CN117420241A - Quality detection method for rush formula particles - Google Patents
Quality detection method for rush formula particles Download PDFInfo
- Publication number
- CN117420241A CN117420241A CN202311604630.5A CN202311604630A CN117420241A CN 117420241 A CN117420241 A CN 117420241A CN 202311604630 A CN202311604630 A CN 202311604630A CN 117420241 A CN117420241 A CN 117420241A
- Authority
- CN
- China
- Prior art keywords
- solution
- rush
- taking
- peak
- methanol
- 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.)
- Pending
Links
- 239000002245 particle Substances 0.000 title claims abstract description 61
- 238000001514 detection method Methods 0.000 title claims abstract description 28
- 238000001228 spectrum Methods 0.000 claims abstract description 28
- 238000005259 measurement Methods 0.000 claims abstract description 24
- 238000004809 thin layer chromatography Methods 0.000 claims abstract description 16
- 238000004811 liquid chromatography Methods 0.000 claims abstract description 14
- 238000010276 construction Methods 0.000 claims abstract description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 141
- 239000000243 solution Substances 0.000 claims description 84
- 238000000034 method Methods 0.000 claims description 55
- 239000013558 reference substance Substances 0.000 claims description 49
- 238000012360 testing method Methods 0.000 claims description 42
- 239000000523 sample Substances 0.000 claims description 40
- 239000000463 material Substances 0.000 claims description 38
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 36
- 239000003814 drug Substances 0.000 claims description 33
- 239000008187 granular material Substances 0.000 claims description 33
- 239000000047 product Substances 0.000 claims description 33
- 239000012488 sample solution Substances 0.000 claims description 33
- 239000002904 solvent Substances 0.000 claims description 33
- 239000000284 extract Substances 0.000 claims description 31
- 238000005303 weighing Methods 0.000 claims description 29
- 230000014759 maintenance of location Effects 0.000 claims description 24
- 239000000706 filtrate Substances 0.000 claims description 22
- 238000001914 filtration Methods 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 20
- 238000001816 cooling Methods 0.000 claims description 20
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 18
- 239000012085 test solution Substances 0.000 claims description 15
- 238000004458 analytical method Methods 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 13
- 239000000126 substance Substances 0.000 claims description 13
- 239000011259 mixed solution Substances 0.000 claims description 12
- 238000000227 grinding Methods 0.000 claims description 11
- 239000012088 reference solution Substances 0.000 claims description 10
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 9
- 235000019253 formic acid Nutrition 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 238000009210 therapy by ultrasound Methods 0.000 claims description 9
- 239000000945 filler Substances 0.000 claims description 8
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 8
- 239000000377 silicon dioxide Substances 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 7
- 238000007689 inspection Methods 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 6
- 238000010992 reflux Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000007598 dipping method Methods 0.000 claims description 5
- 238000007789 sealing Methods 0.000 claims description 5
- 230000001502 supplementing effect Effects 0.000 claims description 5
- NOTFZGFABLVTIG-UHFFFAOYSA-N Cyclohexylethyl acetate Chemical compound CC(=O)OCCC1CCCCC1 NOTFZGFABLVTIG-UHFFFAOYSA-N 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- DHRLEVQXOMLTIM-UHFFFAOYSA-N phosphoric acid;trioxomolybdenum Chemical compound O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.OP(O)(O)=O DHRLEVQXOMLTIM-UHFFFAOYSA-N 0.000 claims description 4
- 238000005507 spraying Methods 0.000 claims description 3
- 238000000691 measurement method Methods 0.000 claims description 2
- 238000000605 extraction Methods 0.000 description 40
- 238000011835 investigation Methods 0.000 description 20
- 238000002360 preparation method Methods 0.000 description 18
- 239000012071 phase Substances 0.000 description 17
- 238000004128 high performance liquid chromatography Methods 0.000 description 10
- 229920002774 Maltodextrin Polymers 0.000 description 8
- 239000005913 Maltodextrin Substances 0.000 description 8
- 229940035034 maltodextrin Drugs 0.000 description 8
- 239000000843 powder Substances 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 210000000692 cap cell Anatomy 0.000 description 6
- 238000013112 stability test Methods 0.000 description 6
- 238000003556 assay Methods 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 239000012925 reference material Substances 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 238000002137 ultrasound extraction Methods 0.000 description 5
- ZZUBHVMHNVYXRR-UHFFFAOYSA-N 3-(4-hydroxyphenyl)-2h-chromen-7-ol Chemical compound C1=CC(O)=CC=C1C1=CC2=CC=C(O)C=C2OC1 ZZUBHVMHNVYXRR-UHFFFAOYSA-N 0.000 description 4
- 241000735470 Juncus Species 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000033228 biological regulation Effects 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 210000002216 heart Anatomy 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000003908 quality control method Methods 0.000 description 3
- 238000010183 spectrum analysis Methods 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 230000004580 weight loss Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 241000219312 Chenopodium Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 244000003187 Juncus effusus Species 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- ADFCQWZHKCXPAJ-GFCCVEGCSA-N equol Chemical compound C1=CC(O)=CC=C1[C@@H]1CC2=CC=C(O)C=C2OC1 ADFCQWZHKCXPAJ-GFCCVEGCSA-N 0.000 description 2
- 235000019126 equol Nutrition 0.000 description 2
- ADFCQWZHKCXPAJ-UHFFFAOYSA-N indofine Natural products C1=CC(O)=CC=C1C1CC2=CC=C(O)C=C2OC1 ADFCQWZHKCXPAJ-UHFFFAOYSA-N 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000000813 small intestine Anatomy 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000011003 system suitability test Methods 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 1
- JPYHHZQJCSQRJY-UHFFFAOYSA-N Phloroglucinol Natural products CCC=CCC=CCC=CCC=CCCCCC(=O)C1=C(O)C=C(O)C=C1O JPYHHZQJCSQRJY-UHFFFAOYSA-N 0.000 description 1
- 208000001431 Psychomotor Agitation Diseases 0.000 description 1
- 206010038743 Restlessness Diseases 0.000 description 1
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 208000002399 aphthous stomatitis Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012490 blank solution Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 239000002024 ethyl acetate extract Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 201000011243 gastrointestinal stromal tumor Diseases 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 206010022437 insomnia Diseases 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 230000027939 micturition Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- QCDYQQDYXPDABM-UHFFFAOYSA-N phloroglucinol Chemical compound OC1=CC(O)=CC(O)=C1 QCDYQQDYXPDABM-UHFFFAOYSA-N 0.000 description 1
- 229960001553 phloroglucinol Drugs 0.000 description 1
- 239000011505 plaster Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000013074 reference sample Substances 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
- 230000005747 tumor angiogenesis Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
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
- G01N30/02—Column 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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
- G01N30/8634—Peak quality criteria
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
-
- 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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/94—Development
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)
- Engineering & Computer Science (AREA)
- Quality & Reliability (AREA)
- Library & Information Science (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention provides a quality detection method of rush formula particles, which comprises the steps of carrying out thin-layer identification, characteristic spectrum construction and total content measurement of the epfurol and the dehydro-epfurol on rush formula particles, limiting the content standard of the formula particles to 1.0-5.6 mg of the total content of the epfurol and the dehydro-epfurol in each 1g of formula particles, wherein the thin-layer identification is carried out by adopting a thin-layer chromatography, and the characteristic spectrum construction and the total content measurement of the epfurol and the dehydro-epfurol are both carried out by adopting a liquid chromatography. According to the quality detection method for the rush formula particles, the quality of the rush formula particles is assessed through the feature map construction and the detection of the total content measurement of the epfurol and the dehydroepfurol, so that the inherent quality of the formula particles can be comprehensively reflected, and the quality of the rush formula particles can be better controlled.
Description
Technical Field
The invention relates to the technical field of quality control of traditional Chinese medicinal materials, in particular to a quality detection method of rush formula particles.
Background
Modern medicines are required to have three characteristics of stability, uniformity, safety and effectiveness, and for Chinese patent medicines, various means are required to be adopted for detection, so that the reliability and stability of detection results are ensured. The traditional Chinese medicine formula granule is a single traditional Chinese medicine product prepared by adopting modern scientific technology and imitating the traditional Chinese medicine decoction decocting way, and refining the traditional Chinese medicine decoction pieces through the processes of leaching, concentrating, drying and the like. The product maintains the property, taste and efficacy of the traditional Chinese medicine decoction pieces, has stable quality, is applied to the formulation of clinical prescriptions of the traditional Chinese medicine, meets the requirements of dialectical treatment and prescription change, and has the advantages of no need of decoction, convenient administration, quick absorption, accurate dosage, safety, cleanness, convenient carrying and the like. The single medicinal materials are subjected to water extraction, concentration, drying, granulation and other procedures to prepare the formula particles, and compared with the traditional medicinal materials, the form characteristics of the formula particles are obviously changed, and the authenticity and quality of the product cannot be obtained by naked eye observation. Therefore, it is necessary to establish a quality evaluation system of the traditional Chinese medicine formula particles so as to comprehensively reflect the inherent quality of the formula particles. Rush is the dry stem pith of rush Juncus effusus L. Harvesting stems in summer to autumn, sun drying, taking out stem marrow, straightening, and bundling into small bundles. Ju Xin Cao is sweet in property, light in nature, slightly cold, and enters heart, lung and small intestine meridians, both clearing damp-heat to treat stranguria, clearing heart and small intestine fire to relieve restlessness, and is commonly indicated for heat stranguria, aphtha and vexation and insomnia. At present, a systematic quality detection method is not formed on the rush formula particles, and the conventional detection means are only adopted to detect the rush formula particles, so that the overall internal quality of the rush formula particles cannot be reflected, and the quality control requirement of the traditional Chinese medicine formula particles cannot be met. Therefore, it is necessary to establish a quality detection method of rush formula particles for controlling the quality of medicinal materials.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provide a rush formula particle quality detection method which comprehensively reflects the internal quality of formula particles and better controls the quality of the formula particles.
In order to achieve the above purpose, the technical scheme of the invention is realized as follows:
a quality detection method of rush formula particles comprises the following steps:
carrying out characteristic spectrum construction and total content measurement of the epsetol and the dehydro-epsetol on the rush formula particles, limiting the content standard of the formula particles to 1.0-5.6 mg of the total content of the epsetol and the dehydro-epsetol in each 1g, and adopting a liquid chromatography for the characteristic spectrum construction and the total content measurement of the epsetol and the dehydro-epsetol;
the determination of the characteristic spectrum by liquid chromatography comprises: taking a solution prepared from a rush control medicinal material as a control medicinal material reference substance solution a, taking a solution prepared from a reference substance of the epfurol and dehydro-epfurol as a reference substance solution a, taking a solution prepared from a rush formula particle sample as a test substance solution a, respectively precisely sucking the control medicinal material reference substance solution a, the reference substance solution a and the test substance solution a, respectively injecting the solutions into a liquid chromatograph, and measuring to obtain the rush-based medicinal material reference substance; wherein the chromatographic conditions adopted are that: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m; mobile phase: using methanol as a mobile phase A and 0.5% formic acid solution as a mobile phase B, and performing gradient elution according to the specification of a table a;
table a gradient elution procedure
| Time (minutes) | Mobile phase a (%) | Mobile phase B (%) |
| 0～10 | 25→44 | 75→56 |
| 10～17 | 44→45 | 56→55 |
| 17～25 | 45→65 | 55→35 |
| 25～35 | 65→70 | 35→30 |
| 35～45 | 70→80 | 30→20 |
| 45～47 | 80→25 | 20→75 |
| 47～57 | 25 | 75 |
Flow rate: 0.8mL/min; column temperature: 30 ℃; sample injection amount: 10. Mu.L; detection wavelength: 282nm.
In one embodiment, the method further comprises identifying the rush formula particles by thin layer chromatography, wherein the thin layer chromatography comprises the following steps:
(1) Preparing a test sample solution b: taking 1.0g of rush formula granule sample, grinding, adding 100ml of methanol, heating and refluxing for 1 hour, cooling, filtering, evaporating filtrate, washing residue with 2ml of diethyl ether, discarding diethyl ether solution, volatilizing, and adding 5ml of methanol to dissolve to obtain a sample solution b;
(2) Preparing a control medicinal material solution b: weighing 1.0g of rush herb reference medicine, grinding, adding 100ml of methanol, heating and refluxing for 1 hour, cooling, filtering, evaporating filtrate, washing residues with 2ml of diethyl ether, discarding diethyl ether liquid, volatilizing, adding 5ml of methanol to dissolve, and preparing reference medicine solution b;
(3) Thin layer chromatography analysis was performed: the thin layer chromatography conditions were as follows: silica gel G thin layer plate; sample application amount: 7 μl of test solution b and 15 μl of control solution b; developing agent: taking cyclohexane-ethyl acetate solution with the volume ratio of 10:7 as a developing agent, and checking: spraying 5% phosphomolybdic acid ethanol solution, and heating at 105deg.C until the color of spots is clear.
In one embodiment, the determination of the characteristic spectrum by liquid chromatography further comprises the steps of:
(1) Preparing a reference substance solution a of a control medicinal material: taking 0.1g of rush herb reference medicine, adding 25ml of water and 25ml of methanol, weighing, performing ultrasonic treatment for 20min, cooling, filtering, and taking the subsequent filtrate as reference medicine reference solution a;
(2) Preparing a reference substance solution a of a reference substance: taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1ml of the mixed solution as the reference substance solution a of the reference substances;
(3) Preparing a test sample solution a: taking appropriate amount of medulla Junci prescription granule, grinding, taking 0.1g, placing into conical flask with plug, adding 25mL of methanol, ultrasonic treating for 30min, cooling, shaking, filtering, and taking the subsequent filtrate as sample solution a.
In one embodiment, the chromatogram of the sample measured by liquid chromatography shows 5 characteristic peaks, and corresponds to the retention time of 5 characteristic peaks in the chromatogram of the reference material; calculating the relative retention time of the peak 1, the peak 2, the peak 5 and the reference peak S in the chromatogram of the sample by taking the peak 3 corresponding to the reference substance of the Eptifhol as the reference peak S; the specified value is: peak 1 relative retention time 0.47, peak 2 relative retention time 0.82, peak 5 relative retention time 1.24.
In one embodiment, the determination of the total content of the epsetol and the dehydroepsetol by liquid chromatography comprises: performing liquid chromatograph analysis, taking a solution prepared from a reference substance of the epfurol and the dehydroepfurol as a reference substance solution c, taking a solution prepared from a rush prescription granule sample as a test substance solution c, respectively precisely sucking the reference substance solution c and the test substance solution c, respectively injecting into the liquid chromatograph, and measuring to obtain the product; wherein the chromatographic conditions adopted are that: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m; mobile phase: methanol-0.5% formic acid solution with the volume ratio of 66:34 is taken as a mobile phase; flow rate: 1.0mL/min; column temperature: 30 ℃; sample injection amount: 10. Mu.L; detection wavelength: 280nm.
In one embodiment, the method for determining the total content of the epsetol and the dehydroepsetol by adopting the liquid chromatography method further comprises the following steps:
(1) Preparing a reference substance solution: taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1ml of the mixed solution as a reference substance solution c;
(2) Preparing a test solution: taking appropriate amount of rush prescription granule, grinding, taking 0.1g, precisely weighing, placing into a conical bottle with a plug, precisely adding 25ml of methanol, sealing, weighing, ultrasonic treating for 20 minutes, cooling, weighing again, supplementing loss weight with methanol, shaking, filtering, and taking the subsequent filtrate as a sample solution c.
In one embodiment, the method further comprises the following detection method, wherein the characteristics of the rush prescription granule are identified, the dry extract yield is checked, the extract is checked, and the extract is measured by a hot dipping method.
In one embodiment, the hot dip method uses ethanol as a solvent and the range of the extract is determined by a hot dip method under the alcohol-soluble extract determination method.
Compared with the prior art, the quality detection method for the rush formula particles has the beneficial effects that: the quality of the rush formula particles is evaluated by carrying out characteristic spectrum construction and total content measurement of the epfurol and the dehydroepfurol on the rush formula particles, a solid foundation is laid for the stable quality of products, a feasible quality standard of the rush formula particles can be established, the effective control of the quality of the rush formula particles is realized, and the chromatographic conditions are adopted for liquid phase analysis, so that a characteristic spectrum with better and clearer separation degree can be obtained, and the method has the advantages of better stability, high precision and better reproducibility. The rush decoction pieces are decocted to prepare rush decoction piece formula granules, and according to the technical requirements of quality control and standard establishment of traditional Chinese medicine formula granules, the total content of the erliflavone and the dehydro-erliflavone is set to be 1.0 mg-5.6 mg.
Drawings
FIG. 1 is an HPLC characteristic spectrum of standard rush decoction; wherein, peak 3 (S): an epfeol; peak 4: dehydro-epfeol; a chromatographic column; CAPCELL PAK AQ C18,4.6 mm. Times.250 mm,5 μm.
FIG. 2 is a thin layer chromatography-based TLC chart of 3 batches of samples of the rush decoction piece formula particles according to an embodiment of the present invention; wherein, group A is negative control 7 μl, group S is rush control medicinal material solution 15 μl, group 1 is test (lot number 220801) solution 7 μl, group 2 is test (lot number 220802) solution 7 μl, and group 3 is test (lot number 220803) solution 7 μl.
Fig. 3 shows HPLC feature patterns of different extraction methods in investigation of feature pattern measurement extraction method according to an embodiment of the present invention.
Fig. 4 shows HPLC profiles at different extraction times in a profile measurement extraction time investigation according to an embodiment of the present invention.
Fig. 5 is a graph showing HPLC profiles of different extraction solvents in a profile measurement of extraction solvents according to an embodiment of the present invention.
FIG. 6 is a graph showing HPLC characteristics of different solvent amounts in a sample size investigation according to an embodiment of the present invention.
FIG. 7 shows HPLC profiles of different solutions in a profile determination specificity study according to an embodiment of the present invention.
FIG. 8 is a graph of the superposition of peaks common to stability tests in a characteristic spectrum measurement according to an embodiment of the present invention.
Fig. 9 is a superposition of 3 batches of feature maps according to an embodiment of the invention.
Fig. 10 is a graph showing the superposition of characteristic maps of 3 batches of rush herb finished product fitting maps and a reference substance of a reference medicinal material in an embodiment of the invention.
FIG. 11 is a graph showing a characteristic map fit of a finished rush product and a reference medicinal material in an embodiment of the present invention; wherein peak 3 (S) is erliflav; peak 4 is dehydro-epfeol; the column was used as CAPCELL PAK AQ C, 4.6 mm. Times.250 mm,5 μm.
FIG. 12 is a graph showing comparison of maltodextrin blanks in assay specificity studies according to an embodiment of the present invention.
Detailed Description
The present invention will be further described in detail below with reference to specific embodiments and with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the present invention more apparent.
Regarding standard rush standard decoction quality standards:
the product is a standard decoction prepared by processing dry stem marrow of Juncus effusus L, a plant of Juncus family, and referring to the management Specification of Chinese medicine decoction Chamber of medical institution.
[ PREPARATION METHOD ] taking medulla Junci decoction pieces, decocting in water, filtering, concentrating the filtrate into fluid extract (dry extract yield of 3.4% -6.2%), freeze drying, and pulverizing.
15 batches of rush decoction pieces are taken, 15 batches of standard decoction dry paste powder are prepared according to the preparation method, dry extract yield is calculated according to the dry paste powder (see table I), average yield is calculated to be 4.87 percent, the allowable range of the paste yield is calculated according to the standard limit (average value 70% -130%), the allowable range of the paste yield is calculated to be 3.41% -6.33 percent, and the allowable range of the standard decoction paste yield of the rush decoction pieces is calculated to be 3.4% -6.2 percent.
Table I rush decoction piece standard decoction extract yield
The results show that the paste yield of 15 batches of standard decoction is 4.54-5.28%, and the paste yield accords with the range of 3.4-6.2% of the planned limit.
[ PROPERTIES ] the product is brown yellow to tan powder; light smell and bland taste.
[ thin layer identification ] taking 1g of the product, adding 100ml of methanol, heating and refluxing for 1 hour, cooling, filtering, evaporating filtrate, washing residue with 2ml of diethyl ether, discarding diethyl ether solution, volatilizing, and adding 5ml of methanol to dissolve to obtain the solution of the test sample. And 1g of rush herb reference medicine is prepared to prepare a reference medicine solution in the same way. According to thin layer chromatography (China Pharmacopeia 2020 edition rule 0502), 15 μl of control medicinal material solution and 7 μl of test sample solution are sucked. Respectively spotting on the same silica gel G thin layer plate, developing with cyclohexane-ethyl acetate (10:7) as developing agent, taking out, air drying, spraying 5% phosphomolybdic acid ethanol solution, and heating at 105deg.C until the spot color is clear. In the chromatogram of the sample, the main spots with the same color appear at the positions corresponding to the chromatogram of the reference medicinal material.
[ characteristic map ] is measured by high performance liquid chromatography (general rule 0512 in the year 2020).
Chromatographic conditions and System applicability test octadecylsilane chemically bonded silica was used as filler (CAPCELL PAK AQ C (4.6 mm. Times.250 mm,5 μm)); using methanol as a mobile phase A and 0.5% formic acid solution as a mobile phase B, and performing gradient elution according to the specification in a table II; the flow rate is 0.8ml/min per minute; the column temperature is 30 ℃; detection wavelength: 282nm. Theoretical plates, the number of theoretical plates should be not less than 1500 calculated as the peak of the epsethoxynol.
Table II:
| time (minutes) | Mobile phase a (%) | Mobile phase B (%) |
| 0～10 | 25→44 | 75→56 |
| 10～17 | 44→45 | 56→55 |
| 17～25 | 45→65 | 55→35 |
| 25～35 | 65→70 | 35→30 |
| 35～45 | 70→80 | 30→20 |
| 45～47 | 80→25 | 20→75 |
| 47～57 | 25 | 75 |
Preparation of reference solution: taking 0.1g of rush herb reference medicine, adding 25ml of methanol, weighing, performing ultrasonic treatment for 20 minutes, cooling, filtering, and taking the subsequent filtrate as a reference substance solution of the reference medicine. And taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1ml of the reference substance solution.
Preparation of test solution: taking appropriate amount of fine powder, taking about 0.1g, placing into conical flask with plug, adding 25mL of methanol, performing ultrasonic treatment (power 300W, frequency 40 kHz) for 30min, cooling, shaking, filtering, and collecting the subsequent filtrate.
Assay: precisely sucking reference solution of reference material, reference solution of reference material and sample solution of test material respectively by 10 μl, and measuring with liquid chromatograph.
As shown in FIG. 1, according to the above-mentioned established characteristic spectrum analysis method, characteristic spectra of 15 batches of standard decoction are measured, and the result analysis is performed, so that 5 common characteristic peaks are calibrated, wherein peak 3 is the equol and peak 4 is the dehydroequol. Calculating the relative retention time of the other 3 characteristic peaks by taking the peak corresponding to the erlifol reference as an S peak, and respectively setting the average value of the relative retention time of 15 batches of sample peaks as a specified value to be: 0.46 (Peak 1), 0.82 (Peak 2), 1.24 (Peak 5), and the relative retention time allowable range was assumed to be + -10% in consideration of multi-factor errors of test operation, instrument, reagent, etc.
[ extract ] it is measured by hot dipping under the condition of alcohol-soluble extract measurement (general rule 2201 of Chinese pharmacopoeia 2020 edition) and ethanol is used as solvent, and the content of ethanol is not less than 21.6%.
[ MEANS FOR SOLVING ] according to high performance liquid chromatography (rule 0512 in the year 2020 edition of Chinese pharmacopoeia).
Chromatographic conditions and system suitability test: octadecylsilane chemically bonded silica as filler (CAPCELL AQ C (4.6 mm. Times.250 mm,5 μm)); methanol-0.5% formic acid (66:34) was used as mobile phase with a flow rate of 0.8ml per minute; the column temperature is 30 ℃; the detection wavelength is 280nm, and the theoretical plate number is not lower than 1500 calculated according to the peak of the Eptifolian.
Preparation of a control solution: taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1 ml.
Preparation of test solution: taking a proper amount of fine powder of the product, taking about 0.1g, precisely weighing, placing into a conical flask with a plug, precisely adding 20ml of methanol, sealing, weighing, performing ultrasonic treatment (with the power of 300W and the frequency of 40 kHz) for 30 minutes, cooling, weighing again, supplementing the weight loss with methanol, shaking uniformly, filtering, and taking a subsequent filtrate.
Assay: respectively precisely sucking 10 μl of the reference solution and the sample solution, and injecting into a liquid chromatograph for measurement.
Phloroglucinol (C) of the standard decoction 17 H 16 O 2 ) Dehydro-epfeol (C) 17 H 14 O 2 ) The allowable range of the total content is as follows: 1.9 mg/g-6.0 mg/g.
Further, this embodiment provides a method for detecting quality of medulla Junci formula granule, comprising identifying characteristics of medulla Junci formula granule, dry extract yield, thin layer identification, characteristic map, extract and erliflav (C) 17 H 16 O 2 ) Dehydro-epfeol (C) 17 H 14 O 2 ) Total content determination, the standard for the content of formulation particles was defined as containing epfolin (C) per 1g 17 H 16 O 2 ) Dehydro-epfeol (C) 17 H 14 O 2 ) The total amount is 1.0 mg-5.6 mg, wherein, the thin layer identification adopts the thin layer chromatography for identification, the characteristic spectrum and the erliflav (C) 17 H 16 O 2 ) Dehydro-epfeol (C) 17 H 14 O 2 ) The total content is determined by liquid chromatography and the extract is determined by hot dipping.
Preparing rush formula particles: the source of the product adopts a standard decoction prepared from the lower source of the rush herb traditional Chinese medicinal material item and the medicinal part according to the technical requirement of the Chinese pharmacopoeia 2020 edition, and comprises the medical plant parts of the family name, the Chinese name, the Latin and the medicinal part, namely: the product is prepared from dry stem and pith of Juncus efugus L. Belonging to Juncus of Juncus by processing and processing according to main quality index of standard decoction.
The specific preparation method comprises the following steps: taking 15000g of rush decoction pieces, adding water for decoction, filtering, concentrating filtrate into clear paste (the paste yield of dry extract is 3.4% -6.2%), adding a proper amount of auxiliary materials, drying (or drying and crushing), adding a proper amount of auxiliary materials, uniformly mixing, granulating and preparing 1000 g.
According to the paste yield of 15 batches of standard decoction, the range of 70% -130% of the average value of the paste yield is defined as the dry extract paste yield.
Inspection of rush formula granule methodology:
1. property investigation
Traits: three samples of rush formula particles were taken one for each, and the morphology, color, odor and mouthfeel of the samples were evaluated and recorded. Particles described as brown to tan; light smell and bland taste.
2. Thin layer authentication
The rush herb reference medicinal material is used as a reference, a thin-layer identification method of the product is established, and 3 batches of pilot sample tests prove that the spots of the test product are clear, and the negative reference sample has no interference, so the test product is assumed to be the product [ identification ]. The test methods and results are as follows:
test by thin layer chromatography (China pharmacopoeia 2020 edition general rule 0502):
sample solution preparation: taking 1g of the product powder, grinding, adding 100ml of methanol, heating and refluxing for 1h, cooling, filtering, evaporating the filtrate, adding 2ml of diethyl ether into the residue for washing, discarding diethyl ether solution, volatilizing, and adding 5ml of methanol for dissolving to obtain a sample solution.
Negative sample solution: a negative sample solution was prepared by the same method as that for maltodextrin 1 g.
Preparing a control medicinal material solution: 1g of rush herb reference medicine is weighed, and a reference medicine solution is prepared by the same method.
According to a thin layer chromatography (four-part rule 0502 of Chinese pharmacopoeia 2020 edition), 7 μl of the sample solution and 15 μl of the control medicinal material solution are sucked and respectively spotted on the same silica gel G thin layer plate, cyclohexane-ethyl acetate (10:7) is used as developing agent, and the mixture is taken out, dried, sprayed with 5% phosphomolybdic acid ethanol solution, and heated at 105deg.C until the spots are clear in color development.
According to the detection method, 3 batches of production sample test sample solution, negative sample (maltodextrin) solution and control medicinal material solution are taken, spotted, and thin-layer identification is carried out. Inspection results show that spots are clear in the chromatogram of the test sample, negative samples have no interference, main spots with the same color appear at the positions corresponding to the chromatograms of the control medicinal materials, and the patterns of 3 batches of TLC are shown in figure 2.
3. Feature map
3.1 instruments, reagents and reagents
3.1.1 Instrument Shimadzu high performance liquid chromatograph (LC-2030 pulses, shimadzu Co., ltd.) Labsolutions chromatography workstation, SPD-10A ultraviolet detector; CAPCELL PAK AQ C18 (4.6 mm. Times.250 mm,5 μm); shimadzu Shim-pack GIST C18-AQ (4.6 mm. Times.250 mm,5 μm); thermostatic waterbath (HMTD-7000, yongguangming medical instruments Co., ltd., beijing); ultrasonic cleaners (KQ-300 DE, kunshan ultrasonic instruments Co., ltd.); one ten-thousandth balance (PX 224ZH, ohus instruments limited); one ten million balance (AWU 220D, japan shimadzu limited).
3.1.2 reagent methanol (Tianjin Co., ltd.) was chromatographic pure ethanol (Tianjin Co., ltd.), formic acid (Tianjin Co., ltd.), and water was ultrapure water (laboratory self-made).
3.1.3 control and control substances including Phlorool (lot number DSTDE0025601, content: 99.24%, chenopodium medical science and technology Co., ltd.), dehydroPhlorool (lot number DSTDQ042801, content: 99.42%, chenopodium medical science and technology Co., ltd.). Control drug (lot number: DSTYD001501, lemeitian medical Desi biological development).
3.2 feature map measuring method
3.2.1 chromatographic conditions and System applicability experiments
Octadecylsilane chemically bonded silica as filler (CAPCELL PAKAQ C (4.6 mm. Times.250 mm,5 μm)); gradient elution was performed as specified in table 1 with methanol as mobile phase a and 0.5% formic acid solution as mobile phase B; the flow rate is 0.8ml per minute; the column temperature is 30 ℃; the detection wavelength was 282nm. The sample loading was 10. Mu.l. The number of theoretical plates should be not less than 1500 in terms of the peak of the epsethoxynol.
TABLE 1 gradient elution procedure
| Time (minutes) | Mobile phase a (%) | Mobile phase B (%) |
| 0～10 | 25→44 | 75→56 |
| 10～17 | 44→45 | 56→55 |
| 17～25 | 45→65 | 55→35 |
| 25～35 | 65→70 | 35→30 |
| 35～45 | 70→80 | 30→20 |
| 45～47 | 80→25 | 20→75 |
| 47～57 | 25 | 75 |
3.2.2 preparation of reference solution
Taking 0.1g of rush herb reference medicine, adding 25ml of methanol, weighing, performing ultrasonic treatment for 20 minutes, cooling, filtering, and taking the subsequent filtrate as a reference substance solution of the reference medicine.
And taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1ml of the reference substance solution.
3.2.3 preparation of sample solutions
Grinding the materials, taking about 0.1g, placing into a conical flask with a plug, adding 25mL of methanol, performing ultrasonic treatment for 30 minutes, cooling, shaking, filtering, and collecting the filtrate.
3.2.4 assay: precisely sucking reference solution of reference material, reference solution of reference material and sample solution of test material respectively by 10 μl, and measuring with liquid chromatograph.
3.3 methodology investigation of feature atlas determination
The method for pre-treating the samples of the characteristic spectrum of the rush prescription granule is examined, and the influence of the extraction method, the extraction time, the extraction solvent and the solvent consumption on the characteristic spectrum of the rush prescription granule is mainly examined.
3.3.1 investigation of extraction method
Test solutions were prepared by different extraction methods, and the measurement was performed according to the method described under "3.2" above. As shown in fig. 3 and table 2, the results show that the main chromatographic peaks of the sample solutions with different extraction modes are consistent in number, and the peak shape difference is not obvious, but the ultrasonic extraction effect is better, so that the ultrasonic is selected as the sample extraction mode.
TABLE 2 comparison of extraction methods
3.3.2 extraction time investigation
Test solutions were prepared at different times, and the measurement was performed according to the method described under "3.2" above. As shown in FIG. 4 and Table 3, the results show that the main chromatographic peaks of the sample solutions with different extraction times are consistent, the difference of peak shapes is not obvious, but the extraction effect is better after 30 minutes of extraction, so that the extraction time of the sample is determined to be 30 minutes.
TABLE 3 comparison of extraction times
3.3.3 investigation of extraction solvent
Test solutions were prepared with different extraction solvents, respectively, and the measurement was performed according to the method described under item "3.2" above. As shown in FIG. 5 and Table 4, the results show that the number of main chromatographic peaks of the sample solutions with different extraction solvents is consistent, the difference of peak shapes is not obvious, but the methanol extraction effect is better, so methanol is selected as the extraction solvent.
TABLE 4 comparison of extraction solvents
3.3.4 investigation of the solvent usage
Test solutions were prepared in different amounts of solvent, and the measurement was performed according to the method described under "3.2" above. As shown in fig. 6 and table 5, the sample solutions extracted with different amounts of solvents had the same number of main chromatographic peaks and had smaller peak shape differences, but the sample extraction effect was better with 25ml of solvent, so 25ml was selected as the amount of solvent for the sample solution preparation.
TABLE 5 comparison of solvent usage
In summary, the preparation method of the sample solution is determined as follows: in summary, the main parameters of the method for preparing the sample solution are determined as follows: taking a proper amount of fine powder, taking about 0.1g, placing into a conical flask with a plug, adding 25mL of methanol, performing ultrasonic extraction for 30 minutes, cooling, shaking uniformly, filtering, and taking a subsequent filtrate.
3.4 feature map analysis method verification
3.4.1 specificity investigation
A test sample was prepared by using about 1.0g of maltodextrin to prepare a maltodextrin blank solution, and the test sample was measured according to the method described in item "3.2".
The results show that: the main characteristic peaks in the chromatogram of the sample are well separated, the maltodextrin blank solvent is free from interference, and the method has good specificity, and is shown in figure 7.
3.4.2 stability test
Approximately 0.1g of rush formula particles (batch number: 220801) are taken and are measured by sampling at 0h, 2h, 4h, 8h, 12h and 24h according to the test method under item 3.2. The measurement results show that the characteristic patterns of the test samples have basically consistent peak shapes and peak numbers (see FIG. 8). The characteristic spectrum has 5 common peaks, the relative retention time and the relative peak area of the peak 1, the peak 3, the peak 5 and the peak S are calculated by taking the peak of the erliflav as a reference peak S, and the RSD value is calculated. The calculation results show that the relative peak area RSD value and the relative retention time RSD value are all within the acceptable range. The details are shown in tables 6-7, and the test shows that the test sample solution is stable within 24 hours.
TABLE 6 stability test characterization profile versus retention time
| Peak number | 0h | 2h | 4h | 8h | 12h | 24h | RSD(％) |
| 1 | 0.465 | 0.465 | 0.465 | 0.465 | 0.465 | 0.465 | 0.00 |
| 2 | 0.824 | 0.824 | 0.824 | 0.824 | 0.824 | 0.825 | 0.05 |
| 3(S) | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.00 |
| 5 | 1.235 | 1.235 | 1.235 | 1.235 | 1.235 | 1.235 | 0.00 |
TABLE 7 stability test characteristic spectrum versus peak area
| Peak number | 0h | 2h | 4h | 8h | 12h | 24h | RSD(％) |
| 1 | 2.894 | 2.926 | 2.936 | 2.908 | 2.898 | 2.883 | 0.63 |
| 2 | 0.438 | 0.440 | 0.442 | 0.439 | 0.437 | 0.435 | 0.54 |
| 3(S) | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.00 |
| 5 | 1.011 | 1.021 | 1.027 | 1.036 | 1.034 | 1.009 | 1.02 |
3.4.3 knots
The characteristic spectrum method meets the requirements through the investigation of specificity and stability, and the results show that the established method can be well used for measuring the characteristic spectrum of the rush formula particles.
3.5 characterization analysis of three batches of finished product feature patterns
3.5.1 feature map determination
According to the proposed characteristic spectrum analysis method, 3 batches of finished product characteristic spectrums are measured, and through the positioning of the epfurol, the result shows that 5 common peaks exist in the characteristic chromatograms of the formula particles and the traditional Chinese medicine decoction pieces used for preparation of the formula particles and correspond to 5 characteristic peak retention times in the chromatogram of the reference substance of the reference medicinal material, wherein the peak corresponding to the reference substance of the epfurol is peak 3, and the characteristic spectrums of the common peaks are shown in figures 9-11 in detail.
3.5.2 determination of characteristic chromatograms relative retention time
And 3 batches of finished product characteristic patterns are measured according to the drawn characteristic pattern analysis method. The results showed that there were 5 common peaks in the profile, and peak 1, peak 2, peak 5 and S peaks were calculated for their relative retention times and ranges using the epfurol peak as reference peak S, as detailed in table 8.
TABLE 8 peak to peak retention times common to batches
| Peak number | Finished product 1 | Finished product 2 | Finished product 3 | Mean value of | Relative retention time range.+ -. 10% |
| 1 | 0.471 | 0.470 | 0.470 | 0.470 | 0.423-0.517 |
| 2 | 0.823 | 0.823 | 0.823 | 0.823 | 0.741-0.905 |
| 3(S) | 1.000 | 1.000 | 1.000 | 1.000 | / |
| 5 | 1.235 | 1.235 | 1.235 | 1.235 | 1.112-1.359 |
In summary, the stability of the standard decoction feature spectrum measurement method established by adopting the high performance liquid chromatography is examined according to the four-part analysis method verification guidelines (general rule 9101) of the Chinese pharmacopoeia 2020 edition, and the stability of the established method meets the requirements. According to the proposed characteristic spectrum analysis method, measuring characteristic spectrums of 3 batches of finished products, analyzing the result, calibrating 5 common characteristic peaks, taking an erliflav peak as a reference peak S, calculating relative retention time of a peak 1, a peak 2, a peak 5 and an S peak, and respectively setting the average value of the relative retention time of 3 batches of sample peaks as a specified value to be: 0.47 (Peak 1), 0.82 (Peak 2), 1.24 (Peak 5). The allowable range of the relative retention time is set to be +/-10% by considering multi-factor errors of test operation, instruments, reagents and the like.
4 test item test
4.1 examination of general rule of granules according to the related requirements under the general rule of 0104 granules of the year 2020 edition of Chinese pharmacopoeia, examination of granularity moisture and solubility of samples (lot numbers: 220801, 220802 and 220803) is carried out, and the results are shown in Table 9, and all the requirements under the general rule of 0104 granules of the year 2020 edition of Chinese pharmacopoeia are met.
Table 9-test sample examination of rush formula particles
| Inspection item | 220801 | 220802 | 220803 |
| Moisture (less than or equal to 8.0 percent) | 6.7％ | 6.7％ | 6.6％ |
| Particle size (less than or equal to 15 percent) | 4％ | 5％ | 5％ |
| Solubility of | Meets the regulations | Meets the regulations | Meets the regulations |
4.2 exogenous toxic and harmful substance detection
The medicinal materials and the decoction pieces are collected in the first edition of Chinese pharmacopoeia 2020, and the standard has no limit requirement of related harmful substances. The rush medicinal materials and the decoction pieces are detected in batches, and the condition that the related harmful substances exceed the standard is not found as a result, so that the related harmful substances are not listed in the inspection items in the rush prescription granule establishment quality standard.
5 extract
5.1 test methods
The extraction solvent of the traditional Chinese medicine formula granule is water for pharmacy, ethanol is used as solvent according to the technical requirement, and the solvent is measured according to a hot dipping method under the alcohol-soluble extract measuring method (2201 in the general rule of the year 2020 of Chinese pharmacopoeia).
5.2 investigation of extract
About 2g each of 3 pilot formulation granule samples were taken and ground and measured according to the law, the results are shown in Table 10.
TABLE 10 results of extract investigation
The mean extract of three pilot samples was 37.38%. According to the specification (each 1g of formula granule is equivalent to 15g of decoction pieces), referring to the lower limit of the alcohol-soluble extract limit (not less than 21.6%) and the allowable range of the ointment yield (3.4% -6.2%) of the standard decoction, calculating the extract limit of the formula granule as 21.6X15X3.4/100=11.02%; the formulated granule extract limit was 21.6x0.7=15.12%, calculated as 70% of the standard soup extract limit, and the alcohol-soluble extract of the product was not less than 15.0% in combination with the test results of the pilot test sample.
6 content determination
6.1 selection basis of the components to be measured
The medulla Junci is dry stem pulp of medulla Junci of Junciaceae, and is distributed in Heilongjiang, jilin, liaoning, hebei, gansu, shanxi, shandong, jiangsu, anhui, zhejiang, jiangxi, fujian, taiwan, henan, hubei, hunan, guangdong, guangxi, sichuan, guizhou, yunnan and Xizang. Is grown at the river side, the side of a pool, a ditch, a paddy field, a grassland and a marsh wet place with the altitude of 1650-3400 meters. Are distributed in warm areas. Medulla Junci is used as a medicament for treating white medulla in stem, and has effects of promoting urination, relieving stranguria, clearing heart fire, and tranquilizing. Modern pharmacological research shows that dehydro-epfurol in rush is used in preparing medicine for resisting tumor and inhibiting tumor angiogenesis, and its acetone, ethanol and ethyl acetate extracts have antioxidant and antimicrobial effects. Therefore, the establishment of the content index of the epfurol and the dehydroepfurol in the rush has certain reference value for the quality detection of rush standard decoction.
6.2 test methods
The measurement is carried out by high performance liquid chromatography (the rule 0512 of the edition of Chinese pharmacopoeia 2020).
Chromatographic conditions and system suitability test: octadecylsilane chemically bonded silica as filler (4.6mm. Times.250 mm,5 μm); methanol-0.5% formic acid (66:34) is taken as a mobile phase; the flow rate is 1.0ml per minute; the column temperature is 30 ℃; the detection wavelength was 280nm.
Preparation of a control solution: taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1 ml.
Preparation of test solution: grinding the product, taking about 0.1g, precisely weighing, placing into a conical flask with a plug, precisely adding 25ml of methanol, sealing, weighing, performing ultrasonic treatment for 20 minutes, cooling, weighing again, supplementing the weight loss with methanol, shaking, filtering, and collecting the filtrate.
Assay: respectively precisely sucking 10 μl of the reference solution and the sample solution, and injecting into a liquid chromatograph for measurement.
6.3 content determination methodology investigation
The method for pre-treating the samples of the characteristic spectrum of the rush prescription granule is studied, and the influence of the extraction method, the extraction solvent, the extraction time and the solvent amount on the characteristic spectrum of the rush standard decoction is mainly studied.
6.3.1 investigation of the extraction method
The sample preparation test solutions were extracted in different ways and assayed according to the method described under "6.2" above. The results show that the two extraction modes have little difference in extraction effect, and the ultrasonic extraction mode is simpler and more convenient, so that the ultrasonic mode is selected to extract the sample, and the details are shown in Table 11.
TABLE 11 comparison of different extraction methods
6.3.2 investigation of extraction solvent
According to the method described in item 6.2, test solutions were prepared by extracting samples with different solvents, respectively, and the measurement was carried out according to the law. The results show that methanol was the most effective solvent extraction, so methanol was selected as the solvent for the extraction of the samples, as detailed in Table 12.
TABLE 12 comparison of different extraction solvents
6.3.3 investigation of extraction time
According to the method described in item 6.2, sample preparation test solutions were prepared by extracting samples at different times, respectively, and measurement was carried out according to law. The results show that the ultrasonic extraction can be basically complete within 10 minutes, so that the ultrasonic extraction time of the sample is determined to be 10 minutes, and the details are shown in Table 13.
TABLE 13 comparison of different extraction times
6.3.4 solvent quantity investigation
Test solutions were prepared with different amounts of solvents and measured according to the method described under "6.2" above. The results showed that the extraction effect was not very different for different amounts of solvent, so 25ml was chosen as the amount of solvent for preparing the test sample solution for error reduction, as detailed in Table 14.
TABLE 14 comparison of different sample volumes
In summary, the main parameters of the method for preparing the sample solution are determined as follows: taking a proper amount of fine powder of the product, taking about 0.1g, precisely weighing, placing into a conical flask with a plug, precisely adding 25mL of methanol, sealing, weighing, performing ultrasonic treatment for 10 minutes, cooling, weighing again, supplementing the weight loss with methanol, shaking uniformly, filtering, and taking the subsequent filtrate.
6.4 content determination analysis method verification
6.4.1 specificity investigation
Maltodextrin was taken at about 0.1g and measured according to the chromatographic conditions set forth under item "6.2" above, and the test showed that: as shown in fig. 12, maltodextrin blank was undisturbed. The method has good specificity.
6.4.2 stability test
A batch of rush formula granule samples (220801) of about 0.1g are taken, samples are respectively taken at 0h, 2h, 4h, 8h, 12h and 24h according to the test method of 6.2, the content of the rush formula granule samples is measured, the average value of the contents of the erfull and the dehydroerfull in the samples is calculated to be 1.6585mg/g and 3.2955mg/g respectively, the total content of the rush formula granule samples is 4.9394mg/g, the RSD value is 0.21%, and the test shows that the test sample solution is stable within 24 hours, and the details are shown in Table 15.
TABLE 15 stability test
6.4.3 knots
In conclusion, the whole analysis method meets the requirements through investigation of specificity and stability, and the results show that the established method can be well used for measuring the contents of the epfurol and the dehydroepfurol.
7 content limitation range establishment
7.1 three pilot sample content determination
The contents of 3 batches of finished products, namely, the contents of the equol and the dehydroequol, were determined according to the above-described proposed content analysis method, and the results are shown in Table 16.
Table 16 results of three batch product measurements
According to the content range, paste yield and specification calculation of the standard rush decoction, the total content range of the epfolin and the dehydroepfolin in the standard rush decoction is as follows: 1.9 mg/g-6.0 mg/g, and the allowable range of the standard decoction plaster yield is 3.4% -6.2%. The specification of the product is 15g of decoction pieces per 1g of particles, and the dosage range of the auxiliary materials is 1-3.4% multiplied by 15=49.0% and 1-6.2% multiplied by 15=7.0% calculated by the specification. Namely, the dosage range of the auxiliary materials is 7.0-49%, so the content range of the calculated formula particles is as follows: 1.9× (1-49.0%) =0.969 mg/g,6.0× (1-7.0%) =5.58 mg/g, i.e. the acceptable range of content standard content is 0.97mg/g to 5.58mg/g. One decimal place is reserved, so the content range of the product is 1.0 mg/g-5.6 mg/g.
The 8 specifications are formulated according to the preparation method feeding amount and the process research making amount, namely each 1g of formula particle is equivalent to 15g of decoction pieces.
9, the storage is formulated according to the basic requirements of variety storage.
Those skilled in the art will appreciate that: the discussion of any of the embodiments above is merely exemplary and is not intended to suggest that the scope of the invention (including the claims) is limited to these examples; any omission, modification, equivalent replacement, improvement, etc. of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. The quality detection method of the rush formula granule is characterized by comprising the following steps of:
carrying out characteristic spectrum construction and total content measurement of the epsetol and the dehydro-epsetol on the rush formula particles, limiting the content standard of the formula particles to 1.0-5.6 mg of the total content of the epsetol and the dehydro-epsetol in each 1g, and adopting a liquid chromatography for the characteristic spectrum construction and the total content measurement of the epsetol and the dehydro-epsetol;
the determination of the characteristic spectrum by liquid chromatography comprises: taking a solution prepared from a rush control medicinal material as a control medicinal material reference substance solution a, taking a solution prepared from a reference substance of the epfurol and dehydro-epfurol as a reference substance solution a, taking a solution prepared from a rush formula particle sample as a test substance solution a, respectively precisely sucking the control medicinal material reference substance solution a, the reference substance solution a and the test substance solution a, respectively injecting the solutions into a liquid chromatograph, and measuring to obtain the rush-based medicinal material reference substance; wherein the chromatographic conditions adopted are that: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m; mobile phase: using methanol as a mobile phase A and 0.5% formic acid solution as a mobile phase B, and performing gradient elution according to the specification of a table a;
table a gradient elution procedure
Flow rate: 0.8mL/min; column temperature: 30 ℃; sample injection amount: 10. Mu.L; detection wavelength: 282nm.
2. The method for detecting the quality of rush formula particles according to claim 1, further comprising the step of identifying rush formula particles by thin-layer chromatography, said thin-layer chromatography comprising the steps of:
(1) Preparing a test sample solution b: taking 1.0g of rush formula granule sample, grinding, adding 100ml of methanol, heating and refluxing for 1 hour, cooling, filtering, evaporating filtrate, washing residue with 2ml of diethyl ether, discarding diethyl ether solution, volatilizing, and adding 5ml of methanol to dissolve to obtain a sample solution b;
(2) Preparing a control medicinal material solution b: weighing 1.0g of rush herb reference medicine, grinding, adding 100ml of methanol, heating and refluxing for 1 hour, cooling, filtering, evaporating filtrate, washing residues with 2ml of diethyl ether, discarding diethyl ether liquid, volatilizing, adding 5ml of methanol to dissolve, and preparing reference medicine solution b;
(3) Thin layer chromatography analysis was performed: the thin layer chromatography conditions were as follows: silica gel G thin layer plate; sample application amount: 7 μl of test solution b and 15 μl of control solution b; developing agent: taking cyclohexane-ethyl acetate solution with the volume ratio of 10:7 as a developing agent, and checking: spraying 5% phosphomolybdic acid ethanol solution, and heating at 105deg.C until the color of spots is clear.
3. The method for detecting the quality of rush formula particles according to claim 1 wherein the step of measuring the characteristic spectrum by liquid chromatography further comprises the steps of:
(1) Preparing a reference substance solution a of a control medicinal material: taking 0.1g of rush herb reference medicine, adding 25ml of water and 25ml of methanol, weighing, performing ultrasonic treatment for 20min, cooling, filtering, and taking the subsequent filtrate as reference medicine reference solution a;
(2) Preparing a reference substance solution a of a reference substance: taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1ml of the mixed solution as the reference substance solution a of the reference substances;
(3) Preparing a test sample solution a: taking appropriate amount of medulla Junci prescription granule, grinding, taking 0.1g, placing into conical flask with plug, adding 25mL of methanol, ultrasonic treating for 30min, cooling, shaking, filtering, and taking the subsequent filtrate as sample solution a.
4. The method for detecting the quality of rush formula particles according to claim 1, wherein the sample chromatogram measured by liquid chromatography shows 5 characteristic peaks corresponding to 5 characteristic peak retention times in a reference chromatogram of a reference medicinal material; calculating the relative retention time of the peak 1, the peak 2, the peak 5 and the reference peak S in the chromatogram of the sample by taking the peak 3 corresponding to the reference substance of the Eptifhol as the reference peak S; the specified value is: peak 1 relative retention time 0.47, peak 2 relative retention time 0.82, peak 5 relative retention time 1.24.
5. The method for detecting the quality of rush formula particles according to claim 1, wherein the step of measuring the total content of the epfurol and the dehydroepfurol by adopting a liquid chromatography method comprises the following steps: performing liquid chromatograph analysis, taking a solution prepared from a reference substance of the epfurol and the dehydroepfurol as a reference substance solution c, taking a solution prepared from a rush prescription granule sample as a test substance solution c, respectively precisely sucking the reference substance solution c and the test substance solution c, respectively injecting into the liquid chromatograph, and measuring to obtain the product; wherein the chromatographic conditions adopted are that: octadecylsilane chemically bonded silica is used as a filler, the column length is 250mm, the inner diameter is 4.6mm, and the particle size is 5 mu m; mobile phase: methanol-0.5% formic acid solution with the volume ratio of 66:34 is taken as a mobile phase; flow rate: 1.0mL/min; column temperature: 30 ℃; sample injection amount: 10. Mu.L; detection wavelength: 280nm.
6. The method for detecting the quality of rush formula particles according to claim 5 wherein the step of measuring the total content of the epfurol and the dehydroepfurol by liquid chromatography further comprises the steps of:
(1) Preparing a reference substance solution: taking a proper amount of the reference substances of the erliflav and the dehydro-erliflav, precisely weighing, and adding methanol to prepare a mixed solution containing 30 mug of the erliflav and 20 mug of the dehydro-erliflav per 1ml of the mixed solution as a reference substance solution c;
(2) Preparing a test solution: taking appropriate amount of rush prescription granule, grinding, taking 0.1g, precisely weighing, placing into a conical bottle with a plug, precisely adding 25ml of methanol, sealing, weighing, ultrasonic treating for 20 minutes, cooling, weighing again, supplementing loss weight with methanol, shaking, filtering, and taking the subsequent filtrate as a sample solution c.
7. The method for detecting the quality of rush formula particles according to any one of claims 1 to 6, further comprising a detection method,
carrying out character identification, dry extract yield inspection and extract inspection on the rush prescription granule, wherein the extract inspection is measured by adopting a hot dipping method.
8. The method for detecting the quality of rush formula granules according to claim 7, wherein the hot dip method uses ethanol as a solvent and the range of extract is measured by the hot dip method under the alcohol-soluble extract measurement method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311604630.5A CN117420241A (en) | 2023-11-27 | 2023-11-27 | Quality detection method for rush formula particles |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311604630.5A CN117420241A (en) | 2023-11-27 | 2023-11-27 | Quality detection method for rush formula particles |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117420241A true CN117420241A (en) | 2024-01-19 |
Family
ID=89526713
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311604630.5A Pending CN117420241A (en) | 2023-11-27 | 2023-11-27 | Quality detection method for rush formula particles |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117420241A (en) |
-
2023
- 2023-11-27 CN CN202311604630.5A patent/CN117420241A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112587642B (en) | Preparation method and detection method of vitality-maintaining pharmaceutical composition | |
| CN109406651B (en) | Quality detection method of medicinal composition for treating restlessness | |
| CN111624271A (en) | Liquid chromatography method for detecting corresponding substance of peony and licorice decoction, standard fingerprint spectrum and application | |
| CN113917041B (en) | Quality detection method for cortex moutan standard decoction | |
| CN113552271A (en) | Quality control method of acanthopanax standard decoction | |
| CN113848278A (en) | Quality control method for standard decoction of radix Cudraniae | |
| CN113933445A (en) | Quality control method for dendrobium standard decoction | |
| CN102274401B (en) | Traditional Chinese medicine preparation for treating gastropathy and preparation method and detection method thereof | |
| CN112773831A (en) | Preparation method and detection method of peony and liquorice pharmaceutical composition | |
| CN101239117A (en) | Quality control method of traditional Chinese medicine formulations for treating gynaecologic disease | |
| CN107884508A (en) | The quality determining method of Yinhuang lung clearing capsule | |
| CN108535399B (en) | Detection method of Fuyankang pills | |
| CN116008419A (en) | Quality detection method for cortex acanthopanacis formula particles | |
| CN117420241A (en) | Quality detection method for rush formula particles | |
| CN113917002B (en) | Construction method of ophiopogon decoction characteristic spectrum | |
| CN115963192A (en) | Quality control method of muscle and bone pain relieving pills | |
| CN100370253C (en) | Method for quality control of Qianbai biyan solid prepn. for treating rhinitis | |
| CN101028474B (en) | Method for inspecting the quality of Chinese preparation with Yang-and kidney tonifying functions | |
| CN110075179B (en) | Aconitum rupestris formula granules, preparation method and detection method thereof | |
| CN118169275A (en) | Quality detection method for loofah sponge formula particles | |
| CN117990847A (en) | Method for detecting quality of fried earthworm formula particles | |
| CN110927303B (en) | HPLC (high performance liquid chromatography) characteristic spectrum of Shuyanqing spray, construction method and application | |
| CN116148390A (en) | Quality detection method for salt orange core formula particles | |
| CN117368370A (en) | Quality detection method for fructus amomi formula particles | |
| CN117607332A (en) | Polyporus umbellatus formula granule quality detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |