CN102134192A - Preparation method for extracting chlorogenci acid from honeysuckle and application of honeysuckle extract - Google Patents
Preparation method for extracting chlorogenci acid from honeysuckle and application of honeysuckle extract Download PDFInfo
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- CN102134192A CN102134192A CN2011100465471A CN201110046547A CN102134192A CN 102134192 A CN102134192 A CN 102134192A CN 2011100465471 A CN2011100465471 A CN 2011100465471A CN 201110046547 A CN201110046547 A CN 201110046547A CN 102134192 A CN102134192 A CN 102134192A
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Abstract
The invention provides a honeysuckle extract and a preparation method for extracting chlorogenci acid. The method has the advantages of good repeatability and good stability; by using the method, the content of an active component can be improved, a large amount of impurities are removed, raw materials are high in yield and low in cost; and the method is suitable for industrial production. The percentage of the main active component chlorogenci acid in the honeysuckle extract is 30%. The preparation process of the honeysuckle extract comprises the following steps: adding water the volume of which is 8-20 times as large as that of taken honeysuckle medicinal material in the taken honeysuckle medicinal to extract for 2-4 times for 0.5-1.5 hours each time; regulating the pH value of the extract to 1-6; and on a large pore absorption resin column, eluting impurities with water firstly, then eluting with 10-70% ethanol, colleting ethanol eluent, recovering the ethanol, concentrating, regulating the pH value of a concentrated liquid to 1-6, adding ethyl acetate for extraction, recovering the ethyl acetate, concentrating and drying. By using the method, the content of the active component in the extract is improved, the curative effect is improved, a large amount of impurities are removed, the problem of large moisture absorption commonly occurring in Chinese patent medicines is solved, and side effects are reduced.
Description
Technical field
The present invention relates to a kind of Flos Lonicerae extract, and from the Chinese medicine Japanese Honeysuckle method of chlorogenic acid extracting, and the application of this extract on formulations such as tablet, particle, capsule and, anti-infectives clearing heat and detoxicating in preparation.
Background technology
This product is the dry flower of caprifoliaceae plant honeysuckle Lonicera japonica Thunb. or the flower that band is just opened.The flowers are in blossom gathers drying before putting for early summer; Or with the smoked after drying of sulphur.The fecund in Shandong, Henan, mostly be cultivation.Main effect is clearing heat and detoxicating, wind-heat dissipating.Be used for the swollen furunculosis of carbuncle, larynx numbness, erysipelas, toxic-heat and blood stasis, common cold due to wind-heat, warm disease heating.
In recent years, Chinese scholars is used modern theory knowledge and has been carried out multianalysis and research, done a large amount of pharmacology clinical positions, from the angle of science, confirmed Japanese Honeysuckle have broad-spectrum antimicrobial, antiviral, antitumor, strengthening immunity, and antipyretic and anti-inflammatory, cholagogic, protect the liver, multiple pharmacological effect such as lipopenicillinase.Because its special drug effect in the struggle of resistance SARS in 2003, is looked at by the expert parent very, great majority all have Japanese Honeysuckle in the prescriptions of traditional Chinese medicine of the national recommendation control SARS of traditional chinese medical science administration, and it has made distinctions won on the battlefield for resisting SARS.The product that contains Japanese Honeysuckle at present mostly is the traditional Chinese medicine product greatly, and the prescription complexity is though the certain curative effect of tool exists also that dose is big, onset is slow, it is inconvenient to use, drug effect is limited and shortcoming such as quality standard control difficulty.
The main active ingredient of Japanese Honeysuckle is a chlorogenic acid, we in extraction based on chlorogenic acid, the preparation method of traditional Flos Lonicerae extract of bibliographical information has water extraction, alcohol extracting method, water extraction and alcohol precipitation method, ethanol extract from water precipitation etc., but the complicated component of traditional Chinese medicine honeysuckle, contain a lot of chlorogenic acids composition in addition, so the content that traditional preparation method obtains is generally lower.
The method of chlorogenic acid extracting of the present invention is easy, cost is low, and the gained chlorogenic acid contents is not less than 30%, is applicable to suitability for industrialized production.
Summary of the invention
The present invention is directed to present technology status, a kind of preparation method of Japanese Honeysuckle chlorogenic acid extracting is provided, and chlorogenic acid contents is higher, simultaneously repeatability, good stability, and raw material green height, cost is low, is fit to suitability for industrialized production.
The present invention also provides the application of this Flos Lonicerae extract.
For solving the problems of the technologies described above, the present invention has studied and defined following technical scheme.
A kind of Flos Lonicerae extract is characterized in that the weight percent of main active ingredient chlorogenic acid in the Flos Lonicerae extract is at least 30%.
Any a kind of preparation that Flos Lonicerae extract of the present invention and pharmaceutically acceptable carrier and/or vehicle are made.
The preparation method of Flos Lonicerae extract of the present invention, its preparation process is as follows:
The extracting honeysuckle medicinal material adds the water extraction 2~4 times of 8~20 times of volumes, each 0.5~1.5 hour, the pH value of regulating extracting solution is 1~6, last macroporous adsorptive resins, first water wash-out impurity, use 10~70% ethanol elution again, collect ethanol eluate, reclaim ethanol, concentrate, regulating concentrated solution pH value is 1~6, adds ethyl acetate extraction, reclaims ethyl acetate, concentrate drying.
The scope of regulating the pH value of extracting solution in the aforesaid method is 2~3; The scope of regulating the pH value of concentrated solution is 2~3.
The application of Flos Lonicerae extract of the present invention in preparation treatment virus infective medicament.
The present invention has following advantage compared to existing technology:
The present invention uses the macroporous resin isolation technique, adopt water body liquid to transfer pH to go up macroporous adsorptive resins, use 10~70% ethanol elution behind the water elution again, use ethyl acetate purified method at last again, obtain good effect, this method is not only easy and simple to handle, has reduced cost, and very is fit to suitability for industrialized production.Studies show that in a large number the macroporous resin of models such as D101, D201, D301 is better to the fractionation by adsorption effect of chlorogenic acid.
The Flos Lonicerae extract that aforesaid method makes with pharmaceutically acceptable carrier and/or mixed with excipients, is made according to a conventional method and is contained the oral preparations that above-mentioned Flos Lonicerae extract is a main active ingredient.
The Flos Lonicerae extract that adopts the present invention to make, its main active ingredient is a chlorogenic acid, measures by high-efficient liquid phase technique, chlorogenic acid contents can account for more than 30% of Flos Lonicerae extract.Pharmacodynamics test proves that Flos Lonicerae extract of the present invention has good antiviral effect generally, compares with traditional preparation, and the effective dose that the present invention treats virus infection is little, and toxic side effect is little.
The present invention is owing to the macroporous adsorbent resin technology that has adopted in the modernization of Chinese medicine technology, the content of the activeconstituents in the extract is improved, improved curative effect, and removed a large amount of impurity, both solve the big problem of moisture absorption of Chinese patent medicine, reduced toxic side effect again.
The present invention has done improvement on the basis of existing technology to the preparation method of Flos Lonicerae extract, at the deficiency in the existing extracting method, utilization macroporous adsorbent resin isolation technique, adopt aqueous extract to transfer pH to go up macroporous adsorptive resins, use 10~70% ethanol elution behind the water elution again, use ethyl acetate purified method at last again, obtain good effect, this method is not only easy and simple to handle, has reduced cost, and very is fit to suitability for industrialized production.
The test of pesticide effectiveness of the Flos Lonicerae extract of the present invention's preparation:
Adopt several animal models to study drug effect of the present invention, and and market on main antiviral cold medicine SHUANGHUANGLIAN KOUFUYE contrast, its method and result are as follows:
1. to the provide protection of influenza virus infecting mouse
1.1 provide protection to the death of influenza virus mouse lung adapted strain infecting mouse
1.1.1 method
Get 114 of ICR mouse, body weight 13~16g, male and female half and half are divided into 6 groups at random, 19 every group.(1) blank group: equivalent NS10mg/kg; (2) model group: equivalent NS10ml/kg; (3) SHUANGHUANLIAN group: 6g/kg; (4) Flos Lonicerae extract I group: 6.6g/kg; (5) Flos Lonicerae extract II group: 13.2g/kg; (6) Flos Lonicerae extract III group: 26.4g/kg.Below respectively organize the equal gastric infusion of mouse.Each organizes administration every day 1 time, successive administration 5 days.Except that the blank group, all the other each groups infect every mouse 50 μ l (3 LD for the mouse collunarium with viral allantoic fluid under the ether light anaesthesia behind the last administration 1h
50Lethal quantity), observe zoogenetic infection sequela and death condition in 14 days.
1.1.2 result
Test-results shows: three dosage groups of Flos Lonicerae extract all can significantly reduce influenza infection dead mouse number, and 26.4g/kg dosage group can obviously prolong the survival fate (P<0.05) of influenza infection mouse.Show that Flos Lonicerae extract has obvious provide protection to the influenza virus infecting mouse.See Table 1.
Table 1 Flos Lonicerae extract is to the provide protection of influenza virus infecting mouse death
Compare with blank group ##p<0.01; * compare with model group p<0.05
1.2 influence to influenza virus Lung Index of mice infected by Influenza virus and tuberculosis change
1.2.1 method
Get 72 of ICR mouse, body weight 18~22g, male and female dual-purpose.Be divided into 6 groups at random, 12 every group.(1) blank group: equivalent NS10mg/kg; (2) model group: equivalent NS 10ml/kg; (3) SHUANGHUANLIAN group: 6g/kg; (4) Flos Lonicerae extract I group: 6.6g/kg; (5) Flos Lonicerae extract II group: 13.2g/kg; (6) Flos Lonicerae extract III group: 26.4g/kg.Below respectively organize the equal gastric infusion of mouse, administration capacity 10ml/kg.Each organizes administration every day 1 time, successive administration 5 days.Except that the blank group, all the other are respectively organized in administration the 1st day, under the ether light anaesthesia, infect every mouse 50 μ l (3 LD for the mouse collunarium with viral allantoic fluid
50Lethal quantity).Continued administration then 4 days.With mouse fasting (can't help water), carry out following test next day after the last administration: take off neck after 1. mouse is weighed and put to death, dissect, get full lung and weigh, calculate each mouse lung index (g/10g body weight) and lung index inhibiting rate [(medicine group-model group)/model group * 100%]; 2. get lung, after fixing with 1% formaldehyde solution, routine is drawn materials, dehydration, and paraffin embedding, film-making is done the pathology histological examination after the HE dyeing, observes the pulmonary lesion degree.
1.2.2 result
1.2.2.1 influence to the influenza virus Lung Index of mice infected by Influenza virus
Experimental result shows: Flos Lonicerae extract 13.2g/kg and 26.4g/kg dosage group all can obviously reduce the lung index value of influenza infection mouse, and lung index inhibiting rate is respectively 15.60% and 24.67%.Show that Flos Lonicerae extract has the effect that alleviates influenza infection mouse lung pathology.See Table 2.
Table 2 Flos Lonicerae extract is to the influence of influenza virus Lung Index of mice infected by Influenza virus
Compare with the blank group ##p<0.01; * compare with model group p<0.05 * * p<0.01
1.2.2.2 influence to the change of influenza virus infecting mouse tuberculosis
Lung's inflammatory lesion with influenza virus is duplicated mainly shows as the pulmonary inflammation pathology, comprises that acute bronchitis reaches inflammation, lobular pneumonia and interstitial pneumonia on every side.Flos Lonicerae extract of the present invention can alleviate pulmonary inflammatory pathology, and each dosage group is compared, and it is best to alleviate pulmonary inflammatory effect with the heavy dose group, is middle dosage group secondly, relatively has significant difference (* p<0.05,0.01) with model group.Show that Flos Lonicerae extract can alleviate the pulmonary infection of influenza virus induced mice, has the effect of resisiting influenza virus.
The influence that table 3 Flos Lonicerae extract becomes influenza virus infecting mouse tuberculosis
Compare with the blank group ##p<0.01; * compare with model group p<0.05 * * p<0.01
Above result shows that the Flos Lonicerae extract that Flos Lonicerae extract of the present invention extracts by this preparation method of above-mentioned description of test has antiviral preferably, has significant therapeutic effect for influenza.
Embodiment
Among the embodiment, traditional Chinese medicine honeysuckle from the Xiushan Mountain, Chongqing City county, all meets " the regulation in one one of the Chinese pharmacopoeia version in 2005 under " Japanese Honeysuckle " respectively.
The measuring method of chlorogenic acid in the extract: photograph high performance liquid chromatography HPLC (" appendix VID of Chinese pharmacopoeia version in 2005) measure.
Chromatographic condition and system suitability test are weighting agent with the octadecylsilane chemically bonded silica; With acetonitrile-0.4% phosphoric acid solution (13: 87) is moving phase; The detection wavelength is 327nm.Number of theoretical plate calculates by the chlorogenic acid peak and is not less than 1000.
It is an amount of that the preparation precision of reference substance solution takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, adds 50% methyl alcohol and make the solution that every 1ml contains 40 μ g, promptly gets (preserving below 10 ℃).
The about 1.0g of this product powder (crossing sieve No. four) is got in the preparation of need testing solution, and accurate the title decides, and puts in the tool plug Erlenmeyer flask, the accurate 50% methyl alcohol 50ml that adds claims to decide weight, supersound process (power 250W, frequency 35kHz) 30 minutes, put coldly, claim to decide weight again, supply the weight that subtracts mistake with 50% methyl alcohol, shake up, filter, precision is measured subsequent filtrate 5ml, puts in the brown measuring bottle of 25ml, adds 50% methyl alcohol to scale, shake up, promptly.
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of assay method inject liquid chromatograph, measure, promptly.
The following example is intended to further describe the present invention, rather than limits the present invention by any way.
Embodiment 1
(place of production: the Xiushan Mountain, Chongqing) 100kg adds water 1000L, 800L respectively and extracts each 1.5 hours 2 times the extracting honeysuckle medicinal material, with extracting solution adjust pH 1, last D101 macroporous adsorptive resins, water fully are eluted to effluent liquid for after clarification and not having reducing sugar reaction, use 10% ethanol elution again, collect ethanol eluate, reclaim ethanol, concentrate, concentrated solution adjust pH 3 adds ethyl acetate extraction, reclaims ethyl acetate, concentrated, dry, get Flos Lonicerae extract 2.24kg.
Measure through the HPLC method, chlorogenic acid content is 72% in the above-mentioned experiment gained Flos Lonicerae extract.
Embodiment 2
(place of production: the Xiushan Mountain, Chongqing) 100kg adds water 1500L, 1000L, 1000L extraction 3 times respectively, each 1.0 hours to the extracting honeysuckle medicinal material, with extracting solution adjust pH 3, last D201 macroporous adsorptive resins, water fully are eluted to effluent liquid for after clarification and not having reducing sugar reaction, use 30% ethanol elution again, collect ethanol eluate, reclaim ethanol, concentrate, concentrated solution adjust pH 1 adds ethyl acetate extraction, reclaims ethyl acetate, concentrated, dry, get Flos Lonicerae extract 2.35kg.
Measure through the HPLC method, chlorogenic acid content is 75.5% in the above-mentioned experiment gained Flos Lonicerae extract.
Embodiment 3
(place of production: the Xiushan Mountain, Chongqing) 100kg adds water 2000L, 800L, 800L, 800L extraction 4 times respectively, each 0.5 hour to the extracting honeysuckle medicinal material, with extracting solution adjust pH 6, last D301 macroporous adsorptive resins, water fully are eluted to effluent liquid for after clarification and not having reducing sugar reaction, use 70% ethanol elution again, collect ethanol eluate, reclaim ethanol, concentrate, concentrated solution adjust pH 6 adds ethyl acetate extraction, reclaims ethyl acetate, concentrated, dry, get Flos Lonicerae extract 2.44kg.
Measure through the HPLC method, chlorogenic acid content is 78% in the above-mentioned experiment gained Flos Lonicerae extract.
Embodiment 4
The preparation of Japanese Honeysuckle particulate:
Get the Flos Lonicerae extract 300g of embodiment 3, starch 700g, mixing is granulated, and sieves, and makes particle 1000g, promptly.
Usage and dosage: oral, each 5g, every day 2 times.
Embodiment 5
The capsular preparation of Japanese Honeysuckle:
Get the Flos Lonicerae extract 300g of embodiment 3, starch 200g, mixing is granulated, and sieves, dry back 1000 of filled capsules (0.5g/ grain), promptly.
Usage and dosage: oral, each 5, every day 2 times.
Embodiment 6
The preparation of Japanese Honeysuckle sheet:
Get the Flos Lonicerae extract 300g of embodiment 3, starch 200g, mixing is granulated, and sieves, and is pressed into 1000 (0.5g/ sheets) after the drying, promptly.
Usage and dosage: oral, each 5, every day 2 times.
Claims (6)
1. a Flos Lonicerae extract is characterized in that the weight percent of main active ingredient chlorogenic acid in the Flos Lonicerae extract is at least 30%.
2. Flos Lonicerae extract according to claim 1 is characterized in that any a kind of preparation that this Flos Lonicerae extract and pharmaceutically acceptable carrier and/or vehicle are made.
3. Flos Lonicerae extract according to claim 2 is characterized in that described preparation is granule, medicinal tea, capsule, tablet.
4. the preparation method of the described Flos Lonicerae extract of claim 1, its preparation process is as follows:
The extracting honeysuckle medicinal material adds the water extraction 2~4 times of 8~20 times of volumes, each 0.5~1.5 hour, the pH value of regulating extracting solution is 1~6, last macroporous adsorptive resins, first water wash-out impurity, use 10~70% ethanol elution again, collect ethanol eluate, reclaim ethanol, concentrate, regulating concentrated solution pH value is 1~6, adds ethyl acetate extraction, reclaims ethyl acetate, concentrate drying.
5. the preparation method of Flos Lonicerae extract according to claim 4 is characterized in that: the scope of regulating the pH value of extracting solution is 2~3; The scope of regulating the pH value of concentrated solution is 2~3.
6. the application of the described Flos Lonicerae extract of claim 1 in preparation treatment virus infective medicament.
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Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102755366A (en) * | 2012-07-19 | 2012-10-31 | 浙江维康药业有限公司 | Honeysuckle soft capsule containing chlorogenic acid compound |
CN102988456A (en) * | 2012-12-17 | 2013-03-27 | 河北神威药业有限公司 | Application of honeysuckle extractive from active components of Qingkailing for preparing multiple-resistant bacteria resistant medicine |
CN106176879A (en) * | 2016-07-27 | 2016-12-07 | 南京正宽医药科技有限公司 | The process of preparing Chinese medicine extracting method of a kind of Flos Lonicerae decoction pieces and honeysuckle mouthwash |
CN108309920A (en) * | 2018-05-15 | 2018-07-24 | 贝德氏(上海)健康科技股份有限公司 | A kind of skin care item for repairing skin barrier |
CN111039786A (en) * | 2020-01-15 | 2020-04-21 | 成都乐美天医药科技有限公司 | Preparation method of high-purity chlorogenic acid |
CN114601857A (en) * | 2020-12-04 | 2022-06-10 | 海图生物科技(上海)有限责任公司 | Honeysuckle extract with high chlorogenic acid content as well as preparation method and application thereof |
-
2011
- 2011-02-28 CN CN2011100465471A patent/CN102134192A/en active Pending
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102755366A (en) * | 2012-07-19 | 2012-10-31 | 浙江维康药业有限公司 | Honeysuckle soft capsule containing chlorogenic acid compound |
CN102988456A (en) * | 2012-12-17 | 2013-03-27 | 河北神威药业有限公司 | Application of honeysuckle extractive from active components of Qingkailing for preparing multiple-resistant bacteria resistant medicine |
CN106176879A (en) * | 2016-07-27 | 2016-12-07 | 南京正宽医药科技有限公司 | The process of preparing Chinese medicine extracting method of a kind of Flos Lonicerae decoction pieces and honeysuckle mouthwash |
CN108309920A (en) * | 2018-05-15 | 2018-07-24 | 贝德氏(上海)健康科技股份有限公司 | A kind of skin care item for repairing skin barrier |
CN111039786A (en) * | 2020-01-15 | 2020-04-21 | 成都乐美天医药科技有限公司 | Preparation method of high-purity chlorogenic acid |
CN114601857A (en) * | 2020-12-04 | 2022-06-10 | 海图生物科技(上海)有限责任公司 | Honeysuckle extract with high chlorogenic acid content as well as preparation method and application thereof |
CN114601857B (en) * | 2020-12-04 | 2023-04-07 | 海图生物科技(上海)有限责任公司 | Honeysuckle extract with high chlorogenic acid content as well as preparation method and application thereof |
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Application publication date: 20110727 |