CN101941961B - Method for extracting and separating kaempferol from impatiens balsamina - Google Patents
Method for extracting and separating kaempferol from impatiens balsamina Download PDFInfo
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Abstract
The invention discloses a method for extracting and separating kaempferol from impatiens balsamina, which relates to a method for extracting and separating active ingredients of traditional Chinese medicaments, in particular to a method for extracting and separating the kaempferol. The method is provided to solve the problems of complicated kaempferol preparation process and high cost of the current kaempferol preparation process. The method comprises the following steps of: firstly, extracting the impatiens balsamina to obtain a total extract; secondly, purifying the total extract by using macroporous resin to obtain a crude kaempferol product; and finally, preparing a pure kaempferol product by using a column chromatography and re-crystallization method. The method can effectively separate the kaempferol monomer from the impatiens balsamina, provides a reference product for performing the quality detection of Chinese medicaments containing the kaempferol and is favorable for developing the activities of the kaempferol.
Description
Technical field
The invention relates to the extraction and separation method of middle pharmaceutically active ingredient, be specifically related to a kind of extraction and separation method of kaempferol.
Background technology
Kaempferol is a kind of flavonols compound.This compound has multiple pharmacologically active, like antioxygenation, anti-inflammatory action etc., is present in plurality of Chinese and the related preparations thereof, like ginkgo and extract thereof and preparation, mulberry leaf, Chinese Bastardtoadflax Herb, pyrrosia lingua, South Dodder Seed Chinese Dodder Seed, Grosvenor Momordica, the treating stranguria particle of money etc.The content of kaempferol is the important detection index of the above-mentioned Chinese medicine and the quality of the pharmaceutical preparations, therefore, in real work, needs the high kaempferol of extraction separation purity as reference substance.Simultaneously, how kaempferol in the process of furtheing investigate for its activity, simplifies extracting and preparing technique, the raising separation purity of kaempferol as a kind of composition with multiple pharmacologically active, is one of key link of preparation process research process.
At present, the preparing kaempferol of report has following method:
1) international monopoly WO2006/093368A1 " MANUFACTURING METHOD OFKAEMPFEROL "; The method of from the seed of tealeaves and leaf, extracting kaempferol is disclosed; This method mainly discloses the main glucosides that adopts acid, alkali or enzyme to two kinds of higher kaempferols of content in tealeaves seed and the leaf; Be camelliaside A, camelliaside B, being hydrolyzed obtains the method for kaempferol.
2) bibliographical information is that (Su Jing, volubility eloquence, Xie Jun etc., the circulation high speed adverse current chromatogram prepares Isorhamnetol and kaempferol from Folium Ginkgo extract, Southwestern University's journal (natural science edition), 2009,31 (10): 96-100) for the method for feedstock production kaempferol with the ginkgo.The main process of this method is that Ginkgo Leaf supersound extraction, the purification of extracting solution chloroform, resin concentration flavone component, total flavones are separated with methyl alcohol-25% hydrochloric acid (4: 1) mixed solution hydrolysis, 3 HSCCC (high speed adverse current chromatogram), obtains kaempferol.
3) bibliographical information other a kind of is that (Xu Tao, Pan see, Yuan Chuanxun etc., and liquid phase chromatography prepares the trifolitin monomer, Anhui chemical industry, 2005, (2): 54-55) for the method for feedstock production kaempferol with the ginkgo.This method is got Semen Ginkgo extrac, n-butanol extraction, butanol extraction liquid concentrate, enriched material is centrifugal with hydrochloric acid hydrolysis, the hydrolyzed solution of 3M, supernatant with resin concentration, the bullion that obtains kaempferol, bullion again with the preparative high-performance liquid chromatographic method separate, lyophilize obtains kaempferol.
It is thus clear that above-mentioned preparing kaempferol is comparatively loaded down with trivial details, is embodied in: 1) need to adopt method for hydrolysis to make the glucosides of kaempferol change kaempferol into; 2) strict to equipment in the preparation of kaempferol rises the preparation cost of kaempferol.
Therefore, need to seek a kind of more convenient method for preparing kaempferol efficiently.
Flower of Garden Balsam is the dry petal of impatiens Flower of Garden Balsam Impatiens balsamina L.; Effect with promoting blood circulation to restore menstrual flow, promoting urination and stopping pain wherein contains multiple flavonoid compound, and document shows that kaempferol is higher flavones ingredient (Hu Xilan, Han Zhaoxiang, Liu Yufen etc. of content wherein; The mensuration of kaempferol in the different extracts of Flower of Garden Balsam; The assay laboratory, 2007,26 (5): 33-35).Therefore the dry petal with Flower of Garden Balsam is that raw material separates the preparation kaempferol, and with respect to the method for above-mentioned employing hydrolysis, method is easy.Based on this, bibliographical information from Flower of Garden Balsam, prepare accordingly kaempferol method (Hu Xilan, Cheng Qingfang, Yin Fujun etc., the extraction of kaempferol, isolation and purification in the Flower of Garden Balsam, the time treasure traditional Chinese medical science traditional Chinese medicines, 2010,21 (4): 932-933).
The concrete steps of this method are:
1) get the dry petal of Flower of Garden Balsam, place cable type extractor according, the sherwood oil soaked overnight, after be back to extracting liquid colourless;
The petal that 2) will extract again places cable type extractor according, the ETHYLE ACETATE soaked overnight, after be back to extracting liquid colourless;
3) acetic acid ethyl acetate extract reclaims solvent, gets the crude extract of kaempferol;
4) adopt polydextran gel (LH-20) to separate, collect the flow point that contains kaempferol, and concentrate corresponding kaempferol extract;
5) adopt preparative high-performance liquid chromatographic that the kaempferol extract is separated the preparation kaempferol.
This method kaempferol preparation cost is higher, improper large-scale industrial production.Reason has: 1) polydextran gel (LH-20) price is more expensive; 2) preparation performance liquid instrument is expensive, and the amount for preparing kaempferol simultaneously is limited.In addition, this method steps is more, and process is comparatively complicated.
Summary of the invention
The objective of the invention is to the deficiency to prior art, providing a kind of is raw material with the Flower of Garden Balsam, the method for extraction separation kaempferol.This method is more easy with respect to existing method, is fit to suitability for industrialized production.
The objective of the invention is to realize through following technical scheme:
A: get exsiccant Flower of Garden Balsam petal, the suitable pulverizing adds 10～100% ethanol or methyl alcohol and extracts, and extracting solution concentrates and reclaims solvent, gets general extractive;
B: general extractive adds water and suitably disperses, last macroporous resin adsorption post, 10～40% ethanol elution removal of impurities are used in the removal of impurities of first water eluant solution again, at last with 40～100% ethanol elution and collect this part elutriant, concentrate and reclaim solvent, the kaempferol bullion;
C: get polymeric amide wet method dress post, the kaempferol crude product solution of 30～100% dissolve with ethanol is added capital, with 30～100% ethanol elutions, collect elutriant, the thin layer inspection is known, and merges stream part of containing kaempferol, reclaims solvent, gets the elaboration of kaempferol;
D: get the elaboration of kaempferol, recrystallization in the mixed solvent of methyl alcohol or ethanol and water composition filters, and drying gets the pure article of kaempferol.
Extracting mode in the aforesaid method in the steps A is any one in decoction, reflux, supersound extraction, microwave extraction, the high pressure extract.
The method of steps A is in the aforesaid method: get exsiccant Flower of Garden Balsam petal 25 weight parts, adding concentration is 40～100% ethanol or methyl alcohol 400～900 parts by volume, heating and refluxing extraction 0.5～2 hour; Filter; The dregs of a decoction successively extract 2～5 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, gets thick paste, i.e. general extractive.
The preferred method of steps A is in the aforesaid method: get exsiccant Flower of Garden Balsam petal 25 weight parts, adding concentration is 80% ethanol 750 parts by volume, heating and refluxing extraction 1 hour; Filter, the dregs of a decoction successively extract 3 times with same method, united extraction liquid; The dregs of a decoction discard, and extracting solution reclaims ethanol, get thick paste; Be general extractive, 6.8 weight parts.
In the aforesaid method among the step B macroporous adsorbent resin be in nonpolar, low-pole or the middle polarity any one.
The method of step B is in the aforesaid method: get general extractive; Add the water dilution of 5～9 times of parts by volume of medicinal material amount; Go up any one macroporous adsorptive resins in nonpolar, low-pole or the middle polarity; Amount of resin is 6～10 times of parts by volume of medicinal material amount, and blade diameter length ratio is 1: 5～8, absorption flow velocity 2～4BV/h; With water elution 5～13BV, flow velocity 2～5BV/h uses ethanol elution 5～10BV of 10～40%, flow velocity 2～5BV/h more earlier; Use ethanol elution 2～5BV of 40～100% at last, flow velocity 2～5BV/h collects 40～100% ethanol eluate, reclaims solvent, gets the kaempferol bullion.
The preferred method of step B is in the aforesaid method: get above-mentioned general extractive 6.8 weight parts, add the water dilution of 6 times of parts by volume of medicinal material amount, and last D101 type macroporous adsorptive resins, amount of resin is 8 times of parts by volume of medicinal material amount; Blade diameter length ratio is 1: 6, and absorption flow velocity 3BV/h uses earlier water elution 9BV, flow velocity 3BV/h; Use 30% ethanol elution 9BV again, flow velocity 3BV/h uses 50% ethanol elution 4BV, flow velocity 2BV/h at last; Collect 50% ethanol eluate, reclaim solvent, get 0.55 weight part kaempferol bullion.
The method of step C is in the aforesaid method: get the kaempferol bullion, fully dissolve with the ethanol of 0.1～1 times of parts by volume 50～90% of medicinal material amount, sample solution; Other gets polymeric amide wet method dress post, and sample solution is added capital, and blade diameter length ratio is 1: 8～15; Aqueous ethanolic solution wash-out with 50～100% is collected elutriant, and the thin layer inspection is known; Merge stream part of containing kaempferol, reclaim the elaboration that solvent gets kaempferol.
Step C preferable methods is in the aforesaid method: get kaempferol bullion 0.55 weight part, fully dissolve with the ethanol of 0.2 times of parts by volume 70% of medicinal material amount, sample solution; Other get it filled 80～100 order polymeric amide of 0.4 times of weight part of material amount, with 70% ethanol wet method dress post, blade diameter length ratio is 1: 10; Sample solution is added capital, and the aqueous ethanolic solution wash-out with 70% is collected elutriant; The thin layer inspection is known, and merges stream part of containing kaempferol, reclaims the elaboration that solvent gets 0.25 weight part kaempferol.
The method of step D is in the aforesaid method: get the elaboration of kaempferol, in 2～8: 4～9 methanol-waters or 2～8: carry out recrystallization in the mixed solvent of 4～9 alcohol-waters, filter, the dry pure article of kaempferol that get.
Step D preferable methods is in the aforesaid method: get the elaboration of 0.25 weight part kaempferol, recrystallization in 3: 7 ethanol-water mixed solvents filters, dry the pure article of 0.15 weight part kaempferol.
Adopt present method to extract the preparation kaempferol, the extraction process route is simple, is suitable for large-scale industrialization production.
Embodiment
Following experimental example is used to further specify but is not limited to the present invention.
Experimental example 1: the purity test of kaempferol
Check purity with HPLC.Instrument: Waters 600 high performance liquid chromatographs.Chromatographic column: YWG-C
18Chromatographic column, 250mm * 4.6mm, 5 μ m.Moving phase: methyl alcohol-0.1% phosphate aqueous solution (50～70: 50～30); Flow velocity: 0.8～1.2mL/min; Column temperature: room temperature; Detect wavelength: 367nm; Sample size: 10 μ l; RT (RT): kaempferol is 5～30min; Color atlas record time: 60min.Area normalization method is surveyed purity, and the purity of kaempferol is 98.8%.
Experimental example 2: the structure determination of kaempferol
Fusing point shows micro melting point apparatus with X-4 type numeral; Ultraviolet (UV) spectrum is with day island proper Tianjin UV-260 type ultraviolet-visible pectrophotometer; Infrared (IR) spectrum is with Perkin-Elmer 983G type Instrument measuring, KBr compressing tablet; Proton nmr spectra and carbon spectrum (
1HNMR with
13CNMR) use the MERCURY-400 nmr determination; Mass spectrum (MS) is used Finnigan-LCQ
DECAThe type mass spectrograph is measured.
The pure article of the kaempferol of above-mentioned process for purification gained are: the crystallization of yellow particle shape, 276～278 ℃ of fusing points.UVλmax(nm)：226，267，349。IRυmax(KBr，cm
-1)：3324(OH)，1662(C＝O)。MS(m/z)：309(M+Na)。
1H?NMR(δ，DMSO-d
6)：12.47(5-OH)，10.85(7-OH)，10.12(3-OH)，9.43(4′-OH)，8.03(2H，d，J＝9.0Hz，C
2′-H，C
6′-H)，6.91(2H，d，J＝9.0Hz，C
3′-H，C
5′-H)，6.43(1H，d，J＝1.8Hz，C
8-H)，6.18(1H，d，J＝1.8Hz，C
6-H)。
13C?NMR(δ，DMSO-d
6)：175.9(C-4)，163.9(C-7)，160.7(C-5)，159.2(C-4′)，156.2(C-9)，146.8(C-2)，135.6(C-3)，129.5(C-2′)，129.5(C-6′)，121.7(C-1′)，115.4(C-3′)，115.4(C-5′)，103.1(C-10)，98.2(C-6)，93.5(C-8)。
Above spectral data is consistent with bibliographical information, and structure is following:
The experiment of experimental example 3 macroporous adsorbent resin enrichment kaempferol bullions
Get exsiccant Flower of Garden Balsam petal 2.5kg, adding concentration is 80% ethanol 75L, and heating and refluxing extraction 1 hour is filtered, and the dregs of a decoction successively extract 3 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 0.68kg.
Get above-mentioned general extractive 0.68kg, add the dilution of 15L water, with D101 type macroporous adsorptive resins, amount of resin is 20L (volume in the water), and blade diameter length ratio is 1: 6, absorption flow velocity 3BV/h; Earlier use water elution 9BV, flow velocity 3BV/h uses 30% ethanol elution 9BV, flow velocity 3BV/h again; Use 50% ethanol elution 4BV at last, flow velocity 2BV/h collects this part elutriant, reclaims solvent, gets 0.055kg kaempferol bullion.
The experiment that experimental example 4 polyamide column chromatography methods and recrystallization method coupling prepare the pure article of kaempferol
The method that adopts polyamide column chromatography and recrystallization coupling for preparing of the pure article of kaempferol prepares.(road and bridge tetramethyl biochemical plastic molding and processing plant in Taizhou city produces the sorbent material that uses during column chromatography, 1kg) as polymeric amide 80～100 orders.Get above-mentioned kaempferol bullion 0.055kg, fully dissolve with the ethanol of 0.5L 70%, sample solution, appearance on the wet method (polymeric amide chromatography post is adorned post with 70% aqueous ethanolic solution wet method, and the diameter height ratio of chromatography column is 1: 10).With 70% aqueous ethanolic solution is the eluent wash-out, collects elutriant, and every part of 400ml collects 60 parts altogether, every part be evaporated to small volume after, detect with the polymeric amide thin layer.The thin layer condition: polyamide layer is prefabricated; Developping agent: chloroform-methanol-formic acid (10: 2: 0.05); Color condition: 1%AlCl
3The ethanolic soln colour developing, uv lamp 365nm wavelength is observed down.Wherein the content of kaempferol is higher in the 25th～40 part, and with its merging, the decompressing and extracting solvent gets yellow kaempferol elaboration 0.025kg.
Get kaempferol elaboration 0.025kg, recrystallization in 3: 7 ethanol-water mixed solvents filters, drying, the pure article of kaempferol of 0.015kg, be the crystallization of yellow particle shape.
Embodiment 1
Get exsiccant Flower of Garden Balsam petal 2.5kg, adding concentration is 90% methyl alcohol 60L, and heating and refluxing extraction 1 hour is filtered, and the dregs of a decoction successively extract 2 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 0.55kg.
Get above-mentioned general extractive 0.55kg, add the dilution of 12.5L water, with AB-8 type macroporous adsorptive resins, amount of resin is 15L (volume in the water), and blade diameter length ratio is 1: 5, absorption flow velocity 2BV/h; Earlier use water elution 5BV, flow velocity 2BV/h uses 20% ethanol elution 6BV, flow velocity 2BV/h again; Use 50% ethanol elution 5BV at last, flow velocity 2BV/h collects this part elutriant, reclaims solvent, gets 0.043kg kaempferol bullion.
The method that adopts polyamide column chromatography and recrystallization coupling for preparing of the pure article of kaempferol prepares.(road and bridge tetramethyl biochemical plastic molding and processing plant in Taizhou city produces the sorbent material that uses during column chromatography, 1kg) as polymeric amide 80～100 orders.Get above-mentioned kaempferol bullion 0.043kg, fully dissolve with the ethanol of 0.5L 70%, sample solution, appearance on the wet method (polymeric amide chromatography post is adorned post with 80% aqueous ethanolic solution wet method, and the diameter height ratio of chromatography column is 1: 10).With 80% aqueous ethanolic solution is the eluent wash-out, collects elutriant, and every part of 200ml collects 80 parts altogether, every part be evaporated to small volume after, detect with the polymeric amide thin layer.The thin layer condition: polyamide layer is prefabricated; Developping agent: chloroform-methanol-formic acid (10: 2: 0.05); Color condition: 1%AlCl
3The ethanolic soln colour developing, uv lamp 365nm wavelength is observed.Wherein the content of kaempferol is higher in the 20th～30 part, and with its merging, the decompressing and extracting solvent gets yellow kaempferol elaboration 0.015kg.
Get kaempferol elaboration 0.015kg, recrystallization in 5: 5 methanol-water mixed solvents filters, drying, the pure article of kaempferol of 0.008kg, be the crystallization of yellow particle shape.
Embodiment 2
Get exsiccant Flower of Garden Balsam petal 2.5kg, adding concentration is 80% ethanol 70L, and heating and refluxing extraction 1.5 hours is filtered, and the dregs of a decoction successively extract 3 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 0.65kg.
Get above-mentioned general extractive 0.65kg, add the dilution of 15L water, with D101 type macroporous adsorptive resins, amount of resin is 20L (volume in the water), and blade diameter length ratio is 1: 7, absorption flow velocity 3BV/h; Earlier use water elution 9BV, flow velocity 3BV/h uses 30% ethanol elution 7BV, flow velocity 2BV/h again; Use 70% ethanol elution 4BV at last, flow velocity 2BV/h collects this part elutriant, reclaims solvent, gets 0.05kg kaempferol bullion.
The method that adopts polyamide column chromatography and recrystallization coupling for preparing of the pure article of kaempferol prepares.(road and bridge tetramethyl biochemical plastic molding and processing plant in Taizhou city produces the sorbent material that uses during column chromatography, 1kg) as polymeric amide 120～200 orders.Get above-mentioned kaempferol bullion 0.05kg, fully dissolve with the ethanol of 0.7L 70%, sample solution, appearance on the wet method (polymeric amide chromatography post is adorned post with 70% aqueous ethanolic solution wet method, and the diameter height ratio of chromatography column is 1: 12).With 70% aqueous ethanolic solution is the eluent wash-out, collects elutriant, and every part of 400ml collects 60 parts altogether, every part be evaporated to small volume after, detect developping agent: chloroform-methanol-formic acid (10: 1: 0.05) with silica gel thin-layer; Color condition: 1%AlCl
3The ethanolic soln colour developing, uv lamp 365nm wavelength is observed.Wherein the content of kaempferol is higher in the 25th～40 part, and with its merging, the decompressing and extracting solvent gets yellow kaempferol elaboration 0.023kg.
Get kaempferol elaboration 0.023kg, recrystallization in 3: 7 ethanol-water mixed solvents filters, drying, the pure article of kaempferol of 0.014kg, be the crystallization of yellow particle shape.
Embodiment 3
Get exsiccant Flower of Garden Balsam petal 2.5kg, adding concentration is 70% ethanol 80L, and heating and refluxing extraction 1 hour is filtered, and the dregs of a decoction successively extract 3 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 0.70kg.
Get above-mentioned general extractive 0.70kg, add the dilution of 20L water, with HPD100 type macroporous adsorptive resins, amount of resin is 22L (volume in the water), and blade diameter length ratio is 1: 7, absorption flow velocity 3BV/h; Earlier use water elution 12BV, flow velocity 3BV/h uses 30% ethanol elution 10BV, flow velocity 4BV/h again; Use 80% ethanol elution 4BV at last, flow velocity 2BV/h collects this part elutriant, reclaims solvent, gets 0.074kg kaempferol bullion.
The method that adopts polyamide column chromatography and recrystallization coupling for preparing of the pure article of kaempferol prepares.(road and bridge tetramethyl biochemical plastic molding and processing plant in Taizhou city produces the sorbent material that uses during column chromatography, 1.3kg) as polymeric amide 80～100 orders.Get above-mentioned kaempferol bullion 0.074kg, fully dissolve with the ethanol of 1L 75%, sample solution, appearance on the wet method (polymeric amide chromatography post is adorned post with 75% aqueous ethanolic solution wet method, and the diameter height ratio of chromatography column is 1: 10).With 75% aqueous ethanolic solution is the eluent wash-out, collects elutriant, and every part of 400ml collects 70 parts altogether, every part be evaporated to small volume after, detect developping agent: chloroform-methanol (7: 3) with silica gel thin-layer; Color condition: 1%AlCl
3The ethanolic soln colour developing, uv lamp 365nm wavelength is observed.Wherein the content of kaempferol is higher in the 20th～35 part, and with its merging, the decompressing and extracting solvent gets yellow kaempferol elaboration 0.022kg.
Get kaempferol elaboration 0.022kg, recrystallization in 4: 6 ethanol-water mixed solvents filters, drying, the pure article of kaempferol of 0.013kg, be the crystallization of yellow particle shape.
Embodiment 4
Get exsiccant Flower of Garden Balsam petal 2.5kg, adding concentration is 60% methyl alcohol 80L, and heating and refluxing extraction 2 hours is filtered, and the dregs of a decoction successively extract 4 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 0.75kg.
Get above-mentioned general extractive 0.75kg, add the dilution of 20L water, with D4020 type macroporous adsorptive resins, amount of resin is 25L (volume in the water), and blade diameter length ratio is 1: 8, absorption flow velocity 4BV/h; Earlier use water elution 13BV, flow velocity 4BV/h uses 30% ethanol elution 10BV, flow velocity 5BV/h again; Use 90% ethanol elution 3BV at last, flow velocity 2BV/h collects this part elutriant, reclaims solvent, gets 0.087kg kaempferol bullion.
The method that adopts polyamide column chromatography and recrystallization coupling for preparing of the pure article of kaempferol prepares.(road and bridge tetramethyl biochemical plastic molding and processing plant in Taizhou city produces the sorbent material that uses during column chromatography, 1.3kg) as polymeric amide 80～100 orders.Get above-mentioned kaempferol bullion 0.087kg, fully dissolve with the ethanol of 1.5L 60%, sample solution, appearance on the wet method (polymeric amide chromatography post is adorned post with 60% aqueous ethanolic solution wet method, and the diameter height ratio of chromatography column is 1: 13).With 60% aqueous ethanolic solution is the eluent wash-out, collects elutriant, and every part of 400ml collects 120 parts altogether, every part be evaporated to small volume after, detect with the polymeric amide thin layer.The thin layer condition: polyamide layer is prefabricated; Developping agent: chloroform-methanol-formic acid (10: 2: 0.05); Color condition: 1%AlCl
3The ethanolic soln colour developing, uv lamp 365nm wavelength is observed.Wherein the content of kaempferol is higher in the 55th～73 part, and with its merging, the decompressing and extracting solvent gets yellow kaempferol elaboration 0.02kg.
Get kaempferol elaboration 0.02kg, recrystallization in 3: 7 methanol-water mixed solvents filters, drying, the pure article of kaempferol of 0.011kg, be the crystallization of yellow particle shape.
Claims (1)
1. the extraction and separation method of a kaempferol, it is characterized in that: this method comprises the steps:
A: get exsiccant Flower of Garden Balsam petal 25 weight parts, suitably pulverize, adding concentration is 80% ethanol 750 parts by volume; Heating and refluxing extraction 1 hour is filtered, and the dregs of a decoction successively extract 3 times with same method; United extraction liquid, the dregs of a decoction discard, and extracting solution reclaims ethanol; Thick paste, i.e. general extractive, 6.8 weight parts;
B: get above-mentioned general extractive 6.8 weight parts, add the water dilution of 6 times of parts by volume of medicinal material amount, last D101 type macroporous adsorptive resins, amount of resin is 8 times of parts by volume of medicinal material amount; Blade diameter length ratio is 1: 6, and absorption flow velocity 3BV/h uses earlier water elution 9BV, flow velocity 3BV/h; Use 30% ethanol elution 9BV again, flow velocity 3BV/h uses 50% ethanol elution 4BV, flow velocity 2BV/h at last; Collect 50% ethanol eluate, concentrate and reclaim solvent, get 0.55 weight part kaempferol bullion;
C: get kaempferol bullion 0.55 weight part, fully dissolve with the ethanol of 0.2 times of parts by volume 70% of medicinal material amount, sample solution; Other get it filled 80～100 order polymeric amide of 0.4 times of weight part of material amount, with 70% ethanol wet method dress post, blade diameter length ratio is 1: 10; Sample solution is added capital, and the aqueous ethanolic solution wash-out with 70% is collected elutriant; The thin layer inspection is known, and merges stream part of containing kaempferol, reclaims the elaboration that solvent gets 0.25 weight part kaempferol;
D: get the elaboration of 0.25 weight part kaempferol, recrystallization in 3: 7 ethanol-water mixed solvents filters, dry the pure article of 0.15 weight part kaempferol.
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| CN102746265A (en) * | 2011-04-21 | 2012-10-24 | 西安通江生物科技有限责任公司 | Method for extracting high purity kaempferol from sophora fruit extraction waste residue |
| CN105267082A (en) * | 2014-07-17 | 2016-01-27 | 伽蓝（集团）股份有限公司 | Impatiens balsamina extract and application thereof |
| TWI672137B (en) * | 2018-06-27 | 2019-09-21 | 鼎赫生物科技股份有限公司 | Method for preparing anti-oxidation component kaempferol by hydrolysis technique in Taiwan native clay |
| CN110642824A (en) * | 2018-06-27 | 2020-01-03 | 鼎赫生物科技股份有限公司 | Method for preparing antioxidant ingredient kaempferol from cortex cinnamomi by hydrolysis technology |
| CN111675618B (en) * | 2020-06-20 | 2023-03-31 | 贵州中医药大学 | Compound in pyrrosia pedunculata and separation and purification method and application thereof |
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| CN1844116A (en) * | 2006-05-10 | 2006-10-11 | 江南大学 | Extraction separation purification and identification of flavonoid monomers in oriental blueberry melanin |
| CN101759678A (en) * | 2009-11-20 | 2010-06-30 | 南京泽朗医药科技有限公司 | Method for preparing kaempferol |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844116A (en) * | 2006-05-10 | 2006-10-11 | 江南大学 | Extraction separation purification and identification of flavonoid monomers in oriental blueberry melanin |
| CN101759678A (en) * | 2009-11-20 | 2010-06-30 | 南京泽朗医药科技有限公司 | Method for preparing kaempferol |
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