CN1785267A - Quality control method of compound gallblader freeflow solid preparation - Google Patents

Quality control method of compound gallblader freeflow solid preparation Download PDF

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CN1785267A
CN1785267A CN 200510003269 CN200510003269A CN1785267A CN 1785267 A CN1785267 A CN 1785267A CN 200510003269 CN200510003269 CN 200510003269 CN 200510003269 A CN200510003269 A CN 200510003269A CN 1785267 A CN1785267 A CN 1785267A
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solution
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methanol
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CN100369614C (en
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叶湘武
王泽坤
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Guizhou Yibai Pharmaceutical Co Ltd
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Guizhou Yibai Pharmaceutical Co Ltd
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Abstract

A quality control method for the Chinese medicing ''Dantong'' in the form of capsule, paticle, or tablet features that a thin-film chromatography is used for qualitative identification to rhubarb, hydroxymethyl coumarin, andrographolide, and other components, and an efficient liquid-phase chromatograph is used to measure the contents of hydroxymethyl coumarin, archin and chrysophanol.

Description

The method of quality control of compound gallblader freeflow solid preparation
Invention field:
The present invention relates to drug world, particularly the method for quality control of compound gallblader freeflow solid preparation.Be specifically related to the method for quality control of fufangdantong capsules, granule and tablet.
Background technology:
Compound gallblader freeflow solid preparation is to record kind compound dantong tablets, capsule change dosage form by pharmacopeia, this solid preparation crude drug is made up of hymecromone, Herba Rabdosiae Lophanthoidis, Herba Artemisiae Scopariae, Herba Andrographis, Radix Et Rhizoma Rhei, be used for acute and chronic cholecystitis clinically, cholangitis, gallbladder, Biliary Calculi concurrent infection, syndrome behind the operation on gallbladder, treatment of diseases such as biliary tract is functional has effect preferably.This side's compatibility is simplified rationally, brings into play curative effect rapidly for making the medicine of taking, and we have successfully developed new solid preparations such as " fufangdantong capsules " with former tablet, capsule by adopting novel adjuvant and new moulding process.This product has clearing heat secreting bile, and the function of relieving spasm to stop pain uses this medicine to bring into play curative effect rapidly, is used for diseases such as biliary tract, bile duct be functional.Existing " compound dantong tablets " reach the quality standard (WS3-B-0973-91) of " fufangdantong capsules " and (WS3-B-0974-91) in differentiate that (1) is the physicochemical identification of rhubarb medicinal material, lack microscopical identification, differentiate in the quality standard that (2) and (3) are the thin layer chromatography discrimination method of hymecromone, wherein method (3) lacks negative control, detect the result and be negative sometimes, impurity disturbs big.Also lack discriminating in the quality standard,, cause certain difficulty to discriminating because this discrimination method lacks specificity to other medical materials or composition (as Herba Andrographis and emodin).In addition, [assay] item also lacks the assay of other main ingredients in the quality standard, can not the better controlled end product quality.
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.Prior art is difficult to the quality of controlling agent owing to lack method of quality control; 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 compound gallblader freeflow solid preparation is controlled, particularly increase assay detection method in the preparation, content with hymecromone and emodin, chrysophanol in this medicine of high effective liquid chromatography for measuring makes sample treatment easier, guarantees this compound preparation higher quality standard level.
For effectively controlling product quality, we are at the above-mentioned deficiency of existing quality standard, improve and set up new compound gallblader freeflow solid preparation method of quality control, this method of quality control is comparatively stable, precision is high, repeatability is good, and differentiate comprehensively, can control product quality preferably, guarantee " homogeneous, stable, effective, controlled " of product quality.
Summary of the invention:
The method of quality control that the objective of the invention is to open compound gallblader freeflow solid preparation.The present invention be directed to compound gallblader freeflow solid preparation and propose method of quality control, this compound gallblader freeflow solid preparation specifically comprises capsule, granule and tablet etc.
Described compound gallblader freeflow solid preparation of the present invention is to be made by the raw material of Chinese medicine of following weight/part:
Hymecromone 100g, Herba Rabdosiae Lophanthoidis 900g, Herba Artemisiae Scopariae 600g, Herba Andrographis 300g, Radix Et Rhizoma Rhei 30g.
The preparation method of compound gallblader freeflow solid preparation of the present invention 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.
Solid 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 solid preparation, as: mannitol, sorbitol, sorbic acid or potassium salt, 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, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, propylene glycol, ethanol, soil temperature 60-80, span-80, Cera Flava, lanoline, liquid paraffin, hexadecanol, gallate ester, agar, triethanolamine, basic amino acid, carbamide, allantoin, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, the phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
Solid preparation of the present invention preferably prepares through following method:
Wherein the capsule method for making is:
The above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, and drying is distributed into 1000, promptly.
Wherein the granule method for making is:
The above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, and drying is distributed into 1000, promptly.
Wherein the tablet method for making is:
The above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, drying, and granulate, tabletting is made 1000, promptly according to a conventional method.
Solid 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.
Method of quality control of the present invention, comprise and adopt thin layer chromatography and physico-chemical method that Radix Et Rhizoma Rhei and effective ingredient, hymecromone and andrographolide are differentiated, adopt the spectrophotometry hymecromone, the content of high effective liquid chromatography for measuring emodin and chrysophanol.
Method of quality control of the present invention may further comprise the steps:
To the observation of [character],
Composition in the content is carried out [discriminating],
Solid preparation is carried out [inspection],
Effective ingredient is carried out [assay].
Described observation to [character] comprises:
When this product is capsule, tan powder or granule; Bitter in the mouth, puckery.
When this product was granule, it is brown to tan granule that content is; It is sweet, little puckery to distinguish the flavor of.
When this product was tablet, content was brown; Bitter in the mouth, puckery.
Describedly composition in the preparation content carried out [discriminating] comprising:
To the discriminating of compositions such as Radix Et Rhizoma Rhei, hymecromone and andrographolide, its method is as follows:
(1) get the content of this product capsule or granule or tablet, porphyrize, put microscopically and observe: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation;
(2) get the content 0.5g of this product capsule or granule or tablet, porphyrize adds ethanol 15ml, put in the water-bath and heated 10-15 minute, filter, filtrate adds hydrochloric acid solution (1mol/L) 5ml, heating is 10-30 minute in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, hydro-oxidation sodium liquid (0.1mol/L) 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off;
(3) get the content 2g of this product capsule or granule or tablet, porphyrize adds chloroform 20-40ml, reflux 10-20 minute, filter, get filtrate 5ml, fling to chloroform, residue dissolves with methanol 5ml, add 1 of ferric chloride test solution, displaing yellow adds 1 of ammonia solution again, show red, and bluish violet fluorescence ring is arranged;
(4) get discriminating (3) filtrate 15ml down, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, dissolves with methanol 2ml, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-acetone (1.2-1.8: 0.8-1.1) be developing solvent, launch, take out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point;
(5) get the content 2g of this product capsule or granule or tablet, porphyrize adds ethanol 20ml, reflux 5-15 minute, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put under the ultra-violet lamp (365nm) and observe, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put ultra-violet lamp (365nm) under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence;
(6) get the content 1g of this product capsule or granule or tablet, add ethanol 30ml, soaked 30 minutes, supersound process 20-40 minute, filter, filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, with chloroform-ethyl acetate-methanol (3.4-4.5: 2.6-3.4: 0.3-0.5) be developing solvent, launch, take out, dry, spray is with 2%3, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution (facing the time spent preparation) is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(7)) get the content 2g of this product capsule or granule or tablet, put in the tool plug conical flask, add the ultrasonic 20-40 of methanol 20ml minute, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 20-40 minute, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, (11-18: 4.3-7.5: upper solution 0.8-1.1) is developing solvent with petroleum ether (30～60 ℃)-Ethyl formate-formic acid, launch, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
Preferably [discriminating] method is:
(1) get the content of this product capsule or granule or tablet, porphyrize, put microscopically and observe: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation;
(2) get the content 0.5g of this product capsule or granule or tablet, porphyrize adds ethanol 15ml, put in the water-bath and heated 10 minutes, filter, filtrate adds hydrochloric acid solution (1mol/L) 5ml, heating is 20 minutes in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, hydro-oxidation sodium liquid (0.1mol/L) 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off;
(3) get the content 2g of this product capsule or granule or tablet, porphyrize adds chloroform 30ml, and reflux 10 minutes filters, get filtrate 5ml, fling to chloroform, residue adds 1 of ferric chloride test solution with methanol 5ml dissolving, displaing yellow adds 1 of ammonia solution again, shows red, and bluish violet fluorescence ring is arranged;
(4) get discriminating (3) filtrate 15ml down, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, dissolves with methanol 2ml, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-acetone (1.5: 1) is developing solvent, launches, and takes out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point;
(5) get the content 2g of this product capsule or granule or tablet, porphyrize adds ethanol 20ml, reflux 10 minutes, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put under the ultra-violet lamp (365nm) and observe, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put ultra-violet lamp (365nm) under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence;
(6) get the content 1g of this product capsule or granule or tablet, add ethanol 30ml, soaked 30 minutes, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent, launch with chloroform-ethyl acetate-methanol (4: 3: 0.4), take out, dry, spray is with 2%3, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution (facing the time spent preparation) is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(7)) get the content 2g of this product capsule or granule or tablet, put in the tool plug conical flask, added methanol 20ml ultrasonic 30 minutes, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 30 minutes, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1) is developing solvent, launch, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
Describedly preparation carried out [inspection] comprising:
For capsule, should meet every regulation relevant under the capsule item (appendix IL of Chinese Pharmacopoeia version in 2000);
For granule, should meet every regulation relevant under the granule item (appendix IC of Chinese Pharmacopoeia version in 2000);
For tablet, should meet every regulation relevant under the tablet item (appendix ID of Chinese Pharmacopoeia version in 2000).
Describedly the preparation effective ingredient carried out [assay] comprise wherein hymecromone and emodin, chrysophanol Determination on content:
Wherein the content assaying method of hymecromone is as follows:
1, the content assaying method of hymecromone in the solid preparation of the present invention: measure according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A).
The preparation lucifuge operation of reference substance solution.Precision takes by weighing the hymecromone reference substance 0.1g that is dried to constant weight at 105 ℃, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and [gets caustic lye of soda (0.1mol/L) 20ml, add ethanol dilution to 1000ml] about 70ml, make dissolving and be diluted to scale, shake up, precision is measured 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and is diluted to scale, shake up, measure immediately.
The preparation lucifuge operation of need testing solution.Get content under the content uniformity item, porphyrize, precision takes by weighing this product an amount of (being equivalent to hymecromone 0.1g approximately), put in the tool plug brown bottle accurate alkaline ethanol liquid 90-110ml, the close plug of adding, jolting was extracted 45-75 minute, filtered, and discarded filtrate just, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, add alkaline ethanol liquid and be diluted to scale, shake up, measure immediately.
Algoscopy is got reference substance solution and need testing solution respectively, measures trap at 372 ± 2nm wavelength place according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A), calculates, promptly.
Said determination trap wavelength preferably is selected in the 372nm place; The preparation of need testing solution preferably adds alkaline ethanol liquid 100ml, and jolting was extracted 60 minutes.
Every of this product contains hymecromone (C 10H 8O 3) should be 90.0%～110.0% of labelled amount.
Wherein as follows to the content assaying method of emodin, chrysophanol:
Measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005).
Chromatographic condition and system suitability test are packed column with octadecylsilane chemically bonded silica; Methanol 0.1% phosphoric acid solution (53-121: 11-22) be mobile phase; Detect wavelength 254 ± 3nm.Number of theoretical plate calculates by the emodin peak should be not less than 1500.
The preparation precision of reference substance solution takes by weighing emodin reference substance 10-14mg, chrysophanol reference substance 14-18mg, puts respectively in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up; Precision is measured emodin and each 2ml of chrysophanol solution respectively, puts in the 25ml measuring bottle, adds methanol to scale, shakes up, and promptly gets (contain among the every 1ml of emodin among 9.6 μ g, the every 1ml of chrysophanol and contain 12.8 μ g).
The about 2.5g of content under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the conical flask, precision adds methanol 50ml, claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 10ml, put in the 50ml round-bottomed flask, fling to methanol, add 2.5mol/L sulfuric acid solution 20ml, supersound process 5,10 minutes adds chloroform 20ml again, reflux 1 hour, cooling moves in the separatory funnel, use the minimum of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution chloroform extraction 3 times, each 10ml, combined chloroform liquid, with anhydrous sodium sulfate dehydration, chloroform solution moves in the 100ml conical flask, flings to chloroform, the residue precision adds methanol 10ml, claims to decide weight, puts slight fever dissolved residue in the water-bath, put cold after, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively above-mentioned two kinds of reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and calculate, promptly.
The preferred proportion numerical range of above-mentioned chromatographic condition mobile phase is methanol 0.1% phosphoric acid solution (83: 17); Detect wavelength and be preferably 254nm; The best precision of the preparation of reference substance solution takes by weighing emodin reference substance 12mg, chrysophanol reference substance 16mg, puts in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale.
Every of this product contains Radix Et Rhizoma Rhei with emodin (C 15H 10O 5) and chrysophanol (C 15H 10O 4) the total amount meter, must not be less than 0.15mg.
Method of quality control of the present invention is comparatively stable to the quality control of product, precision is high, repeatability is good, and differentiates comprehensively, can control product quality preferably, has guaranteed " homogeneous, stable, effective, controlled " of product quality.
The used preparation of experimental example test sample of the present invention all adopts the embodiment of the invention 1 described capsule, can adopt the embodiment of the invention 2 or 3 described granule or tablets, and also available have other preparations that same materials is formed with the solid preparation described in each embodiment.
Experimental example 1 compound gallblader freeflow solid preparation of the present invention is to the research of medical material qualitative identification
1. differentiate: (1) gets this product, puts microscopically and observes: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation.The feature of rhubarb medicinal material is differentiated in this side of being.
(2) it is an amount of to get this product, and porphyrize adds ethanol 15ml, put in the water-bath and heated 10 minutes, filter, filtrate adds hydrochloric acid solution (1mol/L) 5ml, heating is 20 minutes in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, hydro-oxidation sodium liquid (0.1mol/L) 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off.The feature of rhubarb medicinal material is differentiated in this side of being.Through test of many times, the result all is positive.
(3) it is an amount of to get this product, and porphyrize adds chloroform 30ml, and little boiling filters, and gets filtrate 5ml, flings to chloroform, and residue adds 1 of ferric chloride test solution with methanol 5ml dissolving, and displaing yellow adds 1 of ammonia solution again, shows red, and bluish violet fluorescence ring is arranged.The physicochemical characteristics of hymecromone is differentiated in this side of being.Through test of many times, the result all is positive.
(4) get discriminating (3) filtrate 15ml down, fling to chloroform, residue adds methanol 3ml and dissolves, as need testing solution.The negative need testing solution that lacks hymecromone with the method preparation.Other gets hymecromone reference substance 10mg, with methanol 2ml dissolving, product solution in contrast.According to thin layer chromatography (" an appendix VI of Chinese pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-acetone (1.5: 1) is developing solvent, launches, and takes out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point.The qualitative identification of hymecromone in this side of being.Through test of many times, the result all is positive, and negative control is noiseless.
(5) get this product content 2g, porphyrize adds ethanol 20ml, and little boiling filters, filtrate add active carbon a little, warm back filters; Get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, and evaporate to dryness is put under the ultra-violet lamp (365nm) and observed, and shows green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put ultra-violet lamp (365nm) under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence.This differentiates for the material that has the flavone aglycone structure in the side.Through test of many times, the result all is positive.
(6) show that according to documents and materials Herba Andrographis contains compositions such as andrographolide, dehydrorographolide, deoxyandrographolide, owing to be subjected to condition influence such as prescription, production technology, andrographolide might be converted into dehydrorographolide.In process of the test, once with Herba Andrographis medical material in dehydrorographolide and the andrographolide discriminating side, but because the unstability of andrographolide, in production technology, transform, can't detect andrographolide in the preparation, therefore with Herba Andrographis medical material in the dehydrorographolide reference substance discriminating side.Get this product content and add soak with ethanol in right amount, supersound process filters, and filtrate evaporate to dryness, residue add dehydrated alcohol makes dissolving in right amount, filters, and is concentrated into 1ml and makes need testing solution.The negative need testing solution that lacks the Herba Andrographis medical material with the method preparation.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (" an appendix VI of Chinese pharmacopoeia version in 2000 B) test, draw above-mentioned two kinds of each 5ul of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent with chloroform-ethyl acetate-methanol (4: 3: 0.4), launch, take out that dry, spray is with 2%3, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.Through test of many times, the result all is positive, and negative control is noiseless, so list quality standard text of the present invention in.
(7) with rhubarb medicinal material in Radix Et Rhizoma Rhei control medicinal material and the emodin reference substance discriminating side.Get this product content and put in the tool plug conical flask, it is ultrasonic to add methanol, filters the filtrate evaporate to dryness, add water, hydrochloric acid makes dissolving, puts in the water-bath heating 30 minutes, ether extraction is used in cooling immediately, extracting solution evaporate to dryness, residue add an amount of chloroform dissolving, filter, and are concentrated into 1ml and make need testing solution.The negative need testing solution that lacks rhubarb medicinal material with the method preparation.Other gets the Radix Et Rhizoma Rhei control medicinal material and is equipped with control medicinal material solution with legal system.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 the upper solution of petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1), and the speckle colour developing is clear, and negative control is noiseless, so list quality standard text of the present invention in.
2. check: can be by " the regulation in appendix rules of preparations of Chinese pharmacopoeia version in 2000 under capsule (appendix IL) item.
Carry out moisture, content uniformity, disintegration, heavy metal inspection, the inspection of arsenic salt, limit test of microbe (the results are shown in Table 1) respectively.
Table 1 fufangdantong capsules three batch sample check results
Inspection item Check result
20010701 20010711 20010720
Moisture (%) content uniformity disintegration time limited (minute) heavy metal checks that arsenic salt checks microbial limit bacterial population fungi count Escherichia coli full inspection of living 4.8 1/5ths＜5/1000000ths＜1,000,000 up to specification/g＜10/g does not detect 5.0 1/6ths＜5/1000000ths＜1,000,000 up to specification/g＜10/g does not detect 4.7 1/6ths＜5/1000000ths＜1,000,000 up to specification/g＜10/g does not detect
The content assaying method of hymecromone research in experimental example 2 compound gallblader freeflow solid preparations of the present invention
3. the research of assay
3.1 the assay of hymecromone adopts the content of determined by ultraviolet spectrophotometry this product hymecromone.According to documents and materials, the hymecromone content assaying method has ultraviolet spectrophotometry, high performance liquid chromatography etc.In process of the test, we once adopted high performance liquid chromatography that the hymecromone assay is carried out methodological study [its chromatographic condition is: with octadecylsilane chemically bonded silica is filler; Methanol-water (40: 60) is the item that flows; Flow velocity is 1.0ml/min; Temperature is 40 ℃; The detection wavelength is 370nm], but find that this method is very unstable, survey the reference substance solution peak area under the same conditions and differ greatly, can't accurately measure hymecromone content.The back is measured it with ultraviolet spectrophotometry, through methodological study, determines that ultraviolet spectrophotometry is the method one comparatively stable, that precision is high, repeatability is good, can control product quality preferably.Therefore, we, measure the content of hymecromone in the preparation, and method for measuring are studied the hymecromone content assaying method with reference to " fufangdantong capsules " proper mass standard and relevant documents and materials.
3.1.1 instrument and reagent
3.1.1.1 instrument day island proper Tianjin UV-2501 type ultraviolet-visible spectrophotometer; Electronic balance AG135 type (Switzerland Mei Telietuole is many).
3.1.1.2 reagent sodium hydroxide (analytical pure), ethanol (analytical pure); (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the hymecromone reference substance, for assay usefulness, lot number: 10241-200002).
3.1.2 the drafting of absorption spectrum
Getting hymecromone reference substance solution, need testing solution respectively, lack the negative need testing solution of hymecromone, serves as blank scanning with alkaline ethanol liquid in 300～500nm scope.Can get from scanning, the alkaline ethanol solution maximum absorption band of hymecromone reference substance and test sample is 372nm, and negative test liquid only is 0.001 in 372nm place trap, and visual negative test liquid does not have absorption.
3.1.3 linear relationship is investigated
3.1.3.1 the preparation lucifuge of standard solution operation.It is an amount of that precision takes by weighing 105 ℃ of hymecromone reference substances that are dried to constant weight, is mixed with the alkaline ethanol solution of 97.2 μ g/ml.
3.1.3.2 accurate above-mentioned solution 1.0ml, 2.0ml, 3.0ml, 4.0ml, the 5.0ml of drawing, put respectively in the brown measuring bottle of 50ml, add alkaline ethanol liquid and be settled to scale, shake up, make the blank trap (table 2) of measuring in the 372nm place with alkaline ethanol liquid, with trap (A) concentration (μ g/ml) is carried out linear regression and calculate, regression equation is: A=0.1217C-0.0211, r=0.9999.Precision is measured the standard solution 2.5ml of 97.2 μ g/ml, puts in the brown volumetric flask of 50ml, adds alkaline ethanol liquid and is diluted to scale, shakes up, and gets reference substance solution.Measuring trap at the 372nm place is 0.571.One need testing solution gained trap is calculated with regression equation and one point method respectively, and result's relative deviation is 0.022%, so quality standard of the present invention adopts one point method to carry out colorimetric determination.The range of linearity: 1.944～9.720 μ g/ml.
Table 2 hymecromone linear relationship is investigated
Concentration (μ g/ml) Trap (A)
1.944 3.888 5.832 7.776 9.720 0.216 0.449 0.688 0.932 1.157
3.1.4 the selective extraction method of need testing solution extraction time: porphyrize, add the jolting of alkaline ethanol liquid and extract, measurement result sees Table 3.
The investigation (n=3) of table 3 alkaline ethanol liquid different extraction times
Extraction time (min) 15 30 60 90 120
Content (%) RSD (%) 22.62 1.43 23.04 0.98 23.63 0.51 23.60 0.91 23.64 0.73
As seen from Table 3, porphyrize adds alkaline ethanol liquid jolting extraction and the hymecromone in the preparation can be extracted fully in 60 minutes, so select jolting to extract 60 minutes.
3.1.5 precision test
Sample thief (lot number: 20010508) content, preparation method by test liquid under the assay item in the quality standard of the present invention prepares test liquid, do blank in 372nm place replication 5 times with alkaline ethanol liquid, the record trap, result of calculation is listed table 4 in, average content is 23.74%, and RSD is 0.328%.
The precision of hymecromone test in the table 4 preparation test sample
Measure number of times 1 2 3 4 5 Meansigma methods RSD(％)
Content (%) 23.68 23.75 23.64 23.79 23.83 23.74 0.328
3.1.6 repeatability test
Sample thief (lot number: 20010508) content, 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, sentencing alkaline ethanol liquid respectively at 372nm is the blank determination trap, and result of calculation is listed table 5 in, average content is 23.64%, RSD=0.68%.
The repeatability of hymecromone test in the table 5 preparation test sample
Measure number of times 1 2 3 4 5 Meansigma methods RSD(％)
Content (%) 23.72 23.43 23.71 23.82 23.51 23.64 0.68
3.1.7 need testing solution study on the stability
Precision takes by weighing sample (lot number: 20010508) content, preparation method by test liquid under the assay item in the quality standard of the present invention prepares a test liquid, sentencing alkaline ethanol liquid respectively at 0,0.5,1.0,1.5,2.0 hour at 372nm is blank determination trap (table 6), the result shows that the alkaline ethanol solution of hymecromone is stable relatively in 2h.
The study on the stability of hymecromone in the table 6 preparation test sample
Minute (h) 0 0.5 1 1.5 2 RSD(％)
Trap (A) content (%) 0.605 23.69 0.601 23.54 0.598 23.42 0.603 23.61 0.601 23.54 0.433 0.423
3.1.8 recovery test
Adopt the application of sample absorption method, it is an amount of that precision takes by weighing the sample (lot number: 20010508 average contents are 23.64%) of measuring content, put in the tool plug brown bottle, accurate hymecromone reference substance solution (0.4760mg/ml) 100ml that adds, close plug, jolting was extracted 30 minutes, filtered, discard filtrate just, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and is diluted to scale, shake up, make test liquid.Preparing 5 parts respectively, serves as the blank trap of measuring in the 372nm place with alkaline ethanol liquid, and calculate recovery rate the results are shown in Table 7, and average recovery rate is 99.90%, and RSD is 1.47%.
The recovery test of hymecromone in table 7 need testing solution
Experiment number Sample size (mg) Contain hymecromone (mg) Add hymecromone (mg) The amount of recording (mg) The response rate (%) Average recovery rate (%) RSD (％)
1 2 3 4 5 204.2 210.5 207.9 203.0 212.0 48.27 49.76 49.15 47.99 50.12 47.60 95.20 98.00 96.52 96.36 97.01 98.59 101.3 99.52 101.6 98.51 99.90 1.47
3.1.9 sample determination
Preparing test liquid and reference substance solution by quality standard of the present invention, serves as the blank trap of measuring in the 372nm place with alkaline ethanol liquid, is calculated as follows content:
In the formula: Ai: the need testing solution trap;
As: reference substance solution trap;
Cs: reference substance solution concentration (mg/ml);
W 1: average particle heavy (mg);
W 2: sample weighting amount (mg);
W 3: labelled amount (100mg).
Hymecromone assay result (seeing Table 8) in 3 batch samples.
Hymecromone assay result in table 83 batch samples
Lot number Content/labelled amount (%) Average content/labelled amount (%)
1 2
20010701 20010711 20010720 96.1 100.2 94.8 96.7 99.0 95.9 96.4 99.6 95.4
According to methodological study result and 3 batch sample measurement results [sample hymecromone content (C 10H 8O 3) all at labelled amount scope (90.0%～110.0%)] as seen, ultraviolet spectrophotometry is comparatively stable, a perfect method, can accurately determine hymecromone content in the preparation, can guarantee product quality fully, so the assay of hymecromone adopts ultraviolet spectrophotometry in the said preparation.
The content assaying method of emodin, chrysophanol research in experimental example 3 compound gallblader freeflow solid preparations of the present invention
4 Radix Et Rhizoma Rhei assays adopt the content of emodin, chrysophanol in the high effective liquid chromatography for measuring side.According to documents and materials, mainly contain emodin, chrysophanol, glucoside, dianthracene ketoside, rheochrysin, Galla Turcica (Galla Helepensis) glucoside, gallic acid etc. in the Radix Et Rhizoma Rhei.To the content assaying method of emodin, chrysophanol in the Radix Et Rhizoma Rhei, what bibliographical information was more is high performance liquid chromatography, thin layer chromatography scanning, colorimetry, volumetric method etc.With reference to the content assaying method of relevant documents and materials, the content of emodin, chrysophanol in the preparation is measured, and method for measuring is investigated emodin, chrysophanol in the Radix Et Rhizoma Rhei.
4.1 Tianjin, instrument 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; Ultrasonic washing unit (250W, Beijing's armarium two factories).
4.2 reagent methanol (analytical pure), phosphoric acid (analytical pure), pure water, (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the emodin reference substance, use for assay, lot number: 0756-200010), (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the chrysophanol reference substance, for assay usefulness, lot number: 0796-200015)
4.3 chromatographic condition chromatographic column: Description:C 18(200 * 4.6mm); Mobile phase: methanol-0.1% phosphoric acid solution (83: 17); Detect wavelength: 254nm; Flow velocity: 1.0ml/min; Column temperature: 40 ℃; Sample size: 10 μ l.
4.4 system suitability test
The negative need testing solution of getting emodin, chrysophanol mixing reference substance solution, need testing solution and scarce Radix Et Rhizoma Rhei respectively injects chromatograph of liquid, the record chromatograph.Retention time (the t of emodin as can be known R) be about 10.1 minutes, the retention time (t of chrysophanol R) be about 13.5 minutes, the negative sample chromatogram is at emodin, non-false positive peak, place, chrysophanol peak position, emodin, chrysophanol and close impurity peaks separate fully (separating degree＞1.5), and promptly emodin, chrysophanol separate with other components fully under this experimental condition.Theoretical cam curve is calculated with emodin, chrysophanol peak and is respectively 4000,3500.
4.5 linear relationship is investigated
4.5.1 the preparation of standard solution
Precision takes by weighing emodin, the chrysophanol reference substance is an amount of, adds methanol and makes the mixing reference substance solution that every 1ml contains emodin 9.44 μ g, chrysophanol 12.72 μ g.
4.5.2 the drafting of standard curve
The accurate absorption mixed reference substance solution 4,8,10,12,16 μ l, inject chromatograph of liquid respectively, record chromatograph (table 9,10, with peak area A (μ Vs) mass number x (μ g) being carried out linear regression calculates, getting the emodin equation of linear regression is: A=3623736.7585 * X+1192.9500, r=0.9999, the chrysophanol equation of linear regression is: A=6422869.4969 * X+341.4000, r=0.9999, precision is measured reference substance solution 10 μ l and is injected chromatograph of liquid, gained emodin peak area is 343082, the chrysophanol peak area is 813784, 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 emodin 0.46%, chrysophanol 0.24% is so quality standard of the present invention adopts one point method to calculate content.The range of linearity: emodin 0.03776～0.15104 μ g, chrysophanol 0.05088～0.20352 μ g.
Table 9 emodin linear relationship is investigated
Sample size (μ g) Emodin peak area (μ Vs)
0.03776 0.07552 0.09440 0.11328 0.15104 135217 271978 343082 408267 545895
Table 10 chrysophanol linear relationship is investigated
Sample size (μ g) Chrysophanol peak area (μ Vs)
0.05088 0.10176 0.12720 0.15264 0.20352 326163 659450 813784 972814 1311027
4.6 the selection of methanol eddy extraction time is extracted with methanol eddy, mensuration the results are shown in Table 11,12.
The investigation of table 11 reflux extracting time (emodin) (n=3)
Return time 10 20 30 40 50
Content (%) RSD (%) 0.01325 2.59 0.01588 2.04 0.01733 1.66 0.01725 1.88 0.01736 1.97
The investigation of table 12 reflux extracting time (chrysophanol) (n=3)
Return time 10 20 30 40 50
Content (%) RSD (%) 0.02011 2.97 0.02173 1.95 0.02298 1.41 0.02288 1.89 0.02302 2.01
Reflux, extract, 30 minutes does not have significant difference with 40 minutes, 50 minutes result as seen from the table, therefore in quality standard of the present invention reflux extracting time is decided to be 30 minutes.
4.7 the selection of chloroform reflux extracting time chloroform reflux, extract,, mensuration the results are shown in Table 13,14.
The investigation of table 13 reflux extracting time (emodin) (n=3)
Return time 30 45 60 75 90
Content (%) RSD (%) 0.01413 1.94 0.01562 2.08 0.01751 1.52 0.01759 1.56 0.01749 1.97
The investigation of table 14 reflux extracting time (chrysophanol) (n=3)
Return time 30 45 60 75 90
Content (%) RSD (%) 0.02105 2.77 0.02198 1.89 0.02314 1.58 0.02306 1.68 0.02289 2.11
Reflux, extract, 60 minutes does not have significant difference with 75 minutes, 90 minutes result as seen from the table, therefore in quality standard of the present invention reflux extracting time is decided to be 60 minutes.
Investigate and use chloroform extraction 4.8 extract number of times and emodin, the chrysophanol assay result of sample solution, combined chloroform liquid, its extraction time and emodin, chrysophanol assay the results are shown in Table 15,16.
Table 15 extracts the number of times and the emodin content measurement result (n=3) of sample solution
Return time 1 2 3 4 5
Content (%) RSD (%) 0.01478 1.98 0.01624 1.85 0.01769 1.68 0.01757 1.71 0.01772 2.07
Table 16 extracts the number of times and the chrysophanol assay result (n=3) of sample solution
Return time 1 2 3 4 5
Content (%) RSD (%) 0.02043 2.88 0.02209 1.75 0.02331 1.49 0.02318 1.43 0.02328 1.66
Extracting 3 times does not as seen from the table have significant difference with 4 times, 5 times result, therefore in quality standard of the present invention extraction times is decided to be 3 times.
4.9 precision test
Get this product content, prepare test liquid, repeat sample introduction 5 times, measure peak area (table 17,18) by the preparation method of test liquid under the assay item in the quality standard of the present invention.
The precision of emodin test in the table 17 preparation test sample
The sample introduction number of times 1 2 3 4 5 Meansigma methods RSD(％)
Peak area 338815 339075 336719 338475 337926 338202 0.28
The precision of chrysophanol test in the table 18 preparation test sample
The sample introduction number of times 1 2 3 4 5 Meansigma methods RSD(％)
Peak area 734172 731577 733243 735611 732947 733510 0.20
The result as seen, this method precision is good.
4.10 repeatability test
Get this product content, 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 respectively, measure peak area, result of calculation is listed table 19,20 in, and average content is an emodin 0.01765%, RSD is 2.26%, chrysophanol 0.02329%.RSD is that 2.20% repeatability is good.
The repeatability of emodin test in the table 19 preparation test sample
Measure number of times 1 2 3 4 5 Meansigma methods RSD(％)
Content (%) 0.01736 0.01717 0.01799 0.01810 0.01761 0.01765 2.26
The repeatability of chrysophanol test in the table 20 preparation test sample
Measure number of times 1 2 3 4 5 Meansigma methods RSD(％)
Content (%) 0.02242 0.02340 0.02372 0.02360 0.02331 0.02329 2.20
4.11 emodin, chrysophanol stability test in the need testing solution
Get this product content, preparation method by test liquid under the assay item in the quality standard of the present invention prepares test liquid, measure emodin, chrysophanol content (seeing Table 21,22) respectively at 0,2,4,6,8 hour, the result shows that emodin, chrysophanol are stable in 8 hours in the need testing solution.
The stability test of emodin in the table 21 preparation test sample
Time (hour) 0 2 4 6 8 Meansigma methods RSD(％)
Peak area 326319 326296 324953 322144 321964 324335.2 0.66
The stability test of chrysophanol in the table 22 preparation test sample
Time (hour) 0 2 4 6 8 Meansigma methods RSD(％)
Peak area 734459 738402 737697 734572 736597 736345.4 0.24
4.12 recovery test
Adopt the application of sample absorption method, it is an amount of that precision takes by weighing 5 parts of samples (average content is an emodin 0.01765%, chrysophanol 0.02329%) of having measured content respectively, put in the tool plug conical flask, precision adds mixes reference substance solution (wherein every ml contains emodin 4.5ug, chrysophanol 5.6ug) 50ml, claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 10ml, puts in the 50ml round-bottomed flask, fling to methanol, add 2.5mol/L sulfuric acid solution 20ml, supersound process 5 minutes adds chloroform 20ml again, reflux 1 hour, cooling moves in the separatory funnel, uses the minimum of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution chloroform extraction 3 times, each 10ml, combined chloroform liquid, with anhydrous sodium sulfate dehydration, chloroform solution moves in the 100ml conical flask, flings to chloroform, the residue precision adds methanol 10ml, claim to decide weight, put slight fever dissolved residue in the water-bath, put cold after, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, make test liquid.The accurate respectively 10ul of absorption injects chromatograph of liquid, and the record chromatograph is measured content, calculate recovery rate (the results are shown in Table 23,24), and the emodin average recovery rate is 98.82%, and RSD is 1.63%, and the chrysophanol average recovery rate is 99.51%, and RSD is 1.85%.
Emodin is measured recovery test in table 23 need testing solution
Experiment number Sample size (g) Contain emodin (mg) Add emodin (mg) The amount of recording (mg) The response rate (%) Average recovery rate (%) RSD (％)
1 2 3 4 5 1.2731 1.3348 1.3326 1.2697 1.3275 0.2247 0.2356 0.2352 0.2241 0.2343 0.225 0.4455 0.4594 0.4566 0.4518 0.4524 98.13 99.46 98.39 101.2 96.92 98.82 1.63
Chrysophanol is measured recovery test in table 24 need testing solution
Experiment number Sample size (g) Contain chrysophanol (mg) Add chrysophanol (mg) The amount of recording (mg) The response rate (%) Average recovery rate (%) RSD (％)
1 2 3 4 5 1.2164 1.2295 1.2581 1.2414 1.2244 0.2833 0.2864 0.2930 0.2891 0.2852 0.28 0.5701 0.5622 0.5664 0.5694 0.5622 102.4 98.50 97.64 100.1 98.93 99.51 1.85
4.13 sample determination
Prepare test liquid and reference substance solution by quality standard, sample introduction 10 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 solution concentration (μ g/ml)
W: test sample sample weighting amount (g)
W 1: average particle heavy (g)
According to 3 batch samples in strict accordance with the production technology preparation, emodin, chrysophanol content in the working sample, measurement result sees Table 25.
Emodin, chrysophanol assay result in table 25 3 batch samples
Sample Content (mg/ grain) Meansigma methods (mg/ grain)
1 2
20010701 20010711 20010720 0.272 0.262 0.269 0.280 0.254 0.275 0.276 0.258 0.272
Emodin, chrysophanol content limit are calculated as follows in the preparation:
Emodin, chrysophanol total content lower bound=preparation contain Radix Et Rhizoma Rhei (g/g) * medical material and contain emodin, chrysophanol total amount lower bound (pharmacopeia regulation)=(30/420) * 0.5%=3.5714 * 10 -4Every content lower bound=3.5714 * 10 -4* 0.42g/ grain * 10 3=0.15mg/ grain.
So deciding every of this product contains Radix Et Rhizoma Rhei and must not be less than 0.15mg with emodin, chrysophanol total amount.Measurement result per sample, the emodin in 3 batch samples, chrysophanol total content are all in this limits.
The specific embodiment
Further specify the present invention by the following examples, but not as limitation of the present invention.
Following embodiment all can reach the effect of above-mentioned experimental example.
The quality control of embodiment 1 fufangdantong capsules
[prescription] hymecromone 100g, Herba Rabdosiae Lophanthoidis 900g, Herba Artemisiae Scopariae 600g, Herba Andrographis 300g and Radix Et Rhizoma Rhei 30g.
[method for making] above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, and drying incapsulates, and makes 1000, promptly.
[character] this product is a capsule, tan powder or granule; Bitter in the mouth, puckery.
This product content is got in [discriminating] (1), and porphyrize is put microscopically and observed: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation.
(2) get this product content 0.5g, porphyrize adds ethanol 15ml, put in the water-bath and heated 10 minutes, filter, filtrate adds hydrochloric acid solution (1mol/L) 5ml, heating is 20 minutes in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, hydro-oxidation sodium liquid (0.1mol/L) 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off.
(3) get this product content 2g, porphyrize adds chloroform 30ml, and reflux 10 minutes filters, and gets filtrate 5ml, flings to chloroform, and residue adds 1 of ferric chloride test solution with methanol 5ml dissolving, and displaing yellow adds 1 of ammonia solution again, shows red, and bluish violet fluorescence ring is arranged.
(4) get discriminating (3) filtrate 15ml down, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, dissolves with methanol 2ml, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-acetone (1.5: 1) is developing solvent, launches, and takes out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point.
(5) get this product content 2g, porphyrize adds ethanol 20ml, reflux 10 minutes, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put under the ultra-violet lamp (365nm) and observe, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put ultra-violet lamp (365nm) under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence.
(6) get this product content 1g, add ethanol 30ml, soaked 30 minutes, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent, launch with chloroform-ethyl acetate-methanol (4: 3: 0.4), take out, dry, spray is with 2%3, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution (facing the time spent preparation) is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(7) get this product content 2g, put in the tool plug conical flask, added methanol 20ml ultrasonic 30 minutes, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 30 minutes, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (15: 5: 1) is developing solvent, launch, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
[inspection] should meet every regulation relevant under the capsule item (appendix IL of Chinese Pharmacopoeia version in 2000)
The preparation lucifuge operation of [assay] hymecromone reference substance solution.Precision takes by weighing the hymecromone reference substance 0.1g that is dried to constant weight at 105 ℃, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and [gets caustic lye of soda (0.1mol/L) 20ml, add ethanol dilution to 1000ml] about 70ml, make dissolving and be diluted to scale, shake up, precision is measured 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and is diluted to scale, shake up, measure immediately.
The preparation lucifuge operation of need testing solution.Get content under the content uniformity item, porphyrize, precision takes by weighing this product an amount of (being equivalent to hymecromone 0.1g approximately), put in the tool plug brown bottle accurate alkaline ethanol liquid 100ml, the close plug of adding, jolting was extracted 60 minutes, filtered, and discarded filtrate just, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, add alkaline ethanol liquid and be diluted to scale, shake up, measure immediately.
Algoscopy is got reference substance solution and need testing solution respectively, measures trap at 372nm wavelength place according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A), calculates, promptly.
Every of this product contains hymecromone (C 10H 8O 3) should be 90.0%～110.0% of labelled amount.
Emodin, chrysophanol are measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D).
Chromatographic condition and system suitability test are packed column with octadecylsilane chemically bonded silica; Methanol-0.1% phosphoric acid solution (83: 17) is a mobile phase; Detect wavelength: 254nm.Number of theoretical plate calculates by the emodin peak should be not less than 1500.
The preparation precision of reference substance solution takes by weighing emodin reference substance 12mg, chrysophanol reference substance 16mg, puts respectively in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up; Precision is measured emodin and each 2ml of chrysophanol solution respectively, puts in the 25ml measuring bottle, adds methanol to scale, shakes up, and promptly gets (contain among the every 1ml of emodin among 9.6 μ g, the every 1ml of chrysophanol and contain 12.8 μ g).
The about 2.5g of content under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the conical flask, precision adds methanol 50ml, claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 10ml, put in the 50ml round-bottomed flask, fling to methanol, add 2.5mol/L sulfuric acid solution 20ml, supersound process 5 minutes adds chloroform 20ml again, reflux 1 hour, cooling moves in the separatory funnel, uses the minimum of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution chloroform extraction 3 times, each 10ml, combined chloroform liquid is with anhydrous sodium sulfate dehydration, chloroform solution moves in the 100ml conical flask, flings to chloroform, and the residue precision adds methanol 10ml, claim to decide weight, put slight fever dissolved residue in the water-bath, put cold after, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively above-mentioned two kinds of reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and calculate, promptly.Every of this product contains Radix Et Rhizoma Rhei with emodin (C 15H 10O 5) and chrysophanol (C 15H 10O 4) the total amount meter, must not be less than 0.15mg.
The quality control of embodiment 2 compound gallblader freeflow granules
[prescription] hymecromone 100g, Herba Rabdosiae Lophanthoidis 900g, Herba Artemisiae Scopariae 600g, Herba Andrographis 300g and Radix Et Rhizoma Rhei 30g.
[method for making] above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, drying, and granulate, tabletting is made 1000, promptly according to a conventional method.
[character] this product is a granule, and it is brown to tan granule that content is; It is sweet, little puckery to distinguish the flavor of.
[inspection] should meet every regulation relevant under the granule item (appendix IC of Chinese Pharmacopoeia version in 2000);
[method of quality control] discriminating, content assaying method are with embodiment 1, and wherein every of this product contains hymecromone (C 10H 8O 3) should be 90.0%～110.0% of labelled amount.Every of this product contains Radix Et Rhizoma Rhei with emodin (C 15H 10O 5) and chrysophanol (C 15H 10O 4) the total amount meter, must not be less than 0.15mg.
The quality control of embodiment 3 compound gallblader freeflow tablets
[prescription] hymecromone 100g, Herba Rabdosiae Lophanthoidis 900g, Herba Artemisiae Scopariae 600g, Herba Andrographis 300g and Radix Et Rhizoma Rhei 30g.
[method for making] above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, drying, and granulate, tabletting is made 1000, promptly according to a conventional method.
[character] this product is a tablet, and content is brown; Bitter in the mouth, puckery.
[inspection] should meet every regulation relevant under the tablet item (appendix ID of Chinese Pharmacopoeia version in 2000).
[method of quality control] discriminating, content assaying method are with embodiment 1, and wherein every of this product contains hymecromone (C 10H 8O 3) should be 90.0%～110.0% of labelled amount.Every of this product contains Radix Et Rhizoma Rhei with emodin (C 15H 10O 5) and chrysophanol (C 15H 10O 4) the total amount meter, must not be less than 0.15mg.
Embodiment 4
[prescription] hymecromone 100g, Herba Rabdosiae Lophanthoidis 900g, Herba Artemisiae Scopariae 600g, Herba Andrographis 300g and Radix Et Rhizoma Rhei 30g.
[method for making] above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, and drying incapsulates, and makes 1000, promptly.
[inspection] should meet every regulation relevant under the capsule item (appendix IL of Chinese Pharmacopoeia version in 2000);
[character] this product is tan powder or granule; Bitter in the mouth, puckery.
[discriminating]
(1) microscopical identification of rhubarb medicinal material.Get the content of this product capsule or granule or tablet, porphyrize, put microscopically and observe: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation;
(2) physicochemical identification of rhubarb medicinal material.Get the content 0.5g of this product capsule or granule or tablet, porphyrize adds ethanol 15ml, put in the water-bath and heated 10 minutes, filter, filtrate adds hydrochloric acid solution (1mol/L) 5ml, heating is 10 minutes in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, hydro-oxidation sodium liquid (0.1mol/L) 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off;
(3) discriminating of hymecromone.Get the content 2g of this product capsule or granule or tablet, porphyrize adds chloroform 20ml, and reflux 10 minutes filters, get filtrate 5ml, fling to chloroform, residue adds 1 of ferric chloride test solution with methanol 5ml dissolving, displaing yellow adds 1 of ammonia solution again, shows red, and bluish violet fluorescence ring is arranged;
(4) discriminating of hymecromone.Get the filtrate 15ml under discriminating (3) item, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, with methanol 2ml dissolving, product solution in contrast.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-acetone (1.2: 0.8) is developing solvent, launches, and takes out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point;
(5) material that has the flavone aglycone structure among the other side is differentiated.Get the content 2g of this product capsule or granule or tablet, porphyrize adds ethanol 20ml, reflux 5 minutes, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put under the ultra-violet lamp (365nm) and observe, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put ultra-violet lamp (365nm) under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence;
(6) discriminating of Herba Andrographis medical material.Get the content 1g of this product capsule or granule or tablet, add ethanol 30ml, soaked 30 minutes, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent, launch with chloroform-ethyl acetate-methanol (3.4: 2.6: 0.3), take out, dry, spray is with 2%3, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution (facing the time spent preparation) is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(7)) with rhubarb medicinal material in Radix Et Rhizoma Rhei control medicinal material and the emodin reference substance discriminating side.Get the content 2g of this product capsule or granule or tablet, put in the tool plug conical flask, added methanol 20ml ultrasonic 20 minutes, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 20 minutes, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (11: 4.3: 0.8) is developing solvent, launch, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
Assay:
1, hymecromone: measure according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A).
The preparation lucifuge operation of reference substance solution.Precision takes by weighing the hymecromone reference substance 0.1g that is dried to constant weight at 105 ℃, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and [gets caustic lye of soda (0.1mol/L) 20ml, add ethanol dilution to 1000ml] about 70ml, make dissolving and be diluted to scale, shake up, precision is measured 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and is diluted to scale, shake up, measure immediately.
The preparation lucifuge operation of need testing solution.Get content under the content uniformity item, porphyrize, precision takes by weighing this product an amount of (being equivalent to hymecromone 0.1g approximately), put in the tool plug brown bottle accurate alkaline ethanol liquid 90ml, the close plug of adding, jolting was extracted 45 minutes, filtered, and discarded filtrate just, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, add alkaline ethanol liquid and be diluted to scale, shake up, measure immediately.
Algoscopy is got reference substance solution and need testing solution respectively, measures trap at 370nm wavelength place according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A), calculates, promptly.
Every of this product contains hymecromone (C 10H 8O 3) should be 90.0%～110.0% of labelled amount.
2, emodin, chrysophanol:
Measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005).
Chromatographic condition and system suitability test are packed column with octadecylsilane chemically bonded silica; Methanol 0.1% phosphoric acid solution (53: 11) is a mobile phase; Detect wavelength 251nm.Number of theoretical plate calculates by the emodin peak should be not less than 1500.
The preparation precision of reference substance solution takes by weighing emodin reference substance 10mg, chrysophanol reference substance 14mg, puts respectively in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up; Precision is measured emodin and each 2ml of chrysophanol solution respectively, puts in the 25ml measuring bottle, adds methanol to scale, shakes up, and promptly gets (contain among the every 1ml of emodin among 9.6 μ g, the every 1ml of chrysophanol and contain 12.8 μ g).
The about 2.5g of content under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the conical flask, precision adds methanol 50ml, claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 10ml, put in the 50ml round-bottomed flask, fling to methanol, add 2.5mol/L sulfuric acid solution 20ml, supersound process 5 minutes adds chloroform 20ml again, reflux 1 hour, cooling moves in the separatory funnel, uses the minimum of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution chloroform extraction 3 times, each 10ml, combined chloroform liquid is with anhydrous sodium sulfate dehydration, chloroform solution moves in the 100ml conical flask, flings to chloroform, and the residue precision adds methanol 10ml, claim to decide weight, put slight fever dissolved residue in the water-bath, put cold after, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively above-mentioned two kinds of reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and calculate, promptly.
Every of this product contains Radix Et Rhizoma Rhei with emodin (C 15H 10O 5) and chrysophanol (C 15H 10O 4) the total amount meter, must not be less than 0.15mg.
Embodiment 5
[prescription] hymecromone 100g, Herba Rabdosiae Lophanthoidis 900g, Herba Artemisiae Scopariae 600g, Herba Andrographis 300g and Radix Et Rhizoma Rhei 30g.
[method for making] above five tastes, except that hymecromone, Radix Et Rhizoma Rhei, three flavors such as all the other Herba Rabdosiae Lophanthoidiss decoct with water secondary, and amount of water is 12,10 times, each 2 hours, filter, merging filtrate leaves standstill, get supernatant, being condensed into relative density is the thick paste of 1.31 (80 ℃), and drying under reduced pressure is ground into fine powder; Radix Et Rhizoma Rhei powder is broken into fine powder, with hymecromone fine powder and above-mentioned dried cream powder mixing, uses alcohol granulation, and drying incapsulates, and makes 1000, promptly.
[inspection] should meet every regulation relevant under the capsule item (appendix IL of Chinese Pharmacopoeia version in 2000);
[character] this product is tan powder or granule; Bitter in the mouth, puckery.
The microscopical identification of [discriminating] (1) rhubarb medicinal material.Get the content of this product capsule or granule or tablet, porphyrize, put microscopically and observe: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation;
(2) physicochemical identification of rhubarb medicinal material.Get the content 0.5g of this product capsule or granule or tablet, porphyrize adds ethanol 15ml, put in the water-bath and heated 15 minutes, filter, filtrate adds hydrochloric acid solution (1mol/L) 5ml, heating is 30 minutes in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, hydro-oxidation sodium liquid (0.1mol/L) 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off;
(3) discriminating of hymecromone.Get the content 2g of this product capsule or granule or tablet, porphyrize adds chloroform 40ml, and reflux 20 minutes filters, get filtrate 5ml, fling to chloroform, residue adds 1 of ferric chloride test solution with methanol 5ml dissolving, displaing yellow adds 1 of ammonia solution again, shows red, and bluish violet fluorescence ring is arranged;
(4) discriminating of hymecromone.Get the filtrate 15ml under discriminating (3) item, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, with methanol 2ml dissolving, product solution in contrast.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60～90 ℃)-acetone (1.8: 1.1) is developing solvent, launches, and takes out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point;
(5) material that has the flavone aglycone structure among the other side is differentiated.Get the content 2g of this product capsule or granule or tablet, porphyrize adds ethanol 20ml, reflux 15 minutes, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put under the ultra-violet lamp (365nm) and observe, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put ultra-violet lamp (365nm) under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence;
(6) discriminating of Herba Andrographis medical material.Get the content 1g of this product capsule or granule or tablet, add ethanol 30ml, soaked 30 minutes, supersound process 40 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent, launch with chloroform-ethyl acetate-methanol (4.5: 3.4: 0.5), take out, dry, spray is with 2%3, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution (facing the time spent preparation) is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(7)) with rhubarb medicinal material in Radix Et Rhizoma Rhei control medicinal material and the emodin reference substance discriminating side.Get the content 2g of this product capsule or granule or tablet, put in the tool plug conical flask, added methanol 20ml ultrasonic 40 minutes, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 40 minutes, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with petroleum ether (30～60 ℃)-Ethyl formate-formic acid (18: 7.5: 1.1) is developing solvent, launch, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
Assay:
1, hymecromone: measure according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A).
The preparation lucifuge operation of reference substance solution.Precision takes by weighing the hymecromone reference substance 0.1g that is dried to constant weight at 105 ℃, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and [gets caustic lye of soda (0.1mol/L) 20ml, add ethanol dilution to 1000ml] about 70ml, make dissolving and be diluted to scale, shake up, precision is measured 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and is diluted to scale, shake up, measure immediately.
The preparation lucifuge operation of need testing solution.Get content under the content uniformity item, porphyrize, precision takes by weighing this product an amount of (being equivalent to hymecromone 0.1g approximately), put in the tool plug brown bottle accurate alkaline ethanol liquid 110ml, the close plug of adding, jolting was extracted 75 minutes, filtered, and discarded filtrate just, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, add alkaline ethanol liquid and be diluted to scale, shake up, measure immediately.
Algoscopy is got reference substance solution and need testing solution respectively, measures trap at 374nm wavelength place according to spectrophotography (two appendix IV of Chinese Pharmacopoeia version in 2000 A), calculates, promptly.
Every of this product contains hymecromone (C 10H 8O 3) should be 90.0%～110.0% of labelled amount.
2, emodin, chrysophanol:
Measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2005).
Chromatographic condition and system suitability test are packed column with octadecylsilane chemically bonded silica; Methanol 0.1% phosphoric acid solution (121: 22) is a mobile phase; Detect wavelength 257nm.Number of theoretical plate calculates by the emodin peak should be not less than 1500.
The preparation precision of reference substance solution takes by weighing emodin reference substance 14mg, chrysophanol reference substance 18mg, puts respectively in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up; Precision is measured emodin and each 2ml of chrysophanol solution respectively, puts in the 25ml measuring bottle, adds methanol to scale, shakes up, and promptly gets (contain among the every 1ml of emodin among 9.6 μ g, the every 1ml of chrysophanol and contain 12.8 μ g).
The about 2.5g of content under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the conical flask, precision adds methanol 50ml, claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 10ml, put in the 50ml round-bottomed flask, fling to methanol, add 2.5mol/L sulfuric acid solution 20ml, supersound process 10 minutes adds chloroform 20ml again, reflux 1 hour, cooling moves in the separatory funnel, uses the minimum of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution chloroform extraction 3 times, each 10ml, combined chloroform liquid is with anhydrous sodium sulfate dehydration, chloroform solution moves in the 100ml conical flask, flings to chloroform, and the residue precision adds methanol 10ml, claim to decide weight, put slight fever dissolved residue in the water-bath, put cold after, claim to decide weight again, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively above-mentioned two kinds of reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and calculate, promptly.Every of this product contains Radix Et Rhizoma Rhei with emodin (C 15H 10O 5) and chrysophanol (C 15H 10O 4) the total amount meter, must not be less than 0.15mg.

Claims (10)

1, a kind of method of quality control of compound gallblader freeflow solid preparation is characterized in that, may further comprise the steps:
To the observation of character,
Composition in the content is differentiated,
Solid preparation is checked,
Effective ingredient is carried out assay.
2, the method for quality control of claim 1 is characterized in that, described observation to character comprises following content:
When described compound gallblader freeflow solid preparation was capsule, its content was tan powder or granule; Bitter in the mouth, puckery.
When described compound gallblader freeflow solid preparation was granule, it is brown to tan granule that content is; It is sweet, little puckery to distinguish the flavor of.
When described compound gallblader freeflow solid preparation was tablet, content was brown; Bitter in the mouth, puckery.
3, the method for quality control of claim 1 is characterized in that, the composition in the content is differentiated, comprises following content: to the discriminating of Radix Et Rhizoma Rhei, hymecromone and andrographolide composition.
4, the method for quality control of claim 3 is characterized in that, described discriminating step to Radix Et Rhizoma Rhei, hymecromone and andrographolide composition in the compound gallblader freeflow solid preparation content is as follows:
A. get the content of this product capsule or granule or tablet, porphyrize, put microscopically and observe: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation;
B. get the content 0.5g of this product capsule or granule or tablet, porphyrize adds ethanol 15ml, put in the water-bath and heated 10-15 minute, filter, filtrate adds 1mol/L hydrochloric acid solution 5ml, heating is 10-30 minute in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, add 0.1mol/L caustic lye of soda 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off;
C. get the content 2g of this product capsule or granule or tablet, porphyrize adds chloroform 20-40ml, reflux 10-20 minute, filter, get filtrate 5ml, fling to chloroform, residue dissolves with methanol 5ml, add 1 of ferric chloride test solution, displaing yellow adds 1 of ammonia solution again, show red, and bluish violet fluorescence ring is arranged;
D. get the filtrate 15ml that differentiates under a item, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, with methanol 2ml dissolving, product solution in contrast.Test according to an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 1.2-1.8: 0.8-1.1 petroleum ether-acetone is developing solvent, wherein the petroleum ether temperature is 60～90 ℃, launches, and takes out, dry smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point;
E. get the content 2g of this product capsule or granule or tablet, porphyrize adds ethanol 20ml, reflux 5-15 minute, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put the ultra-violet lamp of 365nm and observe down, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put the ultra-violet lamp of 365nm and observe down, show hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence;
F. get the content 1g of this product capsule or granule or tablet, add ethanol 30ml, soaked 30 minutes, supersound process 20-40 minute, filter, filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent, launch with chloroform-ethyl acetate-methanol of 3.4-4.5: 2.6-3.4: 0.3-0.5, take out, dry, spray is to face 2%3 of time spent preparation, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
G. get the content 2g of this product capsule or granule or tablet, put in the tool plug conical flask, add the ultrasonic 20-40 of methanol 20ml minute, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 20-40 minute, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, petroleum ether-Ethyl formate-formic acid upper solution with 11-18: 4.3-7.5: 0.8-1.1 is developing solvent, and wherein the petroleum ether temperature is 60～90 ℃, launches, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
5, the method for quality control of claim 3 is characterized in that, described discriminating step to Radix Et Rhizoma Rhei, hymecromone and andrographolide composition in the compound gallblader freeflow solid preparation content is as follows:
A. get the content of this product capsule or granule or tablet, porphyrize, put microscopically and observe: calcium oxalate cluster crystal diameter 20～160 μ m, have to 190 μ m.The non-lignify of bordered pit, reticulate pattern, screw thread and annular duct.Starch grain is a lot of, simple grain class sphere or polygon, and diameter 3～45 μ m, omphalion is starlike; Composite grain is made up of 2～8 gradation;
B. get the content 0.5g of this product capsule or granule or tablet, porphyrize adds ethanol 15ml, put in the water-bath and heated 10 minutes, filter, filtrate adds 1mol/L hydrochloric acid solution 5ml, heating is 20 minutes in the water-bath, put cold, with ether 10ml jolting, divide and get ether liquid, add 0.1mol/L caustic lye of soda 5ml, jolting, water layer is brownish red, add dilute hydrochloric acid number droplet, redness is taken off;
C. get the content 2g of this product capsule or granule or tablet, porphyrize adds chloroform 30ml, and reflux 10 minutes filters, get filtrate 5ml, fling to chloroform, residue adds 1 of ferric chloride test solution with methanol 5ml dissolving, displaing yellow adds 1 of ammonia solution again, shows red, and bluish violet fluorescence ring is arranged;
D. get the filtrate 15ml that differentiates under the c item, fling to chloroform, residue adds methanol 3ml dissolving, and as need testing solution, other gets hymecromone reference substance 10mg, with methanol 2ml dissolving, product solution in contrast.According to the test of an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with petroleum ether-acetone of 1.5: 1 was developing solvent, and wherein the petroleum ether temperature is 60～90 ℃, launched, take out, dry, smoked iodine vapor.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show identical pale brown mottle point;
E. get the content 2g of this product capsule or granule or tablet, porphyrize adds ethanol 20ml, reflux 10 minutes, filter, filtrate add active carbon a little, warm back filters, get filtrate 2ml, evaporate to dryness adds each 1ml of saturated acetone soln of 1% citric acid acetone soln and boric acid, evaporate to dryness, put under the 365nm ultra-violet lamp and observe, show green-yellow fluorescence.Other get filtrate a little, drip on filter paper, put the 365nm ultra-violet lamp under and observe, apparent hyacinthine fluorescence.Drip 1% aluminum chloride alcoholic solution, show sapphirine fluorescence;
F. get the content 1g of this product capsule or granule or tablet, add ethanol 30ml, soaked 30 minutes, supersound process 30 minutes filters, and filtrate evaporate to dryness, residue add dehydrated alcohol 5ml makes dissolving, filters, and is concentrated into 1ml, as need testing solution.Other gets the dehydrorographolide reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 1mg, in contrast product solution.According to an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be the silica gel G F of adhesive with the sodium carboxymethyl cellulose 254On the lamellae, be developing solvent with chloroform-ethyl acetates of 4: 3: 0.4-methanol, launch, take out, dry that spray is to face 2%3 of time spent preparation, the equivalent mixed liquor of 5-dinitrobenzoic acid alcoholic solution and 2mol/L potassium hydroxide solution is inspected under daylight immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
G. get the content 2g of this product capsule or granule or tablet, put in the tool plug conical flask, added methanol 20ml ultrasonic 30 minutes, filter, the filtrate evaporate to dryness adds water 30ml and makes dissolving, add hydrochloric acid 3ml again, put in the water-bath and heated 30 minutes, immediately cooling, divide 2 extractions with ether, each 20ml merges ether solution, evaporate to dryness, residue add chloroform 4ml makes dissolving, filters, be concentrated into 1ml, as need testing solution.Other gets Radix Et Rhizoma Rhei control medicinal material 0.05g, is equipped with control medicinal material solution with legal system.Get the emodin reference substance again, add methanol and make the solution that every 1ml contains 1mg, in contrast product solution.Test according to an appendix VI of Chinese Pharmacopoeia version in 2000 B thin layer chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with petroleum ether-Ethyl formates of 15: 5: 1-formic acid upper solution is developing solvent, and wherein the petroleum ether temperature is 30～60 ℃, launches, take out, dry, put in the ammonia steam smoked after, inspect under the daylight.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color respectively.
6, the method for quality control of claim 1 is characterized in that, described preparation inspection is comprised: comprise following content:
For capsule, should meet relevant every regulation under an appendix IL of Chinese Pharmacopoeia version in 2000 the capsule item;
For granule, should meet relevant every regulation under an appendix IC of Chinese Pharmacopoeia version in 2000 the granule item;
For tablet, should meet relevant every regulation under an appendix ID of Chinese Pharmacopoeia version in 2000 the tablet item.
7, the method for quality control of claim 1 is characterized in that, effective ingredient is carried out assay comprise wherein hymecromone and emodin, chrysophanol Determination on content.
8, the method for quality control of claim 7 is characterized in that, the assay of described hymecromone to wherein is through following steps:
The preparation lucifuge operation of reference substance solution.Precision takes by weighing the hymecromone reference substance 0.1g that is dried to constant weight at 105 ℃, puts in the brown measuring bottle of 100ml, adds the about 70ml of alkaline ethanol liquid, add ethanol dilution to 1000ml, make dissolving and be diluted to scale, shake up, precision is measured 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, adds alkaline ethanol liquid and is diluted to scale, shake up, measure immediately.Wherein alkaline ethanol liquid is formulated by 0.1mol/L caustic lye of soda 20ml.
The preparation lucifuge operation of need testing solution.Get content under the content uniformity item, porphyrize, precision take by weighing this product an amount of (, be equivalent to hymecromone 0.1g approximately, put in the tool plug brown bottle accurate alkaline ethanol liquid 90-110ml, the close plug of adding, jolting was extracted 45-75 minute, filtered, and discarded filtrate just, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 50ml, adds alkaline ethanol liquid and is diluted to scale, shake up, precision is measured 5ml again, puts in the brown measuring bottle of 100ml, add alkaline ethanol liquid and be diluted to scale, shake up, measure immediately.
Algoscopy is got reference substance solution and need testing solution respectively, measures trap at 372 ± 2nm wavelength place according to two appendix IV of Chinese Pharmacopoeia version in 2000 A spectrophotography, calculates, promptly.
9, the method for quality control of claim 7 is characterized in that, described emodin, chrysophanol Determination on content to wherein is through following steps
According to an appendix VID of Chinese Pharmacopoeia version in 2005 high effective liquid chromatography for measuring.
Chromatographic condition and system suitability test are packed column with octadecylsilane chemically bonded silica; Using methanol 0.1% phosphoric acid solution of ratio 53-121: 11-22 is mobile phase; Detect wavelength 254 ± 3nm.Number of theoretical plate calculates by the emodin peak should be not less than 1500.
The preparation precision of reference substance solution takes by weighing emodin reference substance 10-14mg, chrysophanol reference substance 14-18mg, puts respectively in the 100ml measuring bottle, with dissolve with methanol and be diluted to scale, shakes up; Precision is measured emodin and each 2ml of chrysophanol solution respectively, puts in the 25ml measuring bottle, adds methanol to scale, shakes up, that is, wherein contain among the every 1ml of emodin among 9.6 μ g, the every 1ml of chrysophanol and contain 12.8 μ g.
The about 2.5g of content under the content uniformity item is got in the preparation of need testing solution, and accurate the title decides, and puts in the conical flask, precision adds methanol 50ml, claims to decide weight, reflux 30 minutes, put coldly, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, precision is measured subsequent filtrate 10ml, put in the 50ml round-bottomed flask, fling to methanol, add 2.5mol/L sulfuric acid solution 20ml, supersound process 5,10 minutes adds chloroform 20ml again, reflux 1 hour, cooling moves in the separatory funnel, use the minimum of chloroform washing container, incorporate in the separatory funnel, divide and get chloroform layer, acid solution chloroform extraction 3 times, each 10ml, combined chloroform liquid, with anhydrous sodium sulfate dehydration, chloroform solution moves in the 100ml conical flask, flings to chloroform, the residue precision adds methanol 10ml, claims to decide weight, puts slight fever dissolved residue in the water-bath, put cold after, claim again to decide weight, supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.
Accurate respectively above-mentioned two kinds of reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and calculate, promptly.
10, the method for quality control of claim 7 is characterized in that, the content of hymecromone wherein and emodin, chrysophanol is that every capsules contains hymecromone C 10H 8O 3Should be 90.0%～110.0% of labelled amount; Every capsules contains Radix Et Rhizoma Rhei with emodin C 15H 10O 5)With chrysophanol C 15H 10O 4The total amount meter, must not be less than 0.15mg.
CNB2005100032696A 2005-11-10 2005-11-10 Quality control method of compound gallblader freeflow solid preparation Expired - Fee Related CN100369614C (en)

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* Cited by examiner, † Cited by third party
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CN103512999A (en) * 2012-06-21 2014-01-15 肇庆星湖制药有限公司 Quality detection method for compound baical skullcap root tablet
CN103604901A (en) * 2013-11-18 2014-02-26 广州白云山和记黄埔中药有限公司 Thin-layer identification method of compound andrographis tablet
CN111189944A (en) * 2020-03-11 2020-05-22 济南广盛源生物科技有限公司 Method for simultaneously identifying radix sophorae flavescentis and common andrographis herb in three-flavor bistort rhizome oral liquid
CN115656390A (en) * 2022-12-08 2023-01-31 成都苑东生物制药股份有限公司 Method for measuring content of paracetamol and oxycodone sustained release tablets
CN115920084A (en) * 2023-02-17 2023-04-07 武汉科技大学 Preparation method of hymecromone cyclodextrin inclusion compound

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103512999A (en) * 2012-06-21 2014-01-15 肇庆星湖制药有限公司 Quality detection method for compound baical skullcap root tablet
CN103512999B (en) * 2012-06-21 2015-11-25 肇庆星湖制药有限公司 The quality determining method of Fufang Huangqin Tablets by HPLC
CN103604901A (en) * 2013-11-18 2014-02-26 广州白云山和记黄埔中药有限公司 Thin-layer identification method of compound andrographis tablet
CN111189944A (en) * 2020-03-11 2020-05-22 济南广盛源生物科技有限公司 Method for simultaneously identifying radix sophorae flavescentis and common andrographis herb in three-flavor bistort rhizome oral liquid
CN115656390A (en) * 2022-12-08 2023-01-31 成都苑东生物制药股份有限公司 Method for measuring content of paracetamol and oxycodone sustained release tablets
CN115920084A (en) * 2023-02-17 2023-04-07 武汉科技大学 Preparation method of hymecromone cyclodextrin inclusion compound

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