CN113533613B - Method for identifying specific components in liver soothing granules - Google Patents
Method for identifying specific components in liver soothing granules Download PDFInfo
- Publication number
- CN113533613B CN113533613B CN202110805140.6A CN202110805140A CN113533613B CN 113533613 B CN113533613 B CN 113533613B CN 202110805140 A CN202110805140 A CN 202110805140A CN 113533613 B CN113533613 B CN 113533613B
- Authority
- CN
- China
- Prior art keywords
- solution
- taking
- sample
- liver
- reference substance
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000008187 granular material Substances 0.000 title claims abstract description 15
- 210000004185 liver Anatomy 0.000 title claims description 8
- 239000000463 material Substances 0.000 claims abstract description 16
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 claims abstract description 15
- UILPJVPSNHJFIK-UHFFFAOYSA-N Paeonol Chemical compound COC1=CC=C(C(C)=O)C(O)=C1 UILPJVPSNHJFIK-UHFFFAOYSA-N 0.000 claims abstract description 14
- YKRGDOXKVOZESV-WRJNSLSBSA-N Paeoniflorin Chemical compound C([C@]12[C@H]3O[C@]4(O)C[C@](O3)([C@]1(C[C@@H]42)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C)OC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-WRJNSLSBSA-N 0.000 claims abstract description 13
- YKRGDOXKVOZESV-UHFFFAOYSA-N paeoniflorin Natural products O1C(C)(C2(CC34)OC5C(C(O)C(O)C(CO)O5)O)CC3(O)OC1C24COC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-UHFFFAOYSA-N 0.000 claims abstract description 13
- 238000004809 thin layer chromatography Methods 0.000 claims abstract description 11
- IBFYXTRXDNAPMM-BVTMAQQCSA-N Geniposide Chemical compound O([C@@H]1OC=C([C@@H]2[C@H]1C(=CC2)CO)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O IBFYXTRXDNAPMM-BVTMAQQCSA-N 0.000 claims abstract description 10
- IBFYXTRXDNAPMM-FZEIBHLUSA-N Geniposide Natural products COC(=O)C1=CO[C@@H](O[C@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2O)[C@H]2[C@@H]1CC=C2CO IBFYXTRXDNAPMM-FZEIBHLUSA-N 0.000 claims abstract description 10
- 239000004378 Glycyrrhizin Substances 0.000 claims abstract description 10
- VGLLGNISLBPZNL-RBUKDIBWSA-N arborescoside Natural products O=C(OC)C=1[C@@H]2C([C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)OC=1)=C(CO)CC2 VGLLGNISLBPZNL-RBUKDIBWSA-N 0.000 claims abstract description 10
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229960004949 glycyrrhizic acid Drugs 0.000 claims abstract description 10
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 claims abstract description 10
- 235000019410 glycyrrhizin Nutrition 0.000 claims abstract description 10
- YLTGFGDODHXMFB-UHFFFAOYSA-N isoacetovanillon Natural products COC1=CC=C(C(C)=O)C=C1O YLTGFGDODHXMFB-UHFFFAOYSA-N 0.000 claims abstract description 7
- MLIBGOFSXXWRIY-UHFFFAOYSA-N paeonol Natural products COC1=CC=C(O)C(C(C)=O)=C1 MLIBGOFSXXWRIY-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 51
- 239000013558 reference substance Substances 0.000 claims description 28
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 26
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N EtOH Substances CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 22
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 239000012488 sample solution Substances 0.000 claims description 15
- 239000000523 sample Substances 0.000 claims description 14
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 239000011259 mixed solution Substances 0.000 claims description 7
- 238000002137 ultrasound extraction Methods 0.000 claims description 7
- 238000007605 air drying Methods 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 238000005507 spraying Methods 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 5
- 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 4
- 229960000583 acetic acid Drugs 0.000 claims description 4
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 claims description 4
- 235000019253 formic acid Nutrition 0.000 claims description 4
- 239000012362 glacial acetic acid Substances 0.000 claims description 4
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- OAMZXMDZZWGPMH-UHFFFAOYSA-N ethyl acetate;toluene Chemical compound CCOC(C)=O.CC1=CC=CC=C1 OAMZXMDZZWGPMH-UHFFFAOYSA-N 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 229920006395 saturated elastomer Polymers 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 2
- DQYBDCGIPTYXML-UHFFFAOYSA-N ethoxyethane;hydrate Chemical compound O.CCOCC DQYBDCGIPTYXML-UHFFFAOYSA-N 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 239000013074 reference sample Substances 0.000 claims 1
- 239000012088 reference solution Substances 0.000 claims 1
- 239000000126 substance Substances 0.000 abstract description 7
- 244000303040 Glycyrrhiza glabra Species 0.000 abstract description 6
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 abstract description 6
- 238000000605 extraction Methods 0.000 abstract description 5
- 238000007689 inspection Methods 0.000 abstract description 5
- 235000011477 liquorice Nutrition 0.000 abstract description 5
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 5
- 238000003908 quality control method Methods 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 2
- 230000000857 drug effect Effects 0.000 abstract description 2
- 244000111489 Gardenia augusta Species 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- VYQNWZOUAUKGHI-UHFFFAOYSA-N monobenzone Chemical compound C1=CC(O)=CC=C1OCC1=CC=CC=C1 VYQNWZOUAUKGHI-UHFFFAOYSA-N 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- 244000236658 Paeonia lactiflora Species 0.000 description 3
- 235000008598 Paeonia lactiflora Nutrition 0.000 description 3
- 235000003889 Paeonia suffruticosa Nutrition 0.000 description 3
- 240000005001 Paeonia suffruticosa Species 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- WHBHBVVOGNECLV-OBQKJFGGSA-N 11-deoxycortisol Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 WHBHBVVOGNECLV-OBQKJFGGSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 241000202807 Glycyrrhiza Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- LTINPJMVDKPJJI-UHFFFAOYSA-N iodinated glycerol Chemical compound CC(I)C1OCC(CO)O1 LTINPJMVDKPJJI-UHFFFAOYSA-N 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 241000213006 Angelica dahurica Species 0.000 description 1
- 241000132012 Atractylodes Species 0.000 description 1
- 241000202726 Bupleurum Species 0.000 description 1
- 206010008570 Chloasma Diseases 0.000 description 1
- 244000150195 Cyperus longus Species 0.000 description 1
- 235000018109 Cyperus longus Nutrition 0.000 description 1
- 244000075634 Cyperus rotundus Species 0.000 description 1
- 206010013789 Dry throat Diseases 0.000 description 1
- 235000018958 Gardenia augusta Nutrition 0.000 description 1
- XJMPAUZQVRGFRE-SCHFUKFYSA-N Gardenoside Natural products O=C(OC)C=1[C@H]2[C@H]([C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)OC=1)[C@@](O)(CO)C=C2 XJMPAUZQVRGFRE-SCHFUKFYSA-N 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 235000017443 Hedysarum boreale Nutrition 0.000 description 1
- 235000007858 Hedysarum occidentale Nutrition 0.000 description 1
- 206010027146 Melanoderma Diseases 0.000 description 1
- 208000003351 Melanosis Diseases 0.000 description 1
- 208000019255 Menstrual disease Diseases 0.000 description 1
- 244000246386 Mentha pulegium Species 0.000 description 1
- 235000016257 Mentha pulegium Nutrition 0.000 description 1
- 235000004357 Mentha x piperita Nutrition 0.000 description 1
- YNWAFLRDCQAJNF-UHFFFAOYSA-N O.C(=O)O.CC(=O)C.C(C)(=O)OCC Chemical compound O.C(=O)O.CC(=O)C.C(C)(=O)OCC YNWAFLRDCQAJNF-UHFFFAOYSA-N 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 235000008599 Poria cocos Nutrition 0.000 description 1
- 244000197580 Poria cocos Species 0.000 description 1
- 235000005010 Scirpus paludosus Nutrition 0.000 description 1
- 206010047513 Vision blurred Diseases 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- PQLVXDKIJBQVDF-UHFFFAOYSA-N acetic acid;hydrate Chemical compound O.CC(O)=O PQLVXDKIJBQVDF-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 239000002021 butanolic extract Substances 0.000 description 1
- HYGYSIDIKIGPJA-UHFFFAOYSA-N chloroform;ethyl acetate;methanol Chemical compound OC.ClC(Cl)Cl.CCOC(C)=O HYGYSIDIKIGPJA-UHFFFAOYSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 239000001947 glycyrrhiza glabra rhizome/root Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 235000001050 hortel pimenta Nutrition 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000005906 menstruation Effects 0.000 description 1
- XJMPAUZQVRGFRE-AYDWLWLASA-N methyl (1s,4as,7s,7as)-7-hydroxy-7-(hydroxymethyl)-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4a,7a-dihydro-1h-cyclopenta[c]pyran-4-carboxylate Chemical compound O([C@@H]1OC=C([C@@H]2[C@H]1[C@](C=C2)(O)CO)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O XJMPAUZQVRGFRE-AYDWLWLASA-N 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
- 239000002574 poison Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 description 1
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 description 1
- 235000012141 vanillin Nutrition 0.000 description 1
- 239000002699 waste material Substances 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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a method for identifying specific components in liver-soothing granules, which uses paeonol, geniposide, paeoniflorin and glycyrrhizin as standard substances and liquorice as reference medicinal materials to carry out thin-layer chromatography analysis. The invention can identify 4 traditional Chinese medicines in a specific way by utilizing an extraction method and two thin-layer conditions, has the advantages of simple, convenient and environment-friendly method, reduces the inspection cost of enterprises, is simple to operate and low in cost, and aims to provide references for drug effect substance basis and quality control research of liver-soothing granules, avoid judging the one-sided property of the whole quality of the preparation by only measuring single chemical components, and reduce the possibility of artificial treatment for reaching the standard of the quality.
Description
Technical Field
The invention belongs to the technical field of formula component detection, and particularly relates to a method for identifying specific components in liver-soothing granules.
Background
The liver soothing granule has effects of soothing liver, regulating qi, dispelling melancholy and regulating menstruation, and can be used for treating pain in hypochondrium, chest and abdomen distention, menoxenia, headache, blurred vision, vexation, bitter taste in mouth, dry throat, and facial black spot (chloasma) caused by stagnation of liver-qi. The prescription of the liver-soothing granule is prepared from 10 Chinese medicinal materials including bupleurum root, chinese angelica, white peony root, white atractylodes rhizome, poria cocos, licorice root, peppermint, tree peony bark, cape jasmine and nutgrass galingale rhizome.
The traditional Chinese medicine compound is prepared from a plurality of traditional Chinese medicines, has the principle of overall appearance and diagnosis and treatment, and is difficult to express the overall efficacy of the traditional Chinese medicine compound and can not fully control the quality because of complex ingredients of the traditional Chinese medicine compound and detection of single active ingredients. The identification method of the specific components of the tree peony bark is complex in operation, more reagents are consumed, identification and detection are needed to be carried out respectively, the identification method of the specific components of the tree peony bark is studied in the quality standard of the liver soothing granules, the method is complex, the number of waste products after identification is large, and the enterprise inspection cost is increased. Therefore, a method for identifying specific components in liver-soothing granules is needed.
Disclosure of Invention
The invention provides a method for identifying specific components in liver-soothing granules.
The invention comprises the following steps:
grinding the liver soothing particles to be detected, and adding water and diethyl ether according to the volume ratio of 1:3, carrying out ultrasonic treatment for 20 minutes, sucking the supernatant, volatilizing, and then adding acetone for dissolution to serve as a first sample solution;
b, taking paeonol reference substance, adding acetone to prepare a solution containing 1mg per 1ml, and taking the solution as a primary reference substance solution;
c, performing thin layer chromatography test, namely sucking 5 μl of reference substance solution, respectively spotting 20 μl of test substance solution on the same silica gel G thin layer plate, spreading with toluene and ethyl acetate mixed solution as developing agent, taking out, air drying, spraying mixed solution of hydrochloric acid and ferric trichloride ethanol solution, heating at 105deg.C until the spots develop clearly, and inspecting the chromatograms of the test substance and the reference substance under sunlight;
d, taking the lower liquid in the step A, adding n-butanol for ultrasonic extraction, and filtering to obtain a second sample solution;
e, adding ethanol into paeoniflorin, glycyrrhizin and geniposide reference substances to prepare a mixed reference substance solution containing 2mg of paeoniflorin, 0.5mg of glycyrrhizin and 1mg of geniposide per 1m1, and taking the mixed reference substance solution as a secondary reference substance solution; decocting Glycyrrhrizae radix control material 0.5g with water 25ml for 30 min, filtering, collecting filtrate, concentrating to about 10ml, and preparing Glycyrrhrizae radix medicinal material solution as secondary control medicinal material solution according to the preparation method of the second test sample solution;
f, performing a secondary thin layer chromatography test, sucking 4 μl of the sample solution, 2 μl of the secondary reference substance and the reference medicinal material solution, respectively spotting on a silica gel G thin layer plate, spreading with a mixed solution of ethyl acetate, formic acid, glacial acetic acid and water as a developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until spots are clear, and respectively inspecting the chromatograms of the sample, the reference substance and the reference medicinal material under sunlight and 365nm ultraviolet light.
Further, in the step C, toluene-ethyl acetate (9:1) is used as a developing agent, and a mixed solution of hydrochloric acid and 5% ferric trichloride ethanol solution (0.25 ml: 100 ml) is sprayed after the drying.
Further, in the step D, the lower layer liquid is taken, 25ml of water saturated n-butanol is added, ultrasonic extraction is carried out twice for 15 minutes each time, the n-butanol liquid after ultrasonic extraction is combined, 30ml of n-butanol saturated water is added, shaking washing is carried out, the n-butanol layer is taken, 2ml of ethanol is added into residues after evaporation to dissolve, and the residues are used as a sample solution for secondary thin layer identification.
Further, in step F, the developing agent comprises ethyl acetate, formic acid, glacial acetic acid and water, wherein the volume ratio of the ethyl acetate, the formic acid, the glacial acetic acid and the water is 15:1:1:2.
The method is applied to quality control of liver-soothing granules.
One or more embodiments of the present invention may have the following advantages over the prior art:
the invention can identify 4 traditional Chinese medicines in a specific way by utilizing an extraction method and two thin-layer conditions, has the advantages of simple, convenient and environment-friendly method, reduces the inspection cost of enterprises, is simple to operate and low in cost, and is expected to provide reference for the drug effect substance basis and quality control research of the liver-soothing granules.
The invention avoids the unilateral performance of judging the whole quality of the preparation by only measuring single chemical components, reduces the possibility of artificial treatment for reaching the quality standard, and simultaneously can more comprehensively and scientifically evaluate the quality of the liver-soothing granules by carrying out systematic analysis on a plurality of batches of samples, thereby ensuring the quality and curative effect of the product and providing a good solution for quality control, thereby having great practical application value.
Drawings
FIG. 1 is a comparison of paeonol thin layer chromatography identification patterns;
wherein S. paeonol reference NA. negative samples 1-10, samples of different batches
FIG. 2 is a comparison of thin layer chromatography identification patterns of fructus Gardeniae, glycyrrhrizae radix, radix Paeoniae alba and cortex moutan under sunlight
Wherein S is 1 A mixed reference substance S of glycyrrhizin, paeoniflorin and geniposide 2 Gardenia glycoside reference substance NA 1 Glycyrrhiza negative sample NA 2 Radix Paeoniae alba negative sample NA 3 Radix Paeoniae alba and cortex moutan negative sample NA 4 Gardenia negative samples 1-10. Samples of different batches;
FIG. 3 is a comparison of thin layer chromatography identification patterns of fructus Gardeniae, glycyrrhrizae radix, radix Paeoniae alba and cortex moutan under 365nm ultraviolet light;
wherein S is 1 A mixed reference substance S of glycyrrhizin, paeoniflorin and geniposide 2 Gardenia glycoside reference substance NA 1 Glycyrrhiza negative sample NA 2 Radix Paeoniae alba negative sample NA 3 Radix Paeoniae alba and cortex moutan negative sample NA 4 Gardenia negative samples 1-10. Samples of different batches;
Detailed Description
The drawings and the embodiments of the present invention will be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the invention are shown.
In this example, 20g (sugar-containing type) or 6g (low sugar-containing type) of the sample was ground, 10ml (sugar-containing type) or 5ml (low sugar-containing type) of water was added, 30ml of diethyl ether was further added, the mixture was sealed, and the mixture was sonicated for 20 minutes, and the supernatant was sucked, evaporated, and dissolved in 0.5ml of acetone to give a sample solution. The lower layer is reserved.
And adding acetone into paeonol reference substance to obtain 1mg solution per 1ml, and taking the solution as reference substance solution. According to thin layer chromatography (rule 0502 of Chinese pharmacopoeia 2020 edition), sucking 5 μl of control solution and 20 μl of test solution, respectively spotting on the same silica gel G thin layer plate, spreading with toluene-ethyl acetate (9:1) as developing agent, taking out, air drying, spraying mixed solution of hydrochloric acid and 5% ferric trichloride ethanol solution (0.25 ml: 100 ml), and heating at 105deg.C until the spot color is clear.
As shown in FIG. 1, the spots of the same color appear on the chromatogram of the test sample at the positions corresponding to the chromatogram of the control sample when examined under sunlight. The lower liquid was taken, 25ml of water-saturated n-butanol was added, and ultrasonic extraction was performed twice for 15 minutes each time. Mixing n-butanol solutions, adding n-butanol saturated water 30ml, shaking, washing, collecting n-butanol layer, evaporating to dryness, and dissolving the residue with ethanol 2ml to obtain test solution. And adding ethanol into paeoniflorin, glycyrrhizin and geniposide reference substance to obtain mixed reference substance solution containing paeoniflorin 2 mg/1 m1, glycyrrhizin 0.5mg and geniposide 1 mg. Decocting Glycyrrhrizae radix control material 0.5g with water 25ml for 30 min, filtering, collecting filtrate, concentrating to about 10ml, and preparing Glycyrrhrizae radix control material solution according to sample solution preparation method. According to thin layer chromatography (rule 0502 of Chinese pharmacopoeia 2020 edition), sucking 4 μl of sample solution, 2 μl of reference substance and reference medicinal solution, respectively spotting on the same silica gel G thin layer plate, spreading with ethyl acetate-formic acid-glacial acetic acid-water (15:1:1:2) as developing agent, taking out, air drying, spraying with 10% sulfuric acid ethanol solution, heating to 105 deg.C until spots are clear, and respectively placing under sunlight and ultraviolet lamp (365 nm) for inspection. In the sample chromatogram shown in FIG. 2, spots of the same color appear at the positions corresponding to the control material and the control material chromatogram.
Comparative example
Grinding 15g (sugar-containing) or 5g (low sugar-containing) of the product, adding 20ml of ethyl acetate, reflux-extracting for 30 min, filtering, evaporating the filtrate to dryness, dissolving the residue with 1ml of methanol, and filtering to obtain filtrate as sample solution. Adding methanol into the gardenoside reference substance to obtain a solution containing 1mg per 1ml, and taking the solution as reference substance solution. According to the thin layer chromatography (appendix VIB) test, 5-10 μl of each of the above two solutions is sucked and respectively spotted on the same silica gel G thin layer plate, and the two solutions are spread with ethyl acetate-acetone-formic acid-water (10:6:2:0.5) as a developing agent, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and baked at 105 ℃ for about 10 minutes until spots are clear. Spots of the same color appear in the sample chromatogram at positions corresponding to those of the control chromatogram.
Taking the residue extracted by ethyl acetate under the condition of [ identification ] (1), volatilizing, adding 30ml of ethanol, reflux-extracting for 30 minutes, filtering, evaporating filtrate to dryness, adding 20ml of water into the residue, heating to dissolve, extracting 3 times with water-saturated n-butanol, each time with 0ml, combining n-butanol extract, washing 2 times with water, each time with 10ml, discarding water solution, evaporating n-butanol solution to dryness, and adding 1ml of ethanol into the residue to dissolve, thereby obtaining a sample solution. And adding ethanol into paeoniflorin reference substance to obtain a solution containing 1mg of paeoniflorin per 1m1 as reference substance solution. According to a thin layer chromatography (appendix VIB) test, 5-10 mu l of each of the two solutions is sucked and respectively spotted on the same silica gel G thin layer plate, and the two solutions are spread by using chloroform-methanol-ethyl acetate (8:4:1) as a developing agent, taken out, dried and sprayed with 5% vanillin sulfuric acid solution, and baked for a few minutes at 105 ℃ until spots are clear. Spots of the same color appear in the sample chromatogram at positions corresponding to those of the control chromatogram.
Compared with the comparative example, the invention has the advantages of improved efficiency, requirement of reflux extraction in the original standard, multiple times of extraction, long operation time and complex operation steps. The existing method only needs ultrasonic extraction and combined extraction.
In addition, as the detected medicine has more medicines, the original standard only detects 2 traditional Chinese medicines, and the existing method can detect 4 traditional Chinese medicines. The special property is strong, paeonol can only indicate the quality of moutan bark, paeoniflorin is a common component of the moutan bark and the white peony root, can indicate the quality of the moutan bark and the white peony root, geniposide can only indicate the quality of gardenia, glycyrrhizin can indicate the quality of liquorice, liquorice reference medicinal materials can only indicate the quality of liquorice, the detection method is safe, environment-friendly and low in cost, chloroform used in the identification process of paeoniflorin is an easy-to-poison reagent in the original standard, the solvent is abandoned in the existing preparation method, the safety and environment-friendly performance are ensured, the purchase cost of enterprises is reduced, the identification reliability is improved, the liquorice shows spots with different colors by using sunlight and 365nm ultraviolet light for inspection, and the identification reliability is improved.
It should be noted that the above examples are only for illustrating the technical solution of the present invention and are not limiting thereof. Although the present invention has been described in detail with reference to the examples given, those skilled in the art can make modifications and equivalents to the technical solutions of the present invention as required, without departing from the spirit and scope of the technical solutions of the present invention.
Claims (2)
1. The identification method of the specific components in the liver soothing granules is characterized by comprising the following steps:
A. grinding a to-be-detected product of the liver soothing granules, and adding water and diethyl ether according to a volume ratio of 1:3, carrying out ultrasonic treatment for 20 minutes, sucking the supernatant, volatilizing, adding acetone for dissolution, and taking the mixture as a first sample solution, wherein the lower layer is reserved;
B. taking paeonol reference substance, adding acetone to prepare a solution containing 1mg per 1ml, and taking the solution as a primary reference substance solution;
C. taking 5 μl of primary reference solution and 20 μl of first sample solution, respectively spotting on the same silica gel G thin layer plate, taking toluene-ethyl acetate volume ratio of 9:1 as developing agent, air drying, spraying mixed solution of hydrochloric acid and 5% ferric trichloride ethanol solution of 0.25 ml:100 ml, heating to 105deg.C until the spot color is clear, and inspecting the chromatograms of the sample and the reference sample in sunlight;
D. taking the lower liquid in the step A, adding 25ml of water saturated n-butanol, carrying out ultrasonic extraction twice for 15 minutes each time, combining the n-butanol liquid obtained after ultrasonic extraction, adding 30ml of n-butanol saturated water, shaking and washing, taking an n-butanol layer, evaporating residues, adding 2ml of ethanol to dissolve the residues, and taking the residues as a sample solution for secondary thin layer identification;
E. adding ethanol into paeoniflorin, glycyrrhizin and geniposide reference substance to obtain mixed reference substance solution containing paeoniflorin 2mg, glycyrrhizin 0.5mg and geniposide 1mg per 1ml, and taking as secondary reference substance solution; decocting Glycyrrhrizae radix control material 0.5g with water 25ml for 30 min, filtering, collecting filtrate, concentrating to about 10ml, and preparing Glycyrrhrizae radix medicinal material solution as secondary control medicinal material solution according to the preparation method of the second test sample solution;
F. performing secondary thin layer chromatography test, sucking 4 μl of the sample solution, and 2 μl of the secondary reference and reference medicinal solutions, respectively spotting on silica gel G thin layer plate, spreading with mixed solution of ethyl acetate, formic acid, glacial acetic acid and water at volume ratio of 15:1:1:2 as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until spots are clear, and respectively placing under sunlight and 365nm ultraviolet lamp to inspect chromatograms of the sample, reference and reference medicinal materials.
2. Use of the method according to claim 1 for quality detection of liver-soothing particles.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110805140.6A CN113533613B (en) | 2021-07-16 | 2021-07-16 | Method for identifying specific components in liver soothing granules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110805140.6A CN113533613B (en) | 2021-07-16 | 2021-07-16 | Method for identifying specific components in liver soothing granules |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113533613A CN113533613A (en) | 2021-10-22 |
| CN113533613B true CN113533613B (en) | 2023-11-28 |
Family
ID=78128354
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110805140.6A Active CN113533613B (en) | 2021-07-16 | 2021-07-16 | Method for identifying specific components in liver soothing granules |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113533613B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101569702A (en) * | 2009-05-22 | 2009-11-04 | 湖南新汇制药有限公司 | Quality detection method for houtou jianweiling Chinese medicament |
| CN101732527A (en) * | 2010-01-28 | 2010-06-16 | 昆明中药厂有限公司 | Quality detection method for liver-soothing granules |
-
2021
- 2021-07-16 CN CN202110805140.6A patent/CN113533613B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101569702A (en) * | 2009-05-22 | 2009-11-04 | 湖南新汇制药有限公司 | Quality detection method for houtou jianweiling Chinese medicament |
| CN101732527A (en) * | 2010-01-28 | 2010-06-16 | 昆明中药厂有限公司 | Quality detection method for liver-soothing granules |
Non-Patent Citations (9)
| Title |
|---|
| 丹栀逍遥胶囊质量标准研究;陈东鸿 等;海峡药学(第02期);第41-44页 * |
| 加味逍遥丸的制备与质量控制;石勇强;医学信息(中旬刊)(第01期);第335-337页 * |
| 加味逍遥片的薄层色谱研究;张雅丽 等;中国医院用药评价与分析(第01期);第84-87页 * |
| 多波长HPLC法测定加味逍遥丸中多种成分含量及化学计量学分析;于新兰 等;药物分析杂志(第07期);第1262-1268页 * |
| 广州市药品检验所.《中草药制齐检验技术》.人民卫生出版社,1979,第110页. * |
| 曾元儿.《中国药典2010年版（一部）化学成分分析简明手册》.中山大学出版社,2010,第282-283页. * |
| 舒肝胶囊的质量控制;付超美 等;中国中药杂志(第06期);第344-347页 * |
| 舒肝颗粒的质量标准研究;朱敏 等;中国民族民间医药;第25卷(第15期);第24-27页 * |
| 芍甘胶囊质量控制标准的研究;瞿发林 等;解放军药学学报;第24卷(第02期);第139-142页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113533613A (en) | 2021-10-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111044624B (en) | Quality detection method of Chinese medicinal preparation | |
| CN110455965B (en) | Preparation method of pharmaceutical composition and HPLC fingerprint spectrum establishment method thereof | |
| CN1325072C (en) | Method for preparing traditional Chinese medicine | |
| CN107515266B (en) | Detection method of medicinal preparation of magnolia officinalis seven-ingredient decoction | |
| CN111521725B (en) | One-plate multi-medicinal material multi-information rapid thin-layer identification method of Gentongping tablets | |
| CN102139046B (en) | Quality control method for antidotal forsythia tablet | |
| CN113514597B (en) | Thin-layer chromatography identification method of gastrodia tuber dizziness relieving granule | |
| CN112697949B (en) | Thin-layer identification method for Baoyuan decoction, similar formula extract and preparation thereof | |
| CN113533613B (en) | Method for identifying specific components in liver soothing granules | |
| CN1907429A (en) | Method for preparing and controlling the quality of Chinese medicinal soft capsule | |
| CN101385808B (en) | Detection method of Zhubai tranquilizing pill | |
| CN101279066B (en) | Quality testing method of medicament composition for curing hysteromyoma of gynecology | |
| CN100559180C (en) | The detection method of aqueous donkey-hide gelatin and Chinese angelica preparation | |
| CN107576749A (en) | A kind of detection method of rheumatic cold pain piece | |
| WO2013044570A1 (en) | Detection method of medicine for curing mastitis and hyperplasia of mammary glands | |
| CN101632804B (en) | Quality control method for wind-dispelling heat-dissipating capsules | |
| CN1309372C (en) | Preparation of granular powder for treating hepatitis and quality control method | |
| CN101721625B (en) | Quality control method of shenyankangfu tablet | |
| CN107782840B (en) | Detection method of Shanmei summer-heat clearing granules | |
| CN109959749B (en) | Method for identifying polygonatum odoratum paste by adopting thin-layer chromatography | |
| CN114689783B (en) | Quick thin-layer identification method for poria, cassia, rhizoma atractylodis and sweet soup freeze-dried powder | |
| CN110470783B (en) | Quality detection method of Shuanghuang lipid-lowering granules | |
| CN115837063B (en) | Preparation and detection methods of lung force cough pharmaceutical composition | |
| CN113633733B (en) | Preparation method of gynecological reconstruction pill | |
| CN117665191B (en) | Method for detecting quality of effective components in known cellulitis particles |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |