CN105784913B - Gynaecology break it is red drink capsule quality determining method - Google Patents

Gynaecology break it is red drink capsule quality determining method Download PDF

Info

Publication number
CN105784913B
CN105784913B CN201610220326.4A CN201610220326A CN105784913B CN 105784913 B CN105784913 B CN 105784913B CN 201610220326 A CN201610220326 A CN 201610220326A CN 105784913 B CN105784913 B CN 105784913B
Authority
CN
China
Prior art keywords
solution
reference substance
motherwort
taken
test solution
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201610220326.4A
Other languages
Chinese (zh)
Other versions
CN105784913A (en
Inventor
龚云
张英帅
白璐
刘逆夫
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhuzhou Qianjin Pharmaceutical Co Ltd
Original Assignee
Zhuzhou Qianjin Pharmaceutical Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhuzhou Qianjin Pharmaceutical Co Ltd filed Critical Zhuzhou Qianjin Pharmaceutical Co Ltd
Priority to CN201610220326.4A priority Critical patent/CN105784913B/en
Publication of CN105784913A publication Critical patent/CN105784913A/en
Application granted granted Critical
Publication of CN105784913B publication Critical patent/CN105784913B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/062—Preparation extracting sample from raw material

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to gynaecology break it is red drink capsule quality determining method, including：Discriminating to active constituent motherwort, the radix paeoniae rubrathe, Radix Notoginseng and hairyvein agrimony, the inspection to moisture, content uniformity, disintegration time limited and microbial limit and the measure to paeoniflorin content；Wherein, the discrimination method of motherwort includes the following steps：(1) gynaecology is taken to break red drink 1.4~1.8g of capsule 's content, is dissolved in 25~35ml absolute ethyl alcohols, 80~100 DEG C of refluxing extractions, it lets cool, filtering and collecting filter liquid, it is evaporated, residue obtained plus 7ml~10ml water dissolutions, to take supernatant after 3000~5000 revs/min of centrifugations, it is evaporated, activated carbon oxidation aluminium column is splined on, with eluent is collected after anhydrous ethanol elution, is evaporated, it is residue obtained to be dissolved with absolute ethyl alcohol, supernatant is taken as test solution；(2) preparation of reference substance solution；(3) differentiate according to thin-layered chromatography.Quality testing provided by the invention, accuracy is high, and stability is good, is particularly suitable for high humidity environment, the quality testing for the red drink capsule that can break effective for gynaecology.

Description

Gynaecology break it is red drink capsule quality determining method
Technical field
The present invention relates to the quality testings of Chinese patent drug, and in particular to gynaecology break it is red drink capsule quality determining method.
Background technology
Gynaecology break it is red drink capsule be Zhuzhou Qianjin Pharmacy Co., Ltd production exclusive product, by the radix paeoniae rubrathe, motherwort, The Six-elements such as Radix Notoginseng, hairyvein agrimony, charred RADIX SANGUISORBAE, charred POLLEN TYPHAE medicinal material forms, and extracted manufactured pure Chinese medicine compound preparation has cool blood, The effect of stagnation resolvation, hemostasis.For dysfunctional uterine bleeding, menorrhalgia is shown as, menostaxis, and tcm diagnosis is " leakage Card ", dialectical category blood-head person symptoms include blood volume are more or dripping not cleaning, and color depth is red or purplish red, and matter is sticky, accompanies a small amount of clot, companion There is flushing dizziness, irritable, dry happiness drink, constipation is urinated red.Motherwort in prescription is monarch drug in a prescription, have promoting blood circulation, silt of dispelling, menstruation regulating, Inducing diuresis to remove edema, the effect for shrinking uterus.
Patent document CN101919942A discloses a kind of matter of Chinese medicinal composition capsules agent for treating gynecologic functional uterine bleeding diseases Quantity measuring method, the detection method of specific open active constituent motherwort are：Capsule 's content 0.9g~the 1.5g is taken, adds second Alcohol 22mL~38mL, is heated to reflux 0.5h~1.5h, lets cool, and filtration, filtrate is concentrated into 2.5mL~7.5mL, is added on internal diameter 5mm On activated carbon-alumina column of the mesh neutral alumina 1g~3g of~15mm, activated carbon 0.3g~0.7g, 100 mesh~120, use 15mL~45mL ethanol elutions are collected eluent, are evaporated, and residue adds 0.2mL~0.8mL ethyl alcohol to dissolve, as test solution； Motherwort control medicinal material 1.5g~4.5g is taken, control medicinal material solution is made in the same way of with test solution；Stachydrine hydrochloride is taken to compare Product add ethyl alcohol that the solution of a concentration of 2.5mg/mL~7.5mg/mL is made, as reference substance solution；It is each to draw above-mentioned three kinds of solution 10 μ l, with n-butanol:Hydrochloric acid:Water is 3~5:0.5~1.5:0.2~0.8 mixed solution is solvent, carries out thin-layer chromatography Method is tested, and is sprayed and is developed the color with dilute bismuth potassium iodide test solution, in test sample chromatography, corresponding with control medicinal material chromatography and reference substance chromatography On position, the spot of same color is shown.However, this method is in practical operation, there are the panel time is long, developing time is long, spirit Sensitivity is not high, is influenced the problems such as big by ambient humidity, affect gynaecology break it is red drink capsule production and quality testing.
《Chinese Pharmacopoeia 2015 editions》First discrimination method for disclosing motherwort of page 290；Its disclosed test sample is molten Liquid and preparation method thereof is that fresh goods crushed after being dried is dissolved in absolute ethyl alcohol, and centrifugation takes supernatant as test solution；Separately take hydrochloric acid Stachydrine reference substance adds absolute ethyl alcohol that solution of every lml containing 1mg is made, as reference substance solution；It is according to thin-layered chromatography：It inhales Each 5~10ml of above two solution is taken, is put respectively on same silica gel g thin-layer plate, with acetone-absolute ethyl alcohol-hydrochloric acid (10:6: 1) it is solvent, is unfolded, take out, dry, is heated 15 minutes at 105 DEG C, let cool, spray with dilute bismuth potassium iodide test solution-ferric trichloride Test solution (10:1) mixed solution is clear to spot development.In practical operation, prepare in this way gynaecology break it is red drink capsule confession Test sample solution simultaneously detects, and test sample is difficult to realize efficiently separate, and target blob obscures, and the R between target blobfValue has difference It is different, it is difficult to realize the accurate discriminating to motherwort ingredient.
Invention content
The defects of the purpose of the present invention is overcoming the prior art, provides the quality testing side that red drink capsule is broken in a kind of gynaecology Method, this method specificity is strong, durability, high sensitivity, can be as the important quality examination criteria of said preparation.
Specifically, the present invention provides the quality determining method that red drink capsule is broken in a kind of gynaecology, the method includes to work Property ingredient motherwort, the discriminating of the radix paeoniae rubrathe, Radix Notoginseng and hairyvein agrimony
Wherein, the discrimination method of the active constituent motherwort includes：(1) preparation of test solution；(2) reference substance is molten The preparation of liquid；(3) differentiate according to thin-layered chromatography.
The preparation of step (1) described test solution includes the following steps：Take gynaecology break it is red drink capsule 's content 1.4~ 1.8g is dissolved in 25~35ml absolute ethyl alcohols, and 80~100 DEG C of refluxing extractions let cool, filtering and collecting filter liquid, are evaporated, residue obtained Add 7ml~10ml water dissolutions, to take supernatant after 3000~5000 revs/min of centrifugations, be evaporated, be splined on activated carbon-aluminium oxide Column with eluent is collected after anhydrous ethanol elution, is evaporated, residue obtained to be dissolved with absolute ethyl alcohol, takes supernatant as test sample Solution.
Preferably, the preparation of the test solution includes the following steps：Take gynaecology break red drink capsule 's content 1.5g~ 1.7g, is dissolved in 28ml~32ml absolute ethyl alcohols, 80~100 DEG C of refluxing extraction 50min~70min, let cool, filter after collect filter Liquid, water bath method, residue obtained plus 7ml~9ml water dissolutions with 3800~4200 revs/min of centrifugation 8min~12min, take Clear liquid, water bath method are splined on activated carbon-alumina column, with eluent is collected after anhydrous ethanol elution, are evaporated, residue obtained It is dissolved with absolute ethyl alcohol, takes supernatant as test solution.
It is highly preferred that the preparation of the test solution includes the following steps：Gynaecology is taken to break red drink capsule 's content 1.6g, 30ml absolute ethyl alcohols are dissolved in, 90 DEG C of refluxing extraction 60min are let cool, filtering and collecting filter liquid, water bath method, residue obtained plus 8ml Water dissolution with 4000 revs/min of centrifugation 10min, takes supernatant, the dissolving of 5ml absolute ethyl alcohols is added after water bath method, is splined on nothing Activated carbon-alumina column of water-ethanol wetting, the internal diameter 20mm of the activated carbon-alumina column, filler is by activated carbon 0.5g It is uniformly mixed with 200~300 mesh aluminium oxide 2g；With eluent is collected after 30ml anhydrous ethanol elutions, it is evaporated, it is residue obtained It is dissolved with 1ml absolute ethyl alcohols, takes supernatant as test solution.
In the preparation method of the test solution, in order to realize excellent Sample Purification on Single effect, the activated carbon-oxidation The filler of aluminium column is by activated carbon and 200~300 mesh neutral aluminas with weight ratio 4~6：15~25 are uniformly mixed.
As a preferred embodiment, 15mm or 20mm, preferably internal diameter can be selected in the internal diameter of the activated carbon-alumina column 20mm；The filler of the activated carbon-alumina column can be by 0.4~0.6g of activated carbon and 200~300 1.5~2.5g of mesh aluminium oxide It is uniformly mixed, the activated carbon and 200~300 mesh neutral aluminas are commercially available.
Under the conditions of the specification of above-mentioned activated carbon-alumina column, described with the elution volume of anhydrous ethanol elution is 20ml ~40ml, preferably 28ml~32ml, further preferably 30ml.
Reference substance solution of the present invention includes positive control, i.e. motherwort contrast solution.The motherwort contrast solution Preparation method be：Motherwort medicinal material 2.9g~3.1g is taken, is prepared using the method identical with preparing test solution.The present invention The reference substance solution still further comprises stachydrine hydrochloride reference substance solution and negative control solution；The stachydrine hydrochloride pair Preparation method according to product solution is：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 4.8~5.2mg/ml, Preferably 5mg/ml；The negative control solution be with do not contain the gynaecology of motherwort ingredient (remaining ingredient is identical) break it is red Drink capsule is sample, is prepared using with the identical method of test solution.
The solvent of the present invention used according to thin-layered chromatography discriminating is by acetone, absolute ethyl alcohol and hydrochloric acid with volume ratio 9 ~11：5~7：1 composition, preferably with volume ratio 10：6：1 composition.
It is described to differentiate according to thin-layered chromatography, after being unfolded using solvent, drying, preferably first 10 are heated at 100~110 DEG C ~20min is sprayed after letting cool with 8~12% ethanol solution of sulfuric acid, and 2~4min are dried at 100~110 DEG C, then spray with color developing agent into Row colour developing.
It is described to differentiate the color developing agent used by dilute bismuth potassium iodide solution and 1% ferric trichloride ethanol solution according to thin-layered chromatography With volume ratio 8~12:1 mixes, preferably with volume ratio 10:1 mixes.The configuration method of dilute bismuth potassium iodide solution Referring to《Chinese Pharmacopoeia》2010 editions annex VIB.
As a preferred embodiment of the present invention, the gynaecology discrimination method of motherwort ingredient in red drink capsule that breaks includes Following steps：
(1) preparation of test solution：Gynaecology is taken to break red drink 1.4~1.8g of capsule 's content, it is anhydrous to be dissolved in 25~35ml Ethyl alcohol, 80~100 DEG C of refluxing extraction 50min~70min, lets cool, filtering and collecting filter liquid, water bath method, residue obtained plus 7ml ~10ml water dissolutions with 3000~5000 revs/min of centrifugation 8min~12min, take supernatant, water bath method is splined on activity Charcoal-alumina column, the internal diameter of the activated carbon-alumina column are 15mm or 20mm, filler by 0.4~0.6g of activated carbon and 200~300 1.5~2.5g of mesh aluminium oxide are uniformly mixed；With eluent is collected after 20ml~40ml anhydrous ethanol elutions, steam Dry, residue obtained 0.5ml~1.5ml absolute ethyl alcohols dissolve, and take supernatant as test solution；
(2) preparation of motherwort contrast solution：Motherwort medicinal material 2.9g~3.1g is taken, using with preparing test solution phase Same method prepares motherwort contrast solution；
The preparation of stachydrine hydrochloride reference substance solution：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 4.8~5.2mg/ml；
(3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution and stachydrine hydrochloride reference substance Each 10~15 μ l of solution, parallel point sample is on same silica G plate, using volume ratio as 9~11：5~7：1 acetone-anhydrous second Alcohol-hydrochloric acid is unfolded for solvent, after drying, heats 10~20min at 100~110 DEG C, lets cool, spray with 8~12% sulfuric acid ethyl alcohol Solution dries 2~4min, then spray using volume ratio as 8~12 at 100~110 DEG C:1-1% tri-chlorination of dilute bismuth potassium iodide test solution Iron alcohol mixed solution is inspected under daylight, you can.
Further preferably include the following steps：
(1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.5g~1.7g, be dissolved in 28ml~32ml without Water-ethanol, 80~100 DEG C of refluxing extraction 50min~70min, lets cool, filtering and collecting filter liquid, water bath method, residue obtained to add 7ml~9ml water dissolutions with 3800~4200 revs/min of centrifugation 8min~12min, take supernatant, water bath method is splined on work Property charcoal-alumina column, the internal diameter of the activated carbon-alumina column is 15mm or 20mm, filler by 0.4~0.6g of activated carbon and 200~300 1.5~2.5g of mesh aluminium oxide are uniformly mixed；With eluent is collected after 28ml~32ml anhydrous ethanol elutions, steam Dry, residue obtained 0.8ml~1.2ml absolute ethyl alcohols dissolve, and take supernatant as test solution；
(2) preparation of motherwort contrast solution：Motherwort medicinal material 2.9g~3.1g is taken, using with preparing test solution phase Same method prepares motherwort contrast solution；
The preparation of stachydrine hydrochloride reference substance solution：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 4.8~5.2mg/ml；
(3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution and stachydrine hydrochloride reference substance Each 10~15 μ l of solution, parallel point sample is on same silica G plate, using volume ratio as 9~11：5~7：1 acetone-anhydrous second Alcohol-hydrochloric acid is unfolded for solvent, after drying, heats 10~20min at 100~110 DEG C, lets cool, spray with 8~12% sulfuric acid ethyl alcohol Solution dries 2~4min, then spray using volume ratio as 8~12 at 100~110 DEG C:1-1% tri-chlorination of dilute bismuth potassium iodide test solution Iron alcohol mixed solution is inspected under daylight, you can.
Most preferably include the following steps：
(1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.6g, is dissolved in 30ml absolute ethyl alcohols, 90 DEG C Refluxing extraction 60min is let cool, filtering and collecting filter liquid, water bath method, residue obtained plus 8ml water dissolutions, with 4000 revs/min Centrifuge 10min, take supernatant, after water bath method plus the dissolving of 5ml absolute ethyl alcohols, be splined on the activated carbon that is soaked with absolute ethyl alcohol- Alumina column, the internal diameter 20mm of the activated carbon-alumina column, filler is by activated carbon 0.5g and 200~300 mesh aluminium oxide 2g It is uniformly mixed；It with eluent is collected after 30ml anhydrous ethanol elutions, is evaporated, residue obtained 1ml absolute ethyl alcohols dissolve, and take Supernatant is as test solution；
(2) preparation of motherwort contrast solution：Motherwort medicinal material 3g is taken, using step (1) the method, prepares motherwort Contrast solution；
The preparation of stachydrine hydrochloride reference substance solution：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 5mg/ml；
(3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution and stachydrine hydrochloride reference substance Each 10 μ l of solution, parallel point sample is on same silica G plate, using volume ratio as 10:6:1 acetone-absolute ethyl alcohol-hydrochloric acid is exhibition Agent expansion is opened, after drying, 15min is heated at 105 DEG C, lets cool, spray with 10% ethanol solution of sulfuric acid, 3min is dried at 105 DEG C, then Spray is using volume ratio as 10:1-1% ferric trichloride alcohol mixed solution of dilute bismuth potassium iodide test solution is inspected under daylight, you can.
Through practical discriminating, test sample chromatography and reference substance and control medicinal material chromatography corresponding position that the method obtains show phase With the spot of color, negative control is noiseless, and method is feasible.
In quality determining method of the present invention, to the active constituent radix paeoniae rubrathe, Radix Notoginseng and the discriminating of hairyvein agrimony, to moisture, dress The inspection for measuring difference, disintegration time limited and microbial limit and the measure to paeoniflorin content, it is preferred to use patent document Scheme disclosed in CN101919942A.Specifically：
The discriminating of the active constituent radix paeoniae rubrathe may include following steps：
(1) the capsule 's content 0.5g~1.5g is taken, adds water 12ml~38ml, is ultrasonically treated 15min~45min, filter It crosses, filtrate adds water saturated butanol solution to extract 1~5 time, each 10ml~30ml, merges extracting solution, ammonification test solution 20ml ~60ml is washed, and is discarded washing lotion, is evaporated, adds 1ml~3ml methanol dissolved residues, as test solution；
(2) radix paeoniae rubrathe control medicinal material 1g~3g is taken, adds methanol 15ml~45ml, is ultrasonically treated 0.5h~1.5h, filtration, filtrate It is evaporated, residue adds water 12ml~38ml, and warm makes dissolving, filters, and control medicinal material is made with method extraction with test solution in filtrate Solution；Paeoniflorin reference substance is taken, adds methanol that the solution of a concentration of 1mg/ml~3mg/ml is made, as reference substance solution；
(3) the μ l of above-mentioned each 3 μ l of three kinds of solution~5 are drawn, using chloroform: ethyl acetate: methanol: formic acid is 30~50: 2 ~8: 5~15: 0.1~0.3 mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 4~6% vanillin-sulfuric acid first Alcoholic solution, it is clear to be heated to spot development at 95 DEG C~115 DEG C, in test sample chromatography, with control medicinal material chromatography and reference substance On the corresponding position of chromatography, the spot of same color is shown.
The discriminating of the active constituent Radix Notoginseng may include following steps：
(1) the capsule 's content 0.5g~1.5g is taken, adds water 12ml~38ml, is ultrasonically treated 15min~45min, filter It crosses, filtrate adds water saturated butanol solution to extract 1~5 time, each 10ml~30ml, merges extracting solution, ammonification test solution 20ml ~60ml is washed, and is discarded washing lotion, is evaporated, adds 1ml~3ml methanol dissolved residues, as test solution；
(2) Radix Notoginseng control medicinal material 0.5g~1.5g is taken, water 5 is added to drip~15 drops, stirs evenly, adds water saturated butanol solution 20ml~60ml, is ultrasonically treated 20min~60min, filtration, and filtrate ammonification test solution 20ml~60ml washings discard ammoniacal liquor, add in Aqueous solution 20ml~60ml washings of n-butanol saturation, discard n-butanol aqueous solution, are evaporated, residue adds methanol 1ml~3ml molten Solution, as control medicinal material solution；Ginsenoside Rb1, Rg1 and notoginsenoside R reference substance are taken, adds methanol that each concentration is made to be The mixed solution of 0.5mg/ml~1.5mg/ml is as reference substance solution；
(3) above-mentioned each 1~3 μ l of three kinds of solution are drawn, using in 10 DEG C of overnight chloroforms arranged below: methanol: water as Lower floor's solution of 10~16: 4~10: 1~3 mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 8~12% sulphur Sour ethanol solution, it is clear to be heated to spot development at 95 DEG C~115 DEG C, in test sample chromatography, with control medicinal material chromatography and right According on the corresponding position of product chromatography, the spot of same color is shown；It puts and is inspected under the ultraviolet lamp of 350nm wavelength, show same color Fluorescence spot.
The discriminating of the active constituent hairyvein agrimony may include following steps：
(1) the capsule 's content 0.5g~1.5g is taken, adds 0.2%~0.8% sodium hydroxide solution 15ml~45ml, is surpassed Sonication 10min~30min, filtration, filtrate add hydrochloric acid tune pH value to 2~3, and filtration, filtrate adds ethyl acetate to extract 1~3 time, Each 10ml~30ml, merges extracting solution, is evaporated, and residue adds methanol 1ml~3ml to dissolve, as test solution；
(2) hairyvein agrimony control medicinal material 4g~12g is taken, adds water 25ml~75ml, decocts 0.5h~1.5h, filtration, filtrate is steamed Dry, residue adds 0.2%~0.8% sodium hydroxide solution 15ml~45ml, and warm makes dissolving, filters, filtrate and test solution Control medicinal material solution is made with method extraction；
(3) the μ l of above two solution each 14 μ l~16 are drawn, using water saturated toluene: ethyl acetate: formic acid is 4~8: 2 ~4: 0.5~1.5 mixed solution is solvent, carries out thin-layered chromatography experiment, sprays with 0.5%~1.5% ferric trichloride second Alcoholic solution develops the color, and in test sample chromatography, on position corresponding with control medicinal material chromatography, shows the spot of same color.
Inspection to moisture is specially：The content of capsule is taken, is measured according to aquametry, 9.0% must not be crossed.
Inspection to content uniformity is specially：Compared with mark loading amount, content uniformity limit should indicate every loading amount Within ± the 10% of loading amount, 2 must not be more than, and there must not be 11 times of overrun beyond content uniformity limit.
Inspection to disintegration time limited is specially：Using disintegration time limited inspection technique inspection, regulation should be met.
Inspection to microbial limit is specially：It is examined using bacterium, mould and saccharomycete inspection technique and Control bacteria examination method It looks into, regulation should be met.
Following steps may include to the measure of paeoniflorin content：
(1) preparation of test solution：Capsule 's content 0.05~0.15g is taken, it is accurately weighed, put tool plug conical flask In, precision adds in 45~55% 45~55ml of methanol, weighed weight, after cold soaking is stayed overnight, in 100~300w, 30~50KHz conditions It is lower to be ultrasonically treated 20~40 minutes, it is re-weighed, the weight of less loss is supplied with 45~55% methanol, is shaken up, 0.45um miillpore filters Filtration, filtrate is as test solution；
(2) preparation of reference substance solution：Precision weighs Paeoniflorin reference substance, adds 45~55% methanol that every 1ml is made and contains The solution of 0.05~0.15mg, as reference substance solution；
(3) it is accurate respectively to draw test solution and each 8~12ul of reference substance solution, liquid chromatograph is injected, is measured, meter Calculate to get；Wherein, using octadecylsilane chemically bonded silica as stationary phase, with volume ratio 65~75：The 0.04 of 25~35~ 0.06mol/L potassium dihydrogen phosphates-methanol is mobile phase, and Detection wavelength 230nm, number of theoretical plate is calculated by Paeoniflorin peak should Not less than 2500；Every containing the radix paeoniae rubrathe with Paeoniflorin C23H28O11Meter, must not be less than 12.0mg.
Gynaecology provided by the invention break red drink capsule quality determining method specificity is strong, high sensitivity, favorable reproducibility, and Can be effectively used for gynaecology break it is red drink capsule discriminating；Wherein, specific aim improvement is carried out to the discriminating of main component motherwort, Gained prioritization scheme makes test solution in panel without hangover, same spot RfValue difference is different small, good separating effect, and develop the color spot Clearly, quick, accurate discriminating can be achieved under high humidity environment, stability is good.This method.
Description of the drawings
Fig. 1 is the schematic diagram of identification result described in embodiment 1；Wherein, serial number 1~6 represents successively：1st, lot number 20130701 Sample, 2, the sample of lot number 20130901,3, the sample of lot number 20131101,4, motherwort control, 5, stachydrine hydrochloride pair According to 6, negative control.
Fig. 2 is the schematic diagram of identification result described in comparative example 3；
Fig. 3 is the schematic diagram of identification result described in experimental example 1；Wherein, Fig. 3 A are self-control thin layer silica G plate, and Fig. 3 B are pre- Thin layer silica G plate processed, Fig. 3 C are High Performance Thin silica G plate；In Fig. 3 A~3C, serial number 1~6 represents successively：1st, lot number 20130701 sample, 2, the sample of lot number 20130901,3, the sample of lot number 20131101,4, motherwort control, 5, hydrochloric acid Stachydrine compares, and 6, negative control.
Fig. 4 is the schematic diagram of identification result described in experimental example 2；Wherein, Fig. 4 A are the identification result at 5 DEG C, and Fig. 4 B are 30 Identification result at DEG C；In Fig. 4 A, 4B, serial number 1~6 represents successively：1st, the sample of lot number 20130701,2, lot number 20130901 Sample, 3, the sample of lot number 20131101,4, motherwort control, 5, stachydrine hydrochloride control, 6, negative control.
Fig. 5 is the schematic diagram of identification result described in experimental example 3；Wherein, Fig. 5 A be humidity 30% under identification result, Fig. 5 B For the identification result under humidity 75%；In Fig. 5 A, 5B, serial number 1~6 represents successively：1st, the sample of lot number 20130701,2, lot number 20130901 sample, 3, the sample of lot number 20131101,4, motherwort control, 5, stachydrine hydrochloride control, 6, negative control.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Instrument and reagent involved in following embodiment include：939 full-automatic thin layer making sheet device (Chongqing south bank Bells Moral technical device factory)；Tlc silica gel (chemical pure, Qingdao Marine Chemical Co., Ltd.)；(Zhuzhou silicification glass has activated carbon Limit company)；Neutral alumina (Shanghai receive the dry chemical reagent work of final roasting)；(analysis is pure, and the permanent emerging chemical reagent manufacture in Tianjin has for n-butanol Limit company)；Hydrochloric acid (analyzes pure, Zhuzhou silicification glass Co., Ltd)；Acetone (analyzes pure, Zhuzhou silicification glass Co., Ltd)； Absolute ethyl alcohol (analyzes pure, Zhuzhou silicification glass Co., Ltd)；Water (purified water is prepared before use).
Embodiment 1
The red drink capsule that breaks in accordance with the following methods to gynaecology carries out quality testing：
A, the discriminating of active constituent motherwort：
(1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.6g, is dissolved in 30ml absolute ethyl alcohols, 90 DEG C Refluxing extraction 60min is let cool, filtering and collecting filter liquid, water bath method, residue obtained plus 8ml water dissolutions, with 4000 revs/min Centrifuge 10min, take supernatant, after water bath method plus the dissolving of 5ml absolute ethyl alcohols, be splined on the activated carbon that is soaked with absolute ethyl alcohol- Alumina column, the internal diameter 20mm of the activated carbon-alumina column, filler is by activated carbon 0.5g and 200~300 mesh aluminium oxide 2g It is uniformly mixed；It with eluent is collected after 30ml anhydrous ethanol elutions, is evaporated, residue obtained 1ml absolute ethyl alcohols dissolve, and take Supernatant is as test solution；
(2) preparation of motherwort contrast solution：Motherwort medicinal material 3g is taken, using step (1) the method, prepares motherwort Contrast solution；
The preparation of stachydrine hydrochloride reference substance solution：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 5mg/ml；
(3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution, stachydrine hydrochloride reference substance Solution and each 10 μ l of negative control, parallel point sample is on same silica G plate, using volume ratio as 10:6:1 acetone-anhydrous Ethanol-hydrogen chloride is unfolded for solvent, after drying, heats 15min at 105 DEG C, lets cool, spray with 10% ethanol solution of sulfuric acid, 105 DEG C drying 3min, then sprays using volume ratio as 10:1-1% ferric trichloride alcohol mixed solution of dilute bismuth potassium iodide test solution, under daylight It inspects；
B, the discriminating of the active constituent radix paeoniae rubrathe：
(1) Capsule content 1g is taken, adds water 25ml, is ultrasonically treated 30 minutes, filtration, filtrate adds water saturated n-butanol Extraction 3 times, each 20ml, merges n-butanol liquid, and the 40ml washings of ammonification test solution discard washing lotion, n-butanol liquid is evaporated, and residue adds first Alcohol 2ml dissolves, as test solution；
(2) radix paeoniae rubrathe control medicinal material 2g is taken, adds methanol 30ml, is ultrasonically treated 1 hour, filtration, filtrate is evaporated, and residue adds water 25ml, warm make dissolving, filter, and control medicinal material solution is made with method extraction with test solution in filtrate；Paeoniflorin reference substance is taken, Add methanol that the solution of a concentration of 2mg/ml is made, as reference substance solution；
(3) above-mentioned each 4 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, with chloroform-acetic acid second Ester-methyl alcohol-formic acid (40: 5: 10: 0.2) is solvent, is unfolded, and takes out, dries, spray with 5% vanillin-sulfuric acid methanol solution, 105 DEG C to be heated to spot development clear；
C, the discriminating of active constituent Radix Notoginseng：
(1) Capsule content 1g is taken, adds water 25ml, is ultrasonically treated 30 minutes, filtration, filtrate adds water saturated n-butanol Extraction 3 times, each 20ml, merges n-butanol liquid, and the 40ml washings of ammonification test solution discard washing lotion, n-butanol liquid is evaporated, and residue adds first Alcohol 2ml dissolves, as test solution；
(2) Radix Notoginseng control medicinal material 1g is taken, water 10 is added to drip, is stirred evenly, adds water saturated n-butanol 40ml, is ultrasonically treated 40 points Clock, filtration, filtrate ammonification test solution 40ml washings discard ammoniacal liquor, then add the water 40ml washings of n-butanol saturation, discard aqueous, just Butanol liquid is evaporated, and residue adds methanol 2ml to dissolve, as control medicinal material solution；Take ginsenoside Rb1, Rg1 and notoginsenoside R pair According to product, add methanol that the mixed solution that each concentration is 1mg/ml is made, as reference substance solution；
(3) above-mentioned each 2 μ l of three kinds of solution are drawn, are put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water (13: 7: 2) 10 DEG C of overnight lower floor's solution arranged below are solvent, are unfolded, and take out, dry, spray molten with 10% sulfuric acid ethyl alcohol Liquid, 105 DEG C to be heated to spot development clear, puts and is inspected under ultraviolet lamp (365nm)；
D, the discriminating of active constituent hairyvein agrimony：
(1) Capsule content 1g is taken, adds 0.5% sodium hydroxide solution 30ml, is ultrasonically treated 20 minutes, filtration, filtrate Adding salt acid for adjusting pH value to 2.5, filtration, filtrate adds ethyl acetate to extract 2 times, each 20ml, and combined ethyl acetate liquid is evaporated, Residue adds methanol 2ml to dissolve, as test solution；
(2) hairyvein agrimony control medicinal material 8g is taken, adds water 50ml, is decocted 1 hour, filtration, filtrate is evaporated, and residue adds 0.5% hydrogen Sodium hydroxide solution 30ml, warm make dissolving, filter, and control medicinal material solution is made with method extraction with test solution in filtrate；
(3) draw each 15 μ 1 of above two solution, put respectively on same silica gel g thin-layer plate, with toluene (using water saturation)- Acetic ether-methanoic acid (6: 3: 1) is solvent, is unfolded, and takes out, dries, spray with 1% ferric trichloride ethanol solution, until colour developing is clear It is clear；
E, to the inspection of moisture：The content of capsule is taken, is measured according to aquametry, 9.0% must not be crossed；
F, to the inspection of content uniformity：For every loading amount compared with mark loading amount, content uniformity limit should be in mark loading amount ± 10% within, must not be more than 2 beyond content uniformity limit, and must not have 11 times of overrun；
G, it is specially to the inspection of disintegration time limited：Using《Chinese Pharmacopoeia 2015 editions》Defined disintegration time limited inspection technique inspection It looks into, regulation should be met；
H, to the inspection of microbial limit：Using《Chinese Pharmacopoeia 2015 editions》Defined bacterium, mould and saccharomycete inspection Method and Control bacteria examination method inspection, should meet regulation；
I, to the measure of paeoniflorin content：
(1) preparation of test solution：Take capsule 's content 0.1g, it is accurately weighed, put in tool plug conical flask, precision plus Enter 50% methanol 50ml, weighed weight after cold soaking is stayed overnight, is ultrasonically treated 30 minutes under the conditions of 200w, 40KHz, is re-weighed, and uses 50% methanol supplies the weight of less loss, shakes up, and the filtration of 0.45um miillpore filters, filtrate is as test solution；
(2) preparation of reference substance solution：Precision weighs Paeoniflorin reference substance, adds 50% methanol that every 1ml is made containing 0.1mg's Solution, as reference substance solution；
(3) it is accurate respectively to draw test solution and each 10ul of reference substance solution, liquid chromatograph is injected, is measured, is calculated, To obtain the final product；Wherein, using octadecylsilane chemically bonded silica as stationary phase, with volume ratio 70：30 0.05mol/L potassium dihydrogen phosphates are molten Liquid-methanol is mobile phase, and Detection wavelength 230nm, number of theoretical plate is calculated by Paeoniflorin peak should be not less than 2500；Every contains the radix paeoniae rubrathe With Paeoniflorin C23H28O11Meter, must not be less than 12.0mg.
The present embodiment uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule.
Wherein, the identification result of motherwort ingredient using the present embodiment the method as shown in Figure 1, as shown in Figure 1, carried out Differentiate, 3 groups of test samples show the spot of same color with motherwort to impinging upon same position, and negative control is noiseless, and panel is without dragging Tail, target blob are clear.
According to the radix paeoniae rubrathe, Radix Notoginseng and hairyvein agrimony identification result it is found that each group sample with accordingly to impinging upon same position The spot of aobvious same color, negative control is noiseless, and for panel without hangover, target blob is clear；According to moisture, content uniformity, Disintegration time limited, microbial limit inspection result it is found that each group sample meets regulation；According to the measure knot to paeoniflorin content Fruit is it is found that Paeoniflorin C in each group sample23H28O11Content be no less than 12.0mg.
It is detected according to quality determining method provided in this embodiment, product batch number is respectively 20130701, 20130901st, the gynaecology of 20,131,101 three different batches break it is red drink capsule it is qualified.
Embodiment 2
The red drink capsule that breaks in accordance with the following methods to gynaecology carries out quality testing：
A, the discriminating of active constituent motherwort：
(1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.7g, is dissolved in 32ml absolute ethyl alcohols, 100 DEG C Refluxing extraction 70min is let cool, filtering and collecting filter liquid, water bath method, residue obtained plus 9ml water dissolutions, with 4200 revs/min Centrifuge 12min, take supernatant, after water bath method plus the dissolving of 5ml absolute ethyl alcohols, be splined on the activated carbon that is soaked with absolute ethyl alcohol- Alumina column；The internal diameter 20mm of the activated carbon-alumina column, filler is by activated carbon 0.5g and 200~300 mesh aluminium oxide 2g It is uniformly mixed；It with eluent is collected after 30ml anhydrous ethanol elutions, is evaporated, residue obtained 1ml absolute ethyl alcohols dissolve, and take Supernatant is as test solution；
(2) preparation of motherwort contrast solution：Motherwort medicinal material 3g is taken, using step (1) the method, prepares motherwort Contrast solution；
The preparation of stachydrine hydrochloride reference substance solution：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 5mg/ml；
(3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution, stachydrine hydrochloride reference substance Solution and each 10 μ l of negative control, parallel point sample is on same silica G plate, using volume ratio as 10:6:1 acetone-anhydrous Ethanol-hydrogen chloride is unfolded for solvent, after drying, heats 15min at 105 DEG C, lets cool, spray with 8% ethanol solution of sulfuric acid, 105 DEG C drying 3min, then sprays using volume ratio as 10:1-1% ferric trichloride alcohol mixed solution of dilute bismuth potassium iodide test solution, under daylight It inspects；
B, the discriminating of the active constituent radix paeoniae rubrathe：
(1) take the capsule 's content 0.5g, add water 12ml, be ultrasonically treated 15min, filtration, filtrate add it is water saturated just Butanol solution extracts 3 times, each 10ml, merges extracting solution, and the 20ml washings of ammonification test solution discard washing lotion, are evaporated, add 1ml methanol Dissolved residue, as test solution；
(2) radix paeoniae rubrathe control medicinal material 1g is taken, adds methanol 15ml, is ultrasonically treated 0.5h, filtration, filtrate is evaporated, and residue adds water 12ml, warm make dissolving, filter, and control medicinal material solution is made with method extraction with test solution in filtrate；Paeoniflorin reference substance is taken, Add methanol that the solution of a concentration of 1mg/ml is made, as reference substance solution；
(3) above-mentioned each 4 μ l of three kinds of solution are drawn, using chloroform: ethyl acetate: methanol: formic acid is 30: 2: 5: 0.1 Mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 4% vanillin-sulfuric acid methanol solution, spot is heated at 95 DEG C Colour developing is clear, in test sample chromatography, on position corresponding with control medicinal material chromatography and reference substance chromatography, shows the spot of same color Point；
C, the discriminating of active constituent Radix Notoginseng：
(1) take the capsule 's content 0.5g, add water 12ml, be ultrasonically treated 15min, filtration, filtrate add it is water saturated just Butanol solution extracts 3 times, each 10ml, merges extracting solution, and the 20ml washings of ammonification test solution discard washing lotion, are evaporated, add 1ml methanol Dissolved residue, as test solution；
(2) Radix Notoginseng control medicinal material 0.5g is taken, water 5 is added to drip, is stirred evenly, adds water saturated butanol solution 20ml, is ultrasonically treated 20min, filtration, filtrate ammonification test solution 20ml washings discard ammoniacal liquor, add in the aqueous solution 20ml washings of n-butanol saturation, discard N-butanol aqueous solution is evaporated, and residue adds methanol 1ml to dissolve, as control medicinal material solution；Take ginsenoside Rb1, Rg1 and Radix Notoginseng Saponin(e R1 reference substances, it is the mixed solution of 0.5mg/ml as reference substance solution to add methanol that each concentration is made；
(3) above-mentioned each 2 μ l of three kinds of solution are drawn, using in 10 DEG C of overnight chloroforms arranged below: methanol: water is 10: 4 : lower floor's solution of 1 mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 8% ethanol solution of sulfuric acid, is added at 95 DEG C Heat is clear to spot development, in test sample chromatography, on position corresponding with control medicinal material chromatography and reference substance chromatography, shows identical The spot of color；It puts and is inspected under the ultraviolet lamp of 350nm wavelength, show the fluorescence spot of same color；
D, the discriminating of active constituent hairyvein agrimony：
(1) the capsule 's content 0.5g is taken, adds 0.2% sodium hydroxide solution 15ml, is ultrasonically treated 10min, is filtered, filter Liquid adds hydrochloric acid tune pH value, and to 2, filtration, filtrate adds ethyl acetate to extract 2 times, each 10ml, merges extracting solution, is evaporated, residue adds Methanol 1ml dissolves, as test solution；
(2) hairyvein agrimony control medicinal material 4g is taken, adds water 25ml, decocts 0.5h, filtration, filtrate is evaporated, and residue adds 0.2% hydrogen-oxygen Change sodium solution 15ml, warm makes dissolving, filters, and control medicinal material solution is made with method extraction with test solution in filtrate；
(3) each 15 μ l of above two solution are drawn, using water saturated toluene: ethyl acetate: mixing of the formic acid as 4: 2: 0.5 Solution is solvent, carries out thin-layered chromatography experiment, sprays and is developed the color with 0.4% ferric trichloride ethanol solution, in test sample chromatography, On position corresponding with control medicinal material chromatography, the spot of same color is shown；
E, to the inspection of moisture：The content of capsule is taken, is measured according to aquametry, 9.0% must not be crossed；
F, to the inspection of content uniformity：For every loading amount compared with mark loading amount, content uniformity limit should be in mark loading amount ± 10% within, must not be more than 2 beyond content uniformity limit, and must not have 11 times of overrun；
G, it is specially to the inspection of disintegration time limited：Using《Chinese Pharmacopoeia 2015 editions》Defined disintegration time limited inspection technique inspection It looks into, regulation should be met；
H, to the inspection of microbial limit：Using《Chinese Pharmacopoeia 2015 editions》Defined bacterium, mould and saccharomycete inspection Method and Control bacteria examination method inspection, should meet regulation；
I, to the measure of paeoniflorin content：
(1) preparation of test solution：Capsule 's content 0.05g is taken, it is accurately weighed, it puts in tool plug conical flask, it is accurate 45% methanol 45ml is added in, weighed weight after cold soaking is stayed overnight, is ultrasonically treated 20 minutes under the conditions of 100w, 30KHz, is re-weighed, The weight of less loss is supplied with 45% methanol, is shaken up, the filtration of 0.45um miillpore filters, filtrate is as test solution；
(2) preparation of reference substance solution：Precision weighs Paeoniflorin reference substance, adds 45% methanol that every 1ml is made containing 0.05mg Solution, as reference substance solution；
(3) it is accurate respectively to draw test solution and each 8ul of reference substance solution, liquid chromatograph is injected, is measured, is calculated, To obtain the final product；Wherein, using octadecylsilane chemically bonded silica as stationary phase, with volume ratio 75：25 0.04mol/L potassium dihydrogen phosphates are molten Liquid-methanol is mobile phase, and Detection wavelength 230nm, number of theoretical plate is calculated by Paeoniflorin peak should be not less than 2500；Every contains the radix paeoniae rubrathe With Paeoniflorin C23H28O11Meter, must not be less than 12.0mg.
The present embodiment uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient For identification result it is found that 3 groups of test samples show the spot of same color with motherwort to impinging upon same position, negative control is noiseless, For panel without hangover, target blob is clear.
According to the radix paeoniae rubrathe, Radix Notoginseng and hairyvein agrimony identification result it is found that each group sample with accordingly to impinging upon same position The spot of aobvious same color, negative control is noiseless, and for panel without hangover, target blob is clear；According to moisture, content uniformity, Disintegration time limited, microbial limit inspection result it is found that each group sample meets regulation；According to the measure knot to paeoniflorin content Fruit is it is found that Paeoniflorin C in each group sample23H28O11Content be no less than 12.0mg.
It is detected according to quality determining method provided in this embodiment, product batch number is respectively 20130701, 20130901st, the gynaecology of 20,131,101 three different batches break it is red drink capsule it is qualified.
Embodiment 3
The red drink capsule that breaks in accordance with the following methods to gynaecology carries out quality testing：
A, the discriminating of active constituent motherwort：
(1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.5g, is dissolved in 28ml absolute ethyl alcohols, 80 DEG C Refluxing extraction 50min is let cool, filtering and collecting filter liquid, water bath method, residue obtained plus 7ml water dissolutions, with 3800 revs/min 8min is centrifuged, supernatant is taken, the dissolving of 5ml absolute ethyl alcohols is added after water bath method, is splined on the activated carbon-oxygen soaked with absolute ethyl alcohol Change aluminium column；The internal diameter 20mm of the activated carbon-alumina column, filler are equal by activated carbon 0.5g and 200~300 mesh aluminium oxide 2g It is even to mix；It with eluent is collected after 30ml anhydrous ethanol elutions, is evaporated, residue obtained 1ml absolute ethyl alcohols dissolve, and take Clear liquid is as test solution；
(2) preparation of motherwort contrast solution：Motherwort medicinal material 3g is taken, using step (1) the method, prepares motherwort Contrast solution；
The preparation of stachydrine hydrochloride reference substance solution：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to a concentration of 5mg/ml；
(3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution, stachydrine hydrochloride reference substance Solution and each 10 μ l of negative control, parallel point sample is on same silica G plate, using volume ratio as 10:6:1 acetone-anhydrous Ethanol-hydrogen chloride is unfolded for solvent, after drying, heats 15min at 105 DEG C, lets cool, spray with 10% ethanol solution of sulfuric acid, 105 DEG C drying 3min, then sprays using volume ratio as 10:1-1% ferric trichloride alcohol mixed solution of dilute bismuth potassium iodide test solution, under daylight It inspects；
B, the discriminating of the active constituent radix paeoniae rubrathe：
(1) take the capsule 's content 1.5g, add water 38ml, be ultrasonically treated 45min, filtration, filtrate add it is water saturated just Butanol solution extracts 3 times, each 30ml, merges extracting solution, and the 60ml washings of ammonification test solution discard washing lotion, are evaporated, add 3ml methanol Dissolved residue, as test solution；
(2) radix paeoniae rubrathe control medicinal material 3g is taken, adds methanol 45ml, is ultrasonically treated 1.5h, filtration, filtrate is evaporated, and residue adds water 38ml, warm make dissolving, filter, and control medicinal material solution is made with method extraction with test solution in filtrate；Paeoniflorin reference substance is taken, Add methanol that the solution of a concentration of 3mg/ml is made, as reference substance solution；
(3) above-mentioned each 4 μ l of three kinds of solution are drawn, using chloroform: ethyl acetate: methanol: formic acid is 50: 8: 15: 0.3 Mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 6% vanillin-sulfuric acid methanol solution, spot is heated at 115 DEG C Colour developing is clear, in test sample chromatography, on position corresponding with control medicinal material chromatography and reference substance chromatography, shows the spot of same color Point；
C, the discriminating of active constituent Radix Notoginseng：
(1) take the capsule 's content 1.5g, add water 38ml, be ultrasonically treated 45min, filtration, filtrate add it is water saturated just Butanol solution extracts 3 times, each 30ml, merges extracting solution, and the 60ml washings of ammonification test solution discard washing lotion, are evaporated, add 3ml methanol Dissolved residue, as test solution；
(2) Radix Notoginseng control medicinal material 1.5g is taken, water 15 is added to drip, is stirred evenly, adds water saturated butanol solution 60ml, is ultrasonically treated 60min, filtration, filtrate ammonification test solution 60ml washings discard ammoniacal liquor, add in the aqueous solution 60ml washings of n-butanol saturation, discard N-butanol aqueous solution is evaporated, and residue adds methanol 3ml to dissolve, as control medicinal material solution；Take ginsenoside Rb1, Rg1 and Radix Notoginseng Saponin(e R1 reference substances, it is the mixed solution of 1.5mg/ml as reference substance solution to add methanol that each concentration is made；
(3) above-mentioned each 2 μ l of three kinds of solution are drawn, using in 10 DEG C of overnight chloroforms arranged below: methanol: water is 16: Lower floor's solution of 10: 3 mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 12% ethanol solution of sulfuric acid, 115 It is clear DEG C to be heated to spot development, in test sample chromatography, on position corresponding with control medicinal material chromatography and reference substance chromatography, shows The spot of same color；It puts and is inspected under the ultraviolet lamp of 350nm wavelength, show the fluorescence spot of same color.
D, the discriminating of active constituent hairyvein agrimony：
(1) the capsule 's content 1.5g is taken, adds 0.8% sodium hydroxide solution 45ml, is ultrasonically treated 30min, is filtered, filter Liquid adds hydrochloric acid tune pH value, and to 3, filtration, filtrate adds ethyl acetate to extract 3 times, each 30ml, merges extracting solution, is evaporated, residue adds Methanol 3ml dissolves, as test solution；
(2) hairyvein agrimony control medicinal material 12g is taken, adds water 75ml, decocts 1.5h, filtration, filtrate is evaporated, and residue adds 0.8% hydrogen Sodium hydroxide solution 45ml, warm make dissolving, filter, and control medicinal material solution is made with method extraction with test solution in filtrate；
(3) each 15 μ l of above two solution are drawn, using water saturated toluene: ethyl acetate: mixing of the formic acid as 8: 4: 1.5 Solution is solvent, carries out thin-layered chromatography experiment, sprays and is developed the color with 1.5% ferric trichloride ethanol solution, in test sample chromatography, On position corresponding with control medicinal material chromatography, the spot of same color is shown；
E, to the inspection of moisture：The content of capsule is taken, is measured according to aquametry, 9.0% must not be crossed.
F, to the inspection of content uniformity：For every loading amount compared with mark loading amount, content uniformity limit should be in mark loading amount ± 10% within, must not be more than 2 beyond content uniformity limit, and must not have 11 times of overrun；
G, it is specially to the inspection of disintegration time limited：Using《Chinese Pharmacopoeia 2015 editions》Defined disintegration time limited inspection technique inspection It looks into, regulation should be met；
H, to the inspection of microbial limit：Using《Chinese Pharmacopoeia 2015 editions》Defined bacterium, mould and saccharomycete inspection Method and Control bacteria examination method inspection, should meet regulation；
I, to the measure of paeoniflorin content：
(1) preparation of test solution：Capsule 's content 0.15g is taken, it is accurately weighed, it puts in tool plug conical flask, it is accurate 55% methanol 55ml is added in, weighed weight after cold soaking is stayed overnight, is ultrasonically treated 40 minutes under the conditions of 300w, 50KHz, is re-weighed, The weight of less loss is supplied with 55% methanol, is shaken up, the filtration of 0.45um miillpore filters, filtrate is as test solution；
(2) preparation of reference substance solution：Precision weighs Paeoniflorin reference substance, adds 55% methanol that every 1ml is made containing 0.15mg Solution, as reference substance solution；
(3) it is accurate respectively to draw test solution and each 12ul of reference substance solution, liquid chromatograph is injected, is measured, is calculated, To obtain the final product；Wherein, using octadecylsilane chemically bonded silica as stationary phase, with volume ratio 65：35 0.06mol/L potassium dihydrogen phosphates are molten Liquid-methanol is mobile phase, and Detection wavelength 230nm, number of theoretical plate is calculated by Paeoniflorin peak should be not less than 2500；Every contains the radix paeoniae rubrathe With Paeoniflorin C23H28O11Meter, must not be less than 12.0mg.
The present embodiment uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient For identification result it is found that 3 groups of test samples show the spot of same color with motherwort to impinging upon same position, negative control is noiseless, For panel without hangover, target blob is clear.
According to the radix paeoniae rubrathe, Radix Notoginseng and hairyvein agrimony identification result it is found that each group sample with accordingly to impinging upon same position The spot of aobvious same color, negative control is noiseless, and for panel without hangover, target blob is clear；According to moisture, content uniformity, Disintegration time limited, microbial limit inspection result it is found that each group sample meets regulation；According to the measure knot to paeoniflorin content Fruit is it is found that Paeoniflorin C in each group sample23H28O11Content be no less than 12.0mg.
It is detected according to quality determining method provided in this embodiment, product batch number is respectively 20130701, 20130901st, the gynaecology of 20,131,101 three different batches break it is red drink capsule it is qualified.
Embodiment 4
Compared with Example 1, it differs only in, the step of motherwort ingredient differentiates (1) is specially：Gynaecology is taken to break red Capsule 's content 1.4g is drunk, is dissolved in 25ml absolute ethyl alcohols, 80 DEG C of refluxing extraction 50min are let cool, filtering and collecting filter liquid, water-bath It is evaporated, residue obtained plus 7ml water dissolutions, with 3000 revs/min of centrifugation 8min, takes supernatant, add the anhydrous second of 5ml after water bath method Alcohol dissolves, and is splined on the activated carbon-alumina column soaked with absolute ethyl alcohol, the internal diameter 20mm of the activated carbon-alumina column, Filler is uniformly mixed by activated carbon 0.4g and 200~300 mesh aluminium oxide 1.5g；It is washed with being collected after 20ml anhydrous ethanol elutions De- liquid, is evaporated, and residue obtained 1ml absolute ethyl alcohols dissolve, and take supernatant as test solution；Remaining step and embodiment 1 It is identical.
The present embodiment uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient Identification result is it is found that 3 groups of test samples show the spot of same color with motherwort to impinging upon same position, and negative control is without dry It disturbs；Clear spot degree is slightly worse than Examples 1 to 3.
Embodiment 5
Compared with Example 1, it differs only in, the step of motherwort ingredient differentiates (1) is specially：Gynaecology is taken to break red Capsule 's content 1.8g is drunk, is dissolved in 35ml absolute ethyl alcohols, 100 DEG C of refluxing extraction 70min are let cool, filtering and collecting filter liquid, water-bath It is evaporated, residue obtained plus 10ml water dissolutions, with 5000 revs/min of centrifugation 12min, takes supernatant, add 5ml anhydrous after water bath method Ethyl alcohol dissolves, and is splined on the activated carbon-alumina column soaked with absolute ethyl alcohol, the internal diameter 15mm of the activated carbon-alumina column, Its filler is uniformly mixed by activated carbon 0.6g and 200~300 mesh aluminium oxide 2.5g；With being collected after 40ml anhydrous ethanol elutions Eluent is evaporated, and residue obtained 1ml absolute ethyl alcohols dissolve, and take supernatant as test solution；Remaining step and embodiment 1 is identical.
The present embodiment uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient Identification result is it is found that 3 groups of test samples show the spot of same color with motherwort to impinging upon same position, and negative control is without dry It disturbs；Clear spot degree is slightly worse than Examples 1 to 3.
Comparative example 1
Compared with Example 1, it differs only in, in the step of motherwort ingredient differentiates (1), gynaecology is taken to break red drink glue Intracapsular tolerant 1.2g is dissolved in 30ml absolute ethyl alcohols, and 90 DEG C of refluxing extraction 60min are let cool, filtering and collecting filter liquid, water bath method, Residue obtained plus 7ml water dissolutions with 5500 revs/min of centrifugation 8min, take supernatant；Remaining operation is same as Example 1.
This comparative example uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient Identification result using this comparative example the method it is found that differentiated, the R between target blobfIt is worth variant, it is difficult to accurately sentence Whether the spot that cut-off test product is compareed with motherwort is in same position, and clear spot degree is worse than Examples 1 to 5.
Comparative example 2
Compared with Example 1, it differs only in, in the step of motherwort ingredient differentiates (1), using supersound process side Legal system available test agent solution, specially：The capsule 's content 1.6g is taken, ethyl alcohol 30ml, ultrasonic extraction 45min is added to let cool, Filtering, filtrate water bath method, residue add 8ml water dissolutions, with 4000 revs/min of centrifugation 10min, take supernatant, water-bath is steamed near It is dry, 5ml ethyl alcohol is added to dissolve, it is splined on the activated carbon-alumina column soaked with absolute ethyl alcohol；Remaining step and 1 phase of embodiment Together.
This comparative example uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient Identification result using this comparative example the method it is found that differentiated, the R between target blobfIt is worth variant, it is difficult to accurately sentence Whether the spot that cut-off test product is compareed with motherwort is in same position, and clear spot degree is worse than Examples 1 to 5.
Comparative example 3
Compared with Example 1, it differs only in, in the step of motherwort ingredient differentiates (1), using patent document Test sample processing method disclosed in CN101919942A prepares test sample solution, specially：Take the capsule 's content 1.2g adds ethyl alcohol 30mL, is heated to reflux 1h, lets cool, and filtration, filtrate is concentrated into 5mL, be added on internal diameter 10mm, activated carbon 0.5g, On activated carbon-alumina column of -120 mesh neutral alumina 2g of 100 mesh, with 30mL ethanol elutions, eluent is collected, is evaporated, it is residual Slag adds 0.5mL ethyl alcohol to dissolve, as test solution；Remaining step is the same as embodiment 1.
This comparative example uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule, the mirror of motherwort ingredient The results are shown in Figure 2 (sample and the corresponding position of reference substance are with embodiment 1).As shown in Figure 2, using described in this comparative example Method differentiated, serious hangover occurs in when panel, and test sample is difficult to realize efficiently separate, and target blob obscures；And target spot R between pointfIt is worth variant, it is difficult to which whether the spot that accurate judgement test sample is compareed with motherwort is in same position.
Comparative example 4
Compared with Example 1, it differs only in, in the step of motherwort ingredient differentiates (3), using patent document Thin-layered chromatography is differentiated disclosed in CN101919942A, specially：With n-butyl alcohol-hydrochloric acid-water (4:1:0.5) it is expansion Agent is unfolded, and takes out, dries, and sprays with dilute bismuth potassium iodide solution to spot development, is inspected under daylight；Remaining step is the same as embodiment 1.
This comparative example uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient Identification result using this comparative example the method it is found that differentiated, duration of run need to (Examples 1 to 5 only needs 1 in 6~8 hours ~1.5 hours), color developing agent need to be sprayed repeatedly, and developing time is up to 2-3 days (Examples 1 to 5 can develop the color at once), and is difficult to accurately Judging test sample, whether color is identical with spot that motherwort compares, and clear spot degree is worse than Examples 1 to 5.
Comparative example 5
Using《Chinese Pharmacopoeia 2015 editions》First discrimination method of motherwort disclosed in page 290 breaks to gynaecology red drink glue Capsule is detected；Wherein：
Test solution preparation method is to take the capsule 's content 1.2g, adds ethyl alcohol 30mL, is heated to reflux 1h, lets cool, Filtering, takes filtrate 10ml to be evaporated, and residue adds absolute ethyl alcohol 1ml to make dissolving, and centrifugation takes supernatant as test solution；
Stachydrine hydrochloride reference substance separately is taken, adds absolute ethyl alcohol that solution of every lml containing 1mg is made, as reference substance solution；
It is according to thin-layered chromatography：Each 5~10 μ l of above two solution are drawn, are put respectively on same silica gel g thin-layer plate, with Acetone-absolute ethyl alcohol-hydrochloric acid (10:6:1) for solvent, be unfolded, take out, dry, heat 15 minutes, let cool at 105 DEG C, spray with Dilute bismuth potassium iodide test solution-ferric trichloride test solution (10:1) mixed solution is clear to spot development.
This comparative example uses above method, authenticated the product batch number of Zhuzhou Qianjin Pharmacy Co., Ltd's production respectively The gynaecology of respectively 20130701,20130901,20,131,101 three different batches breaks red drink capsule；By motherwort ingredient For identification result it is found that being differentiated using this comparative example the method, effect is similar with comparative example 3, occurs during panel seriously dragging Tail, test sample are difficult to realize efficiently separate, and target blob obscures；And the R between target blobfIt is worth variant, it is difficult to accurately sentence Whether the spot that cut-off test product is compareed with motherwort is in same position.
Experimental example 1：Different thin layer silica gel plate durabilities are investigated
The method provided using embodiment 1, differentiates motherwort ingredient in the drug of three batches.Wherein, step (3) self-control thin layer silica G plate is used respectively, is bought from the prefabricated sheets silica G plate of Qingdao Haiyang group and High Performance Thin silica G plate Experiment, 10 μ l of point sample amount；Identification result is as shown in Figure 3.
As a result it shows：Using different silica G plates, in test sample chromatography with reference substance and control medicinal material chromatography corresponding position The spot of aobvious same color, negative control is noiseless.Illustrate that this discrimination method has preferable durability to different silica G plates.
Experimental example 2：The effect expedition differentiated under condition of different temperatures
The method provided using embodiment 1, differentiates motherwort ingredient in the drug of three batches.Wherein, step (3) it is unfolded under 5 DEG C and 30 DEG C of environment temperature respectively；Identification result is as shown in Figure 4.
As a result it shows：According to being tested under the conditions of above-mentioned not equality of temperature, in test sample chromatography with reference substance and control medicinal material Chromatography corresponding position shows the spot of same color, and clear spot, and negative control is noiseless, illustrates this discrimination method different There is preferable durability under ambient temperature conditions.
Experimental example 3：The thin-layer chromatography condition durability being unfolded under the conditions of different humidity is investigated
The method provided using embodiment 1, differentiates motherwort ingredient in the drug of three batches.Wherein, step (3) it is unfolded under 30% and 75% ambient humidity respectively；Identification result is as shown in Figure 5.
As a result it shows：According to being tested under above-mentioned different humidity part, in test sample chromatography with reference substance and control medicinal material Chromatography corresponding position shows the spot of same color, and clear spot, and negative control is noiseless, illustrates this discrimination method different There is preferable durability under environmental damp condition.
Scheme disclosed in the embodiment 6,7 and 8 of patent document CN101919942A is respectively adopted in this experimental example, 75% Quality testing is carried out under ambient humidity.
The results show that when comparison Brittle Falsepimpernel Herb is differentiated, the method that patent document CN101919942A is provided is 75% It is difficult to realize efficiently separate under environmental damp condition, in long-time and develop the color；In comparison, the embodiment of the present invention 1~5 provides Method, it is only necessary to be unfolded 1~1.5 hour, 3~6h after spray color developing agent, you can obtain clear and can be used for the spot accurately differentiated Point.
Further relatively it is found that in the environment of the high humility, the method that the embodiment of the present invention 1~5 provides is to benefit Brittle Falsepimpernel Herb can obtain the spot of complete display when being differentiated, the time required to expansion and acquisition is clear available for accurately differentiating Developing time needed for spot is significantly shorter than in comparative example 1~5 to the discriminating of motherwort.
It is influenced by ambient humidity small since the present invention provides discrimination method, especially main component motherwort is differentiated When stability it is high, clear spot, hence it is evident that shorten and differentiate the time, it is throughout the year moist more to be applicable to south China most area The larger environment of rain, humidity, for gynaecology break it is red drink capsule it is quick, stablize differentiate provide important effective means.
Although above having used general explanation, specific embodiment and experiment, the present invention is made to retouch in detail It states, but on the basis of the present invention, it can be made some modifications or improvements, this is apparent to those skilled in the art 's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed Range.

Claims (9)

  1. The quality determining method of red drink capsule 1. gynaecology breaks, which is characterized in that including：To active constituent motherwort, the radix paeoniae rubrathe, Radix Notoginseng With the discriminating of hairyvein agrimony, inspection to moisture, content uniformity, disintegration time limited and microbial limit and to paeoniflorin content It measures；
    The discrimination method of the active constituent motherwort includes the following steps：
    (1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.5g~1.7g, is dissolved in the anhydrous second of 28ml~32ml Alcohol, 80~100 DEG C of refluxing extraction 50min~70min, lets cool, filtering and collecting filter liquid, water bath method, and residue obtained plus 7ml~ 9ml water dissolutions with 3800~4200 revs/min of centrifugation 8min~12min, take supernatant, water bath method, be splined on activated carbon- Alumina column with eluent is collected after anhydrous ethanol elution, is evaporated, residue obtained to be dissolved with absolute ethyl alcohol, takes supernatant conduct Test solution；
    (2) preparation of reference substance solution；
    (3) differentiate according to thin-layered chromatography；
    Step (3) solvent used according to thin-layered chromatography discriminating is by acetone, absolute ethyl alcohol and hydrochloric acid with volume ratio 10：6： 1 composition；
    Step (3) is described to be differentiated according to thin-layered chromatography, after being unfolded using solvent, drying, first heats 10 at 100~110 DEG C ~20min is sprayed after letting cool with 8~12% ethanol solution of sulfuric acid, and 2~4min are dried at 100~110 DEG C, then spray with color developing agent into Row colour developing；
    Step (3) is described to differentiate the color developing agent used by dilute bismuth potassium iodide solution and 1% ferric trichloride ethyl alcohol according to thin-layered chromatography Solution is with volume ratio 10:1 mixes.
  2. 2. according to the method described in claim 1, it is characterized in that, the filler of step (1) activated carbon-alumina column is by living Property charcoal and 200~300 mesh neutral aluminas are with weight ratio 4~6：15~25 are uniformly mixed.
  3. 3. according to the method described in claim 2, it is characterized in that, the filler of step (1) activated carbon-alumina column is by living Property 0.4~0.6g of charcoal and 200~300 1.5~2.5g of mesh aluminium oxide is uniformly mixed.
  4. 4. according to the method in claim 2 or 3, which is characterized in that the internal diameter of step (1) activated carbon-alumina column For 15mm or 20mm；Described with the elution volume of anhydrous ethanol elution is 20ml~40ml.
  5. 5. according to the method described in claim 4, it is characterized in that, the internal diameter of step (1) activated carbon-alumina column is 20mm；Described with the elution volume of anhydrous ethanol elution is 28ml~32ml.
  6. 6. according to the method described in claims 1 to 3 any one, which is characterized in that step (2) described reference substance solution includes Motherwort contrast solution and stachydrine hydrochloride reference substance solution；
    The preparation method of the motherwort contrast solution is：Take motherwort medicinal material 2.9g~3.1g, using with to prepare test sample molten It is prepared by the same method of liquid phase；
    The preparation method of the stachydrine hydrochloride reference substance solution is：Stachydrine hydrochloride reference substance is taken, absolute ethyl alcohol is added to be dissolved to A concentration of 4.8~5.2mg/ml, you can.
  7. 7. according to the method described in claim 1, it is characterized in that, the discrimination method of the active constituent motherwort is including as follows Step：
    (1) preparation of test solution：Gynaecology is taken to break red drink 1.4~1.8g of capsule 's content, is dissolved in 25~35ml absolute ethyl alcohols, 80~100 DEG C of refluxing extraction 50min~70min, let cool, filtering and collecting filter liquid, water bath method, and residue obtained plus 7ml~ 10ml water dissolutions with 3000~5000 revs/min of centrifugation 8min~12min, take supernatant, water bath method, be splined on activated carbon- Alumina column, the internal diameter of the activated carbon-alumina column are 15mm or 20mm, filler by 0.4~0.6g of activated carbon and 200~ 300 1.5~2.5g of mesh aluminium oxide are uniformly mixed；With eluent is collected after 20ml~40ml anhydrous ethanol elutions, it is evaporated, institute It obtains residue 0.5ml~1.5ml absolute ethyl alcohols to dissolve, takes supernatant as test solution；
    (2) preparation of motherwort contrast solution：Motherwort medicinal material 2.9g~3.1g is taken, using identical with preparing test solution Method prepares motherwort contrast solution；
    The preparation of stachydrine hydrochloride reference substance solution：Take stachydrine hydrochloride reference substance, add absolute ethyl alcohol be dissolved to a concentration of 4.8~ 5.2mg/ml；
    (3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution and stachydrine hydrochloride reference substance solution Each 10~15 μ l, parallel point sample is on same silica G plate, using volume ratio as 10：6：1 acetone-absolute ethyl alcohol-hydrochloric acid is exhibition Agent expansion is opened, after drying, 10~20min is heated at 100~110 DEG C, lets cool, spray with 8~12% ethanol solution of sulfuric acid, 100 ~110 DEG C of 2~4min of drying, then spray using volume ratio as 10:1-1% ferric trichloride ethyl alcohol of dilute bismuth potassium iodide test solution mixing is molten Liquid is inspected under daylight, you can.
  8. 8. according to the method described in claim 1, it is characterized in that, the discrimination method of the active constituent motherwort is including as follows Step：
    (1) preparation of test solution：Gynaecology is taken to break red drink capsule 's content 1.5g~1.7g, is dissolved in the anhydrous second of 28ml~32ml Alcohol, 80~100 DEG C of refluxing extraction 50min~70min, lets cool, filtering and collecting filter liquid, water bath method, and residue obtained plus 7ml~ 9ml water dissolutions with 3800~4200 revs/min of centrifugation 8min~12min, take supernatant, water bath method, be splined on activated carbon- Alumina column, the internal diameter of the activated carbon-alumina column are 15mm or 20mm, filler by 0.4~0.6g of activated carbon and 200~ 300 1.5~2.5g of mesh aluminium oxide are uniformly mixed；With eluent is collected after 28ml~32ml anhydrous ethanol elutions, it is evaporated, institute It obtains residue 0.8ml~1.2ml absolute ethyl alcohols to dissolve, takes supernatant as test solution；
    (2) preparation of motherwort contrast solution：Motherwort medicinal material 2.9g~3.1g is taken, using identical with preparing test solution Method prepares motherwort contrast solution；
    The preparation of stachydrine hydrochloride reference substance solution：Take stachydrine hydrochloride reference substance, add absolute ethyl alcohol be dissolved to a concentration of 4.8~ 5.2mg/ml；
    (3) differentiate according to thin-layered chromatography：Take the test solution, motherwort contrast solution and stachydrine hydrochloride reference substance solution Each 10~15 μ l, parallel point sample is on same silica G plate, using volume ratio as 10：6：1 acetone-absolute ethyl alcohol-hydrochloric acid is exhibition Agent expansion is opened, after drying, 10~20min is heated at 100~110 DEG C, lets cool, spray with 8~12% ethanol solution of sulfuric acid, 100 ~110 DEG C of 2~4min of drying, then spray using volume ratio as 10:1-1% ferric trichloride ethyl alcohol of dilute bismuth potassium iodide test solution mixing is molten Liquid is inspected under daylight, you can.
  9. 9. according to the method described in any one of claims 1 to 3,7 or 8, which is characterized in that the mirror of the active constituent radix paeoniae rubrathe Do not include the following steps：
    (1) the capsule 's content 0.5g~1.5g is taken, adds water 12ml~38ml, is ultrasonically treated 15min~45min, is filtered, filter Liquid adds water saturated butanol solution to extract 1~5 time, each 10ml~30ml, merges extracting solution, ammonification test solution 20ml~60ml Washing, discards washing lotion, is evaporated, add 1ml~3ml methanol dissolved residues, as test solution；
    (2) radix paeoniae rubrathe control medicinal material 1g~3g is taken, adds methanol 15ml~45ml, is ultrasonically treated 0.5h~1.5h, filtration, filtrate is steamed Dry, residue adds water 12ml~38ml, and warm makes dissolving, filters, and filtrate extracts that control medicinal material is made is molten with test solution with method Liquid；Paeoniflorin reference substance is taken, adds methanol that the solution of a concentration of 1mg/ml~3mg/ml is made, as reference substance solution；
    (3) the μ l of above-mentioned each 3 μ l of three kinds of solution~5 are drawn, using chloroform: ethyl acetate: methanol: formic acid is 30~50: 2~8: 5~15: 0.1~0.3 mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays molten with 4~6% vanillin-sulfuric acid methanol Liquid, it is clear to be heated to spot development at 95 DEG C~115 DEG C, in test sample chromatography, with control medicinal material chromatography and reference substance chromatography On corresponding position, the spot of same color is shown；
    The discriminating of the active constituent Radix Notoginseng includes the following steps：
    (1) the capsule 's content 0.5g~1.5g is taken, adds water 12ml~38ml, is ultrasonically treated 15min~45min, is filtered, filter Liquid adds water saturated butanol solution to extract 1~5 time, each 10ml~30ml, merges extracting solution, ammonification test solution 20ml~60ml Washing, discards washing lotion, is evaporated, add 1ml~3ml methanol dissolved residues, as test solution；
    (2) take Radix Notoginseng control medicinal material 0.5g~1.5g, add water 5 drip~15 drop, stir evenly, add water saturated butanol solution 20ml~ 60ml, is ultrasonically treated 20min~60min, filtration, and filtrate ammonification test solution 20ml~60ml washings discard ammoniacal liquor, add in n-butanol Aqueous solution 20ml~60ml washings of saturation, discard n-butanol aqueous solution, are evaporated, and residue adds methanol 1ml~3ml to dissolve, as Control medicinal material solution；Ginsenoside Rb1, Rg1 and notoginsenoside R reference substance are taken, it is 0.5mg/ml to add methanol that each concentration is made The mixed solution of~1.5mg/ml is as reference substance solution；
    (3) above-mentioned each 1~3 μ l of three kinds of solution are drawn, using in 10 DEG C of overnight chloroforms arranged below: methanol: water as 10~ Lower floor's solution of 16: 4~10: 1~3 mixed liquor is solvent, carries out thin-layered chromatography experiment, sprays with 8~12% sulfuric acid second Alcoholic solution, it is clear to be heated to spot development at 95 DEG C~115 DEG C, in test sample chromatography, with control medicinal material chromatography and reference substance On the corresponding position of chromatography, the spot of same color is shown；It puts and is inspected under the ultraviolet lamp of 350nm wavelength, show the glimmering of same color Hot spot point；
    The discriminating of the active constituent hairyvein agrimony includes the following steps：
    (1) the capsule 's content 0.5g~1.5g is taken, adds 0.2%~0.8% sodium hydroxide solution 15ml~45ml, at ultrasound 10min~30min, filtration are managed, filtrate adds hydrochloric acid tune pH value, and filtration, filtrate adds ethyl acetate extraction 1~3 time, every time to 2~3 10ml~30ml merges extracting solution, is evaporated, and residue adds methanol 1ml~3ml to dissolve, as test solution；
    (2) hairyvein agrimony control medicinal material 4g~12g is taken, adds water 25ml~75ml, decocts 0.5h~1.5h, filtration, filtrate is evaporated, residual Slag adds 0.2%~0.8% sodium hydroxide solution 15ml~45ml, and warm makes dissolving, filters, and filtrate carries with test solution with method It takes and control medicinal material solution is made；
    (3) the μ l of above two solution each 14 μ l~16 are drawn, using water saturated toluene: ethyl acetate: formic acid is 4~8: 2~4: 0.5~1.5 mixed solution is solvent, carries out thin-layered chromatography experiment, sprays molten with 0.5%~1.5% ferric trichloride ethyl alcohol Liquid develops the color, and in test sample chromatography, on position corresponding with control medicinal material chromatography, shows the spot of same color；
    Inspection to moisture is specially：The content of capsule is taken, is measured according to aquametry, 9.0% must not be crossed；
    Inspection to content uniformity is specially：For every loading amount compared with mark loading amount, content uniformity limit should be in mark loading amount 10% within, must not be more than 2 beyond content uniformity limit, and must not have 11 times of overrun；
    Inspection to disintegration time limited is specially：Using disintegration time limited inspection technique inspection, regulation should be met；
    Inspection to microbial limit is specially：It, should using bacterium, mould and saccharomycete inspection technique and Control bacteria examination method inspection Meet regulation；
    The measure of paeoniflorin content is included the following steps：
    (1) preparation of test solution：Capsule 's content 0.05~0.15g is taken, it is accurately weighed, it puts in tool plug conical flask, essence 45~55% 45~55ml of methanol of close addition, weighed weight after cold soaking is stayed overnight, surpass under the conditions of 100~300w, 30~50KHz Sonication 20~40 minutes, is re-weighed, and the weight of less loss is supplied with 45~55% methanol, is shaken up, the filtration of 0.45um miillpore filters, Filtrate is as test solution；
    (2) preparation of reference substance solution：Precision weighs Paeoniflorin reference substance, add 45~55% methanol be made every 1ml containing 0.05~ The solution of 0.15mg, as reference substance solution；
    (3) it is accurate respectively to draw test solution and each 8~12ul of reference substance solution, liquid chromatograph is injected, is measured, is calculated, To obtain the final product；Wherein, using octadecylsilane chemically bonded silica as stationary phase, with volume ratio 65~75：The 0.04 of 25~35~ 0.06mol/L potassium dihydrogen phosphates-methanol is mobile phase, and Detection wavelength 230nm, number of theoretical plate is calculated by Paeoniflorin peak should Not less than 2500；Every containing the radix paeoniae rubrathe with Paeoniflorin C23H28O11Meter, must not be less than 12.0mg.
CN201610220326.4A 2016-04-11 2016-04-11 Gynaecology break it is red drink capsule quality determining method Active CN105784913B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610220326.4A CN105784913B (en) 2016-04-11 2016-04-11 Gynaecology break it is red drink capsule quality determining method

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610220326.4A CN105784913B (en) 2016-04-11 2016-04-11 Gynaecology break it is red drink capsule quality determining method

Publications (2)

Publication Number Publication Date
CN105784913A CN105784913A (en) 2016-07-20
CN105784913B true CN105784913B (en) 2018-07-06

Family

ID=56396133

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610220326.4A Active CN105784913B (en) 2016-04-11 2016-04-11 Gynaecology break it is red drink capsule quality determining method

Country Status (1)

Country Link
CN (1) CN105784913B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111735889B (en) * 2020-08-19 2021-02-09 广东一方制药有限公司 Quality detection method of dampness-resolving and toxin-vanquishing composition
CN111983106B (en) * 2020-08-19 2021-06-22 广东一方制药有限公司 Quality control method of dampness-resolving and toxin-vanquishing composition

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1973897B (en) * 2006-12-21 2010-04-14 贵州益佰制药股份有限公司 Quality control method for puerperal blood clot dispersing tablet
CN101919942B (en) * 2009-06-10 2012-12-19 株洲千金药业股份有限公司 Traditional Chinese medicine combination for treating gynecologic functional uterine bleeding diseases, preparation method and capsule quality control method thereof
CN102539588A (en) * 2011-11-11 2012-07-04 云南良方制药有限公司 Method for preparing test solution for quality detection of safe stagnation removing preparation
CN104161847B (en) * 2014-07-25 2016-08-24 北京汉典制药有限公司 A kind of quality determining method of the Chinese medicine composition treating diabetic retinopathy
CN104483402A (en) * 2014-12-02 2015-04-01 贵州汇正制药有限责任公司 Quality inspection method of anti-hysteritis soft capsules

Also Published As

Publication number Publication date
CN105784913A (en) 2016-07-20

Similar Documents

Publication Publication Date Title
CN107843677B (en) Radix paeoniae rubra control extract and preparation method and application thereof
CN102139008B (en) Quality control method of Xiaoer daochi tablet
CN106501442B (en) A kind of quality determining method of a kind of reed mentioned in ancient books Huang powder for clearing lung-heat
CN105784913B (en) Gynaecology break it is red drink capsule quality determining method
CN105301168B (en) The detection method of dredging collateral resolving sputum capsule
CN105241997A (en) Thin-layer chromatography identification method of carbon traditional Chinese medicine formula granules
CN103940972B (en) A kind of detection method for the treatment of hypertensive Chinese medicine preparation
CN106770882B (en) A kind of detection method of the Chinese medicine preparation containing ginseng or pseudo-ginseng
CN107315061A (en) A kind of method of quality control for the alizarin root of Dahurian angelica Chinese medicine preparation for treating uterus bleeding
CN102091297A (en) Quality control method for liver health care medicine
CN105911210B (en) Gynaecology break it is red drink capsule in motherwort composition discrimination method
CN107727787A (en) A kind of TLC Identification for differentiating hymsleya amabilis kind
CN103076403B (en) Inspection method for capsule for treating lower urinary tract infection
CN105616946A (en) Preparation for treating cough, preparation method and quality control method thereof
CN114942297A (en) Developing agent for thin-layer identification method of Taohong Siwu decoction and thin-layer identification method
CN101168009B (en) Quality control method of macaque stone bovinebezoar powder
CN113759016B (en) Construction method of thin-layer chromatography, method for simultaneously identifying components of honeysuckle and liquorice and application of method
CN102662027B (en) Thin-layer chromatography identification method of caulis tinosporae sinensis formula granules
CN105738557A (en) Thin-layer chromatography detection method for icmomordin Ic in Fuyang granules
CN101181363A (en) Ginseng rhubarb injection and preparation method thereof
CN112730724A (en) Thin-layer identification method for cynanchum glaucescens formula granules
CN102139048B (en) Quality control method of tablet capable of improving eyesight and clearing heat
CN106770885B (en) A method of indentification by TLC is carried out to licorice ingredient in a kind of reed mentioned in ancient books Huang powder for clearing lung-heat
CN113759050B (en) Preparation method of rhizoma Dioscoreae Septemlobae test sample and thin layer identification method thereof
CN115389694B (en) Thin layer chromatography for simultaneously identifying Notopterygii rhizoma, radix Angelicae Pubescentis, radix Angelicae sinensis and rhizoma Ligustici Chuanxiong in Juanbi decoction

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant