CN102100720A - Ainsliaea fragrans champ caffeoylquinic acid extracts and preparation and application thereof - Google Patents
Ainsliaea fragrans champ caffeoylquinic acid extracts and preparation and application thereof Download PDFInfo
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Abstract
The invention relates to ainsliaea fragrans champ caffeoylquinic acid extracts and preparation and application thereof. The ainsliaea fragrans champ caffeoylquinic acid extracts are prepared by the following steps of: performing refluxing extraction of the ainsliaea fragrans champ by using ethanol, filtering, concentrating and removing ethanol, adjusting the pH value to 1 to 3 by adding hydrochloric acid, applying macroporous adsorbent resin, water-washing, adding ethanol for gradient elution, and collecting eluent, of which the ethanol concentration is 50 to 70 percent; performing condensation and ethanol removal, adjusting the pH value to 1 to 3 by adding hydrochloric acid, using macroporous adsorbent resin, adding water and ethanol for gradient elution respectively, and collecting the eluent, of which the ethanol concentration is 40 to 70 percent; mixing the eluent, and vacuum drying the eluent to obtain the extracts. The UV content of the ainsliaea fragrans champ caffeoylquinic acid extracts is over 70 percent, the HPLC content is over 50 percent, and the content of 3-caffeoylquinic acid and the content of 3,5-dicaffeoylquinic acid are both over 10 percent. The ainsliaea fragrans champ caffeoylquinic acid extracts have the effects of resisting inflammations, bacteria, medicament-resistant strains, well improves pathologically changed tissues of rat suffering from cervicitis, obviously reduces the uterus index and the number of leukocytes and can be independently prepared into the medicaments for treating cervicitis or combined with other medicaments to prepare the medicaments for treating cervicitis.
Description
Technical field
The invention belongs to the field of Chinese medicines, relate to Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract preparation technology and the application in newtype drugs such as exploitation antiinflammatory, antibacterium, overriding resistance bacterial strain, treatment cervicitis disease thereof.
Background technology
Ainsliasa fragrans Champ (
Ainsliaea fragrans Champ.) be Compositae Ainsliaea fragrans Champ. platymiscium, for " Jiangxi Province's Chinese crude drug standard " version in 1996 is recorded.It has effects such as heat-clearing and toxic substances removing, dampness removing, hemostasis, and is among the people with all herbal medicine, cures mainly the asthenia hectic fever due to YIN-deficiency, pulmonary tuberculosis hemoptysis, woman uterine bleeding, jaundice due to damp-heat, edema, swollen ulcer drug, scrofula tuberculosis, traumatic injury, venom.Be distributed in ground such as Jiangsu, Zhejiang, Jiangxi, Fujian.The modern pharmacology test shows; drug activity composition main in the Ainsliasa fragrans Champ mainly contains 3-caffeoyl quinic acid, 3; 5-two coffee acyl base quininic acids, 4; 5-two coffee acyl base quininic acids etc.; have and improve immunologic function, inflammation-inhibiting medium such as histamine, prostaglandin etc. and discharge, the aspect tools such as regulating action, especially overriding resistance bacterial strain of cervicitis rat cervix uteri mucosa PGE2 and T lymphocyte subsets are had certain effect.
The prevalence of cervicitis occupies first of the gynaecopathia, and up to 39.30%, its pathogenic factor mainly contains and causes infection, pathogenic infection behind chemical substance or the mechanical irritation (damage) and cause inflammation.Cervicitis also is one of high risk factor that brings out infertility and cervical cancer.At present, the medicine of treatment cervicitis seldom, the listing medicine mostly to slight simple cervicitis curative effect still can, but not good enough to cervicitis curative effects such as drug resistance pathogenic bacterial infection, degree of depth infection.
At present, in the document of having delivered, the Ainsliasa fragrans Champ extract preparation serves as to investigate index with total phenolic acid or content of total flavone, by UV spectrophotometer measuring content, the extract component complexity, the main pharmacodynamics composition is indeterminate, will cause the content of drug effect components of preparation low to a certain extent, and day clothes dosage is big.For example: (China, the patent No.: ZL200510075384.4), at flavone and lactone composition, carry through water, polystyrene macroporous resin is refining, obtains general flavone content and be 1 ~ 60% extract for a kind of Ainsliasa fragrans Champ extract and preparation method thereof.Show that through this extract of pharmacological research it has anti-inflammatory activity, and can be made into tablet, capsule, granule or oral liquid formulation.Xingsiang Tuerfeng total phenolic acid extract and preparation method thereof (China, application number: 200810111902.7) refining through alcohol extraction, macroporous resin at Xingsiang Tuerfeng total phenolic acid, obtain total phenolic content greater than 60%, chlorogenic acid content is 1 ~ 5% extract.Show that through pharmacological research it is inhibited to pelvic inflammatory disease.A kind of effect section of frangrant ainsliaea herb and its production and application (China; application number: 200810106926.3) at Xingsiang Tuerfeng total phenolic acid; make with extra care through alcohol extraction, macroporous resin; obtain total phenolic content and be 1 ~ 80% extract; contain 0.05 ~ 20% chlorogenic acid; 0.5 ~ 50% 3,5-dicaffeoyl-quinic acid, 4,5-dicaffeoyl-quinic acid.Show that through pharmacological research it has antiinflammatory, antibacterial, antivirus action.And above-mentioned patent research shows that Ainsliasa fragrans Champ extract has antiinflammatory and bacteriostatic functions, but does not see and relate to it to cervicitis and the inhibiting research of fastbacteria.
Summary of the invention
Content of drug effect components at present Ainsliasa fragrans Champ extract is low, day clothes dosage is big; the present invention mainly adopts drug activity composition coffee mesitoyl quinine acid material for investigating index; to the Ainsliasa fragrans Champ medical material extract, purification; obtain UV content greater than 70%; HPLC content is greater than 50% coffee mesitoyl quinine acid material; 3-caffeoyl quinic acid and 3 wherein, 5-dicaffeoylquinic acid content all is higher than 10%.
Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract of the present invention is characterized in that in the extract with the 3-caffeoyl quinic acid, 3, and 4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid are main.
Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract of the present invention is characterized in that extract calculates with the coffee mesitoyl quinine acid composition, and UV content is more than 70%, and HPLC content is more than 50%.3-caffeoyl quinic acid and 3 wherein, 5-dicaffeoylquinic acid content all is higher than 10%.
Extract of the present invention prepares by following steps:
A) get the dry medical material of Ainsliasa fragrans Champ, cutting is 40 ~ 80% ethanol with the concentration of 5 ~ 20 times of volumes 60 ~ 100 ℃ of following heating and refluxing extraction 2 ~ 4 times, each 1-3 hour, filtration, merge extracting solution A;
B) extracting solution A is evaporated to 1/4 ~ 1/8 times through 40 ~ 60 ℃, promptly obtains concentrated solution B;
C) concentrated solution B transfers pH to 1 ~ 3 with dilute hydrochloric acid, and acid concentrated solution through macroporous resin adsorption, is used earlier the deionized water rinsing resin, the reuse water-ethanol is gradient elution successively, the collection concentration of alcohol is the eluent between 50 ~ 70%, after 40 ~ 60 ℃ of concentrating under reduced pressure remove alcohol, gets concentrated solution D;
D) transfer pH extremely to 1 ~ 3 with dilute hydrochloric acid concentrated solution D again, through macroporous resin adsorption, use earlier the deionized water rinsing resin, the reuse water-ethanol is gradient elution successively, the collection concentration of alcohol is the eluent between 40 ~ 70 %, merge, vacuum drying promptly gets Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract.The extract that extracts with said method calculates with the coffee mesitoyl quinine acid composition, and UV content is more than 70%, and HPLC content is more than 50%.3-caffeoyl quinic acid and 3 wherein, 5-dicaffeoylquinic acid content all is higher than 10%.
Ainsliasa fragrans Champ coffee mesitoyl quinine acid preparation method of extract of the present invention is characterized in that extraction solvent is selected 40% ~ 80% ethanol for use among the preparation process a.
Ainsliasa fragrans Champ coffee mesitoyl quinine acid preparation method of extract of the present invention is characterized in that the ethanol of extraction solvent preferred 50% among the preparation process a.
Ainsliasa fragrans Champ coffee mesitoyl quinine acid preparation method of extract of the present invention is characterized in that described macroporous adsorbent resin is selected from AB-8, HPD-400, NKA-9, HPD-826.
Ainsliasa fragrans Champ coffee mesitoyl quinine acid preparation method of extract of the present invention is characterized in that macroporous adsorbent resin HPD-826.
The invention still further relates to a kind of pharmaceutical composition, it is characterized in that comprising Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract of the present invention and acceptable accessories.The dosage form of described pharmaceutical composition is tablet, capsule, granule, powder, pill, extractum, oral liquid or syrup.
The invention still further relates to the application of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract in the medicine of preparation antiinflammatory, antibacterium, overriding resistance bacterial strain.The particularly application in the medicine of the anti-cervicitis of preparation.
At the Ainsliasa fragrans Champ extract complicated component, indeterminate this problem of main pharmacodynamics composition, Ainsliasa fragrans Champ extract preparation technology of the present invention adopts active guiding fractionation method, and in conjunction with HPLC tracking detection target, choose semi-polarity styrene type HPD-826 macroporous adsorbent resin, carry out twice continuous adsorption and resolve, the loss of having avoided target component, the content of active ingredient is improved significantly, and extract obtained possess the advantage that the main pharmacodynamics composition is clearer and more definite, extraction ratio is high.Be in particular in: Ainsliasa fragrans Champ extract of the present invention is not only through UV spectrophotometer measuring coffee mesitoyl quinine acid component content about 70%, and detect the main pharmacodynamics component content greater than 50% through HPLC, 3-caffeoyl quinic acid and 3 wherein, 5-dicaffeoylquinic acid content all is higher than 10%; And wherein mainly contain coffee mesitoyl quinine acid compositions such as 3-caffeoyl quinic acid, 3,4-dicaffeoyl-quinic acid, 3,5-dicaffeoyl-quinic acid, 4,5-dicaffeoyl-quinic acid.
The Ainsliasa fragrans Champ extract of gained of the present invention has antiinflammatory, antibacterium, the drug action of overriding resistance bacterial strain, there is no obvious acute toxicity effect.Be in particular in the granulomatous hypertrophy of rat, mice caused by dimethylbenzene xylene ear swelling are had significant inhibitory effect; To the effect of having clear improvement of cervicitis rat pathological tissues pathological changes, quantity of leucocyte there is the reduction effect; Anti-drug resistance pathogen escherichia coli, infection of staphylococcus aureus mice had anti-infectious function preferably.Solved present Related product to not good enough problems of cervicitis curative effect such as drug resistance pathogenic bacterial infection, degree of depth infection.
Thereby Ainsliasa fragrans Champ coffee mesitoyl quinine acid extraction process of the present invention reduces day clothes dosage at medical material utilization rate, clear and definite main pharmacodynamics composition, and aspects such as raising curative effect have remarkable advantages.
This patent pharmacological tests shows, the coffee mesitoyl quinine acid composition has obvious improvement effect to the cervicitis animal model in the Ainsliasa fragrans Champ medical material, and have effects such as obvious antimicrobial agent, there is not obvious acute toxicity effect, as seen it is developed further into anti-cervicitis medicine, has vast market prospect with the effect of overriding resistance bacterial strain.
Description of drawings
Fig. 1 Ainsliasa fragrans Champ coffee mesitoyl quinine acid composition measurement canonical plotting.
Fig. 2 reference substance HPLC figure.
Fig. 3 Ainsliasa fragrans Champ extract HPLC figure.
Fig. 4 cervicitis rat model histopathologic slide.
The specific embodiment
Content of drug effect components is measured
(1) the coffee mesitoyl quinine acid component content is measured in the Ainsliasa fragrans Champ extract
The reference substance solution preparation
Get 3,5-dicaffeoyl-quinic acid reference substance is an amount of, places brown volumetric flask, adds 50% methanol and makes the solution that concentration is 0.041mg/ml, and is standby.
The drafting of standard curve
Precision pipettes reference substance solution 0.5,1.0,1.5,2.0,2.5,3.0ml, places the 10ml volumetric flask respectively, adds methanol constant volume to scale, shakes up.Absorbance at the 328nm place, with absorbency Y to mass concentration X(mg/ml) the drawing standard curve.Obtain regression equation: Y=0.279X+0.016, R
2=0.999.Standard curve such as accompanying drawing 1.
Sample determination is identical with the assay method of reference substance.
(2) coffee mesitoyl quinine acid active constituent content measuring
Chromatographic condition:Chromatographic column: Diamonsil(5 μ m * 4.6 mm * 250 mm); Column temperature: 30 ℃; Detect wavelength: 328nm; Flow velocity: 1ml/min; Mobile phase: acetonitrile-0.2% phosphoric acid gradient elution (0 min, 20:80; 8min, 21:79; 11min, 23:77).
The preparation of reference substance solution
Precision takes by weighing 3-caffeoyl quinic acid, 3; 5-dicaffeoyl-quinic acid, reference substance are an amount of; add 50% ethanol and make every l mL and contain 3-caffeoyl quinic acid 0.034mg/mL, 3, the mixing reference substance solution of 5-dicaffeoyl-quinic acid 0.0425mg/mL, standby.Reference substance HPLC the results are shown in accompanying drawing 2.
By above-mentioned chromatographic condition, sample is measured, its HPLC the results are shown in accompanying drawing 3.
Get the dry medical material 1000g of Ainsliasa fragrans Champ, cut off (2 ~ 5cm), with 10 times 50% ethanol, 85 ℃ of heating and refluxing extraction 3 times, filtration, merge extracting solution A, 60 ℃ of 1/6 times of being evaporated to extracting solution A promptly obtain concentrated solution B.Concentrated solution B transfers pH to 2 with dilute hydrochloric acid, with the macroporous adsorbent resin AB-8 of acid concentrated solution B through having handled well, use earlier the deionized water rinsing resin, with 2 column volumes of 20% ethanol elution, use 10 column volumes of 50% ethanol elution then, collect concentration of alcohol 50% eluent, remove alcohol through 60 ℃ of concentrating under reduced pressure, get concentrated solution C, concentrated solution C reuse dilute hydrochloric acid is transferred pH to 2, through macroporous adsorbent resin HPD-826, use earlier the deionized water rinsing resin, the reuse water-ethanol is gradient elution successively, and collecting concentration of alcohol is 50% eluent, merges, through vacuum drying, promptly get Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 20g, its UV content 70%, HPLC content 52%.Wherein 3-caffeoyl quinic acid content is 15%, 3, and 5-dicaffeoyl-quinic acid content is 13%.
The dry medical material 1000g of Ainsliasa fragrans Champ, cut off (2 ~ 5cm), with 15 times 60% ethanol, 85 ℃ of heating and refluxing extraction 3 times, filtration, merge extracting solution A, 60 ℃ of 1/7 times of being evaporated to extracting solution A promptly obtain concentrated solution B.Concentrated solution is transferred pH to 1 with the B dilute hydrochloric acid; with the macroporous adsorbent resin HPD-826 of acid concentrated solution B through having handled well; use earlier the deionized water rinsing resin; with 2 column volumes of 30% ethanol elution; use 8 column volumes of 60% ethanol elution then, collect concentration of alcohol 60% eluent, through 60 ℃ of concentrating under reduced pressure; get concentrated solution C; concentrated solution C reuse dilute hydrochloric acid is transferred pH to 1, through macroporous adsorbent resin HPD-826, uses earlier the deionized water rinsing resin; the reuse water-ethanol is gradient elution successively; collecting concentration of alcohol is 60% eluent, merges, through vacuum drying; promptly get Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 16g; its UV content 79%, HPLC content 55%, wherein 3-caffeoyl quinic acid content is 19%; 3,5-dicaffeoyl-quinic acid content is 15%.
The dry medical material 1000g of Ainsliasa fragrans Champ, cut off (2 ~ 5cm), with 20 times 70% ethanol, 85 ℃ of heating and refluxing extraction 3 times, filtration, merge extracting solution A, 60 ℃ of 1/8 times of being evaporated to extracting solution A promptly obtain concentrated solution B.Concentrated solution B transfers pH to 3 with dilute hydrochloric acid, with the macroporous adsorbent resin HPD-826 of acid concentrated solution B through having handled well, use earlier the deionized water rinsing resin, with 2 column volumes of 30% ethanol elution, use 6 column volumes of 70% ethanol elution then, collect the eluent of concentration of alcohol 70%,, get concentrated solution C through 60 ℃ of concentrating under reduced pressure, concentrated solution C reuse dilute hydrochloric acid is transferred pH to 3, through macroporous adsorbent resin HPD-826, use the deionized water rinsing resin earlier, the reuse water-ethanol is gradient elution successively, collect the eluent of concentration of alcohol 70%, merge,, promptly get Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 12g through vacuum drying.The UV content of coffee mesitoyl quinine acid extract is 74%, HPLC content 50%, and wherein 3-caffeoyl quinic acid content is 13%, 3,5-dicaffeoyl-quinic acid content is 10%.
Embodiment 4
Carry out the anti-cervicitis experimentation of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract after adopting the modeling of phenol rubber cement.High, medium and low dosage group of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract and positive group rat filling every day are given corresponding medicine, totally 10 days.After administration in 10 days, each rat eye is got blood and is surveyed total white blood cells, puts to death that animal is got vagina immediately, weigh in the uterus and fixing in formalin solution.
The results are shown in Table 1, positive group (indomethacin) and Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract group all have the exponential effect of the rat model uterus device of reduction; Positive drug and Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract group have significant inhibitory effect to cervicitis rat model total white blood cells; Histopathological examination the results are shown in accompanying drawing 4, and normal group vagina, cervical tissue do not have pathology and change, and the tangible exuviation of model group vagina, the cervix uteri mucosa thickens, the eosinophilic granulocyte is soaked into; Obviously reduce with the eosinophilic granulocyte of the relatively more positive group of model group, epithelium does not have the nothing that comes off and thickens; Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract high dose group the eosinophilic granulocyte obviously reduce, and epithelium does not have the nothing that comes off and thickens; In a small amount of eosinophilic granulocyte in dosage group uterus soak into, vagina epithelium has necessarily and comes off; The low dosage vagina epithelium does not have to be seen obviously and comes off, and soaks into not have obviously with model group comparison cervix uteri eosinophilic granulocyte to change.
Table 1 Ainsliasa fragrans Champ extract is to the influence of cervix uteri rat model organ index and total white blood cells
| Group | N | Dosage (g/kg) | Uterus index (g/kg) | Total white blood cells (* 10 9）/L |
| Normal saline | 8 | — | 3.7782±0.287 | 6.2400±0.114 |
| Model | 8 | — | 4.8686±1.316 | 8.4167±0.700 |
| Indomethacin | 8 | 0.005 | ?3.5672±0.479* | ?6.1167±1.649** |
| The Ainsliaea fragrans Champ. height | 9 | 10 | ?3.2128±0.399* | ?7.6000±0.77* |
| In the Ainsliaea fragrans Champ. | 9 | 5 | 4.1246±0.149 | ?7.1111±1.102** |
| Ainsliaea fragrans Champ. is low | 10 | 2.5 | 4.0980±1.604 | 8.5600±1.442 |
Annotate: compare * p＜0.05, * * p＜0.01 with matched group.
Embodiment 5
Experiment is divided into normal control group, aspirin group, Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract low dose group, middle dosage group, high dose group, after each group is irritated and is given relative medicine, dimethylbenzene is applied to two sides, ear front and back, a mice left side, cut two ears along the auricle baseline, lay round auricle at same position respectively with the 8mm card punch, weigh, calculate the swelling degree.
The results are shown in Table 2, positive drug, Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract object height as can be seen from Table 2, middle dosage group xylol causes mice ear all significant inhibitory effect.And the DeGrain of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract low dose group.
Table 2 Ainsliasa fragrans Champ extract xylol causes the influence of mice ear
Annotate: compare * p＜0.05, * * p＜0.01 with matched group.
Embodiment 6
Experiment is divided into the blank group, high, medium and low dosage group of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract and positive controls, be implanted into cotton balls by the rat body and make inflammatory model, Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract antiinflammatory action is observed in the relatively variation of granuloma induced by implantation of cotton pellets between each group after the administration.
The results are shown in Table 3, positive group (dexamethasone) has significant inhibitory effect (p＜0.01) to the bullate hypertrophy of rat granuloma as can be drawn from Table 3, and suppression ratio is: 44.49%; The dosage group also has obvious suppression effect (p＜0.05) to the granulomatous hypertrophy of rat in the Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract, and suppression ratio is: 24.66%.
Table 3 Ainsliasa fragrans Champ extract is to the bullate influence of rat granuloma
Annotate: compare * p＜0.05, * * p＜0.01 with matched group.
Embodiment 7
Experiment is divided into Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract group, norfloxacin group and acetylspiramycin group, ten gradient concentration band medicine culture medium of each group preparation, afterwards escherichia coli, staphylococcus aureus are inoculated in each group band medicine culture medium, observe the external Chinese People's Anti-Japanese Military and Political College of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract enterobacteria, staphylococcus aureus effect.
The results are shown in Table 5, table 6, this time test Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract as can be seen from table 5,6 and to colibacillary MIC be: 0.25 g crude drug/ml, staphylococcic MIC is to golden yellow color chips: 0.125g crude drug/ml.
Table 4 Ainsliasa fragrans Champ extract variable concentrations is to colibacillary inhibitory action
Table 5 Ainsliasa fragrans Champ extract variable concentrations is to aureus with inhibition
Embodiment 8
Mice 18 ~ 22g, female half and half or same sex, random packet is established administration group (Herba Ainsliaeae Glabrae is high, medium and low), positive drug group (escherichia coli are norfloxacin, and staphylococcus aureus is an acetylspiramycin) and not administration group.After the administration group was irritated stomach for three days on end, the selected mouse death rate that makes was that to cause infection, colibacillary concentration to each group mouse peritoneal injection be 8 * 10 for about 80 ~ 90% bacterium liquid
8Cfu/ml, the concentration of staphylococcus aureus is 3.6 * 10
8Cfu/ml, every 0.5ml, the administration group with infect after, 12,24,48,72h respectively is administered once, observe in 4 days and respectively organize mortality of mice.
The results are shown in Table 6,7, the result shows that the colibacillary survival rate of inoculation in the Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract administration group mice body is 10 ~ 30%, the survival rate of infection of staphylococcus aureus mice is 20 ~ 30%, and the life survival rate of comparing control group mice all has rising trend.
Table 6 Ainsliasa fragrans Champ extract is to the influence of injection pathogenic bacterium mice survival rate in the body
Embodiment 9
Experiment is divided into Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract group, and the lincomycin group is established 10 concentration flat boards for every group.Afterwards escherichia coli, staphylococcus aureus are inoculated in each group band medicine culture medium flat plate, two groups of antibacterials clump counts being no more than on 5 the flat board bacterium colony of picking is transferred on the inclined-plane and preserves, as 2 generation Resistant strain carry out bacteriostatic experiment, two groups of antibacterials clump counts being no more than on 5 the flat board bacterium colony of picking is transferred on the inclined-plane and preserves again, as 3 generation Resistant strain, carry out above experiment repeatedly.The result shows that Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract has overriding resistance effect preferably, to how inhibited for Resistant strain.
The different passage numbers of the golden yellow staphylococcus of table 7 are to the drug resistance measurement result of Ainsliasa fragrans Champ total extract
The different passage numbers of table 8 escherichia coli are to the drug resistance measurement result of Ainsliasa fragrans Champ total extract
Embodiment 10: the preparation of Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract different dosage form
A) pill: get Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract, starch, cross-linking sodium carboxymethyl cellulose pulverize separately and cross 100 mesh sieves, mix homogeneously is used water pill, dry, every ball contains Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 0.1-100mg, is up to the standards, and packing promptly.
B) oral liquid: get Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract, be dissolved in water, it is an amount of to add correctives, add active carbon 5g, heated 30 minutes, and filtered, add water to 1000ml, filter, embedding, sterilization, every ml contains Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 0.1-100mg, be up to the standards, packing promptly.
C) syrup: get Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract, be dissolved in water, an amount of with sucrose 500g and antiseptic, boil and make dissolving, filter, add water to 1000ml, filter embedding, sterilization, every ml contains Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 0.1-100mg, is up to the standards, and packing promptly.
D) powder, tablet, capsule, granule, extractum: contain Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 0.1-100mg in the equal prepared routinely, every or every capsule; The every pouch of powder or granule is that 1g contains Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract 0.1-100mg, and through after the assay was approved, packing promptly.
Claims (9)
1. Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract is characterized in that in the extract pharmacological component with 3-caffeoyl quinic acid, 3,4-dicaffeoylquinic acid, 3, and 5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid are main; Extract calculates with total coffee mesitoyl quinine acid composition, and UV content is more than 70%, and HPLC content is more than 50%, 3-caffeoyl quinic acid and 3 wherein, and 5-dicaffeoylquinic acid content all is higher than 10%.
2. the described Ainsliasa fragrans Champ coffee mesitoyl quinine acid of claim 1 preparation method of extract is characterized in that described extract prepares by following steps:
A) get the dry medical material of Ainsliasa fragrans Champ, cutting is 40 ~ 80% ethanol with the concentration of 5 ~ 20 times of volumes 60 ~ 100 ℃ of following heating and refluxing extraction 2 ~ 4 times, each 1-3 hour, filtration, merge extracting solution A;
B) extracting solution A is evaporated to 1/4 ~ 1/8 times through 40 ~ 60 ℃, promptly obtains concentrated solution B;
C) concentrated solution B transfers pH to 1 ~ 3 with dilute hydrochloric acid, and acid concentrated solution through macroporous resin adsorption, is used earlier the deionized water rinsing resin, the reuse water-ethanol is gradient elution successively, the collection concentration of alcohol is the eluent between 50 ~ 70%, after 40 ~ 60 ℃ of concentrating under reduced pressure remove alcohol, gets concentrated solution D;
D) transfer pH extremely to 1 ~ 3 with dilute hydrochloric acid concentrated solution D again, through macroporous resin adsorption, use earlier the deionized water rinsing resin, the reuse water-ethanol is gradient elution successively, the collection concentration of alcohol is the eluent between 40 ~ 70 %, merge, vacuum drying promptly gets Ainsliasa fragrans Champ coffee mesitoyl quinine acid extract.
3. according to the described Ainsliasa fragrans Champ coffee mesitoyl quinine acid of claim 2a preparation method of extract, it is characterized in that the described concentration of alcohol of step a) is 50%.
4. the described Ainsliasa fragrans Champ coffee mesitoyl quinine acid of claim 2 preparation method of extract is characterized in that described macroporous resin is selected from AB-8, HPD-400, NKA-9 and HPD-826.
5. claim 2 or 4 described Ainsliasa fragrans Champ coffee mesitoyl quinine acid preparation method of extract is characterized in that described macroporous resin is HPD-826.
6. a pharmaceutical composition is characterized in that comprising described extract of claim 1 and acceptable accessories.
7. pharmaceutical composition according to claim 6, the dosage form that it is characterized in that described pharmaceutical composition is tablet, capsule, granule, powder, pill, extractum, oral liquid or syrup.
8. the application of the described Ainsliasa fragrans Champ coffee mesitoyl quinine acid of claim 1 extract in the medicine of preparation antiinflammatory, antibacterium, overriding resistance bacterial strain.
9. the application of the described Ainsliasa fragrans Champ coffee mesitoyl quinine acid of claim 1 extract in the medicine of the anti-cervicitis of preparation.
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| CN104717892A (en) * | 2012-10-16 | 2015-06-17 | 因德纳有限公司 | Process for obtaining caffeoylquinic acids-rich extracts from helianthus annuus |
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| CN103655769A (en) * | 2012-09-12 | 2014-03-26 | 江西本草天工科技有限责任公司 | Preparation method of northeast clematoside extractive and application thereof in antitumor |
| CN104717892A (en) * | 2012-10-16 | 2015-06-17 | 因德纳有限公司 | Process for obtaining caffeoylquinic acids-rich extracts from helianthus annuus |
| KR20160098570A (en) * | 2015-02-09 | 2016-08-19 | 순천향대학교 산학협력단 | COMPOSITION COMPRISING EXTRACT OF Ainsliaea apiculate HAVING ANTI-OXIDANT AND ANTI-INFLAMMATORY ACTIVITY |
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| KR101620384B1 (en) | 2015-02-27 | 2016-05-13 | 순천향대학교 산학협력단 | COSMETIC COMPOSITIONS CONTAINING Ainsliaea apiculata EXTRACT AND PREPARING METHOD OF THE SAME |
| CN105125612A (en) * | 2015-10-23 | 2015-12-09 | 云南云龙制药股份有限公司 | Preparation method and application of frangrant ainsliaea herb soft capsule |
| CN105125612B (en) * | 2015-10-23 | 2017-09-05 | 云南云龙制药股份有限公司 | A kind of preparation method and application of Ainsliaea fragrans Champ soft capsule |
| CN113197927A (en) * | 2021-01-29 | 2021-08-03 | 中国药科大学 | Application of Ainsliaea fragrans extract in preparation of medicine for treating primary dysmenorrhea |
| CN113896750A (en) * | 2021-11-08 | 2022-01-07 | 陈磊 | Grading extraction process of effective components of erigeron breviscapus |
| CN114848649A (en) * | 2022-04-24 | 2022-08-05 | 黑龙江中医药大学 | Pharmaceutical composition containing caffeoylquinic acid compounds and application thereof in treating rheumatoid arthritis |
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