CN104208269A - Application of capsicum polyphenol to pharmacy and preparation method of the same - Google Patents
Application of capsicum polyphenol to pharmacy and preparation method of the same Download PDFInfo
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- CN104208269A CN104208269A CN201410389258.5A CN201410389258A CN104208269A CN 104208269 A CN104208269 A CN 104208269A CN 201410389258 A CN201410389258 A CN 201410389258A CN 104208269 A CN104208269 A CN 104208269A
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Abstract
The invention discloses the application of capsicum polyphenol to pharmacy and a preparation method of the same, especially the application to preparation of antibacterial and anti tumor drugs. Capsicum polyphenol has antibacterial, antitumor, antioxidant, anti-inflammatory and cholesterol rise inhibiting effects, and has wide application prospect. The preparation method of capsicum polyphenol is by crushing capsicum stem and leaf, extracting by an ethanol-ultrasonic extraction method to obtain a polyphenol crude extract, and carrying out high speed counter current chromatography for enrichment. The preparation method has the advantages of short time, high efficiency, low energy consumption, no solid filler in a purification process, mild conditions, low loss and no pollution, can conduct efficient, fast and low cost enrichment of active capsicum polyphenol, and is suitable for industrialized production.
Description
Technical field:
The invention belongs to medical art, be specifically related to the application of a kind of Fructus Capsici active polyphenol in pharmacy and the preparation method of Fructus Capsici active polyphenol thereof.
Technical background:
China is world Fructus Capsici first manufacturing country and major consumers state, plants and eats with a long history.Fructus Capsici is annual herb plant, and the output of its ripe plants stems, leaf and fruit is suitable, namely produces 1 kilogram of pepper fruit, will produce the Fructus Capsici stem and leaf of about 1 kilogram.But, because Caulis Capsici, leaf mouthfeel should not directly eat, fail for a long time to be effectively utilized, be usually taken as waste combustion and also or directly abandon, cause serious environmental pollution and the wasting of resources.Therefore find a kind of high-efficiency and economic, Caulis Capsici, the leaf Application way of environmental protection are extremely urgent.Research finds containing various active composition in Caulis Capsici, leaf, as: capsaicin, polyphenol etc.The flavonoid glycoside that Chinese patent 201210136560.0 extracts from Folium Capsici has alpha-glucosaccharase enzyme inhibition activity, but its content is extremely low, and less than 0.2 ‰, in Folium Capsici, most of active component is not utilized.Plant polyphenol poor heat stability, changeableness, conventional preparation means product easy in inactivation.Chinese patent 201010181740.1 and 201210229217.0 have employed the method for traditional extraction plant polyphenol respectively: percolation and stirring and leaching method, and the two extracts length consuming time, consume energy high, productive rate is low, and obtain polyphenol biological activity poor.The assisted extraction means of rising in recent years, as supercritical fluid extraction and microwave assisting method etc., though polyphenol extraction ratio is improved, but equipment requirements is high, and investment is large, and cost is high, is not suitable with large production requirement.Folium Capsici polyphenol biological activity is good, but its content is on the low side compared with other plant, there is no the method effectively preparing Folium Capsici active polyphenol at present, seriously constrains the exploitation of Folium Capsici resource.There is no the application of Folium Capsici active polyphenol in preparation treatment and prevention antitumor drug, and preparing the report of the application in antibacterial yet.
Summary of the invention:
The object of the present invention is to provide a kind of Fructus Capsici polyphenol in the preparation method of the application prepared in antibacterial and antitumor drug and this Fructus Capsici polyphenol thereof.The method that the inventive method extracts plant polyphenol than traditional experiment room is more superior, and equipment adopts energy-gathered ultrasonic circulation extraction apparatus, can be amplified to suitability for industrialized production; Use high speed adverse current chromatogram enrichment Fructus Capsici polyphenol, whole process does not need to use solid packing, does not have Irreversible Adsorption, and sample nondestructive loses, pollution-free, can efficient, fast, low cost enrichment Fructus Capsici active polyphenol; The Fructus Capsici polyphenol obtained has antibacterial, anti-tumor activity.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
A kind of Fructus Capsici polyphenol is preparing the application in antibacterial.
A kind of Fructus Capsici polyphenol is preparing the application in antitumor drug.
As described in a kind of Fructus Capsici polyphenol preparing the application in antibacterial, wherein said Fructus Capsici polyphenol is with Caulis Capsici, leaf powder for raw material, take ethanol-water solution as solvent, through ultrasonic extraction extract, through high speed adverse current chromatogram enrichment gained.
As described in a kind of Fructus Capsici polyphenol preparing the application in antibacterial, wherein said Fructus Capsici polyphenol is with Caulis Capsici, leaf powder for raw material, by Caulis Capsici, leaf cleaning, drying, pulverize, cross 20 mesh sieves, obtain Fructus Capsici stem and leaf powder, be placed in aeration-drying place preserve stand-by; Then be the ethanol-water solution of 30% ~ 80% by previous step gained Caulis Capsici, leaf powder and volume fraction, pH value is 1 ~ 7, mix in ultrasonic extraction tank, solid-liquid ratio is 1:5 ~ 30, Extracting temperature is 30 DEG C ~ 60 DEG C, ultrasonic power is 200w ~ 1000w, and ultrasonic time and cavitation time slot are 4s-4s ~ 4s-8s, ultrasonic time 20 ~ 80min; Extract and terminate, filter to get filtrate, filtrate is also dry through decompression precipitation, obtained crude extract extractum, and then by high speed adverse current chromatogram enriching and purifying, obtains Fructus Capsici polyphenol; In described high-speed counter-current enriching step, be separated solvent system and be made up of acetate-methanol-water 10:1:10, add acid further and adjust pH value to be 1 ~ 7; Above-mentioned solvent system is placed in stratification after separatory funnel shake well, getting phase solvent is immobile phase, and lower phase solvent is mobile phase, makes to be full of immobile phase in counter-current chromatograph pillar when system temperature 5 ~ 60 DEG C, then make its main frame rotate, then mobile phase is pumped in post; Sample phase mixed solvent dissolving up and down, by injection valve sample introduction; According to detector spectrogram receiving target composition Fructus Capsici polyphenol.
As described in a kind of Fructus Capsici polyphenol preparing the application in antibacterial, the bacterium in wherein said antibacterial refers to bacillus subtilis, staphylococcus aureus, yeast, penicillium sp, escherichia coli, aspergillus niger.
As described in a kind of Fructus Capsici polyphenol preparing the application in antitumor drug, wherein said tumor cell is human glioma U251 cell and human neuroblastoma SH-SY5Y cell.
As described in a kind of Fructus Capsici polyphenol preparing the application in antitumor drug, wherein said Fructus Capsici polyphenol is with Caulis Capsici, leaf powder for raw material, take ethanol-water solution as solvent, through ultrasonic extraction extract, through high speed adverse current chromatogram enrichment gained.
As described in a kind of Fructus Capsici polyphenol preparing the application in antitumor drug, wherein said Fructus Capsici polyphenol be with Caulis Capsici, leaf powder for raw material, Caulis Capsici, leaf are cleaned, dry, pulverize, cross 20 mesh sieves, obtain Fructus Capsici stem and leaf powder, be placed in aeration-drying place and preserve stand-by; Then be the ethanol-water solution of 30% ~ 80% by previous step gained Caulis Capsici, leaf powder and volume fraction, pH value is 1 ~ 7, mix in ultrasonic extraction tank, solid-liquid ratio is 1:5 ~ 30, Extracting temperature is 30 DEG C ~ 60 DEG C, ultrasonic power is 200w ~ 1000w, and ultrasonic time and cavitation time slot are 4s-4s ~ 4s-8s, ultrasonic time 20 ~ 80min; Extract and terminate, filter to get filtrate, filtrate is also dry through decompression precipitation, obtained crude extract extractum, and then by high speed adverse current chromatogram enriching and purifying, obtains Fructus Capsici polyphenol; In described high-speed counter-current enriching step, be separated solvent system and be made up of acetate-methanol-water 10:1:10, adding sour adjust pH is further 1 ~ 7; Above-mentioned solvent system is placed in stratification after separatory funnel shake well, getting phase solvent is immobile phase, and lower phase solvent is mobile phase, makes to be full of immobile phase in counter-current chromatograph pillar when system temperature 5 ~ 60 DEG C, then make its main frame rotate, then mobile phase is pumped in post; Sample phase mixed solvent dissolving up and down, by injection valve sample introduction; According to detector spectrogram receiving target composition Fructus Capsici polyphenol.
Invention also provides a kind of preparation method of Fructus Capsici polyphenol, the method for raw material with Caulis Capsici, leaf powder, by Caulis Capsici, leaf cleaning, drying, is pulverized, is crossed 20 mesh sieves, obtain Fructus Capsici stem and leaf powder, is placed in aeration-drying place and preserves stand-by; Then be the ethanol-water solution of 30% ~ 80% by previous step gained Caulis Capsici, leaf powder and volume fraction, pH value is 1 ~ 7, mix in ultrasonic extraction tank, solid-liquid ratio is 1:5 ~ 30, during extraction, temperature is 30 DEG C ~ 60 DEG C, ultrasonic power is 200w ~ 1000w, and ultrasonic time and cavitation time slot are 4s-4s ~ 4s-8s, ultrasonic time 20 ~ 80min; Extract and terminate, filter to get filtrate, filtrate is also dry through decompression precipitation, obtained crude extract extractum, and then by high speed adverse current chromatogram enriching and purifying, obtains Fructus Capsici polyphenol; In described high-speed counter-current enriching step, be separated solvent system and be made up of acetate-methanol-water 10:1:10, adding sour adjust pH is further 1 ~ 7; Above-mentioned solvent system is placed in stratification after separatory funnel shake well, getting phase solvent is immobile phase, and lower phase solvent is mobile phase, makes to be full of immobile phase in counter-current chromatograph pillar when system temperature 5 ~ 60 DEG C, then make its main frame rotate, then mobile phase is pumped in post; Sample phase mixed solvent dissolving up and down, by injection valve sample introduction; According to detector spectrogram receiving target composition Fructus Capsici polyphenol.
Detailed description of the invention
Further illustrate essentiality content of the present invention with embodiments of the invention below, but do not limit the present invention with this.
Embodiment 1:
In the 50% ethanol injection extracting cylinder that precision weighing 8L configures, be warming up to 60 DEG C.Caulis Capsici, leaf powder are pressed solid-liquid ratio 1:25 and dropped in extracting cylinder, stirs evenly.The condition that setting is extracted: ultrasonic power 800w, ultrasonic time 30min, ultrasonic gap 4s ~ 8s, pH value 2, extracts.After extraction terminates, by extracting solution sucking filtration, filtrate decompression precipitation is also dry, obtain Fructus Capsici polyphenol extract powder 98.5g.
Adopt Shanghai with field TBE-300C semi-preparative high speed adverse current chromatogram system enrichment Fructus Capsici polyphenol, the volume of post is 300mL, and solvent system is acetate-methanol-water 10:1:10, column temperature: 25 DEG C.The enrichment of Fructus Capsici polyphenol is carried out in two steps: the acetate-methanol-water 1. by volume ratio being 10:1:10 is miscible in separatory funnel, shakes up rear static layering.Getting its upper solution (upper phase) is immobile phase, and lower floor's solution (lower phase) is mobile phase.Before sample introduction, be first full of whole pillar with immobile phase, adjustment engine speed is 890rpm, pumps in post with the flow velocity of 5.0mL/min by mobile phase, sets up the dynamic equilibrium of whole system.2. with the Fructus Capsici polyphenol extract 500mg that the upper and lower phase mixed liquor 10mL dissolving of 1:1 obtains, by injection valve sample introduction, then according to detector 254nm uv-spectrogram, receiving target composition Fructus Capsici polyphenol.
Embodiment 2:
In the 60% ethanol injection extracting cylinder that precision weighing 8L configures, be warming up to 50 DEG C.Caulis Capsici, leaf powder are pressed solid-liquid ratio 1:20 and dropped in extracting cylinder, stirs evenly.The condition that setting is extracted: ultrasonic power 600w, ultrasonic time 50min, ultrasonic gap 4s ~ 6s, pH value 3, extracts.After extraction terminates, by extracting solution sucking filtration, filtrate decompression precipitation is also dry, obtain Fructus Capsici polyphenol extract powder, obtain Fructus Capsici polyphenol extract powder 95.5g.
Adopt Shanghai with field TBE-300C semi-preparative high speed adverse current chromatogram system enrichment Fructus Capsici polyphenol, the volume of post is 300mL, and solvent system is acetate-methanol-water 10:1:10, adds glacial acetic acid and adjusts pH4.5, column temperature: 35 DEG C.The enrichment of Fructus Capsici polyphenol is carried out in two steps: the acetate-methanol-water 1. by volume ratio being 10:1:10 is miscible in separatory funnel, adds glacial acetic acid and adjusts pH4.5, shake up rear static layering.Getting its upper solution (upper phase) is immobile phase, and lower floor's solution (lower phase) is mobile phase.Before sample introduction, first be full of whole pillar with immobile phase, adjustment engine speed is 890rpm, with the flow velocity of 7.0mL/min, mobile phase is pumped in post, set up the dynamic equilibrium of whole system 2. with the Fructus Capsici polyphenol extract 500mg that the upper and lower phase mixed liquor 10mL dissolving of 1:1 obtains, by injection valve sample introduction, then according to detector 254nm uv-spectrogram, receiving target composition Fructus Capsici polyphenol.
Embodiment 3:
In the 70% ethanol injection extracting cylinder that precision weighing 8L configures, be warming up to 40 DEG C.Caulis Capsici, leaf powder are pressed solid-liquid ratio 1:10 and dropped in extracting cylinder, stirs evenly.The condition that setting is extracted: ultrasonic power 400w, ultrasonic time 70min, ultrasonic gap 4s ~ 4s, pH value 4, extracts.After extraction terminates, by extracting solution sucking filtration, filtrate decompression precipitation is also dry, obtain Fructus Capsici polyphenol extract powder, obtain Fructus Capsici polyphenol extract powder 92.5g.
Adopt Shanghai with field TBE-300C semi-preparative high speed adverse current chromatogram system enrichment Fructus Capsici polyphenol, the volume of post is 300mL, and solvent system is acetate-methanol-water 10:1:10, adds glacial acetic acid and adjusts pH3.0, column temperature: 35 DEG C.The enrichment of Fructus Capsici polyphenol is carried out in two steps: the acetate-methanol-water 1. by volume ratio being 10:1:10 is miscible in separatory funnel, adds glacial acetic acid and adjusts pH3.0, shake up rear static layering.Getting its upper solution (upper phase) is immobile phase, and lower floor's solution (lower phase) is mobile phase.Before sample introduction, be first full of whole pillar with immobile phase, adjustment engine speed is 890rpm, pumps in post with the flow velocity of 5.0mL/min by mobile phase, sets up the dynamic equilibrium of whole system.2. with the Fructus Capsici polyphenol extract 500mg that the upper and lower phase mixed liquor 10mL dissolving of 1:1 obtains, by injection valve sample introduction, then according to detector 254nm uv-spectrogram, receiving target composition Fructus Capsici polyphenol.
Embodiment 4:
The Determination of Antibacterial Activity of Fructus Capsici active polyphenol:
Test organisms:
Staphylococcus aureus (Staphylococcus coureus), G
+
Escherichia coli (Escherichia coli), G
-
Bacillus subtilis (Bacillus subtills), G
+
Penicillium sp (Penicillium)
Aspergillus niger (Aspergillus niger)
Yeast (yeast)
Culture medium:
Antibacterial: Carnis Bovis seu Bubali cream agar is trained
Mycete: potato dextrose agar
Yeast: yeast extract peptone glucose agar medium
Polyphenol solution preparation:
Precision takes embodiment 1 gained Fructus Capsici active polyphenol, and with ethanol: water: the solvent of dimethyl sulfoxide (DMSO)=2:2:1, with 2 times of dilution methods, from 50mg/mL, be configured to the solution that least concentration is 0.196mg/mL successively, filtration sterilization is for subsequent use.
Collecting cells:
Antibacterial activates on nutrient agar solid medium flat board, get 1-2 single bacterium colony to join in nutrient broth fluid medium and make seed liquor, preference temperature (bacillus cereus 30 DEG C, staphylococcus aureus 37 DEG C, bacillus subtilis spore bacillus 30 DEG C, escherichia coli 37 DEG C), 200rpm cultivates 6-8h, makes antibacterial be in exponential phase.Bacterial concentration is regulated to make the bacteria concentration of antibacterial be 10 by measuring OD value
6cfu/mL.
Experimental technique:
Adopt plate punch method: put into for examination bacterium asepsis ring picking 1-2 ring the physiological saline solution being loaded with 15mL by after activation, in vibrator concussion evenly.To pour in plate after the solid medium heating after sterilizing, each plate be about 20mL.After culture medium solidifying, aseptically add in plate with the bacteria suspension that sterilizing rifle head absorption 0.2mL shakes up, more rapidly bacterium liquid is smeared evenly with the glass spreading rod of sterilizing, make containing bacterium dull and stereotyped.After bacterium liquid is absorbed by agar, in culture medium, 5 equidistant holes are made a call to the sterilizing stainless steel card punch of external diameter 6mm, draw the culture medium of thawing more respectively with micro sample adding appliance, every hole adds a culture medium back cover melted, and negative control made by dropping 0.03mL test(ing) liquid or solvent subsequently.Often kind of sample repeats for 4 times.The culture dish antibacterial processed is put in 37 DEG C of incubators and cultivates 24h, mycete and Leuconostoc mesenteroides 30 DEG C cultivate 48h.Measure the diameter of inhibition zone with slide gauge, and judge that Folium Capsici polyphenol is to the inhibition for examination bacterium with this.Folium Capsici polyphenol bacteriostasis result as table 1.
Table 1 Folium Capsici polyphenol is to the fungistatic effect for examination bacterium
Embodiment 5:
The Determination of Antibacterial Activity of Fructus Capsici active polyphenol: minimal inhibitory concentration (MIC) is tested
Adopt dilution flat band method, in sterile test tube, add 15mL physiological saline solution, respectively depletion Staphylococcus aureus, escherichia coli, bacillus subtilis, yeast, penicillium sp, aspergillus niger 1-2 ring, then be diluted to 10
-3doubly, bacteria suspension is made for subsequent use.100uL bacteria suspension is drawn with liquid-transfering gun, add on the aseptic flat board of labelling sample number into spectrum, add the Fructus Capsici active polyphenol solution of 300uL variable concentrations again, shake up, often kind of bacterium prepares three flat boards, the culture medium being cooled to about 50 DEG C after pouring thawing immediately into is about 20mL, rocks culture dish rapidly on superclean bench, and after making bacteria suspension, sample solution and culture medium mix homogeneously, horizontal cools.Carry out colony counting after cultivating 24 or 48h, to be less than negative control group, the minimum solution concentration with significant difference is the MIC of Fructus Capsici active polyphenol.Fructus Capsici active polyphenol supplies the MIC of examination bacterium in table 2 to difference.
Table 2 Fructus Capsici active polyphenol is to the minimal inhibitory concentration MIC (mg/mL) for examination bacterium
Embodiment 6:
The antitumor cytolytic activity of Folium Capsici polyphenol
Test sample:
Human glioma U251 cell
Human neuroblastoma SH-SY5Y cell
Culture medium:
RPMI1640 culture medium
MEM culture medium
Test method:
Table 3 cell culture medium used and inoculum density
Cellular processes is in table 3, in aseptic 96 well culture plates, every hole adds 100 μ l cell suspending liquids, after cultivating 24h in 37 DEG C of cell culture incubators, culture medium is abandoned in suction, add the Fructus Capsici active polyphenol solution that corresponding culture medium is made into respectively, concentration is respectively 800,700,600,500,400,300,200 μ g/mL, establishes blank control wells simultaneously, often organizes 4 multiple holes.After continuing cultivation 24 and 48h respectively, culture fluid is abandoned in suction, every hole adds the cell culture fluid 100 μ L containing 10 μ L CCK-8, continue to hatch (table 2 corresponding time), microplate reader detects the absorbance (OD value) of 450nm wavelength, adopt SPSS 16.0 statistical analysis, and detecting Fructus Capsici active polyphenol to the inhibited proliferation of human glioma U251 cell, human neuroblastoma SH-SY5Y cell by CCK-8 method, the half-inhibition concentration of polyphenol to U251 cell and SH-SY5Y cell is as shown in table 4.
Table 4 Fructus Capsici active polyphenol is to U251 cell/SH-SYSY half-inhibition concentration (24h/48h)
The present invention, by energy-gathered ultrasonic extraction apparatus and high speed adverse current chromatogram connected applications, carries out Isolation and purification to Fructus Capsici stem and leaf polyphenol, and this method extraction ratio is high, energy-efficient, and mild condition, its lytic activity keeps.Gained Fructus Capsici polyphenol antibacterial activity of the present invention and Anticancer Activity in vitro well, have application prospect more widely.
Extraction time of the present invention is short, and efficiency is high, can be amplified to suitability for industrialized production.Purification process mild condition, efficient low-loss, its lytic activity keeps good, when the purified polyphenol concentration obtained is 50mg/mL, 11.85,11.28,11.08mm, 9.8mm, 9.67mm, 5.92mm the antibacterial circle diameter of bacillus subtilis, staphylococcus aureus, yeast, penicillium sp, escherichia coli, aspergillus niger is respectively:, when concentration is 25mg/mL, antibacterial circle diameter is respectively: 8.98mm, 8.98mm, 8.88mm, 8.80mm, 8.54mm, 5.31mm.Minimal inhibitory concentration is respectively: 0.391mg/mL, 0.781mg/mL, 0.781mg/mL, 1.563mg/mL, 1.563mg/mL, 3.125mg/mL.In 24h and 48h, to the half-inhibition concentration of human glioma U251 cell and human neuroblastoma SH-SY5Y cell be: 0.49mg/mL, 0.63mg/mL and 0.38mg/mL, 0.47mg/mL.
Embodiment 7:
With the Fructus Capsici polyphenol obtained by embodiment 1-3, method is equipped with various pharmaceutic adjuvant and can be made into tablet routinely.
With the Fructus Capsici polyphenol obtained by embodiment 1-3 as medicine activity component, use several excipient as the adjunct ingredient preparing composition of medicine tablet, proportioning makes the tablet samples that every sheet contains extract component 1-100mg according to a certain percentage, and table 5 provides the formula proportion of conventional tablet.
The Fructus Capsici polyphenol of some and excipients are prepared into various dose tablet formulation: by several excipients and crude drug Homogeneous phase mixing, add 1% Carboxymethyl cellulose sodium solution and make soft material in right amount, to sieve granulation, wet grain is dried and the granulate that sieves, tabletting and get final product after adding magnesium stearate and Pulvis Talci mix homogeneously.
The crude drug of the Fructus Capsici polyphenol composition of medicine tablet obtained by table 5 embodiment 1-3 and accessory formula
Embodiment 8:
Fructus Capsici polyphenol obtained by embodiment 1-3 routinely method is equipped with various pharmaceutic adjuvant and can be made into capsule:
Containing the preparation of Fructus Capsici polyphenol as the drug regimen capsule preparations of effective ingredient, use Fructus Capsici polyphenol as active constituents of medicine, use several excipient as the adjunct ingredient preparing composition of medicine capsule, proportioning makes the capsule preparations containing extract component 1-100mg in every capsules according to a certain percentage, and table 6 provides the formula proportion of conventional capsule preparation.
By the method that the Fructus Capsici polyphenol of some and excipients are prepared into capsule preparations be: several excipients is mixed homogeneously with Fructus Capsici polyphenol, add 1% Carboxymethyl cellulose sodium solution appropriate, make wet grain and dry the granulate that sieves, add magnesium stearate mix homogeneously, insert capsule and obtain; Or do not use granulation step, and directly Fructus Capsici polyphenol is mixed homogeneously with several excipients, after sieving, directly incapsulate.
The crude drug of table 6 embodiment 1-3 Fructus Capsici polyphenol composition of medicine capsule preparations and accessory formula
Claims (9)
1. a Fructus Capsici polyphenol is preparing the application in antibacterial.
2. a Fructus Capsici polyphenol is preparing the application in antitumor drug.
3. a kind of Fructus Capsici polyphenol as claimed in claim 1 is preparing the application in antibacterial, it is characterized in that described Fructus Capsici polyphenol is for raw material with Caulis Capsici, leaf powder, take ethanol-water solution as solvent, extract, through high speed adverse current chromatogram enrichment gained through ultrasonic extraction.
4. a kind of Fructus Capsici polyphenol as claimed in claim 1 is preparing the application in antibacterial, it is characterized in that described Fructus Capsici polyphenol be with Caulis Capsici, leaf powder for raw material, Caulis Capsici, leaf are cleaned, dry, pulverize, cross 20 mesh sieves, obtain Fructus Capsici stem and leaf powder, be placed in aeration-drying place and preserve stand-by; Then be the ethanol-water solution of 30% ~ 80% by previous step gained Caulis Capsici, leaf powder and volume fraction, pH value is 1 ~ 7, mix in ultrasonic extraction tank, solid-liquid ratio is 1:5 ~ 30, Extracting temperature is 30 DEG C ~ 60 DEG C, ultrasonic power is 200w ~ 1000w, and ultrasonic time and cavitation time slot are 4s-4s ~ 4s-8s, ultrasonic time 20 ~ 80min; Extract and terminate, filter to get filtrate, filtrate is also dry through decompression precipitation, obtained crude extract extractum, and then by high speed adverse current chromatogram enriching and purifying, obtains Fructus Capsici polyphenol; In described high-speed counter-current enriching step, be separated solvent system and be made up of acetate-methanol-water 10:1:10, adding sour adjust pH is further 1 ~ 7; Above-mentioned solvent system is placed in stratification after separatory funnel shake well, getting phase solvent is immobile phase, and lower phase solvent is mobile phase, makes to be full of immobile phase in counter-current chromatograph pillar when system temperature 5 ~ 60 DEG C, then make its main frame rotate, then mobile phase is pumped in post; Sample phase mixed solvent dissolving up and down, by injection valve sample introduction; According to detector spectrogram receiving target composition Fructus Capsici polyphenol.
5. a kind of Fructus Capsici polyphenol as claimed in claim 1 is preparing the application in antibacterial, it is characterized in that the bacterium in described antibacterial refers to bacillus subtilis, staphylococcus aureus, yeast, penicillium sp, escherichia coli, aspergillus niger.
6. a kind of Fructus Capsici polyphenol as claimed in claim 2 is preparing the application in antitumor drug, it is characterized in that described tumor cell is human glioma U251 cell and human neuroblastoma SH-SY5Y cell.
7. a kind of Fructus Capsici polyphenol as claimed in claim 2 is preparing the application in antitumor drug, it is characterized in that described Fructus Capsici polyphenol is for raw material with Caulis Capsici, leaf powder, take ethanol-water solution as solvent, extract, through high speed adverse current chromatogram enrichment gained through ultrasonic extraction.
8. a kind of Fructus Capsici polyphenol as claimed in claim 2 is preparing the application in antitumor drug, it is characterized in that described Fructus Capsici polyphenol be with Caulis Capsici, leaf powder for raw material, Caulis Capsici, leaf are cleaned, dry, pulverize, cross 20 mesh sieves, obtain Fructus Capsici stem and leaf powder, be placed in aeration-drying place and preserve stand-by; Then be the ethanol-water solution of 30% ~ 80% by previous step gained Caulis Capsici, leaf powder and volume fraction, pH value is 1 ~ 7, mix in ultrasonic extraction tank, solid-liquid ratio is 1:5 ~ 30, Extracting temperature is 30 DEG C ~ 60 DEG C, ultrasonic power is 200w ~ 1000w, and ultrasonic time and cavitation time slot are 4s-4s ~ 4s-8s, ultrasonic time 20 ~ 80min; Extract and terminate, filter to get filtrate, filtrate is also dry through decompression precipitation, obtained crude extract extractum, and then by high speed adverse current chromatogram enriching and purifying, obtains Fructus Capsici polyphenol; In described high-speed counter-current enriching step, be separated solvent system and be made up of acetate-methanol-water 10:1:10, adding sour adjust pH is further 1 ~ 7; Above-mentioned solvent system is placed in stratification after separatory funnel shake well, getting phase solvent is immobile phase, and lower phase solvent is mobile phase, makes to be full of immobile phase in counter-current chromatograph pillar when system temperature 5 ~ 60 DEG C, then make its main frame rotate, then mobile phase is pumped in post; Sample phase mixed solvent dissolving up and down, by injection valve sample introduction; According to detector spectrogram receiving target composition Fructus Capsici polyphenol.
9. a preparation method for Fructus Capsici polyphenol, is characterized in that the method with Caulis Capsici, leaf powder for raw material, by Caulis Capsici, leaf cleaning, drying, pulverizes, crosses 20 mesh sieves, obtain Fructus Capsici stem and leaf powder, is placed in aeration-drying place and preserves stand-by; Then be the ethanol-water solution of 30% ~ 80% by previous step gained Caulis Capsici, leaf powder and volume fraction, pH value is 1 ~ 7, mix in ultrasonic extraction tank, solid-liquid ratio is 1:5 ~ 30, during extraction, temperature is 30 DEG C ~ 60 DEG C, ultrasonic power is 200w ~ 1000w, and ultrasonic time and cavitation time slot are 4s-4s ~ 4s-8s, ultrasonic time 20 ~ 80min; Extract and terminate, filter to get filtrate, filtrate is also dry through decompression precipitation, obtained crude extract extractum, and then by high speed adverse current chromatogram enriching and purifying, obtains Fructus Capsici polyphenol; In described high-speed counter-current enriching step, be separated solvent system and be made up of acetate-methanol-water 10:1:10, adding sour adjust pH is further 1 ~ 7; Above-mentioned solvent system is placed in stratification after separatory funnel shake well, getting phase solvent is immobile phase, and lower phase solvent is mobile phase, makes to be full of immobile phase in counter-current chromatograph pillar when system temperature 5 ~ 60 DEG C, then make its main frame rotate, then mobile phase is pumped in post; Sample phase mixed solvent dissolving up and down, by injection valve sample introduction; According to detector spectrogram receiving target composition Fructus Capsici polyphenol.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106492051A (en) * | 2016-11-03 | 2017-03-15 | 铜陵桂生生态养殖有限公司 | A kind of spot mold of scorpion control agent and preparation method thereof |
| CN109247576A (en) * | 2018-09-05 | 2019-01-22 | 杨鑫 | A kind of extracting method of mulberries polyphenol |
| CN111013185A (en) * | 2019-11-19 | 2020-04-17 | 苏州永健生物医药有限公司 | Method for enriching characteristic phenols in alligator pepper |
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| 冯淑华等主编: "《药物分离纯化技术》", 30 June 2009, 化学工业出版社 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106492051A (en) * | 2016-11-03 | 2017-03-15 | 铜陵桂生生态养殖有限公司 | A kind of spot mold of scorpion control agent and preparation method thereof |
| CN109247576A (en) * | 2018-09-05 | 2019-01-22 | 杨鑫 | A kind of extracting method of mulberries polyphenol |
| CN111013185A (en) * | 2019-11-19 | 2020-04-17 | 苏州永健生物医药有限公司 | Method for enriching characteristic phenols in alligator pepper |
| CN111013185B (en) * | 2019-11-19 | 2022-01-04 | 苏州永健生物医药有限公司 | Method for enriching characteristic phenols in alligator pepper |
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Application publication date: 20141217 |