CN101623394A - Nakedflower beautyberry extract as well as preparation method, preparation and application thereof - Google Patents
Nakedflower beautyberry extract as well as preparation method, preparation and application thereof Download PDFInfo
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Abstract
The invention relates to a nakedflower beautyberry extract as well as a preparation method, a preparation and an application thereof. The nakedflower beautyberry extract is an extract which is extracted from nakedflower beautyberry leaves and contains a nakedflower beautyberry total flavonoid compound and a tannide class compound. The extract is obtained by the steps of extracting the nakedflower beautyberry leaves by water, diluted alcohol or diluted ketone and carrying out macroporous adsorptive resin column chromatography purification on an extracting solution after being concentrated. The nakedflower beautyberry extract has high purity and has the functions of bacteriostasis, hemostasis, pain relieving and convergence.
Description
Technical field
The present invention relates to field of traditional Chinese medicine pharmacy, be specifically related to a kind of Callicarpa nudiflora extract and preparation method thereof, preparation and purposes.
Background technology
Callicarpa nudiflora (CallicarpanudifloraHook.exArn.) is Verenaceae (Verbenaceae) Callicarpa plant, and main product is a kind of Hainan authentic medicinal herbs in China Hainan, and also there is distribution in China Guangdong, Guangxi.Mainly be distributed in India, Vietnam and Malaysia abroad.Its root, leaf can be used as medicine, and antibacterial anti hemorrhagic is arranged, eliminating inflammation and expelling toxin, and dissipating blood stasis for subsidence of swelling, the effect of wind-expelling and dispelling dampness cures mainly diseases such as suppurative inflammation, acute infectious hepatitis, respiratory tract and digestive tract hemorrhage, wound hemorrhage, and burn and traumatic hemorrhage etc. are controlled in external.
At present existing Callicarpa nudiflora sheet, LUOHUAZIZHU SHUAN, Callicarpa nudiflora mixture etc., current technology is that decocting boils twice, reconcentration, filter, be dried to dry extract, the technology simple coarse, but continued to use nearly 30 years, to Chinese medicine extraction, concentrate, the method for technology such as drying and the reasonability of parameter study, and, existing product all with single total flavonoid composition as quality control standard, quality standard remains further to be improved.
Therefore, technical problem to be solved by this invention is the Callicarpa nudiflora leaf is further furtherd investigate, thereby provides a kind of active component content higher, more reasonable, effective Callicarpa nudiflora extract.
Summary of the invention
An object of the present invention is to provide a kind of Callicarpa nudiflora extract, this extract comprises Callicarpa nudiflora total flavonoid chemical compound and tannin compounds.
Content of flavonoids is 50%～90% in the described extract, and the content of tannin is 5%～20%.Wherein, the total flavonoid chemical compound comprises at least a of luteolin, apigenin and luteoloside.
Another object of the present invention provides a kind of preparation method of above-mentioned Callicarpa nudiflora extract.This preparation method comprises the following steps:
1). add entry, rare alcohol or rare ketone solvent in the Callicarpa nudiflora medical material, heating decocts, and pours out decocting liquid, filters to get filtrate;
2). by macroporous adsorbent resin column chromatography, wash gained filtrate with water cylinder, discard water lotion, the ethanol elution of reuse 10%～90% is collected ethanol elution;
3). ethanol elution is concentrated, get Callicarpa nudiflora extract.
Wherein, step 2) macroporous adsorbent resin in is the polystyrene type macroporous adsorbent resin, the preferred JD-1 of described polystyrene type macroporous adsorbent resin, D101, AB-8, X-5 or D402, and the blade diameter length ratio of resin column is 1: 5～1: 10.
Another purpose of the present invention is to disclose the preparation that above-mentioned Callicarpa nudiflora extract and acceptable accessories are prepared from, the preferred spray of said preparation, tablet, capsule, oral liquid or granule.
A further object of the present invention is to disclose above-mentioned Callicarpa nudiflora extract is used for antibacterial, hemostasis, analgesia or astringent medicine in preparation application.
Callicarpa nudiflora extract of the present invention not only contains the total flavonoid composition, and contains the tannin constituents, and total content of effective improves relatively; The preparation technology of extract is also simple relatively, and yield is higher; That the gained Callicarpa nudiflora extract also has is antibacterial significantly, hemostasis, analgesia, astringent effect.
The specific embodiment
Embodiment one
Get 100kg Callicarpa nudiflora medical material, add water 1500kg, soaked 30 minutes, heated and boiled 120 minutes is poured out decocting liquid, and medicinal residues are fried in shallow oil 2 times again, add water 1500kg at every turn, boil 60 minutes, merge three times decocting liquid, filters, and filtrate is 350～450m by specific surface area
2/ g, skeletal density are 1～1.2g/ml, average pore size is 35～65nm, voidage is 60～68% JD-1 (WLD) type polystyrene type macroporous adsorptive resins, the blade diameter length ratio of resin column is 1: 5, discard effluent, water washing cylinder with 5 times of column volumes discards cleaning mixture, and the no flavone to the eluent of the ethanol elution with 10% detects.Collect eluent, being evaporated to relative density behind the recovery ethanol is 1.2, lyophilization, and the check packing promptly gets Callicarpa nudiflora extract.The polystyrene type macroporous adsorptive resins is regenerated with 5% acidic alcohol.
The yield of extract is 2.53%, and wherein, through UV quantitative analysis (analytical method is operated according to known method), general flavone content is 50%, total tannin content 20%.
Embodiment two
Get 100kg Callicarpa nudiflora medical material, add rare pure 1000kg, soaked 50 minutes, heated and boiled 90 minutes is poured out decocting liquid, and medicinal residues are fried in shallow oil 2 times again, add water 1500kg at every turn, boil 90 minutes, merge three times decocting liquid, filters, and filtrate is 350～450m by specific surface area
2/ g, skeletal density are 1～1.2g/ml, average pore size is 35～65nm, voidage is 60～68% D101 type polystyrene type macroporous adsorptive resins, the blade diameter length ratio of resin column is 1: 7, discard effluent, water washing cylinder with 4 times of column volumes discards cleaning mixture, detects with 50% ethanol elution no flavone to the eluent.Collect eluent, being evaporated to relative density behind the recovery ethanol is 1.1, lyophilization, and the check packing promptly gets Callicarpa nudiflora extract.The polystyrene type macroporous adsorptive resins is regenerated with 10% sodium hydroxide lye washing back reuse 5% acidic alcohol.
The yield of Callicarpa nudiflora extract is 3.23%, and wherein, through the UV quantitative analysis, general flavone content is 81.17%, total tannin content 12.23%.
Embodiment three
Get 100kg Callicarpa nudiflora medical material, add rare ketone 2000kg, soaked 40 minutes, heated and boiled 60 minutes is taken out decocting liquid, and medicinal residues are fried in shallow oil 2 times again, add water 1000kg at every turn, boil 60 minutes, merge three times decocting liquid, filter, and filtrate is 350～450m by specific surface area
2/ g, skeletal density are 1～1.2g/ml, average pore size is 35～65nm, voidage is 60～68% AB-8 type polystyrene type macroporous adsorptive resins, the blade diameter length ratio of resin column is 1: 10, discard effluent, water washing cylinder with 6 times of column volumes discards cleaning mixture, detects with 90% ethanol elution no flavone to the eluent.Collect eluent, being evaporated to relative density behind the recovery ethanol is 1.08, lyophilization, and the check packing promptly gets Callicarpa nudiflora extract.The polystyrene type macroporous adsorptive resins is regenerated with 10% sodium hydroxide lye washing back reuse 5% acidic alcohol.
The yield of Callicarpa nudiflora extract is 3.03%, and wherein, through the UV quantitative analysis, general flavone content is 90%, total tannin content 5%.
Embodiment fourPreparation tablets
With Callicarpa nudiflora extract 100g of the present invention (embodiment one method makes), starch 80g, dextrin 5g mix homogeneously adds 10% starch slurry system soft material, granulates with 14 order nylon screens, 60～70 ℃ of aeration-dryings, 16 mesh sieve granulate add magnesium stearate 1.5g, carboxymethyl starch sodium 5g mixing, be pressed into 1000, coating promptly.Become human oral every day 2～5 times, each 1～10.
Embodiment fiveThe capsule preparation
With Callicarpa nudiflora extract 100g of the present invention (embodiment two methods make), starch 78g, magnesium stearate 2g mixing directly is filled to 1000 with Autocapsulefillingmachine, and polishing is promptly.Become human oral every day 2～5 times, each 1～10.
Embodiment sixThe oral liquid preparation
With Callicarpa nudiflora extract 20g of the present invention (embodiment one method makes), mix with Mel 300g, sucrose 50g, sodium benzoate 2g and distilled water 300ml, be heated to 85～90 ℃, stirring makes dissolving, is incubated 30 minutes, filters, the filtrate thin up is to 1000ml, stir evenly, embedding (10 milliliters every), sterilization is promptly.Become human oral every day 2～5 times, each 1～5.
Embodiment sevenThe granule preparation
It is an amount of to get Callicarpa nudiflora extract 10g of the present invention (embodiment two methods make), dextrin 20g, sucrose 100g and ethanol, and mixing is crossed ten mesh sieves and made granule, in 60～70 ℃ of dryings, granulate, packing is promptly, the heavy 5g of every bag becomes human oral every day 2～5 times, each 1～5 bag.
Embodiment eightThe spray preparation
Get Callicarpa nudiflora extract 2g of the present invention (the tripartite method of embodiment makes), xanthan gum 10mg, Chymotrypsin 100IU, normal saline is diluted to 1000ml, under aseptic condition,, Callicarpa nudiflora effective site is mixed with normal saline, xanthan gum, Chymotrypsin according to the requirement of above-mentioned prescription amount, stir, make suspension; After content analysis is qualified, keeping away under the bacterium condition, with above-mentioned solution with the packing of quantitative liquid feeding device, promptly.
Embodiment nine
The relevant test of pesticide effectiveness of Callicarpa nudiflora extract of the present invention is as follows:
1, bacteriostasis
By external bacteriostatic experiment, with the Callicarpa nudiflora extract of minimum inhibitory concentration (MIC) and the present invention of minimal bactericidal concentration (MBC) index evaluation preparation antibacterial action to common bacterial strain.
The preparation of bacterium liquid: the picking slant strains is inoculated in 20% N and soaks in the juice culture medium, 37 ℃ of overnight incubation, and reuse soaks the juice culture medium for 20% N bacterium liquid is diluted to 10cfu/ml, immediately inoculation.
MIC measures: adopt the liquid tube serial dilution, soak the juice culture medium with 20% N sample is done continuous two-fold dilution.1.0ml is added in the sterilization test tube, and every pipe adds the fresh bacterium liquid of 0.05ml standard, and establishes bacterial strain contrast and solvent matched group, cultivates 18～24 hours for 37 ℃, observes the bacterial growth situation, is MIC with the high dilution of asepsis growth.
MBC measures: adopt the plate streaking inocalation method, the picking culture fluid is seeded on the 10% serum MH bouillon agar flat board from the culture medium that is judged to adjacent two pipes of MIC and MIC, cultivated 18～24 hours for 37 ℃, and be MBC with the high dilution of growth clump count≤5.
Above-mentioned result of the test sees Table one.
Table one Callicarpa nudiflora extract in-vitro antibacterial result of the test
The Callicarpa nudiflora extract of the present invention preparation all has in various degree inhibition and killing action to staphylococcus aureus, escherichia coli, Bacillus typhi, streptococcus pneumoniae, group B streptococcus and helicobacter pylori.
2, anastalsis
Three dosage groups of Callicarpa nudiflora extract 250,500,750mg/kg, normal saline group, medicine solvent group and former Callicarpa nudiflora water extract with the present invention's preparation.Divide 6 groups (30 every group, male and female half and half) at random with 180 healthy Kunming white mice, irritate stomach and give relative medicine every day, continuous 7 days.The last administration begins experiment as follows after 1 hour.
(1) to the influence of mice bleeding time (BT)
Adopt one time the tail vein injection administering mode, after the administration 30 minutes with very sharp shears in the mouse tail tip upwards about 1cm place cut off afterbody fast, and clock with stopwatch immediately, the while is constantly inhaled effusive blood with filter paper.Begin to be mice during this period of time and cut the tail bleeding time from cutting tail to stopped bleeding.
(2) to the influence of clotting time of mice (CT)
With adjoining the ball rear vein beard in the slotting people mice of capillary glass-tube, dark about 4～5mm, autoblood flows to the pipe clocking internal, lie against on the table after filling with capillary tube, every 30 seconds about 0.5cm in the two ends that fracture, and slowly draw back to the left and right, examining the place of fractureing and have or not the blood clotting silk to occur, stopwatch is blood coagulation time when the blood streak occurring.
(3) to the influence of mouse blood coagulation system PT, APTT and FIB
Pluck eyeball of mouse and get blood, adopt 2ml blood and place the negative pressure test tube that contains 3.8% sodium citrate anticoagulant, detect prothrombin time (PT), activated partial thrombin time (APTT) and Fibrinogen quantitatively (FIB) with ACT-200 type coagulation function detector.
(4) influence that mouse platelets is counted
Pluck eyeball of mouse and get blood, adopt 0.5ml blood and place the negative pressure test tube that contains the EDTA anticoagulant, detect platelet count with blood cell numeration instrument.
(5) result of the test
By above-mentioned experiment, the experimental data that records mice bleeding time (BT), clotting time (CT), prothrombin time (PT), activated partial thrombin time (APTT) and Fibrinogen quantitative (FIB) sees the following form two.
Table two Callicarpa nudiflora extract of the present invention is to the result of the test of mice anastalsis
(mean ± standard deviation, N=10)
*P＜0.05,
*Compare with the normal control group P＜0.01.
Above result proves that the Callicarpa nudiflora extract of the present invention's preparation all can shorten BT, CT and APTT and reduce FIB content, has tangible anastalsis.
3, analgesic activity
(1) acetic acid induced mice writhing response (turning round the body experiment)
Healthy kunming mice is divided into 6 groups (10 every group) at random, i.e. three dosage groups of Callicarpa nudiflora extract 250,500,750mg/kg, normal saline group, medicine solvent group (2% ethanol) and the former Callicarpa nudiflora water extract group for preparing with the present invention.Irritate stomach and give relative medicine every day, continuous 7 days.After the last administration 1 hour, lumbar injection 0.6% acetum 0.2ml/ respectively of each group, observe and record injection acetic acid after the time and the number of times of writhing response (abdominal part shrinks indent, stretches hind leg, buttocks is raised, crawling) appearred in 15 minutes, experimental result sees Table three.
The influence of table three Callicarpa nudiflora extract Dichlorodiphenyl Acetate induced mice writhing response
(mean ± standard deviation, N=10)
*P＜0.05,
*Compare with the solvent group P＜0.01.
(2) hot plate experiment
With healthy kunming mice 6 groups of the labellings (10 every group) of weighing, promptly with three dosage groups of Callicarpa nudiflora extract 250,500,750mg/kg, normal saline group, medicine solvent group (2% ethanol) and the former Callicarpa nudiflora water extract of the present invention's preparation.Adopting hot plate method, hot plate temperature be (55 ± 0.5) ℃, measures the normal pain reaction (put into hot plate to mice and lick the metapedes time, the person throws aside as if do not lick the rear solid end in 5～30 seconds) of each Mus before the administration earlier.Then, irritate stomach by 0.42g/kg dosage and give relative medicine, continuous 7 days every day.The normal pain reaction of each Mus is measured in the last administration again after 1 hour, and percentage rate is improved in the threshold of pain before and after calculating the mice administration, the results are shown in Table four.
Table four Callicarpa nudiflora extract is to the influence of the mice hot plate threshold of pain
(mean ± standard deviation, N=10)
Annotate: irritate stomach every day and give the mice medicine, continuous 7 days.
*P＜0.05,
*Compare with the solvent group P＜0.01.
Above-mentioned experiment shows that the Callicarpa nudiflora extract of the present invention's preparation all can prolong writhing response time of occurrence and minimizing and turn round the body number of times in the mouse writhing test, and at hot plate test, the Callicarpa nudiflora extract of the present invention's preparation all can increase the threshold of pain and improve percentage rate.
4, astriction
Select 60 of healthy rabbits, be divided into 6 groups (10 every group, male and female half and half) at random, be i.e. three dosage groups of Callicarpa nudiflora extract 250,500,750mg/kg, normal saline group, medicine solvent group (2% ethanol) and the former Callicarpa nudiflora water extract for preparing with the present invention.Before the experiment selected rabbit is all abraded at the back, shape is rounded, and the about 3cm of area diameter causes the local significant rotten to the corn sepage performance that occurs.Observe once partial response situation every day, and measure what and extinction time of wound area, record seepage discharge, calculate healing rate at last, the area that promptly heals/2nd day wound healing area the results are shown in Table five and table six.
Table five Callicarpa nudiflora extract is to the influence of rabbit wound healing area
(mean ± standard deviation, N=10)
*P＜0.05,
*Compare with the normal control group P＜0.01.
Table six Callicarpa nudiflora extract is to the influence of rabbit wound healing rate
(mean ± standard deviation, N=10)
*P＜0.05,
*Compare with the normal control group P＜0.01.
The middle and high dosage group of Callicarpa nudiflora extract of the present invention's preparation all can significantly increase the wound healing area, improves the wound healing rate.
Claims (10)
1. a Callicarpa nudiflora extract is characterized in that this extract comprises Callicarpa nudiflora total flavonoid chemical compound and tannin compounds.
2. Callicarpa nudiflora extract according to claim 1, the content that it is characterized in that total flavonoid chemical compound in the described extract is 50%～90%, the content of tannin is 5%～20%.
3. Callicarpa nudiflora extract according to claim 1 and 2 is characterized in that described total flavonoid chemical compound comprises at least a in luteolin, apigenin and the luteoloside.
4. the preparation method of claim 1 or 2 described Callicarpa nudiflora extracts is characterized in that this method comprises the following steps:
1). add entry, rare alcohol or rare ketone solvent in the Callicarpa nudiflora medical material, heating decocts, and pours out decocting liquid, filters to get filtrate;
2). by macroporous adsorbent resin column chromatography, wash gained filtrate with water cylinder, discard water lotion, the ethanol elution of reuse 10%～90% is collected ethanol elution;
3). ethanol elution is concentrated, get Callicarpa nudiflora extract.
5. the preparation method of Callicarpa nudiflora extract according to claim 4 is characterized in that step 2) described in macroporous adsorbent resin be the polystyrene type macroporous adsorbent resin.
6. the preparation method of Callicarpa nudiflora extract according to claim 5 is characterized in that described polystyrene type macroporous adsorbent resin is JD-1, D101, AB-8, X-5 or D402.
7. the preparation method of Callicarpa nudiflora extract according to claim 4 is characterized in that step 2) described in the blade diameter length ratio of macroporous adsorptive resins be 1: 5～1: 10.
8. preparation that is prepared from by claim 1 or 2 described Callicarpa nudiflora extracts and acceptable accessories.
9. preparation according to claim 8 is characterized in that described preparation is spray, tablet, capsule, oral liquid or granule.
10. claim 1 or 2 described Callicarpa nudiflora extracts are used for the application of antibacterial, hemostasis, analgesia or astringent medicine in preparation.
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| CN101623394B (en) | 2011-12-28 |
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