CN101926889B - Method for detecting white paeony root-medlar particles - Google Patents
Method for detecting white paeony root-medlar particles Download PDFInfo
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Abstract
The invention discloses a method for detecting white paeony root-medlar particles which have the effects of nourishing the liver and promoting the function of gallbladder. The detection method comprises property, check, identification and content measurement items, wherein the identification refers to identification of thin layers of tangerine peel, tuber fleeceflower root, medlar and liquoric root in a preparation; and the content measurement refers to the measurement of the paeoniflorin content of white paeony root in the preparation. Since a scientific and reasonable quality control detection method for effectively controlling the product quality of the particles exists, a specific and practical component identification and content measurement method is provided by researching and selecting the identification check and content measurement of the particles according to the properties and preparation processes of Chinese medicinal plants in a formula. The adopted method has the advantages of establishing a quality control standard for a white paeony root-medlar particle preparation, effectively controlling the quality of the white paeony root-medlar particles and ensuring the clinical effect and safety of a medicament, along with high specificity, precision, repeatability and yield and correct measurement result.
Description
Technical field
The present invention relates to the quality determining method of a kind of Chinese herbaceous peony Qi particle (nourishing the liver choleretic granules) preparation, belong to the technical field of medicine being carried out quality control.
Background technology
Chronic infection of biliary tract, cholelithiasis are a kind of worldwide common diseases, have a strong impact on human beings'health for a long time, and its incidence of disease day by day raises in China, and according to many provinces and cities of China findings of the survey, it is fallen ill up to 18%, surpass American-European 10%.Brainstrust is thought, calculates by 1,000,000,000 populations 5% above middle age, has 2,500 ten thousand chronic infection of biliary tract, cholelithiasis patient at least in China.
The applicant is in application number is 200610021927.9 patent; Medicine of treatment infection of biliary tract and preparation method thereof has been protected in application; Its medicinal granule is preparation like this: root of herbaceous peony 750.0g, dried orange peel 562.5g, prepared fleece flower root 375.0g, fruit of Chinese wolfberry 375.0g, honey-fried licorice root 187.5g; Boiling three times, amount of water are respectively 6 times, 4 times, 4 times amounts.Decoct earlier for the first time and soaked 30 minutes, each boiling reflux 1 hour; Filter, merging filtrate, being evaporated to relative density is the clear cream of 1.06～1.08 (70～80 ℃), is chilled to room temperature; Centrifuging (3500～4000 rev/mins, centrifugal 20 minutes), it is the clear cream of 1.20～1.22 (75～80 ℃) that collection centrifugate and concentrating under reduced pressure become relative density, 60～70 ℃ of drying under reduced pressure; Dry extract is broken into fine powder, adds an amount of dextrin, mixing; Process particle with ethanol, drying is processed 1000g.This Chinese herbaceous peony Qi particle (nourishing the liver choleretic granules) is made up of the root of herbaceous peony, the fruit of Chinese wolfberry, the fleece-flower root, dried orange peel, honey-fried licorice root etc. 5 flavor medicines, the tool nourishing the liver effects such as liver, cholagogic pain relieving that soften.This medicine cures mainly chronic infection of biliary tract card and belongs to the deficiecny of liver-YIN person, and formulation is a granule.Because this medicine is a kind of new drug, still lack a kind of concrete detection method and the quality standard that can effectively control this granule quality of production at present, so can not guarantee validity, the security of clinical drug treatment.
Summary of the invention
The object of the invention; Be still to lack to above-mentioned existing nourishing the liver choleretic granules effectively to control product quality; And no scientific and reasonable quality determining method and control criterion; According to the characteristic and the preparation technology of nourishing the liver choleretic granules side Chinese crude drug, the identification check and the assay of this granule are studied screening repeatedly, the detection method of a kind of Chinese herbaceous peony Qi particle (nourishing the liver choleretic granules) preparation is provided.It is that feasible composition is differentiated, content assaying method, and the discrimination method specificity of being drafted is strong, and the assay accuracy is high; Favorable reproducibility; For production unit, testing agency provide detection index, detection means and technical method etc.,, make controlling of production process rationally strict more can better instruct production; And effectively control the quality of nourishing the liver choleretic granules, thereby guarantee the clinical efficacy and the security of this medicine.
White paeony root-medlar particles of the present invention is to constitute like this: it root of herbaceous peony 750.0g, dried orange peel 562.5g, prepared fleece flower root 375.0g, fruit of Chinese wolfberry 375.0g, honey-fried licorice root 187.5g and and suitably auxiliary material be prepared from.
The preparation method of white paeony root-medlar particles of the present invention: above five tastes medicinal material, boiling three times, amount of water is respectively 6 times, 4 times, 4 times amounts.Decoct earlier for the first time and soaked 30 minutes, each boiling reflux 1 hour; Filter, merging filtrate, being evaporated to relative density is the clear cream of 1.06～1.08 (70～80 ℃), is chilled to room temperature; Centrifuging (3500～4000 rev/mins, centrifugal 20 minutes), it is the clear cream of 1.20～1.22 (75～80 ℃) that collection centrifugate and concentrating under reduced pressure become relative density, 60～70 ℃ of drying under reduced pressure; Dry extract is broken into fine powder, adds an amount of dextrin, and mixing is processed particle with ethanol; Drying is processed 1000g, promptly gets.
Detection method according to the invention: mainly comprise proterties, inspection, discriminating and assay project; Wherein: discriminating be to the thin layer of dried orange peel in the said preparation differentiate, the thin layer of prepared fleece flower root is differentiated, the thin layer of the fruit of Chinese wolfberry is differentiated, the thin layer of contained ammonium glycyrrhetate is differentiated in the honey-fried licorice root; Assay is that the contained content of paeoniflorin of the root of herbaceous peony in the preparation is measured.
The discriminating of dried orange peel is to be reference substance with control medicinal material dried orange peel and reference substance aurantiamarin, and respectively with ethyl acetate: formic acid: water=100: 17: 13 and toluene: ethyl acetate: formic acid: water=20: 10: 1: 1 is that the thin-layer chromatography that the secondary of developping agent launches is differentiated.
The discriminating of prepared fleece flower root is to be reference substance with the control medicinal material fleece-flower root and reference substance archen, and with toluene: ethyl acetate: formic acid=15: 2: 1 is that the thin-layer chromatography of developping agent is differentiated.
The discriminating of the fruit of Chinese wolfberry is to be reference substance with the control medicinal material fruit of Chinese wolfberry, and with sherwood oil (60～90 ℃): ethyl acetate: glacial acetic acid=10: 7: 0.5 is that the thin-layer chromatography of developping agent is differentiated.
The discriminating of honey-fried licorice root is to be contrast with the reference substance ammonium glycyrrhetate, and with normal butyl alcohol: glacial acetic acid: water=6: 1: 3 is that the thin-layer chromatography of developping agent is differentiated.
It is to be contrast with the reference substance Paeoniflorin that content of paeoniflorin is measured, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is the high performance liquid chromatography of moving phase.
Concrete discrimination method comprises following project:
(1) thin layer of dried orange peel is differentiated
These article of getting are an amount of, and porphyrize adds methyl alcohol, and sonicated filters, and filtrating is steamed near and done, and residue adds methyl alcohol makes dissolving, as need testing solution; Other gets the dried orange peel control medicinal material, shines medicinal material solution in pairs with legal system; Get the aurantiamarin reference substance again, add an amount of methyl alcohol and process reference substance solution; Draw above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate with sodium hydroxide solution preparation, with ethyl acetate: methyl alcohol: water=100: 17: 13 is developping agent; Launch, take out, dry; Again with toluene: ethyl acetate: formic acid: water=20: 10: 1: 1 upper solution is a developping agent, launches, and takes out; Dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
(2) thin layer of prepared fleece flower root is differentiated
These article of getting are an amount of, and porphyrize adds methyl alcohol, and sonicated filters, the filtrating evaporate to dryness, and residue is with the methenyl choloride washing for several times, and is closely colourless to methenyl choloride, merges the methenyl choloride washing lotion, concentrates, as need testing solution; Other gets fleece-flower root control medicinal material, shines medicinal material solution in pairs with legal system; Get the archen reference substance again, add an amount of methyl alcohol and process reference substance solution; Draw above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene: ethyl acetate: formic acid=15: 2: 1 is developping agent, launches, and takes out, and dries, and puts under the ultraviolet lamp (365nm) and inspects; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
(3) thin layer of the fruit of Chinese wolfberry is differentiated
These article of getting are an amount of, and porphyrize adds methyl alcohol, and sonicated filters, the filtrating evaporate to dryness, and residue adds water makes dissolving, uses ethyl acetate extraction, merges extract, and evaporate to dryness, residue add ethyl acetate makes dissolving, as need testing solution; Other gets fruit of Chinese wolfberry control medicinal material, adds water, boils, and filters, and filtrating is shone medicinal material solution in pairs with legal system; Draw above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of bonding agent with the sodium carboxymethyl cellulose, with sherwood oil: ethyl acetate: glacial acetic acid=10: 7: 0.5 is a developping agent; Launch, take out, dry; Put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with control medicinal material chromatogram same position on, show the fluorescence spot of same color.
(4) thin layer of honey-fried licorice root is differentiated
These article of getting, porphyrize adds ethanol, and sonicated filters, the filtrating evaporate to dryness; Add water and make dissolving, be transferred in the centrifuge tube, add watery hydrochloric acid again, sonicated leaves standstill, and is centrifugal; Taking precipitate adds Diluted Alcohol and makes dissolving, regulates pH to neutral with sodium bicarbonate solution, and is centrifugal, gets supernatant, as need testing solution; In addition extracting liquorice acid ammonium reference substance adds Diluted Alcohol and processes reference substance solution, draws above-mentioned two kinds of solution, put respectively in same be the silica G F of bonding agent with the sodium carboxymethyl cellulose
254On the thin layer plate, with normal butyl alcohol: glacial acetic acid: water=6: 1: 3 upper solution are developping agent, launch, and take out, and dry, put under the ultraviolet lamp (254nm) and inspect, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.
Concrete content of paeoniflorin is measured:
With octadecylsilane key and silica gel is filling agent, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is moving phase, and the detection wavelength is 230nm, and theoretical cam curve is calculated by the Paeoniflorin peak should be not less than 2000; It is an amount of that precision takes by weighing the Paeoniflorin reference substance, adds methyl alcohol and process reference substance solution; These article of getting, porphyrize, the accurate title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol that adds, close plug is claimed to decide weight, and sonicated is claimed to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and subsequent filtrate is got in filtration, promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects liquid chromatograph, measures, and promptly gets, and these article contain the root of herbaceous peony in Paeoniflorin for every bag, must not be less than 48.0mg.
Discrimination method comprises following project more specifically:
(1) thin layer of dried orange peel is differentiated:
These article of getting are an amount of, and porphyrize is got 3g, adds methyl alcohol 30ml, and sonicated 20 minutes filters, and filtrating is steamed near and done, and residue adds methyl alcohol 3ml makes dissolving, as need testing solution; Other gets dried orange peel control medicinal material 2g, shines medicinal material solution in pairs with legal system; Get the aurantiamarin reference substance again, add an amount of methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; Draw above-mentioned three kinds of each 5ul of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ethyl acetate: methyl alcohol: water=100: 17: 13 is developping agent; Exhibition is taken out to about 3cm, dries; Again with toluene: ethyl acetate: formic acid: water=20: 10: 1: 1 upper solution is a developping agent, and exhibition is taken out to about 8cm; Dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
(2) thin layer of prepared fleece flower root is differentiated
These article of getting are an amount of, and porphyrize is got 10g, adds methyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue is with the methenyl choloride washing for several times, and is closely colourless to methenyl choloride, merges the methenyl choloride washing lotion, is concentrated into about 1ml, as need testing solution; Other gets fleece-flower root control medicinal material 0.5g, shines medicinal material solution in pairs with legal system; Get the archen reference substance again, add an amount of methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) experiment, draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate; With toluene: ethyl acetate: formic acid=15: 2: 1 is developping agent, launches, and takes out; Dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
(3) thin layer of the fruit of Chinese wolfberry is differentiated
These article of getting are an amount of, and porphyrize is got 0.5g, adds methyl alcohol 10ml, sonicated 5 minutes; Filter, the filtrating evaporate to dryness, residue adds water 10ml makes dissolving, with ethyl acetate extraction twice, each 10ml; Combined ethyl acetate extract, evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets fruit of Chinese wolfberry control medicinal material 1g, adds water 30ml, boils 15 minutes, filters, and filtrating is shone medicinal material solution in pairs with legal system; According to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) experiment, draw each 2～3ul of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of bonding agent with the sodium carboxymethyl cellulose; With sherwood oil (60～90 ℃): ethyl acetate: glacial acetic acid=10: 7: 0.5 is a developping agent, launches (30～35 ℃, saturated in advance 20 minutes); Take out; Dry, put under the ultraviolet lamp (365nm) and inspect, in the test sample chromatogram; With control medicinal material chromatogram same position on, show the fluorescence spot of same color.
(4) thin layer of honey-fried licorice root is differentiated
These article of getting 5g, porphyrize adds ethanol 30ml, and sonicated 15 minutes filters; The filtrating evaporate to dryness adds water 20ml and makes dissolving, is transferred in the centrifuge tube, adds watery hydrochloric acid 5ml again, ultrasonic 5 minutes; Leave standstill, centrifugal, taking precipitate adds Diluted Alcohol 1ml and makes dissolving; Regulate pH to neutral with 10% sodium bicarbonate solution, centrifugal, get supernatant, as need testing solution; The sour ammonium reference substance of other extracting liquorice; Add Diluted Alcohol and process the solution that every 1ml contains 1mg, reference substance solution is tested according to thin-layered chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005) the most; Draw above-mentioned two kinds of solution 5ul, put respectively in same be the silica G F of bonding agent with the sodium carboxymethyl cellulose
254On the thin layer plate, with normal butyl alcohol: glacial acetic acid: water=6: 1: 3 upper solution are developping agent, launch, and taking-up is dried, put under the ultraviolet lamp (254nm) and inspect, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
It is to be reference substance with the Paeoniflorin that content of paeoniflorin is measured, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is the high performance liquid chromatography of moving phase.
The contained content of paeoniflorin of the root of herbaceous peony is measured more specifically
With octadecylsilane key and silica gel is filling agent, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is moving phase, and the detection wavelength is 230nm, and theoretical cam curve is calculated by the Paeoniflorin peak should be not less than 2000; It is an amount of that precision takes by weighing the Paeoniflorin reference substance, adds methyl alcohol and process the reference substance solution that 1ml contains 0.05mg; These article of getting, porphyrize is got about 0.2g, and accurate the title, decide; Put in the 50ml tool plug conical flask, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Power 135W and frequency 59hHz sonicated 30 minutes are claimed to decide weight again, supply the weight that subtracts mistake with methyl alcohol, shake up; Filter, get subsequent filtrate, promptly get need testing solution; Accurate respectively reference substance solution and each 10ul of need testing solution of drawing injects liquid chromatograph, measures, and promptly gets, and these article contain the root of herbaceous peony in Paeoniflorin for every bag, must not be less than 48.0mg;
Differentiate and content assaying method science, reasonable, feasible that in order to ensure composition of the present invention the applicant studies the discriminating and the content assaying method of method Chinese medicine article composition, concrete testing data is following:
One, the thin layer Study on Identification of dried orange peel
With reference to " the thin layer discrimination method of the Chinese crude drug dried orange peel that one one of Chinese pharmacopoeia is recorded; With the aurantiamarin is reference substance; With ethyl acetate-methanol-water (100: 17: 13), toluene-ethyl acetate-formic acid-water (20: 10: 1: be that the developping agent secondary launches 1); Put under the ultraviolet lamp (365nm) and inspect, feasible through six lot sample article experimental observations conclusive evidence this law, interference is not seen in the negative control test.Aurantiamarin Rf value is about 0.2; Separate better revenue standard text (8cm, Rf=0.15～0.20) between the spot.
Two, the thin layer Study on Identification of prepared fleece flower root
Process of preparing Chinese medicine processed goods for polygonaceae plant tuber of multiflower knotweed dried root.The fleece-flower root mainly contains anthraquinone components such as archen.This law is reference substance with the archen, toluene: ethyl acetate: formic acid=15: 2: 1 is developping agent, and exhibition is apart from 8cm; Put under the uviol lamp (365nm) and inspect; Clear spot separates better with proximate spot, and the Rf value is about 0.4; Feasible through six lot sample article experimental observations conclusive evidence this law, interference is not seen in the negative control test.The revenue standard text (8cm, Rf=0.4).
Three, the thin layer Study on Identification of the fruit of Chinese wolfberry
With reference to " the Chinese crude drug matrimony vine subitem that records of one of Chinese pharmacopoeia thin layer discrimination method is down got fruit of Chinese wolfberry water extract, processes need testing solution with ethyl acetate jolting extraction; Get control medicinal material solution with fruit of Chinese wolfberry control medicinal material with legal system, point sample is on same silica gel g thin-layer plate, with ethyl acetate: chloroform: methyl alcohol (3: 2: 1) is developping agent; Exhibition is put under the ultraviolet lamp (365nm) and is inspected apart from 12cm, in supplier's article chromatogram; With the corresponding position of control medicinal material chromatogram on; The blue-fluorescence spot that shows same color, Rf is about 0.8, but sample point is not too clear.Adopt Reinecke's salt deposition betaine HCL in the research again, to remove partial impurities, the purifying need testing solution.Concrete operations are following: get fruit of Chinese wolfberry 5g, adding distil water 30ml transfers PH=1 with hydrochloric acid; Add activated charcoal 1.0g and boil, the cold filtration mistake, filtrating adds freshly prepared 2% Reinecke's salt solution 20ml; Stir, be put in the refrigerator 3 hours, filter with the G4 sintered glass funnel; Deposition adds acetone 1ml and makes dissolving after washing with small amount of ice water, makes need testing solution; Other gets fruit of Chinese wolfberry control medicinal material 2g, shines medicinal material solution in pairs with legal system; Get the betaine HCL reference substance again, add proper amount of acetone and process the solution that every 1ml contains 0.5mg betaine HCL reference substance, as reference substance solution.According to thin-layered chromatography, draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, be developping agent with acetone-absolute ethyl alcohol-hydrochloric acid (10: 6: 1), launch, take out, to dry, spray develops the color with Dragendorff's reagent.In the test sample chromatogram, on control medicinal material chromatogram and the corresponding position of reference substance chromatogram, show the spot of same color, blank noiseless.Betaine Rf value is about 0.48, shows the salmon pink spot, and the betaine spot is well separated with the spot of other compositions in the sample.But this method complex operation, and the granule sample filters difficulty.Finally abandon.
After restudy pre-treatment mode and the development system of having examined or check test sample, be specially: get these article fine powder 0.5g, add methyl alcohol 10ml, sonicated 5 minutes; Filter, the filtrating evaporate to dryness, residue adds water 10ml makes dissolving; With ethyl acetate extraction twice, each 10ml merges extract; Evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets fruit of Chinese wolfberry control medicinal material 1g, adds water 30ml, boils 15 minutes, filters, and filtrating is shone medicinal material solution in pairs with legal system; Draw each 2～3ul of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of bonding agent with the sodium carboxymethyl cellulose, with sherwood oil (60～90 ℃): ethyl acetate: glacial acetic acid=10: 7: 0.5 is a developping agent; Launch (30～35 ℃, saturated in advance 20 minutes), exhibition is apart from 8cm; Take out, dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with control medicinal material chromatogram same position on, show the fluorescence spot of same color; Interference is not seen in the negative control test.The revenue standard text (8cm, Rf=0.48).
Four, the thin layer Study on Identification of honey-fried licorice root
Other medicinal materials are bigger to the thin layer discrimination test interference of honey-fried licorice root in the nourishing the liver choleretic granules prescription, with reference to " thin layer discrimination method under Radix Glycyrrhizae item of Chinese pharmacopoeia, with the ether heating and extracting, the extracting n-butyl alcohol purifying; Evaporate to dryness, residue add methyl alcohol makes dissolving, and as need testing solution, point sample is on the silica gel g thin-layer plate of same usefulness 1% NaOH preparation; With ethyl acetate: formic acid: glacial acetic acid: water (15: 1: 1: 2) be developping agent, launch that exhibition is apart from 8cm; Take out, dry, spray is with 10% ethanol solution of sulfuric acid; It is clear to be heated to spot colour developing at 105 ℃, puts under the ultraviolet lamp (365nm) and inspects, in the test sample chromatogram; With control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color, the Rf value is about 0.2.But negative control has certain interference.After restudy pre-treatment mode and the development system of having examined or check test sample, be specially: get these article 5g, porphyrize adds ethanol 30ml, sonicated 15 minutes; Filter, the filtrating evaporate to dryness adds water 20ml and makes dissolving, is transferred in the centrifuge tube, adds watery hydrochloric acid 5ml again; Ultrasonic 5 minutes, leave standstill, centrifugal, taking precipitate adds Diluted Alcohol 1ml and makes dissolving; Regulate pH to neutral with 10% sodium bicarbonate solution, centrifugal, get supernatant, as need testing solution; Extracting liquorice acid ammonium reference substance adds Diluted Alcohol and processes the solution that every 1ml contains 1mg, reference substance solution the most in addition; Draw above-mentioned two kinds of solution 5ul, put respectively in same be on the silica GF254 thin layer plate of bonding agent with the sodium carboxymethyl cellulose, with normal butyl alcohol: glacial acetic acid: water=6: 1: 3 upper solution are developping agent; Launch, taking-up is dried, and puts under the ultraviolet lamp (254nm) and inspects; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color.Interference is not seen in the negative control test.The revenue standard text (8cm, Rf=0.40).
Five, content of paeoniflorin study on determination method
Instrument and reagent
Waters 600 high performance liquid chromatographs (VWD detecting device): U.S. Waters company;
HYPERSIL C18 post (4.6X 30nlln, 5 μ): Dalian Yi Lite scientific instrument company limited;
Methyl alcohol is chromatographically pure, and it is pure that other reagent is analysis;
Paeoniflorin reference substance: Nat'l Pharmaceutical & Biological Products Control Institute.Lot number: 0736-9710,0736-9912 supplies assay to use.
1. chromatographic condition is selected
Chromatographic column: with the octadecylsilane chemically bonded silica is filler, HYPERSIL C
18Post (4.6 * 300mm, 5 μ); Moving phase: methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4; Detect wavelength: 230nm; Column temperature: room temperature; Number of theoretical plate calculates by the Paeoniflorin peak and is not less than 2000.
Under this condition, Paeoniflorin can reach baseline separation with other component, with adjacent chromatographic peak degree of separation greater than 2.Negative control is noiseless.
In the experiment also once with methyl alcohol: water=22: 78 is moving phase, and Paeoniflorin also can reach baseline separation with other component under this condition, with adjacent chromatographic peak degree of separation greater than 2.But retention time is long partially, reaches about 20 minutes, so do not selected for use.
2. the preparation of need testing solution
Chinese herbaceous peony Qi particle adopts water extraction process, and it is testing index that the end product quality standard adopts content of paeoniflorin.Therefore; The applicant is with reference to " disposal route of sample adopts methyl alcohol for extracting solvent, ultrasonic Extraction during the middle Chinese herbaceous peony medicinal material assay of 2005 editions one of Chinese pharmacopoeia; And different ultrasonic times (15,30,60 minutes) are investigated; The result shows that the granule ultrasonic time is 30 minutes, and the extraction ratio of Paeoniflorin is the highest, sees table 1.
Also compared the extraction ratio of different extraction solvents to Paeoniflorin in the granule in the experiment, do not seen significant difference, RSD＜1% is seen table 2.
The comparison of the different ultrasonic times of table 1 (n=3)
The different comparisons of extracting solvent of table 2
Sample preparation methods after preferred is:
These article of getting content, mixing, porphyrize is got about 0.2g, and accurate the title, decide; Put in the 50ml tool plug conical flask, the accurate close plug of methanol solution 25ml that adds is claimed to decide weight, and ultrasonic Extraction 30 minutes claims to decide weight again; Supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, as need testing solution.
3. linear relationship is investigated
Need testing solution preparation: it is an amount of that precision takes by weighing in the phosphorus pentoxide vacuum drying apparatus dry 36 hours Paeoniflorin reference substance, and Paeoniflorin reference substance 5.03mg puts in the 25ml volumetric flask.With dissolve with methanol and be diluted to scale.Be made into the reference substance solution of a series of concentration gradients of 60.3 μ g/ml, 40.2, μ g/ml, 20.1 μ g/ml, 10.0 μ g/ml, 5.03 μ g/ml successively as mother liquor.Draw above-mentioned reference substance solution 1～5 respectively, each 10 μ l measures by above-mentioned chromatographic condition sample introduction.With the peak area integrated value is ordinate (Y), and Paeoniflorin reference substance concentration is horizontal ordinate (X), and the drawing standard curve calculates regression equation:
Y(Area)＝37096X(Amount[ug/ml])-33204，r＝0.9994
Paeoniflorin concentration is good in 5.03～60.3 μ g/ml scope internal linear relation, sees table 3.
Table 3 linear relationship is investigated the result
4. precision test
The same need testing solution 10 μ l of accurate absorption repeat sample introduction 5 times, calculate the peak area relative standard deviation of Paeoniflorin, see table 4.
Table 4 Precision test result
5. replica test
The sample of same lot number, parallel sampling 5 times makes the sample test liquid respectively.The accurate sample test liquid 10 μ l that draw repeat sample introduction, calculate the peak area relative standard deviation of Paeoniflorin, see table 5.
Table 5 replica test result
6. recovery test
Adopt the application of sample absorption method, get the nourishing the liver choleretic granules that oneself knows paeoniflorin content 6.06mg/g, add the Chinese herbaceous peony reference substance respectively, measure by above-mentioned chromatographic condition, with the following formula calculate recovery rate, the result shows that this law has the good recovery, sees table 6.
Table 6 recovery test result
7. content of paeoniflorin is measured in the medicinal material
More because of the kind of Chinese herbaceous peony in the medicinal material, three batches of medicinal materials are through being accredited as the dry root of ranunculaceae plant Chinese herbaceous peony Paeonia lactifloraPall.In the experiment content of paeoniflorin in three batches of Chinese herbaceous peony medicinal materials is measured, all met " the regulation of Chinese pharmacopoeia (an one) (2005 editions) Chinese herbaceous peony medicinal material.The result is following, sees table 7.
Content of paeoniflorin is measured the result in table 7 medicinal material
8. sample determination
The accurate need testing solution 5 μ l that draw measure by above-mentioned chromatographic condition, and the typical curve by proofreading and correct calculates the content in every bag of granule of these article.Three lot sample article are measured the result and are listed in table 8.
Table 8 sample size is measured the result
9. content limit
" 2005 editions one regulation Chinese herbaceous peony medicinal material of Chinese pharmacopoeia contains Paeoniflorin (C
23H
28O
11) must not be less than 1.60%, and the rate of transform of Paeoniflorin is not less than 50% among the preparation technology, and consider the loss that possibly exist in the production run, tentative these article contain Paeoniflorin (C for every bag
23H
28O
11) every bag contain Paeoniflorin and must not be less than 48.0mg.The content of six lot sample article (three batches of lab scales and three batches of pilot scales) all meets this regulation.
The research of the applicant through effective ingredient in the said preparation is carried out; Scientific and reasonable detection method and quality control index have been set up; Form a perfect quality control standard, effective ingredient in the preparation is differentiated, and the content of effective ingredient is detected; Quality to reach said preparation control effectively, and guarantees the clinical efficacy and the security of product.
Favourable beneficial effect fruit of the present invention:
Compared with prior art; Scientific and reasonable detection method and quality control index have been set up in the research of the present invention through effective ingredient in the said preparation is carried out, and form a perfect quality control standard; Effective ingredient in the preparation is differentiated, and the content of effective ingredient is detected.Quality determining method specificity of the present invention is strong, precision is high; Favorable reproducibility; The recovery is high, and measurement result is accurate, can effectively control the quality of Chinese herbaceous peony Qi particle; These article contain Chinese herbaceous peony for every bag and are no less than 48.0mg in Paeoniflorin in settling the standard, thereby have guaranteed the clinical efficacy and peace security of said preparation.
Embodiment
The detection method of this Chinese herbaceous peony Qi particle
[prescription] root of herbaceous peony 750.0g; Dried orange peel 562.5g; Prepared fleece flower root 375.0g; Fruit of Chinese wolfberry 375.0g; Honey-fried licorice root 187.5g.
[method for making] above five tastes medicinal material, boiling three times, amount of water is respectively 6 times, 4 times, 4 times amounts.Decoct earlier for the first time and soaked 30 minutes, each boiling reflux 1 hour; Filter, merging filtrate, being evaporated to relative density is the clear cream of 1.06～1.08 (70～80 ℃), is chilled to room temperature; Centrifuging (3500～4000 rev/mins, centrifugal 20 minutes), it is the clear cream of 1.20～1.22 (75～80 ℃) that collection centrifugate and concentrating under reduced pressure become relative density, 60～70 ℃ of drying under reduced pressure; Dry extract is broken into fine powder, adds an amount of dextrin, and mixing is processed particle with ethanol; Drying is processed 1000g, promptly gets.
[proterties] these article are brown granular, mildly bitter flavor, and gas is fragrant.
[discriminating]
(1) get these article in right amount, porphyrize is got 3g, adds methyl alcohol 30ml, and sonicated 20 minutes filters, and filtrating is steamed to closely doing, and residue adds methyl alcohol 3ml makes dissolving, as need testing solution; Other gets dried orange peel control medicinal material 2g, shines medicinal material solution in pairs with legal system; Get the aurantiamarin reference substance again, add an amount of methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; Draw above-mentioned three kinds of each 5ul of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ethyl acetate: methyl alcohol: water=100: 17: 13 is developping agent; Exhibition is taken out to about 3cm, dries; Again with toluene: ethyl acetate: formic acid: water=20: 10: 1: 1 upper solution is a developping agent, and exhibition is taken out to about 8cm; Dry, spray is put under the ultraviolet lamp (365nm) and is inspected with the aluminium choride test solution; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;
(2) get these article in right amount, porphyrize is got 10g, adds methyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue is with the methenyl choloride washing for several times, and is closely colourless to methenyl choloride, and merging methenyl choloride washing lotion is concentrated into about 1ml, as need testing solution; Other gets fleece-flower root control medicinal material 0.5g, shines medicinal material solution in pairs with legal system; Get the archen reference substance again, add an amount of methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) experiment, draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate; With toluene: ethyl acetate: formic acid=15: 2: 1 is developping agent, launches, and takes out; Dry, put under the ultraviolet lamp (365nm) and inspect; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color.
(3) get these article in right amount, porphyrize is got 0.5g, adds methyl alcohol 10ml, sonicated 5 minutes; Filter, the filtrating evaporate to dryness, residue adds water 10ml makes dissolving, with ethyl acetate extraction twice, each 10ml; Merge extract, evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets fruit of Chinese wolfberry control medicinal material 1g, adds water 30ml, boils 15 minutes, filters, and filtrating is shone medicinal material solution in pairs with legal system; According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) experiment, draw each 2～3ul of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of bonding agent with the sodium carboxymethyl cellulose; With sherwood oil (60～90 ℃): ethyl acetate: glacial acetic acid=10: 7: 0.5 is a developping agent, launches (30～35 ℃, saturated in advance 20 minutes); Take out; Dry, put under the ultraviolet lamp (365nm) and inspect, in the test sample chromatogram; With control medicinal material chromatogram same position on, show the fluorescence spot of same color;
(4) get these article 5g, porphyrize adds ethanol 30ml, and sonicated 15 minutes filters; The filtrating evaporate to dryness adds water 20ml and makes dissolving, is transferred in the centrifuge tube, adds watery hydrochloric acid 5ml again, ultrasonic 5 minutes; Leave standstill, centrifugal, taking precipitate adds Diluted Alcohol 1ml and makes dissolving; Regulate pH to neutral with 10% sodium bicarbonate solution, centrifugal, get supernatant, as need testing solution; Extracting liquorice acid ammonium reference substance adds Diluted Alcohol and processes the solution that every 1ml contains 1mg, reference substance solution the most in addition; According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 B) experiment, draw above-mentioned two kinds of solution 5ul, put respectively in same be on the silica GF254 thin layer plate of bonding agent with the sodium carboxymethyl cellulose; With normal butyl alcohol: glacial acetic acid: water=6: 1: 3 upper solution are developping agent, launch, and taking-up is dried; Put under the ultraviolet lamp (254nm) and inspect; In the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
[inspection] should meet each item regulation (2005 editions appendix IC of Chinese Pharmacopoeia) relevant under the granule item.
[assay]
The contained content of paeoniflorin of the root of herbaceous peony is measured according to high performance liquid chromatography (2005 editions one appendix VI D of Chinese Pharmacopoeia) and is measured.
With octadecylsilane key and silica gel is filling agent, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is moving phase, and the detection wavelength is 230nm, and theoretical cam curve is calculated by the Paeoniflorin peak should be not less than 2000; It is an amount of that precision takes by weighing the Paeoniflorin reference substance, adds methyl alcohol and process the solution that every 1ml contains 0.05mg, promptly gets reference substance solution; These article of getting, porphyrize is got about 0.2g, and accurate the title, decide; Put in the 50ml tool plug conical flask, the accurate methyl alcohol 25ml that adds, close plug claims to decide weight; Sonicated (power 135W, frequency 59kHz) 30 minutes is claimed to decide weight again, supplies the weight that subtracts mistake with methyl alcohol; Shake up, filter, get subsequent filtrate, promptly get need testing solution; Accurate respectively reference substance solution and each 10ul of need testing solution of drawing injects liquid chromatograph, measures, and promptly gets, and these article contain the root of herbaceous peony in Paeoniflorin for every bag, must not be less than 48.0mg.
Claims (2)
1. the detection method of a white paeony root-medlar particles, said Chinese herbaceous peony Qi granular preparation is prepared from suitable auxiliary material root of herbaceous peony 750.0g dried orange peel 562.5g prepared fleece flower root 375.0g fruit of Chinese wolfberry 375.0g honey-fried licorice root 187.5g; This detection method comprises to be differentiated and the assay project; It is characterized in that: said discriminating is that the thin layer of dried orange peel in the said preparation is differentiated, the thin layer of prepared fleece flower root differentiates that the thin layer of the fruit of Chinese wolfberry differentiates that the thin layer of contained ammonium glycyrrhetate is differentiated in the honey-fried licorice root; Said assay is that the contained content of paeoniflorin of the root of herbaceous peony in the preparation is measured; Said detection method comprises following:
(1) thin layer of dried orange peel is differentiated
These article of getting are an amount of, and porphyrize adds methyl alcohol, and sonicated filters, and filtrating is steamed near and done, and residue adds methyl alcohol makes dissolving, as need testing solution; Other gets the dried orange peel control medicinal material, shines medicinal material solution in pairs with legal system; Get the aurantiamarin reference substance again, add an amount of methyl alcohol and process reference substance solution; Draw above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate with sodium hydroxide solution preparation, with ethyl acetate: methyl alcohol: water=100: 17: 13 is developping agent; Launch, take out, dry; Again with toluene: ethyl acetate: formic acid: water=20: 10: 1: 1 upper solution is a developping agent, launches, and takes out; Dry, spray is put under the 365nm ultraviolet lamp and is inspected with the aluminium choride test solution; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;
(2) thin layer of prepared fleece flower root is differentiated
These article of getting are an amount of, and porphyrize adds methyl alcohol, and sonicated filters, the filtrating evaporate to dryness, and residue is with the methenyl choloride washing for several times, and is closely colourless to methenyl choloride, merges the methenyl choloride washing lotion, concentrates, as need testing solution; Other gets fleece-flower root control medicinal material, shines medicinal material solution in pairs with legal system; Get the archen reference substance again, add an amount of methyl alcohol and process reference substance solution; Draw above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate, with toluene: ethyl acetate: formic acid=15: 2: 1 is developping agent, launches, and takes out, and dries, and puts under the 365nm ultraviolet lamp and inspects; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;
(3) thin layer of the fruit of Chinese wolfberry is differentiated
These article of getting are an amount of, and porphyrize adds methyl alcohol, and sonicated filters, the filtrating evaporate to dryness, and residue adds water makes dissolving, uses ethyl acetate extraction, merges extract, and evaporate to dryness, residue add ethyl acetate makes dissolving, as need testing solution; Other gets fruit of Chinese wolfberry control medicinal material, adds water, boils, and filters, and filtrating is shone medicinal material solution in pairs with legal system; Draw above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, with sherwood oil: ethyl acetate: glacial acetic acid=10: 7: 0.5 is a developping agent; Launch, take out, dry; Put under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with control medicinal material chromatogram same position on, show the fluorescence spot of same color;
(4) thin layer of honey-fried licorice root is differentiated
These article of getting, porphyrize adds ethanol, and sonicated filters, the filtrating evaporate to dryness; Add water and make dissolving, be transferred in the centrifuge tube, add watery hydrochloric acid again, sonicated leaves standstill, and is centrifugal; Taking precipitate adds Diluted Alcohol and makes dissolving, regulates pH to neutral with sodium bicarbonate solution, and is centrifugal, gets supernatant, as need testing solution; In addition extracting liquorice acid ammonium reference substance adds Diluted Alcohol and processes reference substance solution, draws above-mentioned two kinds of solution, put respectively in same be the silica G F of binder with the sodium carboxymethyl cellulose
254On the thin layer plate, with normal butyl alcohol: glacial acetic acid: water=6: 1: 3 upper solution are developping agent, launch, and take out, and dry, put under the 254nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
(5) content of paeoniflorin is measured
With octadecylsilane key and silica gel is filling agent, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is moving phase, and the detection wavelength is 230nm, and theoretical cam curve is calculated by the Paeoniflorin peak should be not less than 2000; It is an amount of that precision takes by weighing the Paeoniflorin reference substance, adds methyl alcohol and process reference substance solution; These article of getting, porphyrize, the accurate title, decide, and puts in the tool plug conical flask, the accurate methyl alcohol that adds, close plug is claimed to decide weight, and sonicated is claimed to decide weight again, supplies the weight that subtracts mistake with methyl alcohol, shakes up, and subsequent filtrate is got in filtration, promptly gets need testing solution; Accurate respectively reference substance solution and the need testing solution drawn injects liquid chromatograph, measures, and promptly gets, and these article contain the root of herbaceous peony in Paeoniflorin for every bag, must not be less than 48.0mg.
2. according to the detection method of the said Chinese herbaceous peony Qi of claim 1 particle, it is characterized in that:
Detection method comprises more specifically:
(1) thin layer of dried orange peel is differentiated
These article of getting are an amount of, and porphyrize is got 3g, adds methyl alcohol 30ml, and sonicated 20 minutes filters, and filtrating is steamed near and done, and residue adds methyl alcohol 3ml makes dissolving, as need testing solution; Other gets dried orange peel control medicinal material 2g, shines medicinal material solution in pairs with legal system; Get the aurantiamarin reference substance again, add an amount of methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; Draw above-mentioned each 5u 1 of three kinds of solution, put respectively on the silica gel g thin-layer plate of same usefulness 0.5% sodium hydroxide solution preparation, with ethyl acetate: methyl alcohol: water=100: 17: 13 is developping agent; Open up to 3cm, take out, dry; Again with toluene: ethyl acetate: formic acid: water=20: 10: 1: 1 upper solution is a developping agent, opens up to 8cm, takes out; Dry, spray is put under the 365nm ultraviolet lamp and is inspected with the aluminium choride test solution; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;
(2) thin layer of prepared fleece flower root is differentiated
These article of getting are an amount of, and porphyrize is got 10g, adds methyl alcohol 30ml, and sonicated 30 minutes filters, the filtrating evaporate to dryness, and residue is with the methenyl choloride washing for several times, and is closely colourless to methenyl choloride, merges the methenyl choloride washing lotion, is concentrated into 1ml, as need testing solution; Other gets fleece-flower root control medicinal material 0.5g, shines medicinal material solution in pairs with legal system; Get the archen reference substance again, add an amount of methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; Draw above-mentioned three kinds of each 5ul of solution, put respectively on same silica gel g thin-layer plate, with toluene: ethyl acetate: formic acid=15: 2: 1 is developping agent; Launch; Take out, dry, put under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with control medicinal material chromatogram and the corresponding position of reference substance chromatogram on, show the fluorescence spot of same color;
(3) thin layer of the fruit of Chinese wolfberry is differentiated
These article of getting are an amount of, and porphyrize is got 0.5g, adds methyl alcohol 10ml, sonicated 5 minutes; Filter, the filtrating evaporate to dryness, residue adds water 10ml makes dissolving, with ethyl acetate extraction twice, each 10ml; Combined ethyl acetate extract, evaporate to dryness, residue add ethyl acetate 1ml makes dissolving, as need testing solution; Other gets fruit of Chinese wolfberry control medicinal material 1g, adds water 30ml, boils 15 minutes, filters, and filtrating is shone medicinal material solution in pairs with legal system; Draw each 2～3ul of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of binder with the sodium carboxymethyl cellulose, with the sherwood oil of 60～90 ℃ of boiling points: ethyl acetate: glacial acetic acid=10: 7: 0.5 is a developping agent; In 30～35 ℃ saturated in advance 20 minutes, launch, take out, dry; Put under the 365nm ultraviolet lamp and inspect; In the test sample chromatogram, with control medicinal material chromatogram same position on, show the fluorescence spot of same color;
(4) thin layer of honey-fried licorice root is differentiated
These article of getting 5g, porphyrize adds ethanol 30ml, and sonicated 15 minutes filters; The filtrating evaporate to dryness adds water 20ml and makes dissolving, is transferred in the centrifuge tube, adds watery hydrochloric acid 5ml again, ultrasonic 5 minutes; Leave standstill, centrifugal, taking precipitate adds Diluted Alcohol 1ml and makes dissolving; Regulate pH to neutral with 10% sodium bicarbonate solution, centrifugal, get supernatant, as need testing solution; In addition extracting liquorice acid ammonium reference substance adds Diluted Alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution, draws above-mentioned two kinds of each 5ul of solution, put respectively in same be the silica G F of binder with the sodium carboxymethyl cellulose
254On the thin layer plate, with normal butyl alcohol: glacial acetic acid: water=6: 1: 3 upper solution are developping agent, launch, and taking-up is dried, put under the 254nm ultraviolet lamp and inspect, in the test sample chromatogram, with the corresponding position of reference substance chromatogram on, show the spot of same color;
(5) content of paeoniflorin is measured
With octadecylsilane key and silica gel is filling agent, and with methyl alcohol: 0.05mol/L potassium dihydrogen phosphate: acetic acid: isopropyl alcohol=67: 173: 4: 4 is moving phase, and the detection wavelength is 230nm, and theoretical cam curve is calculated by the Paeoniflorin peak should be not less than 2000; It is an amount of that precision takes by weighing the Paeoniflorin reference substance, adds methyl alcohol and process the reference substance solution that every 1ml contains 0.05mg; These article of getting, porphyrize is got 0.2g, and accurate the title, decide, and puts in the 50ml tool plug conical flask; The accurate methyl alcohol 25ml that adds, close plug is claimed to decide weight, and power 135W and frequency 59kHz sonicated 30 minutes claim to decide weight again; Supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate, promptly get need testing solution; Accurate respectively reference substance solution and each 10ul of need testing solution of drawing injects liquid chromatograph, measures, and promptly gets, and these article contain the root of herbaceous peony in Paeoniflorin for every bag, must not be less than 48.0mg.
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| CN106198834A (en) * | 2016-06-24 | 2016-12-07 | 广西灵峰药业有限公司 | The quality of production control method of juice mended by the one seed ginseng stilbene Radix Polygoni Multiflori |
| CN112666268B (en) * | 2019-10-16 | 2023-03-10 | 石家庄以岭药业股份有限公司 | Method for identifying various components and measuring content of traditional Chinese medicine composition |
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| CN115902084B (en) * | 2022-11-04 | 2024-08-20 | 杭州市中医院 | Method for detecting quality of spleen-strengthening miscarriage-preventing particles |
| CN115792077B (en) * | 2023-01-05 | 2024-01-02 | 江苏海洋大学 | Quality detection method of compound weight-losing and lipid-lowering oral liquid |
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| 张嗣博等.养肝利胆颗粒对豚鼠胆囊胆固醇结石成石率和血浆胆囊收缩素含量的影响.《中西医结合学报》.2008,第6卷(第4期),第405-408页. * |
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