CN1857435A

CN1857435A - Quality control method for Huganning preparation

Info

Publication number: CN1857435A
Application number: CN 200610200296
Authority: CN
Inventors: 叶湘武; 安崇惠; 闫文超; 杨坤; 武燕; 徐裕彬
Original assignee: Guizhou Yibai Pharmaceutical Co Ltd
Current assignee: Guizhou Yibai Pharmaceutical Co Ltd
Priority date: 2006-03-30
Filing date: 2006-03-30
Publication date: 2006-11-08
Anticipated expiration: 2026-03-30
Also published as: CN100535659C

Abstract

The quality control method for Huganning preparation includes character identification and checking and content measurement, the identification includes the tin-layer chromatography identification of giant knotweed and glossy ganoderma in the preparation, and the content measurement is to measure the content of emodin and polydatin in the giant knotweed. Compared with available technology, the present invention has the advantages of high precision, high repeatability, high stability, high recovery rate and raised quality control standard and can ensure the clinical curative effect of Huganning preparation.

Description

The method of quality control of HUGANNING ZHIJI

Technical field: the present invention relates to a kind of method of quality control of HUGANNING ZHIJI, belong to the technical field of medicine being carried out quality control.

Background technology: HUGANNING ZHIJI has clearing away heat-damp and promoting diuresis, the liver benefiting blood stasis dispelling, and Shugan Zhitong, the digestant effect of regulating the flow of vital energy is mainly used in diseases such as acute hepatitis and chronic hepatitis, liver cirrhosis, and its determined curative effect is reliable, has no side effect.Wherein protect proheparinum tablet and be embodied in the 13 in the Chinese traditional patent formulation preparation promulgated by the ministries or commissions of the Central Government, use for many years clinically, obtained than satisfactory therapeutic effects.But find that after deliberation existing HUGANNING ZHIJI exists that quality control standard is simple, the uppity shortcoming of product quality, in process of producing product, can not effectively control the quality of said preparation with this method of quality control, thereby will influence its clinical efficacy.

Summary of the invention:

The objective of the invention is to: the method for quality control that a kind of HUGANNING ZHIJI is provided, the present invention is directed to simple, the uppity shortcoming of product quality of the peaceful tablet quality control criterion of original hepatoprotective, method of quality control to this preparation is studied, having worked out the thin layer chromatography of the assay of emodin and polygonin in the Rhizoma Polygoni Cuspidati and Rhizoma Polygoni Cuspidati, Ganoderma differentiates, improve the quality control standard of HUGANNING ZHIJI, thereby guaranteed the clinical efficacy of said preparation.

HUGANNING ZHIJI of the present invention is made by Herba Sedi 500～1000g, Rhizoma Polygoni Cuspidati 200～800g, Radix Salviae Miltiorrhizae 100～500g and Ganoderma 100～500g, and its preparation method is: get Herba Sedi, be ground into fine powder, remaining Herba Sedi decocts with water, collecting decoction filters, and filtrate is condensed into thick paste; Get Ganoderma, add alcohol dipping, incline that it is standby to get supernatant, medicinal residues flood respectively with 65～90% ethanol, 30～65% ethanol successively, respectively incline and get supernatant, last press residue is collected press juice, merges with three supernatant, filter, filtrate recycling ethanol also is condensed into thick paste, and is standby; Radix Salviae Miltiorrhizae and Rhizoma Polygoni Cuspidati be according to the percolation under Chinese Pharmacopoeia fluid extract and the extractum item, makes solvent soaking percolation (flow velocity per minute 8ml) slowly after 4 hours with 75～95% ethanol, and the soluble component of waiting is filtered out fully, and diacolation liquid recycling ethanol also is condensed into thick paste; The medicinal residues of Radix Salviae Miltiorrhizae, Rhizoma Polygoni Cuspidati and Ganoderma decoct with water, and collecting decoction filters, and filtrate is condensed into thick paste; Get above-mentioned four kinds of thick pastes, add the Herba Sedi fine powder, mixing, drying under reduced pressure adds suitable adjuvant then and makes different preparations.

Method of quality control of the present invention mainly comprise in character, discriminating, inspection and the assay project partly or entirely; Wherein differentiate the thin layer chromatography discriminating that comprises Rhizoma Polygoni Cuspidati in the preparation and Ganoderma; Assay is the assay to contained emodin of Rhizoma Polygoni Cuspidati in the preparation and polygonin.

The discrimination method of Rhizoma Polygoni Cuspidati is to be contrast with Rhizoma Polygoni Cuspidati control medicinal material, emodin reference substance and physcione reference substance respectively, and with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is the thin layer chromatography of developing solvent.

The discrimination method of Ganoderma is to be contrast with the Ganoderma control medicinal material, and with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is the thin layer chromatography of developing solvent.

Described discrimination method comprises the part or all of of following project:

(1) gets this preparation or its content, add the methanol supersound process, filter the filtrate evaporate to dryness, residue adds the 2.5mol/L sulfuric acid solution, heating hydrolysis, put cold, with chloroform extraction 1～5 time, merge chloroform liquid, evaporate to dryness, residue add chloroform makes dissolving, as need testing solution; Other gets the Rhizoma Polygoni Cuspidati control medicinal material, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add dissolve with methanol respectively, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take out, dry, put under the 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation.

(2) get this preparation or its content, add ethanol, reflux filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution; Other gets the Ganoderma control medicinal material, shines medical material solution in pairs with legal system; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

Discrimination method comprises the part or all of of following project more specifically:

(1) gets this preparation or its content 0.5～5g, add methanol 10～100ml, supersound process 5～100 minutes, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 1～20ml, heating hydrolysis 10～100 minutes, put cold, with chloroform extraction 1～5 time, merge chloroform liquid, evaporate to dryness, residue adds chloroform 0.5～3ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01～0.5g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2～3mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation.

(2) get this preparation or its content 1～10g, add ethanol 10～100ml, reflux 10～100 minutes filters, and filtrate evaporate to dryness, residue add methanol 0.5～5ml makes dissolving, as need testing solution; Other gets Ganoderma control medicinal material 0.5～5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 2～15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

The emodin content assay method is to be contrast with the emodin reference substance in the Rhizoma Polygoni Cuspidati, and with methanol or acetonitrile: 0.05～5% phosphoric acid solution=10～90: 90～10 is the high performance liquid chromatography of mobile phase.

The content assaying method of polygonin is to be contrast with the polygonin reference substance in the Rhizoma Polygoni Cuspidati, and with methanol or acetonitrile: 0.05～5% phosphoric acid=10～90: 90～10 is the high performance liquid chromatography of mobile phase.

Described content assaying method comprises the part or all of of following project:

(1) emodin shines the Chinese Pharmacopoeia high effective liquid chromatography for measuring:

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol or acetonitrile: 0.05～5% phosphoric acid solution=10～90: 90～10 is mobile phase; Detect wavelength 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000;

The preparation of reference substance solution: it is 12～48 hours emodin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds dissolve with methanol, promptly;

The preparation of need testing solution: get this preparation or its content under the content uniformity item, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol that adds claims to decide weight, supersound extraction, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate and is put in the flask, volatilize solvent, add chloroform and 0.5～5mol/L sulfuric acid solution, put reflux in 80 ℃ of water-baths, put cold, in the dislocation separatory funnel, with a small amount of chloroform washing container, incorporate in the separatory funnel, divide and get the chloroform layer, acid solution chloroform extraction 1～5 time, merge chloroform extraction liquid, decompression and solvent recovery is to doing, and residue adds methanol makes dissolving and standardize solution, shake up, filter, get subsequent filtrate, promptly;

Algoscopy: accurate reference substance solution and the need testing solution drawn, inject chromatograph of liquid, measure, promptly;

This capsule contains Rhizoma Polygoni Cuspidati with emodin C ₁₅H ₁₀O ₅Meter must not be less than 1.5mg/g; This tablet contains Rhizoma Polygoni Cuspidati with emodin C ₁₅H ₁₀O ₅Meter must not be less than 0.5mg/g; This granule contains Rhizoma Polygoni Cuspidati with emodin C ₁₅H ₁₀O ₅Meter must not be less than 0.1mg/g.

(2) polygonin is according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring, and lucifuge is operated:

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol or acetonitrile: 0.05～5% phosphoric acid=10～90: 90～10 is mobile phase; Detect wavelength 306nm; Number of theoretical plate calculates by the polygonin peak should be not less than 3000;

The preparation of reference substance solution: it is 12～48 hours polygonin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds the Diluted Alcohol dissolving, promptly;

The preparation of need testing solution: get this preparation or its content under the content uniformity item, the accurate title, decide, and the accurate Diluted Alcohol that adds claims to decide weight, reflux is cooled to room temperature, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly;

This capsule contains Rhizoma Polygoni Cuspidati with polygonin C ₂₀H ₂₂O ₈Meter must not be less than 0.5mg/g; This tablet contains Rhizoma Polygoni Cuspidati with polygonin C ₂₀H ₂₂O ₈Meter must not be less than 0.2mg/g; This granule contains Rhizoma Polygoni Cuspidati with polygonin C ₂₀H ₂₂O ₈Meter must not be less than 0.054mg/g.

Content assaying method comprises the part or all of of following project more specifically:

The preparation of reference substance solution: it is 12～48 hours emodin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds methanol and makes the solution that every 1ml contains 30～100 μ g, promptly;

The preparation of need testing solution: get this preparation or its content 0.1～3g under the content uniformity item, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 10～200ml that adds claims to decide weight, at power 250W, supersound extraction is 10～100 minutes under the frequency 33kHz, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 5～50ml, puts in the flask, volatilize solvent, add chloroform 5～50ml and 0.5～5mol/L sulfuric acid solution, 5～50ml, put in 80 ℃ of water-baths reflux 0.5～5 hour, put cold, in the dislocation separatory funnel, with a small amount of chloroform washing container, incorporate in the separatory funnel, divide and get the chloroform layer, acid solution chloroform extraction 1～5 time, merge chloroform extraction liquid, decompression and solvent recovery is to doing, and residue adds methanol makes dissolving, be transferred in the 10ml measuring bottle, add methanol and be diluted to scale, shake up, filter, get subsequent filtrate, promptly;

Algoscopy: accurate reference substance solution and each 5～25 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

The preparation of reference substance solution: it is 12～48 hours polygonin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds Diluted Alcohol and makes the solution that every 1ml contains 10～50 μ g, promptly;

The preparation of need testing solution: get this preparation or its content 0.05～5g under the content uniformity item, the accurate title, decide, and the accurate Diluted Alcohol 10～200ml that adds claims to decide weight, reflux 10～100 minutes, be cooled to room temperature, claim again to decide weight, supply the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly;

Algoscopy: accurate reference substance solution and each 10～25 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

Described method of quality control comprises:

Character:

For capsule: content is tan granule; Bitter in the mouth, little acid, puckery;

For tablet: medicine shows sepia; Bitter in the mouth, little acid, puckery;

For granule: medicine is tan granule; Little sweet, the little acid, puckery of distinguishing the flavor of;

Differentiate:

Check: should meet every regulation relevant under Chinese Pharmacopoeia capsule, tablet or the granule item.

Assay:

We find after deliberation, adopt the quality of following method of quality control with this preparation of easier control, are more conducive to guarantee the clinical efficacy of preparation, so described method of quality control also can comprise:

Character:

For capsule: content is tan granule; Bitter in the mouth, little acid, puckery;

For tablet: medicine shows sepia; Bitter in the mouth, little acid, puckery;

Differentiate: (1) gets this preparation or its content 0.5～5g, adds methanol 10～100ml, supersound process 5～100 minutes, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 1～20ml, heating hydrolysis 10～100 minutes, put cold, with chloroform extraction 1～5 time, merge chloroform liquid, evaporate to dryness, residue adds chloroform 0.5～3ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01～0.5g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2～3mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation.

Assay:

In order to ensure method of quality control science of the present invention, reasonable, feasible, the discriminating of each medical material and content assaying method are studied among the applicant the other side, and concrete testing data is as follows:

One, emodin content study on determination method

1. the selection of mobile phase:

Mobile phase 1: the mixed solution with methanol, 0.1% phosphoric acid different proportion is a mobile phase;

Mobile phase 2: the mixed solution with acetonitrile, 0.1% phosphoric acid different proportion is a mobile phase;

Mobile phase 3: the mixed solution with methanol, 1% perchloric acid different proportion is a mobile phase.

Result: with methanol or acetonitrile: 0.05～5% phosphoric acid solution=10～90: 90～10 is mobile phase, and the negative sample chromatogram is at non-false positive peak, place, emodin peak position; Particularly under methanol-0.1% phosphoric acid (82: 18) mobile phase condition, the emodin chromatographic peak separates best with other chromatographic peak.

2. need testing solution preparation method research:

Method one: the thing 0.5g that gets it filled, the accurate title, decide, and adds methanol 50ml, supersound extraction (power 250W, frequency 33kHz) 45 minutes filters, get subsequent filtrate 20ml, volatilize solvent, add chloroform 25ml and 2.5mol/L sulfuric acid solution 20ml, put in 80 ℃ of water-baths reflux 2 hours, and be cooled to room temperature, divide and get chloroform liquid, acid solution 3 (20ml of chloroform extraction, 10ml 10ml), merges chloroform extraction liquid, decompression and solvent recovery is to doing, residue adds methanol makes dissolving, is transferred in the 10ml measuring bottle, adds methanol and is diluted to scale, shake up, filter, get subsequent filtrate, promptly.

Method two, the thing 0.5g that gets it filled accurate claim surely, and accurate chloroform 50ml and the 2.5mol/L sulfuric acid solution 20ml of adding claims decide weight, put in 80 ℃ of water-baths reflux 2 hours, are cooled to room temperature, and weight decided in title again, supplies the weight that subtracts mistake with chloroform, shakes up.Divide and get chloroform liquid, precision is measured 20ml, and evaporate to dryness, residue add methanol makes dissolving, is transferred in the 10ml measuring bottle, adds methanol and is diluted to scale, shakes up, promptly.

Draw each 10 μ l of above-mentioned two kinds of need testing solutions respectively, inject chromatograph of liquid, press the text chromatographic condition, measure in accordance with the law, the results are shown in following table.

Different Extraction Method is investigated the result

Extracting method	Sample weighting amount (g)	Emodin extraction ratio (mg/g)
Extracting method	Sample weighting amount (g)	Emodin extraction ratio (mg/g)	Method one	0.5006	3.091
Method two	0.5011	2.893	Method one	0.5006	3.091

Result of the test shows, employing method one, and the emodin extraction ratio is higher.

3. precision test

Accurate reference substance solution (49.25 μ g/ml) the 10 μ l that draw, continuous sample introduction 5 times calculates relative standard deviation:

Experiment number	1	2	3	4	5	Meansigma methods	RSD％
Experiment number	1	2	3	4	5	Meansigma methods	RSD％	Peak area	1778.4	1782.2	1781.6	1783.7	1788.8	1782.9	0.21

Result of the test shows that this method precision is good.

4. stability test

Get same need testing solution, measured in 0,2,4,6,8 hour in accordance with the law respectively at the preparation back.

Experiment number	0	2	4	6	8	Meansigma methods	RSD％
Experiment number	0	2	4	6	8	Meansigma methods	RSD％	Peak area	2145.8	2089.4	2142.8	2091.7	2139.3	2121.8	1.35

The result shows that need testing solution is stable in 8 hours.

5. replica test

Preparation method by test liquid under the assay item among the present invention takes by weighing same lot number sample 0.5g respectively, and 6 parts, measure, calculate relative standard deviation, the result is as follows.

Number of times	Sample weighting amount (g)	Content (mg/g)	Average content (mg/g)	RSD％
Number of times	Sample weighting amount (g)	Content (mg/g)	Average content (mg/g)	RSD％	1	0.4974	3.054	3.097	1.87
2	0.5007	3.034			1	0.4974	3.054
2	0.5007	3.034	3	0.4936	3.185
4	0.4989	3.147	3	0.4936	3.185
4	0.4989	3.147	5	0.5003	3.070
6	0.4902	3.091	5	0.5003	3.070

The result shows that this method repeatability is good.

6. recovery test

Take application of sample absorption method (adding) by 1: 1, get the about 0.25g of same lot number sample of known content, the accurate title, decide, respectively put in the 50ml tool plug bottle respectively accurate emodin reference substance solution (each 2ml of 3.91mg → 10ml), the accurate methanol 48ml that adds of adding, other operate same text, and measure, calculate recovery rate the results are shown in following table as follows. in accordance with the law

Sequence number	Sample weighting amount (g)	Content in the sample (mg)	Addition (mg)	Measure total amount (mg)	The response rate (%)	Meansigma methods (%)	RSD (％)
Sequence number	Sample weighting amount (g)	Content in the sample (mg)	Addition (mg)	Measure total amount (mg)	The response rate (%)	Meansigma methods (%)	RSD (％)	1	0.2523	0.781	0.782	1.571	101.0	101.1	2.45
2	0.2514	0.779	0.782	1.564	100.4			1	0.2523	0.781	0.782	1.571	101.0
2	0.2514	0.779	0.782	1.564	100.4	3	0.2646	0.819	0.782	1.625	103.0
4	0.2605	0.807	0.782	1.625	104.6	3	0.2646	0.819	0.782	1.625	103.0
4	0.2605	0.807	0.782	1.625	104.6	5	0.2539	0.786	0.782	1.568	99.9
6	0.2491	0.771	0.782	1.534	97.5	5	0.2539	0.786	0.782	1.568	99.9

Result of the test shows that the average average recovery of emodin is between 95%～105%, and application of sample reclaims good.

Two, polygonin content assaying method research

1. the selection of mobile phase:

Mobile phase 1: the mixed solution with the acetonitrile-water different proportion is a mobile phase;

Mobile phase 2: the mixed solution with methanol-0.1% phosphoric acid different proportion is a mobile phase;

Mobile phase 3: the mixed solution with methanol-0.05% phosphoric acid different proportion is a mobile phase;

Mobile phase 4: the mixed solution with methanol-5% phosphoric acid different proportion is a mobile phase;

Mobile phase 5: the mixed solution with acetonitrile-0.1% phosphoric acid different proportion is a mobile phase;

Mobile phase 6: the mixed solution with acetonitrile alcohol-0.05% phosphoric acid different proportion is a mobile phase;

Mobile phase 7: the mixed solution with acetonitrile-0.5% phosphoric acid different proportion is a mobile phase.

Result: with methanol or acetonitrile: 0.05～5% phosphoric acid=10～90: 90～10 is mobile phase, and the negative sample chromatogram is at non-false positive peak, place, polygonin peak position; Particularly under acetonitrile-0.1% phosphoric acid (16: 84) mobile phase condition, the polygonin chromatographic peak all can reach better with other chromatographic peak and separate (separating degree is 3.69), and peak shape symmetry, sharp-pointed, retention time suit (about 10 minutes).

2. need testing solution preparation method research:

Because the polygonin phenolic hydroxy group sees that oxidation/polyreaction easily takes place light, so following operation is lucifuge.

(1) adopt reflux, operate as follows: the about 0.15g of the thing of getting it filled, the accurate title, decide, and the accurate Diluted Alcohol 50ml that adds claims to decide weight, and heating and refluxing extraction is cooled to room temperature, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and gets supernatant, promptly.

(2) adopt the supersound process method, operate as follows: the about 0.15g of the thing of getting it filled, accurately claim surely, accurate adding Diluted Alcohol 50ml claims to decide weight, and supersound process is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with Diluted Alcohol, shakes up, and gets supernatant, promptly.

Draw each 20 μ l of above-mentioned two kinds of need testing solutions respectively, inject chromatograph of liquid, press the text chromatographic condition, measure in accordance with the law.

Extracting method	Sample weighting amount (g)	Extraction ratio (mg/g)
Extracting method	Sample weighting amount (g)	Extraction ratio (mg/g)	(1) reflux	0.1513	6.731
(2) supersound process	0.1522	6.706	(1) reflux	0.1513	6.731

Result of the test shows, adopts the extracting method of reflux, and the polygonin extraction ratio is higher.

3. precision test

Accurate polygonin reference substance solution (23.2 μ g/ml) the 20 μ l that draw, continuous sample introduction 5 times calculates relative standard deviation.

Experiment number	Peak area	Meansigma methods	RSD％
Experiment number	Peak area	Meansigma methods	RSD％	1	1676.6	1673.2	0.14
2	1672.0			1	1676.6
2	1672.0	3	1670.3
4	1674.5	3	1670.3
4	1674.5	5	1672.7

Result of the test shows that this method precision is good.

4. stability test

Get same need testing solution, measured in 0,2,4,6,8 hour respectively at the preparation back in accordance with the law, the result is as follows.

Time (hr)	Peak area	Meansigma methods	RSD％
Time (hr)	Peak area	Meansigma methods	RSD％	0	1474.3	1465.6	0.84
2	1464.6			0	1474.3
2	1464.6	4	1476.6
6	1466.8	4	1476.6
6	1466.8	8	1445.5

The result shows that need testing solution is stable in 8 hours.

5. replica test

Preparation method by test liquid under the assay item among the present invention takes by weighing same lot number sample 0.15g respectively, and 6 parts, measure, calculate relative standard deviation, the result is as follows.

Number of times	Sample weighting amount (g)	Content (mg/g)	Average content (mg/g)	RSD％
Number of times	Sample weighting amount (g)	Content (mg/g)	Average content (mg/g)	RSD％	1	0.1483	6.769	6.731	0.68
2	0.1497	6.710			1	0.1483	6.769
2	0.1497	6.710	3	0.1482	6.741
4	0.1486	6.692	3	0.1482	6.741
4	0.1486	6.692	5	0.1478	6.795
6	0.1496	6.677	5	0.1478	6.795

The result shows that this method repeatability is good.

6. recovery test

Take application of sample absorption method (adding) by 1: 1, get the about 0.075g of same lot number sample of known content, the accurate title, decide, respectively accurate polygonin reference substance solution (each 1ml of 5.08mg → 10ml) that adds, other are measured by need testing solution preparation method under the assay item of the present invention and chromatographic condition, calculate recovery rate as follows, result such as following table.

Sequence number	Sample weighting amount (g)	Content in the sample (mg)	Addition (mg)	Measure total amount (mg)	The response rate (%)	Meansigma methods (%)	RSD (％)
Sequence number	Sample weighting amount (g)	Content in the sample (mg)	Addition (mg)	Measure total amount (mg)	The response rate (%)	Meansigma methods (%)	RSD (％)	1	0.0750	0.505	0.508	1.004	98.3	97.8	1.09
2	0.0744	0.501	0.508	1.004	99.1			1	0.0750	0.505	0.508	1.004	98.3
2	0.0744	0.501	0.508	1.004	99.1	3	0.0748	0.503	0.508	0.999	97.5
4	0.0753	0.507	0.508	0.994	95.9	3	0.0748	0.503	0.508	0.999	97.5
4	0.0753	0.507	0.508	0.994	95.9	5	0.0751	0.505	0.508	1.004	98.1
6	0.0744	0.501	0.508	0.999	98.1	5	0.0751	0.505	0.508	1.004	98.1

Result of the test shows that the average average recovery of polygonin is between 95%～105%, and application of sample reclaims good.

Three, the thin layer Study on Identification of Rhizoma Polygoni Cuspidati

The preparation of reference substance solution: get emodin reference substance and physcione reference substance, add methanol respectively and make the solution that every 1ml contains 1mg, in contrast product solution.

The preparation of control medicinal material solution: get the Rhizoma Polygoni Cuspidati control medicinal material, make control medicinal material solution according to the need testing solution preparation method.

The preparation of negative controls: get the pharmaceutical decocting piece of other except that Rhizoma Polygoni Cuspidati in the prescription, prepare by preparation technology and lack the Rhizoma Polygoni Cuspidati negative sample, with the preparation method preparation identical, promptly with need testing solution.

The preparation method one of need testing solution: the thing 1.5g that gets it filled adds methanol 50ml, supersound process, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 5ml, heating hydrolysis 30 minutes, put cold, with chloroform extraction 2 times, combined chloroform liquid, evaporate to dryness, residue adds chloroform makes dissolving, as need testing solution.

Need testing solution preparation method two: the thing 1.5g that gets it filled, add 0.25mol/L sulfuric acid solution 5ml, heating hydrolysis 30 minutes is put coldly, uses chloroform extraction, combined chloroform liquid, evaporate to dryness, residue adds dissolve with methanol, as need testing solution.

Developing solvent is selected: respectively with the mixed solution of 30～60 ℃ of petroleum ether, butyl acetate, methanol, glacial acetic acid different proportion; Mixed solution with 30～60 ℃ of petroleum ether, Ethyl formate, formic acid different proportion is developing solvent.

The result: employing method one preparation need testing solution, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, good separating effect, clear spot, negative control is noiseless, the method favorable reproducibility.Best developing solvent is the upper solution of petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1).

Four, the thin layer Study on Identification of Ganoderma

The preparation of control medicinal material solution: get the Ganoderma control medicinal material, make control medicinal material solution according to the need testing solution preparation method.

The preparation of negative controls: get the pharmaceutical decocting piece of other except that Ganoderma in the prescription, prepare by preparation technology and lack the Ganoderma negative sample, with the preparation method preparation identical, promptly with need testing solution.

Need testing solution preparation method one: get this product content, add ethanol, reflux filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution.

Need testing solution preparation method two: get this product content, add methanol, reflux filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution.

Developing solvent is selected: respectively with the mixed solution of 60～90 ℃ of petroleum ether, Ethyl formate, formic acid different proportion; Mixed solution with 30～60 ℃ of petroleum ether, butyl acetate, methanol, glacial acetic acid different proportion is developing solvent.

The result: employing method one preparation need testing solution, with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, good separating effect, clear spot, negative control is noiseless, the method favorable reproducibility.Best developing solvent is the upper solution of petroleum ether (60～90 ℃)-Ethyl formate-formic acid (15: 5: 1).

Compared with prior art, method of quality control of the present invention research has also been worked out the assay of emodin and polygonin in the Rhizoma Polygoni Cuspidati and the thin layer chromatography of Rhizoma Polygoni Cuspidati, Ganoderma is differentiated, the method precision height that is adopted, favorable reproducibility, good stability, response rate height has improved the quality control standard of HUGANNING ZHIJI, thereby has guaranteed the clinical efficacy of said preparation.

The specific embodiment:

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

Embodiments of the invention 1: described capsule method of quality control comprises:

Character: content is tan granule; Bitter in the mouth, little acid, puckery;

Differentiate: (1) gets this product content 1.5g, adds methanol 50ml, supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 5ml, heating hydrolysis 30 minutes, put cold, with chloroform extraction 2 times, each 5ml, merge chloroform liquid, evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.1g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 1mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 5 μ l of need testing solution and control medicinal material solution, each 1 μ l of reference substance solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (30～60 ℃): Ethyl formate: the upper solution of formic acid=15: 5: 1 is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation.

(2) get this product content 3.5g, add ethanol 30ml, reflux 30 minutes filters, and filtrate evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets Ganoderma control medicinal material 2g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃): Ethyl formate: the upper solution of formic acid=15: 5: 1 is developing solvent, launches, take out, dry, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

Check: should meet every regulation relevant under the Chinese Pharmacopoeia capsule item.

Assay:

(1) emodin is according to the Chinese Pharmacopoeia high effective liquid chromatography for measuring

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol: 0.1% phosphoric acid solution=82: 18 is a mobile phase; Detect wavelength 254nm.Number of theoretical plate calculates by the emodin peak should be not less than 3000.

The preparation of reference substance solution: it is that 24 hours emodin reference substance of desiccant drying under reduced pressure is an amount of that precision takes by weighing with the phosphorus pentoxide, adds methanol and makes the solution that contains 55 μ g among every 1ml, promptly.

The preparation of need testing solution: get this product content 0.5g under the content uniformity item, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, supersound extraction (power 250W, frequency 33kHz) 45 minutes, put coldly, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 20ml, puts in the flask, volatilizes solvent, add chloroform 25ml and 2.5mol/L sulfuric acid solution 20ml, put in 80 ℃ of water-baths reflux 2 hours, put cold, in the dislocation separatory funnel, with a small amount of chloroform washing container, incorporate in the separatory funnel, divide and get the chloroform layer, acid solution 3 (20ml of chloroform extraction, 10ml 10ml), merges chloroform extraction liquid, decompression and solvent recovery is to doing, residue adds methanol makes dissolving, is transferred in the 10ml measuring bottle, adds methanol and is diluted to scale, shake up, filter, get subsequent filtrate, promptly.

Algoscopy: accurate reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.

This capsule contains Rhizoma Polygoni Cuspidati with emodin C ₁₅H ₁₀O ₅Meter must not be less than 1.5mg/g.

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With acetonitrile: 0.1% phosphoric acid=16: 84 is mobile phase; Detect wavelength 306nm, number of theoretical plate calculates by the polygonin peak should be not less than 3000.

The preparation of reference substance solution: it is that 24 hours polygonin reference substance of desiccant drying under reduced pressure is an amount of that precision takes by weighing with the phosphorus pentoxide, adds Diluted Alcohol and makes the solution that every 1ml contains 20 μ g, promptly.

The preparation of need testing solution: get this product content 0.15g under the content uniformity item, the accurate title, decide, and the accurate Diluted Alcohol 50ml that adds claims to decide weight, reflux 30 minutes is cooled to room temperature, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly.

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

This capsule contains Rhizoma Polygoni Cuspidati with polygonin C ₂₀H ₂₂O ₈Meter must not be less than 0.5mg/g.

Embodiments of the invention 2: described tablet quality control method can comprise:

Character: medicine shows sepia; Bitter in the mouth, little acid, puckery;

Differentiate: (1) gets this preparation 5g, adds methanol 100ml, supersound process 100 minutes, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 20ml, heating hydrolysis 100 minutes, put cold, with chloroform extraction 5 times, merge chloroform liquid, evaporate to dryness, residue adds chloroform 3ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.5g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 3mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=5: 10: 0.5 is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

(2) get this preparation 10g, add ethanol 100ml, reflux 100 minutes filters, and filtrate evaporate to dryness, residue add methanol 5ml makes dissolving, as need testing solution; Other gets Ganoderma control medicinal material 5g, shines medical material solution in pairs with legal system; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw each 15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=30: 1: 5 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

Check: should meet every regulation relevant under the Chinese Pharmacopoeia tablet item;

Assay:

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With acetonitrile: 0.05% phosphoric acid solution=90: 10 is a mobile phase; Detect wavelength 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000;

The preparation of reference substance solution: it is 48 hours emodin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds methanol and makes the solution that every 1ml contains 100 μ g, promptly;

The preparation of need testing solution: get this preparation 3g under the content uniformity item, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 200ml that adds claims to decide weight, at power 250W, supersound extraction is 100 minutes under the frequency 33kHz, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 50ml, put in the flask, volatilize solvent, add chloroform 50ml and 5mol/L sulfuric acid solution 50ml, put in 80 ℃ of water-baths reflux 5 hours, put cold, in the dislocation separatory funnel, with a small amount of chloroform washing container, incorporate in the separatory funnel, divide and get the chloroform layer, acid solution chloroform extraction 5 times, merge chloroform extraction liquid, decompression and solvent recovery is to doing, and residue adds methanol makes dissolving, be transferred in the 10ml measuring bottle, add methanol and be diluted to scale, shake up, filter, get subsequent filtrate, promptly;

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

This tablet contains Rhizoma Polygoni Cuspidati with emodin C ₁₅H ₁₀O ₅Meter must not be less than 0.5mg/g.

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol: 5% phosphoric acid=90: 10 is mobile phase; Detect wavelength 306nm; Number of theoretical plate calculates by the polygonin peak should be not less than 3000;

The preparation of reference substance solution: it is 48 hours polygonin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds Diluted Alcohol and makes the solution that every 1ml contains 50 μ g, promptly;

The preparation of need testing solution: get this preparation 5g under the content uniformity item, the accurate title, decide, and the accurate Diluted Alcohol 200ml that adds claims to decide weight, reflux 100 minutes is cooled to room temperature, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly;

Algoscopy: accurate reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

This tablet contains Rhizoma Polygoni Cuspidati with polygonin C ₂₀H ₂₂O ₈Meter must not be less than 0.2mg/g.

Embodiments of the invention 3: described granule method of quality control can comprise:

Character: medicine is tan granule; Little sweet, the little acid, puckery of distinguishing the flavor of;

Differentiate: (1) gets this preparation 0.5g, add methanol 10ml, supersound process 5 minutes filters, the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 1ml, and heating hydrolysis 10 minutes is put cold, with chloroform extraction 1 time, chloroform liquid evaporate to dryness, residue add chloroform 0.5ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=30: 1: 5 is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

(2) get this preparation 1g, add ethanol 10ml, reflux 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution; Other gets Ganoderma control medicinal material 0.5g, shines medical material solution in pairs with legal system; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=5: 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 the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

Check: should meet every regulation relevant under the Chinese Pharmacopoeia granule item;

Assay:

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With acetonitrile: 5% phosphoric acid solution=10: 90 is a mobile phase; Detect wavelength 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000;

The preparation of reference substance solution: it is 12 hours emodin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds methanol and makes the solution that every 1ml contains 30 μ g, promptly;

The preparation of need testing solution: get this preparation 0.1g under the content uniformity item, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 10ml that adds claims to decide weight, at power 250W, supersound extraction is 10 minutes under the frequency 33kHz, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 5ml, put in the flask, volatilize solvent, add chloroform 5ml and 0.5mol/L sulfuric acid solution 5ml, put in 80 ℃ of water-baths reflux 0.5 hour, put cold, in the dislocation separatory funnel, with a small amount of chloroform washing container, incorporate in the separatory funnel, divide and get the chloroform layer, acid solution chloroform extraction 1 time, chloroform extraction liquid decompression and solvent recovery is to doing, residue adds methanol makes dissolving, is transferred in the 10ml measuring bottle, adds methanol and is diluted to scale, shake up, filter, get subsequent filtrate, promptly;

Algoscopy: accurate reference substance solution and each 25 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

This granule contains Rhizoma Polygoni Cuspidati with emodin C ₁₅H ₁₀O ₅Meter must not be less than 0.1mg/g.

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol: 0.05% phosphoric acid=10: 90 is mobile phase; Detect wavelength 306nm; Number of theoretical plate calculates by the polygonin peak should be not less than 3000;

The preparation of reference substance solution: it is 12 hours polygonin reference substance of desiccant drying under reduced pressure that precision takes by weighing with the phosphorus pentoxide, adds Diluted Alcohol and makes the solution that every 1ml contains 10 μ g, promptly;

The preparation of need testing solution: get this preparation 0.05g under the content uniformity item, the accurate title, decide, and the accurate Diluted Alcohol 10ml that adds claims to decide weight, reflux 10 minutes is cooled to room temperature, claims to decide weight again, supplies the weight that subtracts mistake with Diluted Alcohol, shake up, filter, get subsequent filtrate, promptly;

This granule contains Rhizoma Polygoni Cuspidati with polygonin C ₂₀H ₂₂O ₈Meter must not be less than 0.054mg/g.

Embodiments of the invention 4: described capsule method of quality control can comprise:

Character: content is tan granule; Bitter in the mouth, little acid, puckery;

Differentiate: (1) gets this preparation content 0.5g, add methanol 10ml, supersound process 5 minutes filters, the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 1ml, and heating hydrolysis 10 minutes is put cold, with chloroform extraction 1 time, chloroform liquid evaporate to dryness, residue add chloroform 0.5ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=30: 1: 0.5 is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

(2) get this preparation content 1g, add ethanol 10ml, reflux 10 minutes filters, and filtrate evaporate to dryness, residue add methanol 0.5ml makes dissolving, as need testing solution; Other gets Ganoderma control medicinal material 0.5g, shines medical material solution in pairs with legal system; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=5: 10: 5 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.

Embodiments of the invention 5: described tablet quality control method can comprise:

Character: medicine shows sepia; Bitter in the mouth, little acid, puckery;

Differentiate: get this preparation 0.5g, add methanol 100ml, supersound process 5 minutes filters, the filtrate evaporate to dryness, residue adds 2.5mol/L sulfuric acid solution 1ml, and heating hydrolysis 10 minutes is put cold, with chloroform extraction 1 time, chloroform liquid evaporate to dryness, residue add chloroform 0.5ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: the upper solution of formic acid=30: 1: 5 is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

Assay:

Emodin shines the Chinese Pharmacopoeia high effective liquid chromatography for measuring:

Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol: 5% phosphoric acid solution=30: 70 is a mobile phase; Detect wavelength 254nm; Number of theoretical plate calculates by the emodin peak should be not less than 3000;

Claims

1. the method for quality control of a HUGANNING ZHIJI, described preparation comprises capsule, tablet and granule, it is characterized in that: described method of quality control mainly comprise in character, discriminating, inspection and the assay project partly or entirely; Wherein differentiate the thin layer chromatography discriminating that comprises Rhizoma Polygoni Cuspidati in the preparation and Ganoderma; Assay is the assay to contained emodin of Rhizoma Polygoni Cuspidati in the preparation and polygonin.

2. according to the method for quality control of the described HUGANNING ZHIJI of claim 1, it is characterized in that: the discrimination method of Rhizoma Polygoni Cuspidati is to be contrast with Rhizoma Polygoni Cuspidati control medicinal material, emodin reference substance and physcione reference substance respectively, and with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is the thin layer chromatography of developing solvent.

3. according to the method for quality control of the described HUGANNING ZHIJI of claim 1, it is characterized in that: the discrimination method of Ganoderma is to be contrast with the Ganoderma control medicinal material, and with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is the thin layer chromatography of developing solvent.

4. according to the method for quality control of claim 1,2 or 3 described HUGANNING ZHIJI, it is characterized in that: described discrimination method comprises the part or all of of following project:

(1) gets this preparation or its content, add the methanol supersound process, filter the filtrate evaporate to dryness, residue adds the 2.5mol/L sulfuric acid solution, heating hydrolysis, put cold, with chloroform extraction 1～5 time, merge chloroform liquid, evaporate to dryness, residue add chloroform makes dissolving, as need testing solution; Other gets the Rhizoma Polygoni Cuspidati control medicinal material, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add dissolve with methanol respectively, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take out, dry, put under the 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

5. according to the method for quality control of the described HUGANNING ZHIJI of claim 4, it is characterized in that: discrimination method comprises the part or all of of following project more specifically:

(1) gets this preparation or its content 0.5～5g, add methanol 10～100ml, supersound process 5～100 minutes, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 1～20ml, heating hydrolysis 10～100 minutes, put cold, with chloroform extraction 1～5 time, merge chloroform liquid, evaporate to dryness, residue adds chloroform 0.5～3ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01～0.5g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2～3mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

6. according to the method for quality control of the described HUGANNING ZHIJI of claim 1, it is characterized in that: the emodin content assay method is to be contrast with the emodin reference substance in the Rhizoma Polygoni Cuspidati, and with methanol or acetonitrile: 0.05～5% phosphoric acid solution=10～90: 90～10 is the high performance liquid chromatography of mobile phase.

7. according to the method for quality control of the described HUGANNING ZHIJI of claim 1, it is characterized in that: the content assaying method of polygonin is to be contrast with the polygonin reference substance in the Rhizoma Polygoni Cuspidati, and with methanol or acetonitrile: 0.05～5% phosphoric acid=10～90: 90～10 is the high performance liquid chromatography of mobile phase.

8. according to the method for quality control of claim 1,6 or 7 described HUGANNING ZHIJI, it is characterized in that: described content assaying method comprises the part or all of of following project:

This capsule contains Rhizoma Polygoni Cuspidati in emodin C15H10O5, must not be less than 1.5mg/g; This tablet contains Rhizoma Polygoni Cuspidati in emodin C15H10O5, must not be less than 0.5mg/g; This granule contains Rhizoma Polygoni Cuspidati in emodin C15H10O5, must not be less than 0.1mg/g;

This capsule contains Rhizoma Polygoni Cuspidati in polygonin C20H22O8, must not be less than 0.5mg/g; This tablet contains Rhizoma Polygoni Cuspidati in polygonin C20H22O8, must not be less than 0.2mg/g; This granule contains Rhizoma Polygoni Cuspidati in polygonin C20H22O8, must not be less than 0.054mg/g.

9. according to the method for quality control of the described HUGANNING ZHIJI of claim 8, it is characterized in that: content assaying method comprises the part or all of of following project more specifically:

10. according to the method for quality control of the described HUGANNING ZHIJI of claim 1, it is characterized in that: described method of quality control comprises:

Character:

For capsule: content is tan granule; Bitter in the mouth, little acid, puckery;

For tablet: medicine shows sepia; Bitter in the mouth, little acid, puckery;

Differentiate:

(2) get this preparation or its content, add ethanol, reflux filters, and filtrate evaporate to dryness, residue add methanol makes dissolving, as need testing solution; Other gets the Ganoderma control medicinal material, shines medical material solution in pairs with legal system; According to the test of Chinese Pharmacopoeia thin layer chromatography, draw above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

Check: should meet every regulation relevant under Chinese Pharmacopoeia capsule, tablet or the granule item;

Assay:

11. the method for quality control according to the described HUGANNING ZHIJI of claim 10 is characterized in that: described method of quality control comprises:

Character:

For capsule: content is tan granule; Bitter in the mouth, little acid, puckery;

For tablet: medicine shows sepia; Bitter in the mouth, little acid, puckery;

Differentiate: (1) gets this preparation or its content 0.5～5g, adds methanol 10～100ml, supersound process 5～100 minutes, filter, filtrate evaporate to dryness, residue add 2.5mol/L sulfuric acid solution 1～20ml, heating hydrolysis 10～100 minutes, put cold, with chloroform extraction 1～5 time, merge chloroform liquid, evaporate to dryness, residue adds chloroform 0.5～3ml makes dissolving, as need testing solution; Other gets Rhizoma Polygoni Cuspidati control medicinal material 0.01～0.5g, shines medical material solution in pairs with legal system; Get emodin and physcione reference substance again, add methanol respectively and make the solution that every 1ml contains 0.2～3mg, in contrast product solution; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw need testing solution, control medicinal material solution and reference substance solution, put respectively on same silica gel g thin-layer plate, with 30～60 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take 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; Put in the ammonia steam smoked after, inspect under the daylight, show identical punctation;

(2) get this preparation or its content 1～10g, add ethanol 10～100ml, reflux 10～100 minutes filters, and filtrate evaporate to dryness, residue add methanol 0.5～5ml makes dissolving, as need testing solution; Other gets Ganoderma control medicinal material 0.5～5g, shines medical material solution in pairs with legal system; Test according to the Chinese Pharmacopoeia thin layer chromatography, draw each 2～15 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 60～90 ℃ of petroleum ether: Ethyl formate: formic acid=5～30: 1～10: 0.5～5 upper solution is developing solvent, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;

Assay: