CN101987154B - Thin-layer chromatographic detection method for cornel in gynecological recovery pills - Google Patents
Thin-layer chromatographic detection method for cornel in gynecological recovery pills Download PDFInfo
- Publication number
- CN101987154B CN101987154B CN2009100656965A CN200910065696A CN101987154B CN 101987154 B CN101987154 B CN 101987154B CN 2009100656965 A CN2009100656965 A CN 2009100656965A CN 200910065696 A CN200910065696 A CN 200910065696A CN 101987154 B CN101987154 B CN 101987154B
- Authority
- CN
- China
- Prior art keywords
- solution
- weight portions
- preparation
- woman
- fruit
- 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
- 238000001514 detection method Methods 0.000 title claims abstract description 21
- 239000006187 pill Substances 0.000 title claims abstract 4
- 238000011084 recovery Methods 0.000 title abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 63
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 26
- WCGUUGGRBIKTOS-GPOJBZKASA-N (3beta)-3-hydroxyurs-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C WCGUUGGRBIKTOS-GPOJBZKASA-N 0.000 claims abstract description 22
- 229940096998 ursolic acid Drugs 0.000 claims abstract description 22
- PLSAJKYPRJGMHO-UHFFFAOYSA-N ursolic acid Natural products CC1CCC2(CCC3(C)C(C=CC4C5(C)CCC(O)C(C)(C)C5CCC34C)C2C1C)C(=O)O PLSAJKYPRJGMHO-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 19
- 238000005303 weighing Methods 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 15
- 239000000741 silica gel Substances 0.000 claims abstract description 6
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 81
- 238000002360 preparation method Methods 0.000 claims description 77
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 70
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 54
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 54
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 45
- 238000012360 testing method Methods 0.000 claims description 40
- 239000013642 negative control Substances 0.000 claims description 38
- 239000013558 reference substance Substances 0.000 claims description 37
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 36
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 36
- 230000007935 neutral effect Effects 0.000 claims description 36
- 235000013399 edible fruits Nutrition 0.000 claims description 34
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 31
- 239000007788 liquid Substances 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 27
- 229960000583 acetic acid Drugs 0.000 claims description 18
- 239000012362 glacial acetic acid Substances 0.000 claims description 18
- 238000005406 washing Methods 0.000 claims description 18
- 238000004587 chromatography analysis Methods 0.000 claims description 16
- 239000000463 material Substances 0.000 claims description 11
- 239000000377 silicon dioxide Substances 0.000 claims description 10
- 244000061458 Solanum melongena Species 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- 239000003480 eluent Substances 0.000 claims description 9
- 238000011049 filling Methods 0.000 claims description 9
- 238000012856 packing Methods 0.000 claims description 9
- 238000010992 reflux Methods 0.000 claims description 9
- 239000007921 spray Substances 0.000 claims description 9
- 125000003944 tolyl group Chemical group 0.000 claims description 9
- 241000132012 Atractylodes Species 0.000 claims description 3
- 241000756943 Codonopsis Species 0.000 claims description 3
- 241000112528 Ligusticum striatum Species 0.000 claims description 3
- 244000197580 Poria cocos Species 0.000 claims description 3
- 235000008599 Poria cocos Nutrition 0.000 claims description 3
- 241000405414 Rehmannia Species 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000001117 sulphuric acid Substances 0.000 claims description 3
- 235000011149 sulphuric acid Nutrition 0.000 claims description 3
- 241000233948 Typha Species 0.000 claims 2
- 238000004458 analytical method Methods 0.000 abstract description 8
- KIWBPDUYBMNFTB-UHFFFAOYSA-N Ethyl hydrogen sulfate Chemical compound CCOS(O)(=O)=O KIWBPDUYBMNFTB-UHFFFAOYSA-N 0.000 abstract 1
- 238000005507 spraying Methods 0.000 abstract 1
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- HLXRWTJXGMHOFN-XJSNKYLASA-N Verbenalin Chemical compound O([C@@H]1OC=C([C@H]2C(=O)C[C@H](C)[C@H]21)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HLXRWTJXGMHOFN-XJSNKYLASA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 239000011800 void material Substances 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 206010000087 Abdominal pain upper Diseases 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 208000034656 Contusions Diseases 0.000 description 1
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 description 1
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 description 1
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 208000010513 Stupor Diseases 0.000 description 1
- 244000134857 Typha capensis Species 0.000 description 1
- HLXRWTJXGMHOFN-UHFFFAOYSA-N Verbenalin Natural products C12C(C)CC(=O)C2C(C(=O)OC)=COC1OC1OC(CO)C(O)C(O)C1O HLXRWTJXGMHOFN-UHFFFAOYSA-N 0.000 description 1
- 208000019790 abdominal distention Diseases 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 208000034526 bruise Diseases 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940074391 gallic acid Drugs 0.000 description 1
- 235000004515 gallic acid Nutrition 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229940100243 oleanolic acid Drugs 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention discloses a thin-layer chromatographic detection method for cornel in gynecological recovery pills, and belongs to the technical field of thin-layer chromatographic detection. The method comprises the following steps of: (1) weighing ursolic acid, and adding methanol into the ursolic acid to prepare contrast solution; (2) preparing trial solution; (3) preparing negative contrast solution; and (4) performing chromatographic identification, namely sucking the solution prepared in the steps (1), (2) and (3) to apply samples on a silica gel plate, performing development in a developing agent, spraying ethanol sulfate solution for color development, and performing contrastive analysis. The method is simple in operation, strong in feasibility and good in repeatability.
Description
Technical field
The invention belongs to thin-layer chromatography detection technique field, be specifically related to a kind of method of utilizing thin-layer chromatography to detect ursolic acid in the fruit of medicinal cornel.
Background technology
Woman's health ball is a kind of Chinese patent drug, has nourishing qi and blood, and the effect of regulating qi to disperse stagnation is used for the insufficiency of vital energy and blood in postpartum, adds the stasis of blood in the void, the rib abdominal distention and pain of cold-heat jumble, stomachache, head-body-pain, lochiorrhea, bruise stupor, constipation, diseases such as agalasisa.Ministerial drug in the fruit of medicinal cornel side of being contains ursolic acid, oleanolic acid, and cornin, malic acid, gallic acid etc. can consolidating channel for hemostasis, availablely controls diseases such as human female void, menorrhalgia.Existing quality standard adopts the dual wavelength scanning method to measure content of ursolic acid in the fruit of medicinal cornel, complicated operation, poor repeatability." Chinese pharmacopoeia adopts tlc scanning determination fruit of medicinal cornel medicinal material content of ursolic acid, and the running time is long, and step is loaded down with trivial details, and the colour developing error is big, and stability and repeatability are relatively poor relatively.
Summary of the invention
The technical matters that the present invention will solve is that fruit of medicinal cornel medicinal material ursolic acid is identified more complicated, provides a kind of simple to operate, the detection method that feasibility is strong.
Content of the present invention is: the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, and testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing the ursolic acid reference substance, add methyl alcohol and process the solution that every 1ml methyl alcohol contains the 1mg ursolic acid;
(2) preparation of need testing solution: take by weighing woman's health ball 5～10g, pulverize, add ethanol 20～30ml, reflux 1～2h filters; Filtrating evaporate to dryness, residue add water 15～20ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 2～4 times; Each 20～30ml merges n-butanol extracting liquid, with water washing 2～4 times, each 15～20ml; The layer that anhydrates that inclines, with the n-butanol layer evaporate to dryness, residue adds ethanol 1～2ml makes it dissolving, adds 0.5～1g neutral alumina; Mix thoroughly in the water-bath, drying, the neutral alumina column top that fills in advance of packing into is with methyl alcohol 40～50ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5～1ml ethyl acetate makes it dissolving, as the test sample analytic liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on silica gel plate of drawing step (1), (2), (3) preparation; In developping agent, launch; Spray develops the color with ethanol solution of sulfuric acid; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said step (3) neutral alumina column internal diameter is 10～15mm, and the neutral alumina of filling is 1～2g, 200～300 orders.
The point sample amount of said step (4) reference substance solution is 2～3 μ l, and the point sample amount of need testing solution and negative control solution is 4～6 μ l, and the exhibition distance is 6～10cm.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=(20～30): (6～10): 1.
Said silica gel plate is the silica G plate.
Described ethanol solution of sulfuric acid concentration is 30%, and the preparation method is: in beaker, add 70ml ethanol, measure the 30ml concentrated sulphuric acid and slowly add along walls of beaker, the limit edged stirs.
Described colour developing is that 30% ethanol solution of sulfuric acid is sprayed on the plate, and 90～110 ℃ are heated to the aubergine clear spot, inspect under the daylight.
Among the present invention, the negative control test has been done in the evaluation of the fruit of medicinal cornel, and the result shows that other medicines do not disturb the evaluation of ursolic acid in the fruit of medicinal cornel in woman's health ball.
The invention has the beneficial effects as follows simple to operately, cheap, feasibility is strong, and good reproducibility helps promoting.
Embodiment
Through embodiment the present invention is elaborated below, but and do not limit the present invention in any way.
Embodiment 1
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 10mg, add methyl alcohol 10ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 8g, pulverize, add ethanol 25ml, reflux 1h filters; Filtrating evaporate to dryness, residue add water 15ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 3 times; Each 20ml merges n-butanol extracting liquid, and each 15 with water washing 3 times, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 1ml makes it dissolving, adds the 0.5g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 10mm, and the neutral alumina of filling is 1g, and 200 orders are with methyl alcohol 40ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 2 μ l, and the point sample amount of need testing solution and negative control solution is 4 μ l, and the exhibition distance is 6cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 105 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=20: 6: 1.
Embodiment 1
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 10mg, add methyl alcohol 10ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 8g, pulverize, add ethanol 25ml, reflux 1h filters; Filtrating evaporate to dryness, residue add water 15ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 3 times; Each 20ml merges n-butanol extracting liquid, with water washing 3 times, and each 15ml, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 1ml makes it dissolving, adds the 0.5g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 10mm, and the neutral alumina of filling is 1g, and 200 orders are with methyl alcohol 40ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 2 μ l, and the point sample amount of need testing solution and negative control solution is 4 μ l, and the exhibition distance is 6cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 105 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=20: 6: 1.
Embodiment 2
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 10mg, add methyl alcohol 10ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 6g, pulverize, add ethanol 20ml, reflux 1h filters; Filtrating evaporate to dryness, residue add water 16ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 2 times; Each 20ml merges n-butanol extracting liquid, with water washing 3 times, and each 20ml, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 2ml makes it dissolving, adds the 0.5g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 12mm, and the neutral alumina of filling is 1g, and 200 orders are with methyl alcohol 40ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 2 μ l, and the point sample amount of need testing solution and negative control solution is 5 μ l, and the exhibition distance is 6cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 95 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=25: 6: 1.
Embodiment 3
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 10mg, add methyl alcohol 10ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 5g, pulverize, add ethanol 20ml, reflux 1h filters; Filtrating evaporate to dryness, residue add water 15ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 2 times; Each 25ml merges n-butanol extracting liquid, with water washing 3 times, and each 15ml, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 1ml makes it dissolving, adds the 0.5g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 10mm, and the neutral alumina of filling is 1g, and 250 orders are with methyl alcohol 42ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 2 μ l, and the point sample amount of need testing solution and negative control solution is 6 μ l, and the exhibition distance is 6cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 100 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=24: 8: 1.
Embodiment 4
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 20mg, add methyl alcohol 20ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 9g, pulverize, add ethanol 30ml, reflux 2h filters; Filtrating evaporate to dryness, residue add water 20ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 3 times; Each 20ml merges n-butanol extracting liquid, with water washing 3 times, and each 15ml, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 1ml makes it dissolving, adds the 1g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 15mm, and the neutral alumina of filling is 1g, and 300 orders are with methyl alcohol 45ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 2 μ l, and the point sample amount of need testing solution and negative control solution is 6 μ l, and the exhibition distance is 8cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 105 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=30: 8: 1.
Embodiment 5
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 20mg, add methyl alcohol 20ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 8g, pulverize, add ethanol 28ml, reflux 2h filters; Filtrating evaporate to dryness, residue add water 18ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 4 times; Each 25ml merges n-butanol extracting liquid, with water washing 3 times, and each 20ml, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 1ml makes it dissolving, adds the 1g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 15mm, and the neutral alumina of filling is 1g, and 300 orders are with methyl alcohol 45ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 3 μ l, and the point sample amount of need testing solution and negative control solution is 6 μ l, and the exhibition distance is 10cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 105 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=22: 7: 1.
Embodiment 6
The thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball, testing process is operated as follows:
(1) preparation of reference substance solution: take by weighing ursolic acid reference substance 20mg, add methyl alcohol 20ml, shake up;
(2) preparation of need testing solution: take by weighing woman's health ball 7g, pulverize, add ethanol 28ml, reflux 2h filters; Filtrating evaporate to dryness, residue add water 18ml makes it dissolving, washs to ether layer colourlessly with ether, and the ether layer that inclines is with normal butyl alcohol washing water layer 3 times; Each 25ml merges n-butanol extracting liquid, with water washing 3 times, and each 15ml, the layer that anhydrates inclines; With the n-butanol layer evaporate to dryness, residue adds ethanol 2ml makes it dissolving, adds the 1g neutral alumina, mixes drying in the water-bath thoroughly; The neutral alumina column top that fills in advance of packing into, the neutral alumina column internal diameter is 15mm, and the neutral alumina of filling is 1g, and 300 orders are with methyl alcohol 42ml wash-out; Collect eluent, evaporate to dryness, residue add 0.5ml ethyl acetate makes it dissolving, as sample analysis liquid;
(3) preparation of negative control solution: be equipped with woman's health ball (not containing the fruit of medicinal cornel) by the recipe quantity medicinal material with legal system, set by step (2) preparation negative control analytic liquid;
(4) chromatogram identification: solution point sample on the silica G plate of drawing step (1), (2), (3) preparation; The point sample amount of reference substance solution is 2 μ l, and the point sample amount of need testing solution and negative control solution is 6 μ l, and the exhibition distance is 8cm; In developping agent, launch; Spray is 30% ethanol solution of sulfuric acid with volume fraction, and 105 ℃ of hot blasts are heated to the aubergine clear spot, inspect under the daylight; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation analyze.
Said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=22: 7: 1.
In the foregoing description, concentration is that the preparation method of 30% ethanol solution of sulfuric acid is: in beaker, add 70ml ethanol, measure the 30ml concentrated sulphuric acid and slowly add along walls of beaker, the limit edged stirs.
In the foregoing description; Chromatogram recognition result: at the reference substance solution spot and R f value place of step (1) preparation; The negative control solution immaculate of step (3) preparation, other medicines do not disturb the evaluation of ursolic acid in the fruit of medicinal cornel in this explanation woman health ball, and the need testing solution of step (2) preparation is spottiness at Rf value place; The existence of ursolic acid in the proof woman health ball, and then proof thin-layered chromatography detection method of the present invention has detected the fruit of medicinal cornel in woman's health ball.
In the foregoing description, ursolic acid is bought in Nat'l Pharmaceutical & Biological Products Control Institute, and silica G is bought in Haiyang Chemical Plant, Qingdao, and woman's health ball is by Yumi Pharmaceutical Co., Ltd., Zhengzhou; Prescription and recipe quantity are: the bighead atractylodes rhizome 12 weight portions, Radix Codonopsis 12 weight portions, Poria cocos 12 weight portions, rhizoma atractylodis 12 weight portions; Ligusticum wallichii 12 weight portions, prepared rhizome of rehmannia 12 weight portions, radix cyathulae 12 weight portions; Cattail pollen 12 weight portions, rhizoma cyperi 12 weight portions, the fruit of medicinal cornel (steaming) 28 weight portions.
Claims (6)
1. the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball is characterized in that testing process operates as follows:
(1) preparation of reference substance solution: take by weighing the ursolic acid reference substance, add methyl alcohol and process the solution that every 1ml methyl alcohol contains the 1mg ursolic acid;
(2) preparation of need testing solution: take by weighing woman's health ball 5～10g, pulverize, add ethanol 20～30ml, reflux 1～2h filters, the filtrating evaporate to dryness; Residue adds water 15～20ml makes it dissolving, wash to ether layer with ether colourless, the ether layer that inclines, with normal butyl alcohol washing water layer 2～4 times, each 20～30ml; Merge n-butanol extracting liquid, with water washing 2～4 times, each 15～20ml, the layer that anhydrates that inclines is with the n-butanol layer evaporate to dryness; Residue adds ethanol 1～2ml makes it dissolving, adds 0.5～1g neutral alumina, mixes drying, the neutral alumina column top that fills in advance of packing in the water-bath thoroughly; With methyl alcohol 40～50ml wash-out, collect eluent, evaporate to dryness, residue add 0.5～1ml ethyl acetate makes it dissolving, as the test sample analytic liquid; Said woman's health ball prescription and recipe quantity are: the bighead atractylodes rhizome 12 weight portions, Radix Codonopsis 12 weight portions, Poria cocos 12 weight portions, rhizoma atractylodis 12 weight portions, Ligusticum wallichii 12 weight portions; Prepared rhizome of rehmannia 12 weight portions, radix cyathulae 12 weight portions, cattail pollen 12 weight portions, rhizoma cyperi 12 weight portions steam the fruit of medicinal cornel 28 weight portions;
(3) preparation of negative control solution: contrast pill by the recipe quantity medicinal material fully with legal system, (2) prepare the negative control analytic liquid set by step, and said contrast pill prescription and recipe quantity are: the bighead atractylodes rhizome 12 weight portions; Radix Codonopsis 12 weight portions, Poria cocos 12 weight portions, rhizoma atractylodis 12 weight portions; Ligusticum wallichii 12 weight portions, prepared rhizome of rehmannia 12 weight portions, radix cyathulae 12 weight portions; Cattail pollen 12 weight portions, rhizoma cyperi 12 weight portions;
(4) chromatogram identification: solution point sample on silica gel plate of drawing step (1), (2), (3) preparation; In developping agent, launch; Spray develops the color with ethanol solution of sulfuric acid; Need testing solution and the negative control solution of step (3) preparation and the same Rf value of the reference substance solution spot place of step (1) preparation according to step (2) preparation; Analyze, said developping agent is the mixed liquor of toluene, ethyl acetate, glacial acetic acid, and the volume ratio of each component is a toluene in the mixed liquor: ethyl acetate: glacial acetic acid=(20～30): (6～10): 1.
2. the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball according to claim 1 is characterized in that: said step (2) neutral alumina column internal diameter is 10～15mm, and the neutral alumina of filling is 1～2g, 200～300 orders.
3. the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball according to claim 1; It is characterized in that: the point sample amount of said step (4) reference substance solution is 2～3 μ l; The point sample amount of need testing solution and negative control solution is 4～6 μ l, and the exhibition distance is 6～10cm.
4. the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball according to claim 1, it is characterized in that: said silica gel plate is the silica G plate.
5. the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball according to claim 1; It is characterized in that: described ethanol solution of sulfuric acid concentration is 30%; The preparation method is: in beaker, add 70ml ethanol, measure the 30ml concentrated sulphuric acid and slowly add along walls of beaker, the limit edged stirs.
6. the thin-layered chromatography detection method of the fruit of medicinal cornel in woman's health ball according to claim 1, it is characterized in that: described colour developing is that 30% ethanol solution of sulfuric acid is sprayed on the silica gel plate, 90～110 ℃ are heated to the aubergine clear spot, inspect under the daylight.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100656965A CN101987154B (en) | 2009-08-07 | 2009-08-07 | Thin-layer chromatographic detection method for cornel in gynecological recovery pills |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100656965A CN101987154B (en) | 2009-08-07 | 2009-08-07 | Thin-layer chromatographic detection method for cornel in gynecological recovery pills |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101987154A CN101987154A (en) | 2011-03-23 |
| CN101987154B true CN101987154B (en) | 2012-03-21 |
Family
ID=43744073
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100656965A Active CN101987154B (en) | 2009-08-07 | 2009-08-07 | Thin-layer chromatographic detection method for cornel in gynecological recovery pills |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101987154B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103207148A (en) * | 2013-04-17 | 2013-07-17 | 西安碑林药业股份有限公司 | Quality detection method for Chinese herbal preparation for treating glaucoma and early-middle period cataract |
| CN104950068B (en) * | 2015-06-03 | 2016-07-06 | 贵阳中医学院第二附属医院 | The kidney invigorating is dispeled the wind the quality determining method of climacteric mixture |
| CN108562682B (en) * | 2018-04-01 | 2020-10-27 | 浙江贝尼菲特药业有限公司 | Thin-layer identification method for dogwood fruit formula particles and alcohol dogwood fruit formula particles |
-
2009
- 2009-08-07 CN CN2009100656965A patent/CN101987154B/en active Active
Non-Patent Citations (2)
| Title |
|---|
| 中华人民共和国卫生部药典委员会.妇康丸.《卫生部颁药品标准（中药成方制剂第十一册）》.1996,75. * |
| 刘莉,等.降糖宁胶囊薄层鉴别方法的研究.《黑龙江医药》.2007,第20卷(第1期),18-19. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101987154A (en) | 2011-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111044624B (en) | Quality detection method of Chinese medicinal preparation | |
| CN102998411B (en) | A kind of detection method of ageratum dripping pill | |
| CN103197027A (en) | Quality control method of astragalus-leech capsules capable of regulating collaterals | |
| CN104165962B (en) | The quality determining method of WEINAIAN sheet | |
| CN101537088B (en) | Method for controlling quality of Longdanxiegan Capsule | |
| CN105301168B (en) | The detection method of dredging collateral resolving sputum capsule | |
| CN101028388B (en) | Quality inspection of Chinese-medicinal preparation for treating shortsighness and asthenopia | |
| CN102133363A (en) | Quality detection method for chinarue cough particle | |
| CN110579561A (en) | quality control method of kidney tonifying and pulse harmonizing preparation | |
| CN101987154B (en) | Thin-layer chromatographic detection method for cornel in gynecological recovery pills | |
| CN104849369A (en) | Quality detection method of ephedra sinica-aconitum napellus-liquorice medicine | |
| CN102139067A (en) | Quality control method for antipyretic and antitoxic tablet | |
| CN100401061C (en) | Quality control method of kidney beneficial bone fortifying capsule | |
| CN109557192A (en) | A kind of raw white kalgan lamb skin takes the detection method of liquid | |
| CN100408069C (en) | Quality control method of oral preparation for yin enriching kidney supplementing | |
| CN101385808B (en) | Detection method of Zhubai tranquilizing pill | |
| CN114544852B (en) | Improved urine retention capsule quality detection method | |
| CN102429998A (en) | Detection method for blood benefiting paste | |
| CN104833754A (en) | Method for quality detection of monkshood-radix glycyrrhizae medicament | |
| CN101987115A (en) | Jade screen oral preparation as well as preparation method and detection method thereof | |
| CN103675192A (en) | Detection method of inflammation-diminishing compound pierasma quassioides benn capsule | |
| CN101385834B (en) | Medicine composition for treating urinary system disease and quality control method thereof | |
| CN102552751A (en) | Quality detection method for medicine (named as Er bao concentrated decoction) | |
| CN100334446C (en) | Method for detecting constituents of liquorice-ginseng capsule | |
| CN101703728B (en) | Quality detection method for stomach warming and soothing capsules |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent for invention or patent application | ||
| CB02 | Change of applicant information |
Address after: Xinmi City, Henan province 452392 Zheng Deng Lu 168 Applicant after: Henan Kangqi Pharmaceutical Co.,Ltd. Address before: Xinmi City, Henan province 452392 Zheng Deng Lu 168 Applicant before: Zhengzhou Yumi Pharmaceutical Co., Ltd. |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: APPLICANT; FROM: YUMI PHARMACEUTICAL CO., LTD., ZHENGZHOU TO: HE'NAN KANGQI PHARMACEUTICAL CO., LTD. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: HENAN TAIFENG PHARMACEUTICAL CO., LTD. Free format text: FORMER NAME: HE'NAN KANGQI PHARMACEUTICAL CO., LTD. |
|
| CP03 | Change of name, title or address |
Address after: Xinmi City, Henan province 452392 Zhengzhou City No. 168 Deng Lu Zheng Patentee after: Henan Taifeng Pharmaceutical Co., Ltd. Address before: Xinmi City, Henan province 452392 Zheng Deng Lu 168 Patentee before: Henan Kangqi Pharmaceutical Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20181017 Address after: 475000 intersection of seven Avenue and Longhai two road in Kaifeng, Henan Patentee after: HENAN TAIFENG BIOTECHNOLOGY CO., LTD. Address before: 452392 zhengdeng Road, Xinmi, Zhengzhou, Henan 168 Patentee before: Henan Taifeng Pharmaceutical Co., Ltd. |