CN1785281A

CN1785281A - Quality control method of spirit quieting oral liquid preparation

Info

Publication number: CN1785281A
Application number: CN 200510003272
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: 2005-11-10
Filing date: 2005-11-10
Publication date: 2006-06-14
Anticipated expiration: 2025-11-10
Also published as: CN1785281B

Abstract

A quality control method for the Chinese medicine ''Calming oral liquid'' features that a thin-layer chromatography is used for identification to white atractylodes rhizome, fleece flower stem and hairyvien agrimonia herb, and an efficient liquid-phase chromatography is used to measure the content of glycyrrhizic acid.

Description

A kind of method of quality control of spirit quieting oral liquid preparation

Technical field:

The present invention relates to a kind of method of quality control of the spirit quieting oral liquid preparation of making by Chinese crude drug Ganoderma 50g, the Rhizoma Atractylodis Macrocephalae (stir-fry) 100g, Fructus Ligustri Lucidi (system) 100g, Cortex Albiziae 150g, Caulis Polygoni Multiflori 150g, Herba Agrimoniae 250g, Herba Ecliptae 100g and Radix Glycyrrhizae 50g, particularly with the content of glycyrrhizic acid in this medicine of high effective liquid chromatography for measuring with differentiate the spirit quieting oral liquid preparation method of quality control of the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae with thin layer chromatography.

Background technology:

" Drug Standard of Ministry of Public Health of the Peoples Republic of China " (Chinese patent medicine prescribed preparation) the 6th (WS3-B-1130-92) announced treatment anemia body void, dizzy, the syrup of calming the nerves of diseases such as insomnia, this pharmaceutical formulation is made up of Ganoderma 50g, the Rhizoma Atractylodis Macrocephalae (stir-fry) 100g, Fructus Ligustri Lucidi (system) 100g, Cortex Albiziae 150g, Caulis Polygoni Multiflori 150g, Herba Agrimoniae 250g, Herba Ecliptae 100g, Radix Glycyrrhizae 50g, and above-mentioned raw materials is made syrup 1000ml altogether.The content of above-mentioned standard does not illustrate the syrupy method of quality control of calming the nerves.

Chinese medicine preparation for a long time, difficult quality is effectively controlled, the quality of effectively controlling tcm product is a great problem, the method for available technology adopting is imperfect, the minority specific aim is not strong, uses these methods to be difficult to reach the purpose of real control of quality.More than calm the nerves the syrup said preparation owing to lack method of quality control, be difficult to control the quality of said preparation; Brought difficulty for normal production and operation, utilize more known technology to be difficult to the method for quality control that the said preparation product is provided, the present invention is directed to the deficiencies in the prior art, adopt new means that the quality of spirit quieting oral liquid preparation is controlled, particularly adopt with the content of glycyrrhizic acid in this medicine of high effective liquid chromatography for measuring with thin layer chromatography and differentiate the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae in the finished product, solved the difficult problem of said preparation quality control.The present invention is directed to the characteristics and the prescription of the present invention of spirit quieting oral liquid preparation, a kind of method of quality control of practicality is provided.

Summary of the invention:

The object of the present invention is to provide a kind of method of quality control of spirit quieting oral liquid preparation, in process of production product quality is monitored, improve the accuracy of quality-monitoring standard to guarantee said preparation.

The present invention be directed to spirit quieting oral liquid preparation and propose method of quality control, described spirit quieting oral liquid preparation comprises the oral liquid of the syrup of calming the nerves, ANSHEN KOUFUYE and the following formulation of any usefulness.

Ganoderma 50g, the Rhizoma Atractylodis Macrocephalae (stir-fry) 100g, Fructus Ligustri Lucidi (system) 100g, Cortex Albiziae 150g, Caulis Polygoni Multiflori 150g, Herba Agrimoniae 250g, Herba Ecliptae 100g, Radix Glycyrrhizae 50g.

Spirit quieting oral liquid preparation of the present invention, its preparation method can adopt following method:

By the raw material of Chinese medicine of above-mentioned prescription is processed through extraction or other modes, make pharmaceutically active substance, subsequently, be raw material with this material, add the medicine acceptable carrier when needing, make capsule, granule, tablet 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 material of extractum form, can be that dry extract also can be a fluid extract, make different concentration according to the different needs decision of preparation.

Spirit quieting oral liquid 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 spirit quieting oral liquid 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.

Spirit quieting oral liquid preparation of the present invention preferably prepares through following method:

The syrup method for making of wherein calming the nerves is:

More than eight flavor Chinese crude drugs, Ganoderma decocts with water secondary, 5 hours for the first time, 3 hours for the second time, collecting decoction filtered, it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, it is standby to get supernatant; Seven flavors such as all the other Rhizoma Atractylodis Macrocephalaes decoct with water secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, and it is standby to get supernatant.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, promptly.

Wherein the ANSHEN KOUFUYE method for making is:

More than eight flavors, Ganoderma decocts with water secondary, 5 hours for the first time, 3 hours for the second time, collecting decoction filtered, it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, it is standby to get supernatant; Seven flavors such as all the other Rhizoma Atractylodis Macrocephalaes decoct with water secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, and it is standby to get supernatant.Add simple syrup 264ml, sodium benzoate 2.59,, add water to 1000ml, stir evenly, filter with above-mentioned two kinds of concentrated solution mixings, embedding, every bag of capacity is 10ml, promptly.

Spirit quieting oral liquid preparation of the present invention, in its Chinese medicine preparation, part Chinese medicine can be replaced with the Chinese medicine of equivalent effect, replaces the back curative effect and can not change.

The present invention is achieved by following technical proposals: the one content of glycyrrhizic acid in the high effective liquid chromatography for measuring spirit quieting oral liquid preparation; Its dual-purpose thin layer chromatography is differentiated the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and the Herba Agrimoniae in the spirit quieting oral liquid preparation.

The method of quality control of concrete spirit quieting oral liquid preparation can be achieved by following steps:

Step with the content of glycyrrhizic acid in the high effective liquid chromatography for measuring spirit quieting oral liquid preparation:

The preparation of monoammonium glycyrrhizinate reference substance solution,

The preparation of spirit quieting oral liquid preparation sample solution,

Measure above solution with high performance liquid chromatograph, get chromatogram,

Draw the reference substance standard curve, according to spirit quieting oral liquid preparation sample peak area, with standard curve, calculate the content of monoammonium glycyrrhizinate in this sample.

Concrete determination step is as follows:

Test apparatus, reagent, reagent

Tianjin, island LC-10ATvp high performance liquid chromatograph, the LC-10ATvp infusion pump; SPD-M10Avp secondary array detector; The SCL-10Avp system controller; CTO-10ASvp thermocolumn case; The CLASS-VP6.1 workstation data processing system; Microsyringe (25 μ l), HAMILTON company.

(Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the monoammonium glycyrrhizinate reference substance, for assay usefulness, lot number: 0731-200204).Methanol (chromatographically pure), ammonium acetate (analytical pure), glacial acetic acid (analytical pure), water for injection.

The content of measuring monoammonium glycyrrhizinate according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D) may further comprise the steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution; The detection wavelength is 250 ± 3nm; Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000.

B. the preparation extracting liquorice of reference substance solution acid mono-ammonium reference substance is an amount of, accurately claims surely, puts in the 50ml measuring bottle, with the mobile phase dissolving and be diluted to scale, shakes up, promptly.

C. the preparation precision of need testing solution is measured this product 2.5ml under the loading amount item, puts in the 10ml measuring bottle, with mobile phase dissolving and be diluted to scale, shakes up, and gets supernatant and filters (0.45 μ m), 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-2.5% glacial acetic acid aqueous solution, and (26-53: 58-69), preferred proportion is eyeball-2.5% glacial acetic acid aqueous solution (37: 63); The best precision of the preparation of reference substance solution takes by weighing monoammonium glycyrrhizinate reference substance 10mg, put in the 50ml measuring bottle, with mobile phase dissolving and be diluted to scale, shake up, make every 1ml contain monoammonium glycyrrhizinate reference substance 0.2-0.3mg, amounting to glycyrrhizic acid is 0.1959-0.2939mg; The preparation of need testing solution is preferably got supernatant and is filtered 0.45 μ m; Accurate respectively reference substance solution and the need testing solution 5-10 μ l injecting chromatograph drawn measured, promptly.The every 1ml of this product contains Radix Glycyrrhizae with glycyrrhizic acid (C ₄₂H ₆₂O ₁₆) meter, must not be less than 0.50mg.

In the method for quality control of the present invention, differentiate that with thin layer chromatography the step of the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae in the spirit quieting oral liquid preparation is as follows:

Thin layer chromatography is differentiated the step of the Chinese crude drug Rhizoma Atractylodis Macrocephalae (an appendix VI of Chinese Pharmacopoeia version in 2000 B):

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material, add alcohol reflux, filter, filtrate evaporate to dryness, residue add the dissolving of water slight fever, use water saturated n-butanol extraction, merge n-butyl alcohol liquid, and evaporate to dryness, residue add methanol makes dissolving, in contrast medical material solution.

B. the preparation of need testing solution: get this product, put in the water-bath and steam, add ethanol to an amount of, stir, leave standstill, get supernatant, evaporate to dryness, residue add the dissolving of water slight fever, use ethyl acetate extraction, abandon ethyl acetate extraction liquid, the water saturated n-butanol extraction of water layer merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol makes dissolving, filters, and filtrate is as need testing solution.

C. according to thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid is developing solvent, launches, and takes out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.

The processing of above-mentioned this Rhizoma Atractylodis Macrocephalae thin layer discriminating sample can be adopted ethyl acetate extraction, and developing solvent is n-butyl alcohol-acetone-water-glacial acetic acid, and (7: 2: 1: be best 1), developer was ninhydrin solution the best to n-butyl alcohol-acetone-water-glacial acetic acid ratio.

Thin layer chromatography is differentiated the step of Chinese crude drug Caulis Polygoni Multiflori (an appendix VI of Chinese Pharmacopoeia version in 2000 B):

A. the preparation of control medicinal material solution: get the Caulis Polygoni Multiflori control medicinal material, add dehydrated alcohol, supersound process filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, in contrast medical material solution.

B. get the emodin reference substance, add methanol and make solution, in contrast product solution.

C. the preparation of need testing solution: get this product, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol, supersound process filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution.

D. according to the thin layer chromatography test, draw above-mentioned need testing solution and control medicinal material solution, reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with normal hexane-ethyl acetate-formic acid, launch, take out, dry, put under the ultra-violet lamp and inspect.

The processing of above-mentioned this Caulis Polygoni Multiflori thin layer discriminating sample can be adopted dehydrated alcohol extraction, and developing solvent is normal hexane-ethyl acetate-formic acid, and normal hexane-ethyl acetate-formic acid ratio (30: 10: 0.5) is best, and developing the color down at ultra-violet lamp (365nm) is the best.

Thin layer chromatography is differentiated the step of Chinese crude drug Herba Agrimoniae (an appendix VI of Chinese Pharmacopoeia version in 2000 B):

A. the preparation of need testing solution: get this product, add water, use ether extraction, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value, uses ethyl acetate extraction, merges ethyl acetate extraction liquid, and evaporate to dryness, residue add methanol 1 makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get the Herba Agrimoniae control medicinal material, add water and be heated to and boil, and keep little and boil, put coldly, filter, filtrate is from " using ether extraction ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

C. according to the thin layer chromatography test, draw need testing solution, control medicinal material solution is put respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-Ethyl formate-formic acid, launches, and takes out, and dries, and spray is with the ferric chloride alcoholic solution.

The processing of above-mentioned this Herba Agrimoniae thin layer discriminating sample can be adopted ether and ethyl acetate extraction successively, and adds hydrochloric acid regulating solution pH value to 2; Developing solvent is chloroform-Ethyl formate-formic acid, and chloroform-Ethyl formate-formic acid ratio (5: 5: 1) is best, and developer is ferric chloride alcoholic solution the best of 1%.

Preferred Chinese crude drug Rhizoma Atractylodis Macrocephalae 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 control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 1-3g, add ethanol 40-100ml reflux, extract, 1-1.5 hour, filter, filtrate evaporate to dryness, residue add the dissolving of water 20-50ml slight fever, with water saturated n-butanol extraction 1-3 time, each 10-30ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 0.5-3ml makes dissolving, in contrast medical material solution.

B. the preparation of need testing solution: get this product 10-50ml, put in the water-bath and steam, add ethanol 15-40ml to about 10ml, stir, left standstill 1-2 hour, get supernatant, evaporate to dryness, residue add the dissolving of water 10-30ml slight fever, use ethyl acetate extraction 2-4 time, each 15-20ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 2-4 time, each 15-20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 2-6ml makes dissolving, filters, and filtrate is as need testing solution.

C. test according to thin layer chromatography, draw each 5-10 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

Preferred Chinese crude drug Caulis Polygoni Multiflori 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 control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.1-0.2g, add dehydrated alcohol 15-20ml, supersound process 15-30 minute, filter, filtrate evaporate to dryness, residue add methanol 0.5-3ml makes dissolving, in contrast medical material solution.

B. get the emodin reference substance, add methanol and make the solution that every 1ml contains 0.8-1.2mg, in contrast product solution.

C. the preparation of need testing solution: get this product 5-10ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 10-30ml, supersound process 10-30 minute, filter, filtrate evaporate to dryness, residue add methanol 0.5-3ml makes dissolving, as need testing solution.

D. test according to thin layer chromatography, draw above-mentioned need testing solution and control medicinal material solution 5-10 μ l, reference substance solution 0.5-2.5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Preferred Chinese crude drug Herba Agrimoniae 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 5-10ml, add water 10-30ml, use ether extraction 1-3 time, each 10-25ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 1-3, uses ethyl acetate extraction 1-2 time, each 15-30ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 0.5-3ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 1.0-2.0g, add water 100-200ml and be heated to and boil, and kept little 1.5-2.5 of boiling hour, put cold, filter, filtrate is from " using ether extraction 1-3 time ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

C. according to the thin layer chromatography test, draw need testing solution 6-12 μ l, control medicinal material solution 5-10 μ l, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-Ethyl formate-formic acid (5: 5: 1), launches, take out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

More preferably Chinese crude drug Rhizoma Atractylodis Macrocephalae 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 control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 1.5g, add ethanol 50ml reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add the dissolving of water 30ml slight fever, with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 1ml makes dissolving, in contrast medical material solution.

B. the preparation of need testing solution: get this product 20ml, put in the water-bath and steam, add ethanol 30ml to about 10ml, stir, left standstill 1 hour, get supernatant, evaporate to dryness, residue add the dissolving of water 30ml slight fever, use ethyl acetate extraction 3 times, each 20ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 3ml makes dissolving, filters, and filtrate is 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 in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.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 Caulis Polygoni Multiflori 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 control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.1g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.

B. get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.

C. the preparation of need testing solution: get this product 5ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 30ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.

D. test according to thin layer chromatography, draw above-mentioned need testing solution and control medicinal material solution 8 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

More preferably Chinese crude drug Herba Agrimoniae 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 5ml, add water 15ml, use ether extraction 3 times, each 25ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 2, uses ethyl acetate extraction 2 times, each 30ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 2.0g, add water 150ml and be heated to and boil, and keep little and boiled 2 hours, put coldly, filter, filtrate is from " usefulness ether extraction 3 times ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

C. according to the thin layer chromatography test, draw need testing solution 6 μ l, control medicinal material solution 10 μ l, putting respectively on same silica gel g thin-layer plate, is developing solvent with chloroform-Ethyl formate-formic acid (5: 5: 1), launches, take out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

The TCL that the invention provides the Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae differentiates.Increased the quality determining method to glycyrrhizic acid 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 index of the stability of oral formulations quality control and investigation technology.

The used preparation of experimental example test sample of the present invention all adopts the embodiment of the invention 1 described syrup, can adopt the embodiment of the invention 2 described oral liquids, and also available have other preparations that same materials is formed with the preparation described in each embodiment.

The methodological study of experimental example one assay method of the present invention

1 linear relationship is investigated

1.1 the preparation of standard solution

Precision takes by weighing monoammonium glycyrrhizinate reference substance 9.44mg, puts in the 50ml measuring bottle, with mobile phase dissolving and be diluted to scale, shakes up, and promptly gets (every 1ml contains monoammonium glycyrrhizinate reference substance 0.1888mg, and amounting to glycyrrhizic acid is 0.1849mg).

1.2 the drafting of standard curve

The accurate monoammonium glycyrrhizinate reference substance solution 2 of drawing, 4,6,8,10 μ l, inject chromatograph of liquid respectively, record chromatograph (table 1), with peak area A (μ Vs) mass number x (μ g) being carried out linear regression calculates, getting equation of linear regression is: A=545709.30 * X-6565.30, r=0.9999 (), precision is measured reference substance solution 5 μ l and is injected chromatograph of liquid, the gained peak area is 497831, one need testing solution sample introduction is analyzed the gained peak area calculate content with regression equation and one point method respectively, result's relative deviation is 0.0318%, so quality standard of the present invention adopts one point method to calculate content.The glycyrrhizic acid range of linearity: 0.3698 μ g～1.849 μ g.

Table 1 glycyrrhizic acid linear relationship is investigated

Glycyrrhizic acid sample size (μ g)	Glycyrrhizic acid peak area (μ Vs)
Glycyrrhizic acid sample size (μ g)	Glycyrrhizic acid peak area (μ Vs)	?0.3698 ?0.7396 ?1.1094 ?1.4792	199155 393751 597459 797622

?1.849

1006236

The test of 2 precision

Precision is measured this product 25ml, 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 2).

The precision of glycyrrhizic acid test in the table 2 preparation test sample

The sample introduction number of times	1	2	3	4	5	Meansigma methods	?RSD(％)
The sample introduction number of times	1	2	3	4	5	Meansigma methods	?RSD(％)	Peak area	506706	500356	501886	500419	506534	503180	?0.64

The result as seen, this method precision is good.

The test of 3 repeatability

Precision is measured this product 25ml respectively, and 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, and result of calculation is listed table 3 in, and the glycyrrhizic acid average content is 0.9392mg/ml, and RSD is 1.58%.The result as seen, this method repeatability is good.

The repeatability of glycyrrhizic acid test in the table 3 preparation test sample

The sample introduction number of times	1	2	3	4	5	Meansigma methods	RSD(％)
The sample introduction number of times	1	2	3	4	5	Meansigma methods	RSD(％)	Content (mg/ml)	0.7622	0.7645	0.7392	0.7508	0.7401	0.7514	1.58

4. the stability test of glycyrrhizic acid in the need testing solution

Precision is measured this product 25ml, preparation method by test liquid under the assay item in the quality standard of the present invention prepares test liquid, respectively at 0,2,4,6,8 hour mensuration glycyrrhizic acid peak area (seeing Table 4), the result showed that glycyrrhizic acid is stable in 8 hours in the need testing solution.

The stability test of glycyrrhizic acid in the table 4 preparation test sample

Time (hour)	0	2	4	6	8	Meansigma methods	?RSD(％)
Time (hour)	0	2	4	6	8	Meansigma methods	?RSD(％)	Peak area	492033	482523	484747	489104	486687	487019	?0.76

5. recovery test

Adopt the application of sample absorption method, precision is measured 5 parts of sample (average content is 0.7514mg/ml) 1.25ml that measured content respectively, put in the 10ml measuring bottle, precision adds the monoammonium glycyrrhizinate reference substance solution, and (every 1ml contains monoammonium glycyrrhizinate reference substance 0.181mg, amounting to glycyrrhizic acid is 0.1773mg) 5ml, be diluted to scale with mobile phase, shake up, filter (0.45um), make test liquid.The accurate respectively 5ul of absorption injects chromatograph of liquid, and the record chromatograph is measured content, calculate recovery rate (the results are shown in Table 5), and average recovery rate is 99.33%, RSD is 2.03%.

Glycyrrhizic acid is measured recovery test in table 5 need testing solution

Experiment number	Sample size (ml)	Contain glycyrrhizic acid (mg)	Add glycyrrhizic acid (mg)	The amount of recording (mg)	The response rate (%)	Average recovery rate (%)	RSD (％)
Experiment number	Sample size (ml)	Contain glycyrrhizic acid (mg)	Add glycyrrhizic acid (mg)	The amount of recording (mg)	The response rate (%)	Average recovery rate (%)	RSD (％)	1 2 3 4 5	1.25 1.25 1.25 1.25 1.25	0.9393 0.9393 0.9393 0.9393 0.9393	? ? 0.8865 ? ?	1.8335 1.7926 1.8254 1.8122 1.8355	100.9 96.25 99.95 98.47 101.1	? ? 99.33 ? ?	? ? 2.03 ? ?

6. sample determination

Prepare need testing solution and reference substance solution by quality standard of the present invention, sample introduction 5 μ l write down chromatograph respectively, measure peak area, are calculated as follows content:

In the formula: Ai: the need testing solution peak area

As: reference substance solution peak area

Cs: reference substance (glycyrrhizic acid) solution concentration (mg/ml)

10: constant volume (ml)

2.5: sampling amount (ml)

According to preparing 3 batch samples in strict accordance with production technology, glycyrrhizic acid content in the working sample, measurement result sees Table 6.

Glycyrrhizic acid content measurement result in table 63 batch samples

Sample	Content (mg/ml)		Meansigma methods (mg/ml)
	Content (mg/ml)			1	2
	030910 030917 030921	0.748 0.722 0.770		1	2	0.760 0.708 0.762	0.743 0.715 0.766

From three batches of data that contain survey, the decoction yield minimum of this preparation technology's glycyrrhizic acid is 50.39%, is 54.80% to the maximum, and therefore the yield lower bound being fixed tentatively is 50%, and the glycyrrhizic acid content limit is calculated as follows in the preparation:

Glycyrrhizic acid content limit (mg/ml)=preparation contains licorice medicinal materials (g/ml) * medical material and contains glycyrrhizic acid yield * 10 in glycyrrhizic acid limit (pharmacopeia regulation) * preparation ³=(50/1000) * 2.0% * 50% * 10 ³=0.50mg/ml.So the tentative every 1ml of this product contains Radix Glycyrrhizae and must not be less than 0.50mg in glycyrrhizic acid.Measurement result per sample, the glycyrrhizic acid content in 3 batch samples is all in this limits.

The research of qualitative [discriminating] of two pairs of medical materials of experimental example of the present invention

The research of medical material qualitative identification:

(1) with Rhizoma Atractylodis Macrocephalae medical material in the Rhizoma Atractylodis Macrocephalae control medicinal material discriminating side.Method one: get this product, put to steam in the water-bath near and do, added the ethyl acetate supersound process 30 minutes, filter, filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Method two: get this product, put to steam in the water-bath and do, add and leave standstill filtration, the filtrate evaporate to dryness after ethanol stirs near, residue adds the dissolving of water slight fever, use ethyl acetate extraction, abandon ethyl acetate extraction liquid, the water saturated n-butanol extraction of water layer, evaporate to dryness, residue adds methanol 3ml makes dissolving, filters, and filtrate is as need testing solution.Prepare the negative need testing solution that lacks Rhizoma Atractylodis Macrocephalae medical material respectively with method.Other gets Rhizoma Atractylodis Macrocephalae control medicinal material, makes control medicinal material solution by correlation method respectively.Respectively with chloroform-methanol-glacial acetic acid (10: 1.5: 0.2), n-butyl alcohol-acetone-water (4: 3: 1) and n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch.The result shows, selecting method two, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: be that the unfolded separating degree of developing solvent is good 1), speckle colour developing was clear, the noiseless (see figure 1) of negative control, the method favorable reproducibility is so list quality standard text of the present invention in.

(2) with Caulis Polygoni Multiflori medical material in Caulis Polygoni Multiflori control medicinal material and the emodin reference substance discriminating side.Get this product, evaporate to dryness is put coldly, filters after adding the dehydrated alcohol supersound process, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.The negative need testing solution that lacks the Caulis Polygoni Multiflori medical material with the method preparation.Other gets Caulis Polygoni Multiflori control medicinal material 0.1g, adds dehydrated alcohol 20ml, and supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Upper solution with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) and petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1) is developing solvent respectively, launches.The result shows, is that the unfolded separating degree of developing solvent is good with normal hexane-ethyl acetate-formic acid (30: 10: 0.5), and the speckle colour developing is clear, the noiseless (see figure 2) of negative control, and favorable reproducibility is so list quality standard text of the present invention in.

(3) with Herba Agrimoniae medical material in the Herba Agrimoniae control medicinal material discriminating side.Method one: get this product, use chloroform extraction behind the thin up, merge extractive liquid,, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Method two: get this product, thin up is used ether extraction, and water layer is transferred pH1～2 with hydrochloric acid, adds ethyl acetate extraction, and extracting solution evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Method three: get this product, put and steam in the water-bath, add and leave standstill filtration after ethanol stirs to partly near, filtrate evaporate to dryness, residue add water slight fever dissolving, add 5% ethanol solution of sulfuric acid and put on the boiling water bath and refluxed 30 minutes, add water 30ml, put in the water-bath and to steam, put coldly, use ether extraction to about 30ml, extracting solution adds water washing, evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.The negative need testing solution that lacks the Herba Agrimoniae medical material respectively with corresponding method preparation.Other gets the Herba Agrimoniae control medicinal material, prepares control medicinal material solution with corresponding method respectively.Be developing solvent with chloroform-methanol-glacial acetic acid (9: 1: 0.5), chloroform-Ethyl formate-formic acid (5: 5: 1), launch that spray is with different developers respectively.The result shows, selecting method two is that the unfolded separating degree of developing solvent is good with chloroform-Ethyl formate-formic acid (5: 5: 1), and the speckle colour developing is clear, the noiseless (see figure 3) of negative control, and the method favorable reproducibility is so list quality standard text of the present invention in.

(4) Ganoderma and Herba Ecliptae medical material are through test of many times in the side, and all because of feminine gender has interference, separating degree is bad etc., and factor is not set up exclusive thin layer chromatography discrimination method, so exclude the quality standard text.In addition this product adopts the extraction process that decocts, and ursolic acid in the Fructus Ligustri Lucidi can not be proposed, and Fructus Ligustri Lucidi, Cortex Albiziae is unmatchful according to medical material, so do not set up exclusive thin layer chromatography discrimination method.

The specific embodiment

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

The embodiment 1 syrupy quality control of calming the nerves

Get Chinese crude drug Ganoderma 50g, the Rhizoma Atractylodis Macrocephalae (stir-fry) 100g, Fructus Ligustri Lucidi (system) 100g, Cortex Albiziae 150g, Caulis Polygoni Multiflori 150g, Herba Agrimoniae 250g, Herba Ecliptae 100g, Radix Glycyrrhizae 50g, above-mentioned raw materials, Ganoderma decocts with water secondary, 5 hours for the first time, 3 hours for the second time, collecting decoction, filter, it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, and it is standby to get supernatant; Seven flavors such as all the other Rhizoma Atractylodis Macrocephalaes decoct with water secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, and it is standby to get supernatant.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, and thin up stirs evenly to 1000ml, filter, fill, every bag of capacity 10ml, promptly.

[character] this product is the thick liquid of brownish black; Sweet and the little hardship of distinguishing the flavor of.

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

1, measure the content of monoammonium glycyrrhizinate with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D):

Instrument, reagent, Tianjin, reagent island LC-10ATvp high performance liquid chromatograph, the LC-10ATvp infusion pump; SPD-M10Avp secondary array detector; The SCL-10Avp system controller; CTO-10ASvp thermocolumn case; The CLASS-VP6.1 workstation data processing system; Microsyringe (25 μ l), HAMILTON company;

Methanol (chromatographically pure), ammonium acetate (analytical pure), glacial acetic acid (analytical pure), water for injection;

(Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the monoammonium glycyrrhizinate reference substance, for assay usefulness, lot number: 0731-200204).

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Second eyeball-2.5% glacial acetic acid aqueous solution (37: 63) is a mobile phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000.

B. the about 10mg of the preparation extracting liquorice of reference substance solution acid mono-ammonium reference substance accurately claims surely, puts in the 50ml measuring bottle, dissolves and is diluted to scale with mobile phase, shakes up, and promptly gets (every 1ml contains monoammonium glycyrrhizinate reference substance 0.2mg, and amounting to glycyrrhizic acid is 0.1959mg).

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

The every 1ml of this product contains Radix Glycyrrhizae with glycyrrhizic acid (C ₄₂H ₆₂O ₁₆) meter, must not be less than 0.50mg.

2, differentiate this medicine Chinese crude drug Rhizoma Atractylodis Macrocephalae with thin layer chromatography

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material (available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number 22082-200305) 1.5g adds ethanol 50ml reflux, extract, 1 hour, filters, the filtrate evaporate to dryness, residue adds water 30ml slight fever dissolving, with water saturated n-butanol extraction 3 times, and 20ml at every turn, merge n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 10 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

3, differentiate this medicine Chinese crude drug Caulis Polygoni Multiflori with thin layer chromatography

A. the preparation of control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.1g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.

B. the preparation of need testing solution: get this product 5ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 30ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes 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 need testing solution and control medicinal material solution 8 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Can the processing of above-mentioned this Caulis Polygoni Multiflori thin layer discriminating sample be adopted ethanol?, developing solvent is normal hexane-ethyl acetate-formic acid, normal hexane-ethyl acetate-formic acid ratio (30: 10: 0.5) is best, develops the color down for best at ultra-violet lamp (365nm).

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

A. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 2.0g, add water 150ml and be heated to and boil, and keep little and boiled 2 hours, put coldly, filter, filtrate is from " usefulness ether extraction 3 times ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

B. the preparation of need testing solution: get this product 5ml, add water 15ml, use ether extraction 3 times, each 25ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 2, uses ethyl acetate extraction 2 times, each 30ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw need testing solution 6 μ l, control medicinal material solution 10 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-Ethyl formate-formic acid (5: 5: 1) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

The quality control of embodiment 2 ANSHEN KOUFUYE

Get Chinese crude drug Ganoderma 50g, the Rhizoma Atractylodis Macrocephalae (stir-fry) 100g, Fructus Ligustri Lucidi (system) 100g, Cortex Albiziae 150g, Caulis Polygoni Multiflori 150g, Herba Agrimoniae 250g, Herba Ecliptae 100g, Radix Glycyrrhizae 50g, above-mentioned raw materials, Ganoderma decocts with water secondary, 5 hours for the first time, 3 hours for the second time, collecting decoction, filter, it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, and it is standby to get supernatant; Seven flavors such as all the other Rhizoma Atractylodis Macrocephalaes decoct with water secondary, and 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and it is 1.05 (90 ℃) that filtrate is concentrated into relative density, leaves standstill 24 hours, and it is standby to get supernatant.Add simple syrup 264ml, sodium benzoate 2.59,, add water to 1000ml, stir evenly, filter with above-mentioned two kinds of concentrated solution mixings, embedding, every bag of capacity is 10ml, promptly.

[character] this product is the supernatant liquid of brownish black; It is sweet and little puckery to distinguish the flavor of.

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

Embodiment 3

Get Ganoderma 50g, the Rhizoma Atractylodis Macrocephalae (stir-fry) 100g, Fructus Ligustri Lucidi (system) 100g, Cortex Albiziae 150g, Caulis Polygoni Multiflori 150g, Herba Agrimoniae 250g, Herba Ecliptae 100g, Radix Glycyrrhizae 50g,

[method for making], [character], [inspection] are all identical with embodiment 1.Its method of quality control is:

1, measure the content of monoammonium glycyrrhizinate with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D), can carry out according to the following steps:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution (26: 58); The detection wavelength is 247nm; Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000.

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

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 1, add ethanol 40ml reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add the dissolving of water 20ml slight fever, with water saturated n-butanol extraction 1 time, each 10ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 0.5ml makes dissolving, in contrast medical material solution.

B. the preparation of need testing solution: get this product 10ml, put in the water-bath and steam, add ethanol 15ml to about 10ml, stir, left standstill 1 hour, get supernatant, evaporate to dryness, residue add the dissolving of water 10ml slight fever, use ethyl acetate extraction 2 times, each 15ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 2 times, each 15ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 2ml makes dissolving, filters, and filtrate is as need testing solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

A. the preparation of control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.1g, add dehydrated alcohol 15ml, supersound process 15 minutes filters, and filtrate evaporate to dryness, residue add methanol 0.5ml makes dissolving, in contrast medical material solution.

D. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned need testing solution and control medicinal material solution 5 μ l, reference substance solution 0.5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

A. the preparation of need testing solution: get this product 5ml, add water 10ml, use ether extraction 1 time, each 10ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 1, uses ethyl acetate extraction 1 time, each 15ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 1.0g, add water 100ml and be heated to and boil, and keep little and boiled 1.5 hours, put cold, filter, filtrate is from " using ether extraction 1 time ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw need testing solution 6 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-Ethyl formate-formic acid (5: 5: 1) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

Embodiment 4

[method for making], [character], [inspection] are all identical with embodiment 2.Its method of quality control is:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution (53: 69); The detection wavelength is 253nm; Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000.

B. the about 10mg of the preparation extracting liquorice of reference substance solution acid mono-ammonium reference substance accurately claims surely, puts in the 50ml measuring bottle, dissolves and is diluted to scale with mobile phase, shakes up, and promptly gets (every 1ml contains monoammonium glycyrrhizinate reference substance 0.3mg, and amounting to glycyrrhizic acid is 0.2939mg).

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

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 3g, add ethanol 100ml reflux, extract, 1.5 hours, filter, filtrate evaporate to dryness, residue add the dissolving of water 50ml slight fever, with water saturated n-butanol extraction 3 times, each 30ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 3ml makes dissolving, in contrast medical material solution.

B. the preparation of need testing solution: get this product 50ml, put in the water-bath and steam, add ethanol 40ml to about 10ml, stir, left standstill 2 hours, get supernatant, evaporate to dryness, residue add the dissolving of water 30ml slight fever, use ethyl acetate extraction 4 times, each 20ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 4 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 6ml makes dissolving, filters, and filtrate is as need testing solution.

A. the preparation of control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.2g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, in contrast medical material solution.

B. get the emodin reference substance, add methanol and make the solution that every 1ml contains 1.2mg, in contrast product solution.

C. the preparation of need testing solution: get this product 10ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 30ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 3ml makes dissolving, as need testing solution.

D. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned need testing solution and control medicinal material solution 10 μ l, reference substance solution 2.5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

A. the preparation of need testing solution: get this product 10ml, add water 30ml, use ether extraction 3 times, each 25ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 3, uses ethyl acetate extraction 2 times, each 30ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 3ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 2.0g, add water 200ml and be heated to and boil, and keep little and boiled 2.5 hours, put cold, filter, filtrate is from " using ether extraction 3 times ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw need testing solution 12 μ l, control medicinal material solution 10 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-Ethyl formate-formic acid (5: 5: 1) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

Embodiment 5

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

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution (30: 59); The detection wavelength is 251nm; Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000.

B. the about 10mg of the preparation extracting liquorice of reference substance solution acid mono-ammonium reference substance accurately claims surely, puts in the 50ml measuring bottle, dissolves and is diluted to scale with mobile phase, shakes up, and promptly gets (every 1ml contains monoammonium glycyrrhizinate reference substance 0.25mg, and amounting to glycyrrhizic acid is 0.2646mg).

D. accurate respectively reference substance solution and the need testing solution 8 μ l of drawing of algoscopy inject chromatograph of liquid, measure;

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 2g, add ethanol 70ml reflux, extract, 1.5 hours, filter, filtrate evaporate to dryness, residue add the dissolving of water 35ml slight fever, with water saturated n-butanol extraction 2 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 2ml makes dissolving, in contrast medical material solution.

B. the preparation of need testing solution: get this product 30ml, put in the water-bath and steam, add ethanol 25ml to about 10ml, stir, left standstill 1.5 hours, get supernatant, evaporate to dryness, residue add the dissolving of water 20ml slight fever, use ethyl acetate extraction 3 times, each 20ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 4ml makes dissolving, filters, and filtrate is as need testing solution.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 8 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.

A. the preparation of control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.15g, add dehydrated alcohol 20ml, supersound process 25 minutes filters, and filtrate evaporate to dryness, residue add methanol 2.5ml makes dissolving, in contrast medical material solution.

B. get the emodin reference substance, add methanol and make the solution that every 1ml contains 1.0mg, in contrast product solution.

C. the preparation of need testing solution: get this product 8ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 20ml, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1.5ml makes dissolving, as need testing solution.

D. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned need testing solution and control medicinal material solution 8 μ l, reference substance solution 1.5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

A. the preparation of need testing solution: get this product 8ml, add water 20ml, use ether extraction 2 times, each 20ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 2, uses ethyl acetate extraction 1 time, each 20ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 1.5ml makes dissolving, as need testing solution.

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 2.5g, add water 150ml and be heated to and boil, and keep little and boiled 2 hours, put coldly, filter, filtrate is from " usefulness ether extraction 2 times ", and remaining step is made control medicinal material solution by the need testing solution preparation method.

C. test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw need testing solution 9 μ l, control medicinal material solution 8 μ l, put respectively on same silica gel g thin-layer plate, with chloroform-Ethyl formate-formic acid (5: 5: 1) is developing solvent, launches, and takes out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

Claims

1, a kind of method of quality control of spirit quieting oral liquid preparation is characterized in that, may further comprise the steps:

The content of glycyrrhizic acid in the high effective liquid chromatography for measuring spirit quieting oral liquid preparation; With the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and the Herba Agrimoniae in the thin layer chromatography discriminating spirit quieting oral liquid preparation.

2, the method for quality control of claim 1 is characterized in that, the content of glycyrrhizic acid in the described high effective liquid chromatography for measuring spirit quieting oral liquid preparation, and the process following steps:

The preparation of a monoammonium glycyrrhizinate reference substance solution,

The preparation of b spirit quieting oral liquid preparation sample solution,

C measures above solution with high performance liquid chromatograph, gets chromatogram,

D draws the reference substance standard curve, according to spirit quieting oral liquid preparation sample peak area, with standard curve, calculates the content of monoammonium glycyrrhizinate in this sample.

3, the method for quality control of claim 1 is characterized in that, and is described with the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae in the thin layer chromatography discriminating spirit quieting oral liquid preparation, the process following steps:

A. the preparation of the Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae control medicinal material solution

B. the preparation of spirit quieting oral liquid preparation need testing solution

C. test according to thin layer chromatography.

4, the method for quality control of claim 2 is characterized in that, the condition of described high performance liquid chromatography is:

A. chromatographic condition: with octadecylsilane chemically bonded silica is filler; Mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution; The detection wavelength is 250 ± 3nm; Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000;

B. the preparation extracting liquorice of reference substance solution acid mono-ammonium reference substance is an amount of, accurately claims surely, puts in the 50ml measuring bottle, with the mobile phase dissolving and be diluted to scale, shakes up, promptly;

C. the preparation precision of need testing solution is measured this product 2.5ml under the loading amount item, puts in the 10ml measuring bottle, with mobile phase dissolving and be diluted to scale, shakes up, and gets supernatant and filters through 0.45 μ m filter membrane, promptly;

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

5, the method for quality control of claim 2 is characterized in that, the numerical range of described mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution 26-53: 58-69.

6, the method for quality control of claim 5 is characterized in that, the numerical range of described mobile phase is second eyeball-2.5% glacial acetic acid aqueous solution 37: 63.

7, the method for quality control of claim 5 is characterized in that, the process following steps:

Precision takes by weighing monoammonium glycyrrhizinate reference substance 10mg, puts in the 50ml measuring bottle, with mobile phase dissolving and be diluted to scale, shakes up, and makes every 1ml contain monoammonium glycyrrhizinate reference substance 0.2-0.3mg, and amounting to glycyrrhizic acid is 0.1959-0.2939mg; Need testing solution is got supernatant and is filtered 0.45 μ m; Accurate respectively reference substance solution and the need testing solution 5-10 μ l injecting chromatograph drawn measured, that is, the every 1ml of test sample contains Radix Glycyrrhizae with glycyrrhizic acid C ₄₂H ₆₂O ₁₆Meter must not be less than 0.50mg.

8, the method for quality control of claim 3 is characterized in that, and is described with the Chinese crude drug Rhizoma Atractylodis Macrocephalae, Caulis Polygoni Multiflori and Herba Agrimoniae in the thin layer chromatography discriminating spirit quieting oral liquid preparation, the process following steps:

Differentiate the step of the Chinese crude drug Rhizoma Atractylodis Macrocephalae:

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material, add alcohol reflux, filter, filtrate evaporate to dryness, residue add the dissolving of water slight fever, use water saturated n-butanol extraction, merge n-butyl alcohol liquid, and evaporate to dryness, residue add methanol makes dissolving, in contrast medical material solution;

B. the preparation of need testing solution: get this product, put in the water-bath and steam, add ethanol to an amount of, stir, leave standstill, get supernatant, evaporate to dryness, residue add the dissolving of water slight fever, use ethyl acetate extraction, abandon ethyl acetate extraction liquid, the water saturated n-butanol extraction of water layer merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol makes dissolving, filters, and filtrate is as need testing solution;

C. according to thin layer chromatography test, draw above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid is developing solvent, launches, and takes out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing;

Differentiate the step of Chinese crude drug Caulis Polygoni Multiflori:

A. the preparation of control medicinal material solution: get the Caulis Polygoni Multiflori control medicinal material, add dehydrated alcohol, supersound process filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, in contrast medical material solution;

B. get the emodin reference substance, add methanol and make solution, in contrast product solution;

C. the preparation of need testing solution: get this product, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol, supersound process filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution;

D. according to the thin layer chromatography test, draw above-mentioned need testing solution and control medicinal material solution, reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, be developing solvent with normal hexane-ethyl acetate-formic acid, launch, take out, dry, put under the ultra-violet lamp and inspect;

Differentiate the step of Chinese crude drug Herba Agrimoniae:

A. the preparation of need testing solution: get this product, add water, use ether extraction, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value, uses ethyl acetate extraction, merges ethyl acetate extraction liquid, and evaporate to dryness, residue add methanol 1 makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get the Herba Agrimoniae control medicinal material, add water and be heated to and boil, and keep little and boil, put coldly, filter, filtrate is from " using ether extraction ", and remaining step is made control medicinal material solution by the need testing solution preparation method;

9, the method for quality control of claim 7 is characterized in that, described step is:

To the Rhizoma Atractylodis Macrocephalae:

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 1-3g, add ethanol 40-100ml reflux, extract, 1-1.5 hour, filter, filtrate evaporate to dryness, residue add the dissolving of water 20-50ml slight fever, with water saturated n-butanol extraction 1-3 time, each 10-30ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 0.5-3ml makes dissolving, in contrast medical material solution;

B. the preparation of need testing solution: get this product 10-50ml, put in the water-bath and steam, add ethanol 15-40ml to about 10ml, stir, left standstill 1-2 hour, get supernatant, evaporate to dryness, residue add the dissolving of water 10-30ml slight fever, use ethyl acetate extraction 2-4 time, each 15-20ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 2-4 time, each 15-20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 2-6ml makes dissolving, filters, and filtrate is as need testing solution;

C. test according to thin layer chromatography, draw each 5-10 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid (7: 2: 1: 1) be developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

To Caulis Polygoni Multiflori

A. the preparation of control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.1-0.2g, add dehydrated alcohol 15-20ml, supersound process 15-30 minute, filter, filtrate evaporate to dryness, residue add methanol 0.5-3ml makes dissolving, in contrast medical material solution;

B. get the emodin reference substance, add methanol and make the solution that every 1ml contains 0.8-1.2mg, in contrast product solution;

C. the preparation of need testing solution: get this product 5-10ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 10-30ml, supersound process 10-30 minute, filter, filtrate evaporate to dryness, residue add methanol 0.5-3ml makes dissolving, as need testing solution;

D. test according to thin layer chromatography, draw above-mentioned need testing solution and control medicinal material solution 5-10 μ l, reference substance solution 0.5-2.5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid (30: 10: 0.5) is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

To Herba Agrimoniae

A. the preparation of need testing solution: get this product 5-10ml, add water 10-30ml, use ether extraction 1-3 time, each 10-25ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 1-3, uses ethyl acetate extraction 1-2 time, each 15-30ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 0.5-3ml makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 1.0-2.0g, add water 100-200ml and be heated to and boil, and kept little 1.5-2.5 of boiling hour, put cold, filter, filtrate is from " using ether extraction 1-3 time ", and remaining step is made control medicinal material solution by the need testing solution preparation method;

C. according to the thin layer chromatography test, draw need testing solution 6-12 μ l, control medicinal material solution 5-10 μ l, put respectively on same silica gel g thin-layer plate, be developing solvent with chloroform-Ethyl formate-formic acid at 5: 5: 1, launches, take out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.

10, the method for quality control of claim 8 is characterized in that, described step is:

To the Rhizoma Atractylodis Macrocephalae:

A. the preparation of control medicinal material solution: get Rhizoma Atractylodis Macrocephalae control medicinal material 1.5g, add ethanol 50ml reflux, extract, 1 hour, filter, filtrate evaporate to dryness, residue add the dissolving of water 30ml slight fever, with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 1ml makes dissolving, in contrast medical material solution;

B. the preparation of need testing solution: get this product 20ml, put in the water-bath and steam, add ethanol 30ml to about 10ml, stir, left standstill 1 hour, get supernatant, evaporate to dryness, residue add the dissolving of water 30ml slight fever, use ethyl acetate extraction 3 times, each 20ml abandons ethyl acetate extraction liquid, and water layer is with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, evaporate to dryness, residue adds methanol 3ml makes dissolving, filters, and filtrate is 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 in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-acetone-water-glacial acetic acid 7: 2: 1: 1 was developing solvent, launch, take out, dry, spray is with ninhydrin solution, and it is clear that hot blast blows to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;

To Caulis Polygoni Multiflori

A. the preparation of control medicinal material solution: get Caulis Polygoni Multiflori control medicinal material 0.1g, add dehydrated alcohol 20ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, in contrast medical material solution;

B. get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution;

C. the preparation of need testing solution: get this product 5ml, put evaporate to dryness in the water-bath, put coldly, add dehydrated alcohol 30ml, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution;

D. test according to thin layer chromatography, draw above-mentioned need testing solution and control medicinal material solution 8 μ l, reference substance solution 2 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with normal hexane-ethyl acetate-formic acid is developing solvent at 30: 10: 0.5, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

To Herba Agrimoniae

A. the preparation of need testing solution: get this product 5ml, add water 15ml, use ether extraction 3 times, each 25ml, abandon ether solution, aqueous solution adds hydrochloric acid and regulates pH value to 2, uses ethyl acetate extraction 2 times, each 30ml, merge ethyl acetate extraction liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution;

B. the preparation of control medicinal material solution: get Herba Agrimoniae control medicinal material 2.0g, add water 150ml and be heated to and boil, and keep little and boiled 2 hours, put coldly, filter, filtrate is from " usefulness ether extraction 3 times ", and remaining step is made control medicinal material solution by the need testing solution preparation method;

C. according to the thin layer chromatography test, draw need testing solution 6 μ l, control medicinal material solution 10 μ l, put respectively on same silica gel g thin-layer plate, be developing solvent with chloroform-Ethyl formate-formic acid at 5: 5: 1, launches, take out, dry, spray ferric chloride alcoholic solution with 1%.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph principal spot on, show the speckle of same color.