutellarin, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 3ul of solution, ribbon is put on same polyamide membrane respectively, with glacial acetic acid-ethanol 2.4-7.5: 0.5-2.5 is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution, put under the daylight lamp and inspect with 2%; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

3. the method for claim 1 is characterized in that, wherein said content Herba Polygoni Orientalis and Herba Erigerontis is differentiated, may further comprise the steps:

Herba Polygoni Orientalis is differentiated:

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, with methanol-water-glacial acetic acid is developing solvent at 4: 1: 1, launches, and takes out, dry, spray aluminum chloride alcoholic solution, put under the ultra-violet lamp 365mn and inspect with 2%; In the test sample chromatograph, with contrast medicine chromatograph corresponding position on, show the fluorescence speckle of same color;

Or follow these steps to carry out:

C. according to the thin layer chromatography test, draw need testing solution 10ul, reference substance solution 3ul, put respectively on same silica gel g thin-layer plate, with chloroform: ethyl acetate: formic acid=5: 3: 1 is developing solvent, launches, take out, dry, spray ferric chloride alcoholic solution with 2%; It is clear to be heated to speckle colour developing at 105 ℃, puts under the daylight lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Herba Erigerontis is differentiated:

B. the preparation of reference substance solution: get the scutellarin reference substance, add methanol and make the solution that every 1ml contains 1mg, brilliant in contrast solution;

C. according to the thin layer chromatography test, draw above-mentioned two kinds of each 2-3ul of solution, ribbon is put on same polyamide membrane respectively, with the developing solvent in 4: 1 of glacial acetic acid-ethanol, launches, and takes out, and dries, and sprays the ferric chloride alcoholic solution with 2%, puts under the daylight lamp and inspects; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

4. the method for claim 1 is characterized in that, described compound oriental smartweed preparation is oral liquid, syrup, granule, freeze-dried powder or injection.

5. the method for claim 1 is characterized in that, described compound oriental smartweed preparation is made by following parts by weight of Chinese traditional medicine raw material: Herba Polygoni Orientalis 5000-10000 part and Herba Erigerontis 1000-2000 part.

6. the method for claim 1 is characterized in that, described compound oriental smartweed preparation is made by following parts by weight of Chinese traditional medicine raw material: 1500 parts of 7500 parts of Herba Polygoni Orientaliss and Herba Erigerontiss.