CN111366673B - Identification method of traditional Chinese medicine preparation qingjiang tablet - Google Patents
Identification method of traditional Chinese medicine preparation qingjiang tablet Download PDFInfo
- Publication number
- CN111366673B CN111366673B CN201811599671.9A CN201811599671A CN111366673B CN 111366673 B CN111366673 B CN 111366673B CN 201811599671 A CN201811599671 A CN 201811599671A CN 111366673 B CN111366673 B CN 111366673B
- Authority
- CN
- China
- Prior art keywords
- solution
- methanol
- taking
- chloroform
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 239000003814 drug Substances 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 14
- LQGUBLBATBMXHT-UHFFFAOYSA-N chrysophanol Chemical compound C1=CC=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O LQGUBLBATBMXHT-UHFFFAOYSA-N 0.000 claims abstract description 42
- 238000004809 thin layer chromatography Methods 0.000 claims abstract description 36
- CRDNMYFJWFXOCH-YPKPFQOOSA-N (3z)-3-(3-oxo-1h-indol-2-ylidene)-1h-indol-2-one Chemical compound N/1C2=CC=CC=C2C(=O)C\1=C1/C2=CC=CC=C2NC1=O CRDNMYFJWFXOCH-YPKPFQOOSA-N 0.000 claims abstract description 32
- VWDXGKUTGQJJHJ-UHFFFAOYSA-N Catenarin Natural products C1=C(O)C=C2C(=O)C3=C(O)C(C)=CC(O)=C3C(=O)C2=C1O VWDXGKUTGQJJHJ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000010282 Emodin Substances 0.000 claims abstract description 26
- RBLJKYCRSCQLRP-UHFFFAOYSA-N Emodin-dianthron Natural products O=C1C2=CC(C)=CC(O)=C2C(=O)C2=C1CC(=O)C=C2O RBLJKYCRSCQLRP-UHFFFAOYSA-N 0.000 claims abstract description 26
- YOOXNSPYGCZLAX-UHFFFAOYSA-N Helminthosporin Natural products C1=CC(O)=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O YOOXNSPYGCZLAX-UHFFFAOYSA-N 0.000 claims abstract description 26
- NTGIIKCGBNGQAR-UHFFFAOYSA-N Rheoemodin Natural products C1=C(O)C=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1O NTGIIKCGBNGQAR-UHFFFAOYSA-N 0.000 claims abstract description 26
- RHMXXJGYXNZAPX-UHFFFAOYSA-N emodin Chemical compound C1=C(O)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O RHMXXJGYXNZAPX-UHFFFAOYSA-N 0.000 claims abstract description 26
- VASFLQKDXBAWEL-UHFFFAOYSA-N emodin Natural products OC1=C(OC2=C(C=CC(=C2C1=O)O)O)C1=CC=C(C=C1)O VASFLQKDXBAWEL-UHFFFAOYSA-N 0.000 claims abstract description 26
- PKUBGLYEOAJPEG-UHFFFAOYSA-N physcion Natural products C1=C(C)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O PKUBGLYEOAJPEG-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000000463 material Substances 0.000 claims abstract description 25
- NZPQWZZXRKZCDU-UHFFFAOYSA-N chrysophanol Natural products Cc1cc(O)c2C(=O)c3c(O)cccc3Oc2c1 NZPQWZZXRKZCDU-UHFFFAOYSA-N 0.000 claims abstract description 21
- CRDNMYFJWFXOCH-BUHFOSPRSA-N Couroupitine B Natural products N\1C2=CC=CC=C2C(=O)C/1=C1/C2=CC=CC=C2NC1=O CRDNMYFJWFXOCH-BUHFOSPRSA-N 0.000 claims abstract description 16
- KFFCKOBAHMGTMW-LGQRSHAYSA-N Forsythin Chemical compound C1=C(OC)C(OC)=CC=C1[C@H]1[C@@H](CO[C@@H]2C=3C=C(OC)C(O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)=CC=3)[C@@H]2CO1 KFFCKOBAHMGTMW-LGQRSHAYSA-N 0.000 claims abstract description 16
- CRDNMYFJWFXOCH-UHFFFAOYSA-N isoindigotin Natural products N1C2=CC=CC=C2C(=O)C1=C1C2=CC=CC=C2NC1=O CRDNMYFJWFXOCH-UHFFFAOYSA-N 0.000 claims abstract description 16
- 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 12
- 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 12
- 240000004980 Rheum officinale Species 0.000 claims abstract description 10
- 235000008081 Rheum officinale Nutrition 0.000 claims abstract description 10
- 244000303040 Glycyrrhiza glabra Species 0.000 claims abstract description 7
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 claims abstract description 7
- 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 6
- 235000011477 liquorice Nutrition 0.000 claims abstract description 6
- NOOLISFMXDJSKH-UHFFFAOYSA-N DL-menthol Natural products CC(C)C1CCC(C)CC1O NOOLISFMXDJSKH-UHFFFAOYSA-N 0.000 claims abstract description 5
- QQILFGKZUJYXGS-UHFFFAOYSA-N Indigo dye Chemical compound C1=CC=C2C(=O)C(C3=C(C4=CC=CC=C4N3)O)=NC2=C1 QQILFGKZUJYXGS-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000010231 banlangen Substances 0.000 claims abstract description 5
- NOOLISFMXDJSKH-UTLUCORTSA-N (+)-Neomenthol Chemical compound CC(C)[C@@H]1CC[C@@H](C)C[C@@H]1O NOOLISFMXDJSKH-UTLUCORTSA-N 0.000 claims abstract description 4
- 241000405414 Rehmannia Species 0.000 claims abstract description 4
- 229940041616 menthol Drugs 0.000 claims abstract description 4
- 241000205585 Aquilegia canadensis Species 0.000 claims abstract 2
- 241000255789 Bombyx mori Species 0.000 claims abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 228
- 239000000243 solution Substances 0.000 claims description 133
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 106
- 239000000706 filtrate Substances 0.000 claims description 77
- 238000001914 filtration Methods 0.000 claims description 74
- 239000013558 reference substance Substances 0.000 claims description 67
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 57
- 239000012488 sample solution Substances 0.000 claims description 54
- 239000002904 solvent Substances 0.000 claims description 51
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 50
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 40
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 40
- 238000001704 evaporation Methods 0.000 claims description 40
- 238000010438 heat treatment Methods 0.000 claims description 40
- 238000010992 reflux Methods 0.000 claims description 29
- 238000005303 weighing Methods 0.000 claims description 26
- 238000000227 grinding Methods 0.000 claims description 25
- 239000003795 chemical substances by application Substances 0.000 claims description 24
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 23
- 239000012085 test solution Substances 0.000 claims description 23
- COHYTHOBJLSHDF-UHFFFAOYSA-N Indigo Chemical compound N1C2=CC=CC=C2C(=O)C1=C1C(=O)C2=CC=CC=C2N1 COHYTHOBJLSHDF-UHFFFAOYSA-N 0.000 claims description 21
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 20
- 239000003208 petroleum Substances 0.000 claims description 20
- 239000000523 sample Substances 0.000 claims description 18
- 238000005406 washing Methods 0.000 claims description 18
- 238000009210 therapy by ultrasound Methods 0.000 claims description 17
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 16
- 238000005507 spraying Methods 0.000 claims description 15
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 14
- 239000004952 Polyamide Substances 0.000 claims description 14
- 239000011259 mixed solution Substances 0.000 claims description 14
- 229920002647 polyamide Polymers 0.000 claims description 14
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 12
- 235000000177 Indigofera tinctoria Nutrition 0.000 claims description 11
- 238000001816 cooling Methods 0.000 claims description 11
- 239000003480 eluent Substances 0.000 claims description 11
- 229940097275 indigo Drugs 0.000 claims description 11
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 10
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 10
- 239000000377 silicon dioxide Substances 0.000 claims description 10
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 claims description 10
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 claims description 10
- 235000012141 vanillin Nutrition 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 8
- 235000019253 formic acid Nutrition 0.000 claims description 8
- 239000000945 filler Substances 0.000 claims description 7
- 238000001228 spectrum Methods 0.000 claims description 7
- IBPFZCOJYVDKGN-UHFFFAOYSA-N OC.OC=O.ClC(Cl)Cl.CCOC(C)=O Chemical compound OC.OC=O.ClC(Cl)Cl.CCOC(C)=O IBPFZCOJYVDKGN-UHFFFAOYSA-N 0.000 claims description 6
- SIHHLZPXQLFPMC-UHFFFAOYSA-N chloroform;methanol;hydrate Chemical compound O.OC.ClC(Cl)Cl SIHHLZPXQLFPMC-UHFFFAOYSA-N 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 239000011347 resin Substances 0.000 claims description 6
- 229920005989 resin Polymers 0.000 claims description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 239000011248 coating agent Substances 0.000 claims description 5
- 238000000576 coating method Methods 0.000 claims description 5
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 claims description 5
- 239000012088 reference solution Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 230000001502 supplementing effect Effects 0.000 claims description 5
- 240000005001 Paeonia suffruticosa Species 0.000 claims description 3
- 235000003889 Paeonia suffruticosa Nutrition 0.000 claims description 3
- 238000003556 assay Methods 0.000 claims 5
- 238000007605 air drying Methods 0.000 claims 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims 3
- 239000002245 particle Substances 0.000 claims 2
- 238000001179 sorption measurement Methods 0.000 claims 2
- 235000001453 Glycyrrhiza echinata Nutrition 0.000 claims 1
- 235000017382 Glycyrrhiza lepidota Nutrition 0.000 claims 1
- 239000002021 butanolic extract Substances 0.000 claims 1
- 229940010454 licorice Drugs 0.000 claims 1
- 238000005259 measurement Methods 0.000 claims 1
- 238000011003 system suitability test Methods 0.000 claims 1
- 238000003908 quality control method Methods 0.000 abstract description 6
- 235000006484 Paeonia officinalis Nutrition 0.000 abstract description 4
- 241001106477 Paeoniaceae Species 0.000 abstract description 4
- 240000002948 Ophiopogon intermedius Species 0.000 abstract description 3
- 241000935235 Fritillaria meleagris Species 0.000 abstract 1
- 240000007171 Imperata cylindrica Species 0.000 abstract 1
- 238000011835 investigation Methods 0.000 abstract 1
- 238000002156 mixing Methods 0.000 description 12
- 239000000284 extract Substances 0.000 description 6
- 210000003608 fece Anatomy 0.000 description 5
- 239000003463 adsorbent Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 241000628997 Flos Species 0.000 description 3
- 201000007100 Pharyngitis Diseases 0.000 description 3
- 206010068319 Oropharyngeal pain Diseases 0.000 description 2
- 241000219061 Rheum Species 0.000 description 2
- 235000009411 Rheum rhabarbarum Nutrition 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000007888 film coating Substances 0.000 description 2
- 238000009501 film coating Methods 0.000 description 2
- 238000000691 measurement method Methods 0.000 description 2
- 238000003825 pressing Methods 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 206010013954 Dysphoria Diseases 0.000 description 1
- 241001598107 Imperata Species 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 208000016150 acute pharyngitis Diseases 0.000 description 1
- 206010001093 acute tonsillitis Diseases 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 210000002784 stomach Anatomy 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
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)
- Investigating Or Analysing Biological Materials (AREA)
- Medicinal Preparation (AREA)
Abstract
A method for identifying a traditional Chinese medicine preparation qingjiang tablet is characterized by providing a thin layer chromatography for identifying indigoid indirubin, forsythin, paeoniflorin and liquorice in the traditional Chinese medicine preparation by using a thin layer chromatography for identifying the total amount of rheum officinale according to emodin and chrysophanol in the traditional Chinese medicine preparation, wherein the qingjiang tablet is prepared from Chinese medicinal materials such as silkworm excrement, rheum officinale, indigo naturalis, radix scrophulariae, fructus gleditsiae, red paeony root, radix isatidis, dwarf lilyturf tuber, fructus forsythiae, moutan bark, rehmannia root, liquorice, cogongrass rhizome, honeysuckle, menthol and fritillary bulb. The method can effectively control quality of QINGJIANGPING tablet, and can be used as index for quality control and process reliability investigation.
Description
Technical Field
The invention relates to a method for identifying a traditional Chinese medicine preparation qingjiang tablet, belonging to the field of pharmacy.
Background
The qingjiang tablet performs the national medicine standard, has the functions of clearing heat and detoxicating, relieving sore throat and pain, is used for treating sore throat, fever and dysphoria, constipation, infantile acute pharyngitis, acute tonsillitis and the like caused by lung and stomach heat accumulation. The medicine is prepared from Chinese medicinal materials of faeces Bombycis, radix et rhizoma Rhei, indigo naturalis, radix scrophulariae, fructus Gleditsiae Abnormalis, radix Paeoniae Rubra, radix Isatidis, radix Ophiopogonis, fructus forsythiae, cortex moutan, rehmanniae radix, glycyrrhrizae radix, lalang grass rhizome, flos Lonicerae, mentholum, and Bulbus Fritillariae Cirrhosae; wherein, the red paeony root, the natural indigo, the liquorice and the rhubarb are all main medicinal herbs, have the effects of clearing heat and purging fire, cooling blood and detoxifying, and play a main treatment role. The existing national medicine standard lacks the monitoring standard aiming at the medicinal taste in the quality control method of qingjiang tablets, and has limitations. The invention overcomes the defects of the prior art, perfects the quality control standard of the qingjiang tablet, increases the thin layer chromatography qualitative identification method and the high performance liquid chromatography content measurement method of rheum officinale, and further perfects the quality control standard of the qingjiang tablet.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, perfects and formulates a quality control method of qingjiang tablets, and adds a thin-layer chromatography qualitative identification method of the compound preparation and a high-efficiency liquid chromatography content measurement method of rheum officinale, so that the quality detection method is more perfect, and the quality of qingjiang tablets can be effectively controlled.
The invention is realized by the following technical scheme:
1. the indigo blue and indirubin contained in qingjiang tablet are identified by thin layer chromatography:
a. preparing reference solution by adding chloroform into indigo reference and indirubin reference to obtain mixed solution containing indigo 0.5-2.0 mg and indirubin 0.25-1.0 mg per 1 ml;
b. preparation of sample solution comprises grinding 2.0-3.0g of QINGJIANGPING, adding 40-60ml of petroleum ether (30-60deg.C), ultrasonic treating for 40-50 min, filtering, discarding the filtrate, volatilizing solvent from the residue, adding 40-60ml of chloroform, ultrasonic treating for 40-50 min, filtering, and concentrating the filtrate to about 2ml to obtain sample solution;
c. the thin layer chromatography test comprises the steps of sucking 4-6 μl of the reference substance solution and 2-5 μl of the sample solution, respectively spotting on the same silica gel G thin layer plate or any thin layer plate, taking toluene-chloroform-acetone (5:4:1) as a developing agent or taking toluene-chloroform-acetone (6:5:2) as a developing agent, developing, taking out, airing, and displaying spots with the same color on the positions corresponding to the chromatograph of the reference substance in the sample chromatograph;
2. the forsythin contained in qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into the control solution containing 0.5-2.0 mg of forsythin per 1 ml;
b. preparing test solution by grinding 2.0-3.0g of Qingjiang tablet, adding 40-60ml of petroleum ether (30-60 ℃) for ultrasonic treatment for 40-50 min, filtering, discarding filtrate, volatilizing solvent from residue, adding 40-60ml of chloroform for ultrasonic treatment for 40-50 min, filtering, discarding filtrate, volatilizing solvent from residue, adding 40-50ml of methanol for ultrasonic treatment for 40-50 min, filtering, evaporating filtrate to dryness, adding 10-30 ml of residue to dissolve, centrifuging, collecting supernatant, passing through D101 type macroporous adsorbent resin column (column inner diameter is 1.5cm, column height is 16 cm), eluting with 30% ethanol and 70% ethanol respectively, evaporating to dryness, adding 2-3ml of methanol into residue to dissolve, and collecting the residue as test solution;
c. the thin layer chromatography test comprises the steps of sucking 4-6 μl of the reference substance solution, and 5-10 μl of the sample solution, respectively spotting on the same silica gel G thin layer plate or any thin layer plate, developing with chloroform-methanol-anhydrous formic acid (17:2:1.5) as developing agent or with chloroform-methanol-anhydrous formic acid (15:2:1) as developing agent, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until the color of spots is clear, and developing spots with the same color on the positions corresponding to the color of the reference substance in the color spectrum of the sample;
3. the paeoniflorin contained in the qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into paeoniflorin control solution, and preparing 0.5-2.0 mg of solution containing forsythin per 1ml as control solution;
b. preparing test solution by grinding 2.0-3.0g of QINGJIANGPING, adding 40-60ml of petroleum ether (30-60deg.C), ultrasonic treating for 40-50 min, filtering, discarding filtrate, volatilizing solvent from residue, adding 40-60ml of chloroform, ultrasonic treating for 40-50 min, filtering, discarding filtrate, volatilizing solvent from residue, adding 40-50ml of methanol, heating and refluxing for 50-70 min, filtering, evaporating filtrate, adding 10-30 ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50-ml of water), eluting with 50-70ml of water, collecting eluate, evaporating to dryness, adding 2-3ml of methanol to dissolve the residue to obtain test solution;
c. the thin layer chromatography test comprises the steps of sucking 4-6 μl of the reference substance solution, and 5-10 μl of the sample solution, respectively spotting on the same silica gel G thin layer plate or any thin layer plate, taking chloroform-ethyl acetate-methanol-formic acid (40:5:10:0.2) as a developing agent, or taking chloroform-ethyl acetate-methanol-formic acid (40:6:10:0.2) as a developing agent, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the chromatogram of the reference substance in the chromatogram of the sample;
4. identifying Glycyrrhrizae radix control materials contained in QINGJIANG tablet by thin layer chromatography:
a. preparing control medicinal material solution by adding methanol 20-40ml into Glycyrrhrizae radix control medicinal material 1g, refluxing under heating for 50-70 min, filtering, evaporating filtrate to dryness, dissolving residue in water 5-15 ml, extracting with water saturated n-butanol for 3 times under shaking for 5-15 ml each time, mixing n-butanol extractive solutions, washing with water 2 times, 5-15 ml each time, discarding water solution, evaporating n-butanol solution to dryness, dissolving residue in methanol 2-3ml, and collecting control medicinal material solution;
b. preparing test solution by grinding 2.0-3.0g of QINGJIANGPING, adding 40-60ml of petroleum ether (30-60deg.C), ultrasonic treating for 40-50 min, filtering, discarding filtrate, volatilizing solvent from residue, adding 40-60ml of chloroform, ultrasonic treating for 40-50 min, filtering, discarding filtrate, volatilizing solvent from residue, adding 40-50ml of methanol, heating and refluxing for 50-70 min, filtering, evaporating filtrate, adding 10-30 ml of residue to dissolve, adding onto polyamide column (80-100 mesh, 3g, 1.5cm inner diameter, pre-washing with 50-ml of water), eluting with 50-70ml of water, discarding eluent, eluting with 80-120ml of ethanol, collecting eluent, evaporating to dryness, adding 2-3ml of methanol to dissolve the residue to obtain test solution;
c. the thin layer chromatography test comprises the steps of sucking 1-3 μl of the reference substance solution, and 5-10 μl of the sample solution, respectively spotting on the same silica gel G thin layer plate or any thin layer plate, using chloroform-methanol-water (40:10:1) as developing agent or using chloroform-methanol-water (40:8:1) as developing agent, developing, taking out, airing, spraying 10% sulfuric acid ethanol solution, heating at 105 ℃ until the color of spots is clear, and displaying fluorescent spots with the same color on the positions corresponding to the color of the reference substance chromatogram in the sample chromatogram;
5. the total amount of emodin and chrysophanol contained in the Chinese medicinal material rheum officinale of qingjiang tablet is measured by high performance liquid chromatography:
a. chromatographic conditions and system applicability test, namely, octadecylsilane chemically bonded silica is taken as a filler, methanol-0.1% phosphoric acid aqueous solution is taken as a mobile phase, the ratio of the octadecylsilane chemically bonded silica to the mobile phase is 85:15, the detection wavelength is 254nm, and the theoretical plate number is not lower than 6000 according to the emodin peak;
b. preparing a reference substance solution, namely taking a proper amount of emodin reference substance and chrysophanol reference substance, precisely weighing, adding methanol to prepare a mixed solution containing 10-20 mu g of emodin and 20-40 mu g of chrysophanol per 1ml, and taking the mixed solution as the reference substance solution;
c. preparing a sample solution, namely taking 2.0-3.0g of clear descending tablet, removing coating, grinding, precisely weighing about 0.30-0.50g, precisely weighing, placing into a conical flask with a plug, precisely adding 20-30ml of methanol, heating and refluxing for 1 hour by the weighed weight, cooling, weighing again, supplementing the lost weight with methanol, shaking uniformly, filtering, precisely weighing 8-12ml of continuous filtrate, placing into a flask, recovering solvent under reduced pressure until dryness, adding 8-12ml of hydrochloric acid (22-100) solution into residues, performing ultrasonic treatment for 2 minutes, adding 8-12ml of chloroform, heating and refluxing for 1 hour, cooling, placing into a separating funnel, washing a container with a small amount of chloroform, introducing into the separating funnel, separately taking a chloroform layer, extracting an acid liquor with chloroform for 3 times, 8-12ml each time, combining the chloroform solution, recovering solvent under reduced pressure until dryness, adding methanol into a 10ml measuring flask for dissolution, adding methanol into a scale bottle, adding the methanol into the scale, filtering, and taking the filtrate as the sample solution;
d. measuring by high performance liquid chromatography, namely precisely sucking 4-6 mu l of reference substance solution and 8-12 mu l of sample solution respectively, injecting into a high performance liquid chromatograph, and measuring;
the content of radix et rhizoma Rhei should be no less than 0.72mg/g based on total amount of emodin and chrysophanol.
The invention can be realized by the following optimal technical scheme:
1. the indigo blue and indirubin contained in qingjiang tablet are identified by thin layer chromatography:
a. preparing reference solution by adding chloroform into indigo reference and indirubin reference to obtain mixed solution containing 1.0mg of indigo and 0.5mg of indirubin per 1 ml;
b. preparation of sample solution comprises grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, and concentrating the filtrate to about 2ml to obtain sample solution;
c. sucking 5 mu l of the reference substance solution, respectively dropping 2-5 mu l of the sample solution on the same silica gel G thin layer plate, taking toluene-chloroform-acetone (5:4:1) as developing agent, developing, taking out, airing, and displaying spots with the same color on the positions corresponding to the reference substance chromatograph in the sample chromatograph;
2. the forsythin contained in qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into the control solution containing 1.0mg of forsythin per 1ml, and taking the control solution as control solution;
b. preparing test solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 45ml of methanol, ultrasonic treating for 45 min, filtering, evaporating filtrate to dryness, dissolving residue with 20. 20ml water, centrifuging, collecting supernatant, passing through D101 macroporous adsorbent resin column (column inner diameter of 1.5cm, column height of 16 cm), eluting with 30% ethanol and 70% ethanol respectively, evaporating to dryness, and dissolving residue with 2ml of methanol to obtain test solution;
c. sucking 5 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-methanol-anhydrous formic acid (17:2:1.5) as developing agent, developing, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the reference substance chromatogram in the sample chromatogram;
3. the paeoniflorin contained in the qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into paeoniflorin control solution to obtain 1.0mg of solution containing forsythin per 1ml, and taking the solution as control solution;
b. preparing test solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 40ml of methanol, refluxing under heating for 1 hr, filtering, evaporating filtrate, adding 20ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50ml of water), eluting with 60ml of water, collecting eluate, evaporating to dryness, and adding 2ml of methanol to dissolve residue to obtain test solution;
c. sucking 5 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-ethyl acetate-methanol-formic acid (40:5:10:0.2) as developing agent, developing, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the reference substance chromatograph in the sample chromatograph;
4. identifying Glycyrrhrizae radix control materials contained in QINGJIANG tablet by thin layer chromatography:
a. preparing control medicinal material solution by adding methanol 30ml into Glycyrrhrizae radix control medicinal material 1g, refluxing under heating for 1 hr, filtering, evaporating filtrate, dissolving residue in water 10ml, extracting with water saturated n-butanol for 3 times under shaking 10ml each time, mixing n-butanol extractive solutions, washing with water 2 times 10ml each time, discarding water solution, evaporating n-butanol solution, and dissolving residue in methanol 2ml to obtain control medicinal material solution;
b. preparation of sample solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 40ml of methanol, refluxing under heating for 1 hr, filtering, evaporating filtrate, adding 20ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50ml of water), eluting with 60ml of water, removing eluent, eluting with 100ml of ethanol, collecting eluent, evaporating to dryness, and adding 2ml of methanol to residue to dissolve to obtain sample solution;
c. the thin layer chromatography test comprises the steps of sucking 2 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-methanol-water (40:10:1) as developing agent, developing, taking out, airing, spraying 10% sulfuric acid ethanol solution, heating at 105 ℃ until spots develop clearly, and developing fluorescent spots with the same color on the positions corresponding to the reference medicine chromatogram in the sample chromatogram;
5. the total amount of emodin and chrysophanol contained in the Chinese medicinal material rheum officinale of qingjiang tablet is measured by high performance liquid chromatography:
a. chromatographic conditions and system applicability test, namely, octadecylsilane chemically bonded silica is taken as a filler, methanol-0.1% phosphoric acid aqueous solution is taken as a mobile phase, the ratio of the octadecylsilane chemically bonded silica to the mobile phase is 85:15, the detection wavelength is 254nm, and the theoretical plate number is not lower than 6000 according to the emodin peak;
b. preparing a reference substance solution, namely taking a proper amount of emodin reference substance and chrysophanol reference substance, precisely weighing, adding methanol to prepare a mixed solution containing 15 mug of emodin and 30 mug of chrysophanol per 1ml, and taking the mixed solution as the reference substance solution;
c. preparing a sample solution, namely taking 2.5g of clear descending tablet, removing coating, grinding, precisely weighing about 0.40g, precisely weighing and placing into a conical flask with a plug, precisely adding 25ml of methanol, heating and refluxing for 1 hour, cooling, then weighing, supplementing the reduced weight with methanol, shaking uniformly, filtering, precisely weighing 10ml of continuous filtrate, placing into a flask, recovering solvent under reduced pressure until the solvent is dry, adding 10ml of hydrochloric acid (22-100) solution into residues, performing ultrasonic treatment for 2 minutes, adding 10ml of chloroform again, heating and refluxing for 1 hour, cooling, placing into a separating funnel, washing a container with a small amount of chloroform, merging into the separating funnel, taking a chloroform layer separately, extracting 3 times by shaking with chloroform, 10ml each time, merging the chloroform solution, recovering the solvent under reduced pressure until the solvent is dry, transferring the residues into a 10ml measuring flask, adding methanol to the scale, shaking uniformly, filtering, and taking the continuous filtrate as the sample solution;
d. measuring by high performance liquid chromatography, namely precisely sucking 5 mu l of reference substance solution and 10 mu I of sample solution respectively, injecting into the high performance liquid chromatograph, and measuring;
the content of radix et rhizoma Rhei should be no less than 0.72mg/g based on total amount of emodin and chrysophanol.
The qingjiang tablet of the invention implements the national drug standard, which refers to the national food and drug administration standard (YBZ 02002004) implemented by qingjiang tablet. Legal unit names and symbols used in the claims and specification, including volumetric units milliliter (ml) and microliter (μl); comprises the mass units of gram (g), milligrams (mg) and micrograms (mug).
The invention increases the qualitative identification method of the qingjiang tablet by thin layer chromatography and the high performance liquid chromatography content determination method of rhubarb, is simple and easy to implement, and can be used as the quality control index of the qingjiang tablet.
Detailed Description
Example 1:
pulverizing faeces Bombycis 41.68g, radix et rhizoma Rhei 41.68g, indigo naturalis 20.84g, bulbus Fritillariae Cirrhosae 5.20g into fine powder; extracting 41.68g of radix scrophulariae and 41.68g of fructus Gleditsiae Abnormalis with 80% ethanol twice, wherein the extraction temperature is 78-82 deg.C, adding 4 times of 80% ethanol for the first time, reflux extracting for 1.5 hr, adding 3 times of 80% ethanol for the second time, reflux extracting for 1.5 hr, mixing ethanol extractive solutions, recovering ethanol, and concentrating to obtain extract with relative density of 1.30-1.35 (60+ -5deg.C; taking 41.68g of red paeony root, 41.68g of radix isatidis, 41.68g of dwarf lilyturf tuber, 41.68g of weeping forsythiae capsule, 27.08g of tree peony bark, 41.68g of rehmannia root and 13.76g of liquorice, extracting twice with water, wherein the first time is 1.5 hours, the second time is 1.5 hours, the water is added for 8 times, the water is added for 7 times, the water temperature is 98-102 ℃ for each time, filtering the extracting solution, and combining the filtrates; extracting rhizoma Imperatae 41.68g and flos Lonicerae 41.68g with water twice for 30 min for 10 times and 8 times for 15 min, filtering the extractive solution, and mixing filtrates; concentrating the two filtrates under reduced pressure to obtain extract with relative density of 1.35-1.40 (60+ -5deg.C; adding fine powder of faeces Bombycis and adjuvants into the concentrated extract, mixing, granulating, drying, adding menthol 0.104g, mixing, and pressing into 1000 coated film coating each tablet 0.25 g; the identification method of the medicine comprises the following steps:
1. the indigo blue and indirubin contained in qingjiang tablet are identified by thin layer chromatography:
a. preparing reference solution by adding chloroform into indigo reference and indirubin reference to obtain mixed solution containing 1.0mg of indigo and 0.5mg of indirubin per 1 ml;
b. preparation of sample solution comprises grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, and concentrating the filtrate to about 2ml to obtain sample solution;
c. sucking 5 mu l of the reference substance solution, respectively dropping 2-5 mu l of the sample solution on the same silica gel G thin layer plate, taking toluene-chloroform-acetone (5:4:1) as developing agent, developing, taking out, airing, and displaying spots with the same color on the positions corresponding to the reference substance chromatograph in the sample chromatograph;
2. the forsythin contained in qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into the control solution containing 1.0mg of forsythin per 1ml, and taking the control solution as control solution;
b. preparing test solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 45ml of methanol, ultrasonic treating for 45 min, filtering, evaporating filtrate to dryness, dissolving residue with 20. 20ml water, centrifuging, collecting supernatant, passing through D101 macroporous adsorbent resin column (column inner diameter of 1.5cm, column height of 16 cm), eluting with 30% ethanol and 70% ethanol respectively, evaporating to dryness, and dissolving residue with 2ml of methanol to obtain test solution;
c. sucking 5 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-methanol-anhydrous formic acid (17:2:1.5) as developing agent, developing, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the reference substance chromatogram in the sample chromatogram;
3. the paeoniflorin contained in the qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into paeoniflorin control solution to obtain 1.0mg of solution containing forsythin per 1ml, and taking the solution as control solution;
b. preparing test solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 40ml of methanol, refluxing under heating for 1 hr, filtering, evaporating filtrate, adding 20ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50ml of water), eluting with 60ml of water, collecting eluate, evaporating to dryness, and adding 2ml of methanol to dissolve residue to obtain test solution;
c. sucking 5 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-ethyl acetate-methanol-formic acid (40:5:10:0.2) as developing agent, developing, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the reference substance chromatograph in the sample chromatograph;
4. identifying Glycyrrhrizae radix control materials contained in QINGJIANG tablet by thin layer chromatography:
a. preparing control medicinal material solution by adding methanol 30ml into Glycyrrhrizae radix control medicinal material 1g, refluxing under heating for 1 hr, filtering, evaporating filtrate, dissolving residue in water 10ml, extracting with water saturated n-butanol for 3 times under shaking 10ml each time, mixing n-butanol extractive solutions, washing with water 2 times 10ml each time, discarding water solution, evaporating n-butanol solution, and dissolving residue in methanol 2ml to obtain control medicinal material solution;
b. preparation of sample solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 40ml of methanol, refluxing under heating for 1 hr, filtering, evaporating filtrate, adding 20ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50ml of water), eluting with 60ml of water, removing eluent, eluting with 100ml of ethanol, collecting eluent, evaporating to dryness, and adding 2ml of methanol to residue to dissolve to obtain sample solution;
c. the thin layer chromatography test comprises the steps of sucking 2 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-methanol-water (40:10:1) as developing agent, developing, taking out, airing, spraying 10% sulfuric acid ethanol solution, heating at 105 ℃ until spots develop clearly, and developing fluorescent spots with the same color on the positions corresponding to the reference medicine chromatogram in the sample chromatogram;
5. the total amount of emodin and chrysophanol contained in the Chinese medicinal material rheum officinale of qingjiang tablet is measured by high performance liquid chromatography:
a. chromatographic conditions and system applicability test, namely, octadecylsilane chemically bonded silica is taken as a filler, methanol-0.1% phosphoric acid aqueous solution is taken as a mobile phase, the ratio of the octadecylsilane chemically bonded silica to the mobile phase is 85:15, the detection wavelength is 254nm, and the theoretical plate number is not lower than 6000 according to the emodin peak;
b. preparing a reference substance solution, namely taking a proper amount of emodin reference substance and chrysophanol reference substance, precisely weighing, adding methanol to prepare a mixed solution containing 15 mug of emodin and 30 mug of chrysophanol per 1ml, and taking the mixed solution as the reference substance solution;
c. preparing a sample solution, namely taking 2.5g of clear descending tablet, removing coating, grinding, precisely weighing about 0.40g, precisely weighing and placing into a conical flask with a plug, precisely adding 25ml of methanol, heating and refluxing for 1 hour, cooling, then weighing, supplementing the reduced weight with methanol, shaking uniformly, filtering, precisely weighing 10ml of continuous filtrate, placing into a flask, recovering solvent under reduced pressure until the solvent is dry, adding 10ml of hydrochloric acid (22-100) solution into residues, performing ultrasonic treatment for 2 minutes, adding 10ml of chloroform again, heating and refluxing for 1 hour, cooling, placing into a separating funnel, washing a container with a small amount of chloroform, merging into the separating funnel, taking a chloroform layer separately, extracting 3 times by shaking with chloroform, 10ml each time, merging the chloroform solution, recovering the solvent under reduced pressure until the solvent is dry, transferring the residues into a 10ml measuring flask, adding methanol to the scale, shaking uniformly, filtering, and taking the continuous filtrate as the sample solution;
d. measuring by high performance liquid chromatography, namely precisely sucking 5 mu l of reference substance solution and 10 mu I of sample solution respectively, injecting into the high performance liquid chromatograph, and measuring;
the content of radix et rhizoma Rhei should be no less than 0.72mg/g based on total amount of emodin and chrysophanol.
Example 2:
pulverizing faeces Bombycis 41.68g, radix et rhizoma Rhei 41.68g, indigo naturalis 20.84g, bulbus Fritillariae Cirrhosae 5.20g into fine powder; extracting 41.68g of radix scrophulariae and 41.68g of fructus Gleditsiae Abnormalis with 80% ethanol twice, wherein the extraction temperature is 78-82 deg.C, adding 4 times of 80% ethanol for the first time, reflux extracting for 1.5 hr, adding 3 times of 80% ethanol for the second time, reflux extracting for 1.5 hr, mixing ethanol extractive solutions, recovering ethanol, and concentrating to obtain extract with relative density of 1.30-1.35 (60+ -5deg.C; taking 41.68g of red paeony root, 41.68g of radix isatidis, 41.68g of dwarf lilyturf tuber, 41.68g of weeping forsythiae capsule, 27.08g of tree peony bark, 41.68g of rehmannia root and 13.76g of liquorice, extracting twice with water, wherein the first time is 1.5 hours, the second time is 1.5 hours, the water is added for 8 times, the water is added for 7 times, the water temperature is 98-102 ℃ for each time, filtering the extracting solution, and combining the filtrates; extracting rhizoma Imperatae 41.68g and flos Lonicerae 41.68g with water twice for 30 min for 10 times and 8 times for 15 min, filtering the extractive solution, and mixing filtrates; concentrating the two filtrates under reduced pressure to obtain extract with relative density of 1.35-1.40 (60+ -5deg.C; adding fine powder of faeces Bombycis and adjuvants into the concentrated extract, mixing, granulating, drying, adding menthol 0.104g, mixing, and pressing into 2000 coated film coating each tablet 0.125 g; the identification method of the medicine comprises the following steps:
1. the indigo blue and indirubin contained in qingjiang tablet are identified by thin layer chromatography:
a. preparing reference solution by adding chloroform into indigo reference and indirubin reference to obtain mixed solution containing 1.0mg of indigo and 0.5mg of indirubin per 1 ml;
b. preparation of sample solution comprises grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, and concentrating the filtrate to about 2ml to obtain sample solution;
c. sucking 5 mu l of the reference substance solution, respectively dropping 2-5 mu l of the sample solution on the same silica gel G thin layer plate, taking toluene-chloroform-acetone (5:4:1) as developing agent, developing, taking out, airing, and displaying spots with the same color on the positions corresponding to the reference substance chromatograph in the sample chromatograph;
2. the forsythin contained in qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into the control solution containing 1.0mg of forsythin per 1ml, and taking the control solution as control solution;
b. preparing test solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 45ml of methanol, ultrasonic treating for 45 min, filtering, evaporating filtrate to dryness, dissolving residue with 20. 20ml water, centrifuging, collecting supernatant, passing through D101 macroporous adsorbent resin column (column inner diameter of 1.5cm, column height of 16 cm), eluting with 30% ethanol and 70% ethanol respectively, evaporating to dryness, and dissolving residue with 2ml of methanol to obtain test solution;
c. sucking 5 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-methanol-anhydrous formic acid (17:2:1.5) as developing agent, developing, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the reference substance chromatogram in the sample chromatogram;
3. the paeoniflorin contained in the qingjiang tablet is identified by thin layer chromatography:
a. preparing control solution by adding methanol into paeoniflorin control solution to obtain 1.0mg of solution containing forsythin per 1ml, and taking the solution as control solution;
b. preparing test solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 40ml of methanol, refluxing under heating for 1 hr, filtering, evaporating filtrate, adding 20ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50ml of water), eluting with 60ml of water, collecting eluate, evaporating to dryness, and adding 2ml of methanol to dissolve residue to obtain test solution;
c. sucking 5 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-ethyl acetate-methanol-formic acid (40:5:10:0.2) as developing agent, developing, taking out, airing, spraying 5% vanillin sulfuric acid solution, heating at 105 ℃ until spots develop clearly, and developing spots with the same color on the positions corresponding to the reference substance chromatograph in the sample chromatograph;
4. identifying Glycyrrhrizae radix control materials contained in QINGJIANG tablet by thin layer chromatography:
a. preparing control medicinal material solution by adding methanol 30ml into Glycyrrhrizae radix control medicinal material 1g, refluxing under heating for 1 hr, filtering, evaporating filtrate, dissolving residue in water 10ml, extracting with water saturated n-butanol for 3 times under shaking 10ml each time, mixing n-butanol extractive solutions, washing with water 2 times 10ml each time, discarding water solution, evaporating n-butanol solution, and dissolving residue in methanol 2ml to obtain control medicinal material solution;
b. preparation of sample solution by grinding 2.5g of QINGJIANGPING, adding 50ml of petroleum ether (30-60deg.C), ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 50ml of chloroform, ultrasonic treating for 45 min, filtering, removing filtrate, volatilizing solvent from residue, adding 40ml of methanol, refluxing under heating for 1 hr, filtering, evaporating filtrate, adding 20ml of residue to dissolve, adding on polyamide column (80-100 mesh, 3g, inner diameter of 1.5cm, pre-washing with 50ml of water), eluting with 60ml of water, removing eluent, eluting with 100ml of ethanol, collecting eluent, evaporating to dryness, and adding 2ml of methanol to residue to dissolve to obtain sample solution;
c. the thin layer chromatography test comprises the steps of sucking 2 mu l of the reference substance solution, respectively pointing 5-10 mu l of the sample solution on the same silica gel G thin layer plate, taking chloroform-methanol-water (40:10:1) as developing agent, developing, taking out, airing, spraying 10% sulfuric acid ethanol solution, heating at 105 ℃ until spots develop clearly, and developing fluorescent spots with the same color on the positions corresponding to the reference medicine chromatogram in the sample chromatogram;
5. the total amount of emodin and chrysophanol contained in the Chinese medicinal material rheum officinale of qingjiang tablet is measured by high performance liquid chromatography:
a. chromatographic conditions and system applicability test, namely, octadecylsilane chemically bonded silica is taken as a filler, methanol-0.1% phosphoric acid aqueous solution is taken as a mobile phase, the ratio of the octadecylsilane chemically bonded silica to the mobile phase is 85:15, the detection wavelength is 254nm, and the theoretical plate number is not lower than 6000 according to the emodin peak;
b. preparing a reference substance solution, namely taking a proper amount of emodin reference substance and chrysophanol reference substance, precisely weighing, adding methanol to prepare a mixed solution containing 15 mug of emodin and 30 mug of chrysophanol per 1ml, and taking the mixed solution as the reference substance solution;
c. preparing a sample solution, namely taking 2.5g of clear descending tablet, removing coating, grinding, precisely weighing about 0.40g, precisely weighing and placing into a conical flask with a plug, precisely adding 25ml of methanol, heating and refluxing for 1 hour, cooling, then weighing, supplementing the reduced weight with methanol, shaking uniformly, filtering, precisely weighing 10ml of continuous filtrate, placing into a flask, recovering solvent under reduced pressure until the solvent is dry, adding 10ml of hydrochloric acid (22-100) solution into residues, performing ultrasonic treatment for 2 minutes, adding 10ml of chloroform again, heating and refluxing for 1 hour, cooling, placing into a separating funnel, washing a container with a small amount of chloroform, merging into the separating funnel, taking a chloroform layer separately, extracting 3 times by shaking with chloroform, 10ml each time, merging the chloroform solution, recovering the solvent under reduced pressure until the solvent is dry, transferring the residues into a 10ml measuring flask, adding methanol to the scale, shaking uniformly, filtering, and taking the continuous filtrate as the sample solution;
d. measuring by high performance liquid chromatography, namely precisely sucking 5 mu l of reference substance solution and 10 mu I of sample solution respectively, injecting into the high performance liquid chromatograph, and measuring;
the content of radix et rhizoma Rhei should be no less than 0.72mg/g based on total amount of emodin and chrysophanol.
Claims (1)
1. The identification method of the Qingjiang tablet of the traditional Chinese medicine preparation, wherein the traditional Chinese medicine preparation formula consists of 41.68g of silkworm excrement, 41.68g of rheum officinale, 20.84g of indigo naturalis, 41.68g of radix scrophulariae, 41.68g of fructus gleditsiae, 41.68g of radix paeoniae rubra, 41.68g of radix isatidis, 41.68g of radix ophiopogonis, 41.68g of fructus forsythiae, 27.08g of tree peony bark, 41.68g of rehmannia, 13.76g of liquorice, 41.68g of rhizoma imperatae, 41.68g of honeysuckle, 0.104g of menthol and 5.20g of bulbus fritillariae cirrhosae, and the method is characterized by comprising the following steps of:
(1) The indigo blue and indirubin contained in qingjiang tablet are identified by thin layer chromatography:
a. preparation of a control solution: adding chloroform into indigo reference substance and indirubin reference substance to obtain mixed solution containing 1.0mg of indigo and 0.5mg of indirubin per 1ml, and taking as reference substance solution;
b. preparation of test solution: taking 2.5g of qingjiang tablet, grinding, adding 50ml of petroleum ether at 30-60 ℃, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing solvent from dregs, adding 50ml of chloroform, carrying out ultrasonic treatment for 45 minutes, filtering, concentrating the filtrate to 2ml, and taking the filtrate as a sample solution;
c. thin layer chromatography assay: sucking 5 μl of the reference substance solution, respectively spotting 2-5 μl of the sample solution on the same silica gel G thin layer plate, spreading with toluene-chloroform-acetone as developing agent, taking out, air drying, and displaying spots of the same color on the corresponding position of the reference substance chromatogram in the sample chromatogram; the proportion of toluene-chloroform-acetone is 5:4:1;
(2) The forsythin contained in qingjiang tablet is identified by thin layer chromatography:
a. preparation of a control solution: taking a forsythin reference substance, adding methanol to prepare a solution containing 1.0mg of forsythin per 1ml, and taking the solution as a reference substance solution;
b. preparation of test solution: taking 2.5g of qingjiang tablet, grinding, adding 50ml of petroleum ether at 30-60 ℃, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing a solvent from dregs, adding 50ml of chloroform, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing the solvent from dregs, adding 45ml of methanol, carrying out ultrasonic treatment for 45 minutes, filtering, evaporating filtrate to dryness, adding 20ml of water into residues to dissolve, centrifuging, taking supernatant, passing through a D101 macroporous adsorption resin column, eluting with 60ml of 30% ethanol and 60% ethanol respectively, evaporating to dryness, and adding 2ml of methanol into residues to dissolve to obtain a sample solution; the inner diameter of the D101 macroporous adsorption resin column is 1.5cm, and the height of the column is 16cm;
c. thin layer chromatography assay: sucking 5 μl of the reference substance solution, respectively spotting 5-10 μl of the sample solution on the same silica gel G thin layer plate, developing with chloroform-methanol-anhydrous formic acid as developing agent, taking out, air drying, spraying 5% vanillin sulfuric acid solution, heating at 105deg.C until the color of the spot is clear, and displaying the same color spot on the position corresponding to the color spectrum of the reference substance in the color spectrum of the sample; the ratio of the chloroform to the methanol to the anhydrous formic acid is 17:2:1.5;
(3) The paeoniflorin contained in the qingjiang tablet is identified by thin layer chromatography:
a. preparation of a control solution: taking paeoniflorin reference substance, adding methanol to prepare 1.0mg of solution containing paeoniflorin per 1ml of solution as reference substance solution;
b. preparation of test solution: taking 2.5g of qingjiang tablet, grinding, adding 50ml of petroleum ether at 30-60 ℃, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing solvent from dregs, adding 50ml of chloroform, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing solvent from dregs, adding 40ml of methanol, heating and refluxing for 1 hour, filtering, evaporating filtrate, adding 20ml of residue into water to dissolve, adding the residue into a polyamide column, eluting with 60ml of water, collecting eluent, evaporating to dryness, and adding 2ml of methanol into the residue to dissolve to serve as a sample solution; the particle size specification of the polyamide column is 80-100 meshes, the filler amount is 3g, the inner diameter is 1.5cm, and the polyamide column is pre-washed by 50ml of water;
c. thin layer chromatography assay: sucking 5 μl of the reference substance solution, respectively spotting 5-10 μl of the sample solution on the same silica gel G thin layer plate, developing with chloroform-ethyl acetate-methanol-formic acid as developing agent, taking out, air drying, spraying 5% vanillin sulfuric acid solution, heating at 105deg.C until the color of the spot becomes clear, and making the spot with the same color appear on the position corresponding to the color spectrum of the reference substance in the color spectrum of the sample solution; the ratio of the chloroform to the ethyl acetate to the methanol to the formic acid is 40:5:10:0.2;
(4) Identifying Glycyrrhrizae radix control materials contained in QINGJIANG tablet by thin layer chromatography:
a. preparation of control medicinal material solution: taking 1g of licorice reference medicine, adding 30ml of methanol, heating and refluxing for 1 hour, filtering, evaporating filtrate to dryness, adding 10ml of water into residues to dissolve, extracting 3 times by shaking with water saturated n-butanol, 10ml each time, combining n-butanol extract, washing 2 times with water, 10ml each time, discarding water solution, evaporating n-butanol solution to dryness, adding 2ml of methanol into residues to dissolve, and taking the residues as reference medicine solution;
b. preparation of test solution: taking 2.5g of qingjiang tablet, grinding, adding 50ml of petroleum ether at 30-60 ℃, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing solvent from dregs, adding 50ml of chloroform, carrying out ultrasonic treatment for 45 minutes, filtering, discarding filtrate, volatilizing solvent from dregs, adding 40ml of methanol, heating and refluxing for 1 hour, filtering, evaporating filtrate to dryness, adding 20ml of residue into water to dissolve, adding the residue onto a polyamide column, eluting with 60ml of water, discarding eluent, eluting with 100ml of ethanol, collecting eluent, evaporating to dryness, adding 2ml of methanol into the residue to dissolve, and taking the residue as a sample solution; the particle size specification of the polyamide column is 80-100 meshes, the filler amount is 3g, the inner diameter is 1.5cm, and the polyamide column is pre-washed by 50ml of water;
c. thin layer chromatography assay: absorbing 2 μl of the reference substance solution, respectively spotting 5-10 μl of the sample solution on the same silica gel G thin layer plate, spreading with chloroform-methanol-water as developing agent, taking out, air drying, spraying 10% sulfuric acid ethanol solution, heating at 105deg.C until the color of the spot is clear, and displaying fluorescent spots of the same color on the position corresponding to the color spectrum of the reference substance in the color spectrum of the sample solution; the ratio of the chloroform to the methanol to the water is 40:10:1;
(5) The total amount of emodin and chrysophanol contained in the Chinese medicinal material rheum officinale of qingjiang tablet is measured by high performance liquid chromatography:
a. chromatographic conditions and system suitability test: octadecylsilane chemically bonded silica is used as a filler, methanol-0.1% phosphoric acid aqueous solution is used as a mobile phase, and the ratio of the octadecylsilane chemically bonded silica to the mobile phase is 85:15, the detection wavelength is 254nm, and the theoretical plate number is not lower than 6000 according to the emodin peak;
b. preparation of a control solution: taking appropriate amount of emodin reference substance and chrysophanol reference substance, precisely weighing, adding methanol to obtain mixed solution containing emodin 15 μg and chrysophanol 30 μg per 1ml, and taking as reference substance solution;
c. preparation of test solution: taking 2.5g of qingjiang tablet, removing the coating, grinding, precisely weighing 0.40g, precisely weighing, placing into a conical bottle with a plug, precisely adding 25ml of methanol, weighing, heating and refluxing for 1 hour, cooling, weighing again, supplementing the weight of the liquid with methanol, shaking uniformly, filtering, precisely weighing 10ml of subsequent filtrate, placing into a flask, recovering solvent under reduced pressure to dryness, adding 22ml of hydrochloric acid into 100ml of hydrochloric acid solution, adding 10ml of hydrochloric acid solution into the residue, performing ultrasonic treatment for 2 minutes, adding 10ml of chloroform, heating and refluxing for 1 hour, cooling, placing into a separating funnel, washing a container with a small amount of chloroform, merging the chloroform solution into the separating funnel, taking a chloroform layer separately, shaking and extracting 3 times with chloroform, 10ml each time, recovering solvent under reduced pressure, dissolving the residue with methanol, transferring into a 10ml measuring bottle, adding methanol to a scale, shaking uniformly, filtering, and taking the subsequent filtrate as a sample solution;
d. high performance liquid chromatography assay: precisely sucking 5 μl of reference solution and 10 μl of sample solution, respectively, and injecting into high performance liquid chromatograph for measurement;
the content of radix et rhizoma Rhei should be no less than 0.72mg/g based on total amount of emodin and chrysophanol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811599671.9A CN111366673B (en) | 2018-12-26 | 2018-12-26 | Identification method of traditional Chinese medicine preparation qingjiang tablet |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811599671.9A CN111366673B (en) | 2018-12-26 | 2018-12-26 | Identification method of traditional Chinese medicine preparation qingjiang tablet |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111366673A CN111366673A (en) | 2020-07-03 |
| CN111366673B true CN111366673B (en) | 2024-03-29 |
Family
ID=71206023
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811599671.9A Active CN111366673B (en) | 2018-12-26 | 2018-12-26 | Identification method of traditional Chinese medicine preparation qingjiang tablet |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111366673B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1616013A (en) * | 2004-09-17 | 2005-05-18 | 鲁南制药股份有限公司 | Chinese medicine composition for treating dizziness and its preparing and quality control method |
| CN1799584A (en) * | 2005-01-07 | 2006-07-12 | 黄伟民 | Chinese medicinal composition, its preparation process and quality control method |
| CN1824192A (en) * | 2005-12-29 | 2006-08-30 | 贵州益佰制药股份有限公司 | Quality control method of oral preparation for heat clearing and exterior resolution |
-
2018
- 2018-12-26 CN CN201811599671.9A patent/CN111366673B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1616013A (en) * | 2004-09-17 | 2005-05-18 | 鲁南制药股份有限公司 | Chinese medicine composition for treating dizziness and its preparing and quality control method |
| CN1799584A (en) * | 2005-01-07 | 2006-07-12 | 黄伟民 | Chinese medicinal composition, its preparation process and quality control method |
| CN1824192A (en) * | 2005-12-29 | 2006-08-30 | 贵州益佰制药股份有限公司 | Quality control method of oral preparation for heat clearing and exterior resolution |
Non-Patent Citations (6)
| Title |
|---|
| HPLC 法测定通腑胶囊中大黄素、大黄酸、大黄酚、大黄素甲醚及芦荟大黄素的含量;樊磊磊;中国医药指;第9卷(第12期);11-12 * |
| 中华人民共和国卫生部药典委员会.中华人民共和国卫生部 药品标准 中药成方制剂 第十七册.1998,29-30. * |
| 凉血胶囊的薄层色谱鉴别;具善姬 等;中国药业;第21卷(第22期);51 * |
| 清毒祛痨丸的薄层色谱鉴别及没食子酸的含量测定;杨荣艳 等;中国现代中药;第12卷(第8期);33-36 * |
| 灵益胶囊质量标准的研究;马双成 等;中国中药杂志;第23 卷(第07期);408-410 * |
| 补肺康复片的质量标准研究;李卫红 等;中医药导报;第11卷(第05期);58-60 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111366673A (en) | 2020-07-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111044624B (en) | Quality detection method of Chinese medicinal preparation | |
| CN1325072C (en) | Method for preparing traditional Chinese medicine | |
| CN1768854B (en) | Chinese medicinal capsule for treating ulcerative colitis | |
| CN102488863A (en) | Chinese herbal medicine compound with anticancer effect, preparation method and detection method thereof | |
| CN101757099B (en) | Desmodium-capillary artemisia cholecystagogue, preparation method and quality control method thereof | |
| CN101011527B (en) | Improved preparation of Juhong pill, its preparation method and quality inspection method | |
| CN101002909A (en) | Antiviral traditional Chinese medicine, its preparing method and quality control method | |
| CN100438903C (en) | Quality control method of fuyuekang Chinese medicinal preparation | |
| CN102091168A (en) | Quality control method for Chinese medicine preparation Xuefuzhuyu capsule | |
| CN102078443B (en) | Medicine composition, application and preparation thereof | |
| CN102579734A (en) | Traditional Chinese medicine composition of bone healing medicine, preparing method thereof and detecting method thereof | |
| CN104274721A (en) | Preparation method of cold heat clearing granules | |
| CN106620610A (en) | Preparation method of liquorice heart fire purging granule | |
| CN102240366B (en) | Quality detection method for preparation for vision rehabilitation | |
| CN101011566A (en) | Medicine composition for treating chronic pelvic inflammatory disease, preparing method and quality controlling method thereof | |
| CN111366673B (en) | Identification method of traditional Chinese medicine preparation qingjiang tablet | |
| CN101716270B (en) | Method for detecting quality of traditional Chinese herbal medicament compound preparation for invigorating blood and regulating menses | |
| CN1879850A (en) | Capsule with hairy vein agrimony and arenobufagin, its preparation process and quality control method | |
| CN112516219A (en) | Preparation process and quality control method of traditional Chinese medicine granules for treating systemic lupus erythematosus | |
| CN100515476C (en) | Compound capsule with pseudo-ginseng and Chinese fanpalm seed, its preparation process and quality control method | |
| CN101700362A (en) | Preparation method and quality detection method of traditional Chinese medicine preparation for recovering sight | |
| CN105521439A (en) | Preparation method of pharmaceutical preparation for promoting blood circulation and removing stasis, diminishing inflammation and relieving pain | |
| CN107961321B (en) | Extraction process of liuhe pill raw material medicine | |
| CN105535146B (en) | Pretreatment method of radix puerariae, radix scutellariae and rhizoma coptidis composition medicinal materials | |
| CN101474332A (en) | Danxiang capsule and preparation method thereof |
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 |