CN101721455B - Manyflower tickclove herb extract, and preparation method and application thereof - Google Patents
Manyflower tickclove herb extract, and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a manyflower tickclove herb extract, and a preparation method and application thereof, which relate to Chinese medicinal extracts. The extract is a manyflower tickclove herb extract solution prepared by the following steps of: taking a Chinese medicament manyflower tickclove herb as a raw material and leaching the raw material with ethanol; recycling a solvent to obtain a general ethanol extract; extracting the water solution of the general ethanol extract orderly with petroleum ether and ethyl acetate for impurity removal and then extracting the water solution with normal butanol to produce a normal butanol extract liquor; and recycling a normal butanol extract liquor solvent to obtain a normal butanol extract, wherein the general ethanol extract and the normal butanol extract both show the characteristics of liver protection and liver damage protective treatment. The manyflower tickclove herb extract can be applied to the preparation of medicaments for the liver protection and the liver damage protective treatment.
Description
Technical field: the present invention relates to a kind of extract of Chinese medicine, specifically the extract of a kind of hungry Hirudo also relates to the preparation method of extract and the application of starving Hirudo simultaneously.
Background technology
Hepatic disease is one of the most common disease that influences human health.Various harmful factors cause hepatic injury, mainly contain viral hepatitis, alcoholic hepatitis and drug induced hepatitis etc., and its clinical manifestation is hepatic necrosis, fatty liver, cholestasis, liver cirrhosis and hepatocarcinoma etc.At present, the treatment to hepatic injury is still global severe problem.
Inventor of the present invention is engaged in the extraction separation and the activity research of Chinese herbal medicine effective ingredients always, special extraction and the activity research of paying attention to hepatic injury is had the effective ingredient of protective effect.Patent of invention " Folium Nelumbinis extract " application number 2009100643537 is exactly the extract that protects the liver with liver protection effect that has that from numerous Chinese herbal medicine, screens and extract with " a kind of desertliving cistanche alcohol extract and preparation method thereof " application number 200810231291X.Inventor of the present invention discovers that manyflower tickclover herbal extract has equally and protects the liver and liver protection effect.
Put down in writing according to " Chinese medicine dictionary "; Herba desmodii multifloi has another name called red palm grass (" the mount emei medicine is planted "); Radix Sophorae Tonkinensis, mashed cajaput, Radix Campylotropis Hirtella (Herba Myrsines Africanae) (" Guizhou medical herbs "); Beggarweed, sticking body grass (" Guangxi Chinese herbal medicine ") are the Herb of leguminous plant mountain Whitmania plant Herba desmodii multifloi Herba Desmodii Multiflori.Have promoting blood circulation and stopping pain, removing toxic substances and promoting subsidence of swelling, heat-clearing and toxic substances removing helps digestion, and the analgesic effect is used for stomachache, infantile malnutrition, parotitis, lymphadenitis, venom, epigastric pain, women's emaciation due to blood disorder, lumbar sprain, wound, urethritis etc.But in all Chinese medicine data, find to starve Hirudo or its extract as yet and be used in treatment hepatopathy or tool and protect the liver record and report with liver protection effect character.
Summary of the invention
The object of the present invention is to provide the extract of a kind of hungry Hirudo.
Simultaneously, the present invention also aims to provide the preparation method of extract of a kind of hungry Hirudo.
The present invention also aims to provide the application of extract aspect the medicine of preparation control hepar damnification of a kind of hungry Hirudo; And the application aspect the liver-protective medicine of preparation.
To achieve these goals, a kind of manyflower tickclover herbal extract of the present invention, its extract are with the Chinese medicine Herba desmodii multifloi to be raw material; Through the ethanol lixiviate and a kind of Herba desmodii multifloi ethanol extract; Reclaim behind the solvent the ethanol total extract, after the ethanol total extract is used petroleum ether, ethyl acetate, extract and separate impurity respectively after with aqueous dispersion successively, the reuse n-butanol extraction; After reclaiming the n-butyl alcohol extract solvent; Get n-butyl alcohol extract, ethanol total extract and n-butyl alcohol extract are all the gluey extractum of brown, and the alcoholic solution of ethanol total extract and n-butyl alcohol extract scans through uv-spectrophotometric; Absworption peak is arranged between 240-280nm, and Herba desmodii multifloi ethanol total extract, n-butyl alcohol extract all show the characteristic of hepatoprotective and control hepatic injury.
The preparation method for distilling of a kind of manyflower tickclover herbal extract of the present invention, preparation according to the following steps:
(1) exsiccant Herba desmodii multifloi root, stem or leaf are pulverized the alcohol solution dipping heating and refluxing extraction 2 hours of 60～80% (V/V) that add 2-4 times of raw material weight; Take out alcohol extract; The alcoholic solution that adds 60～80% (V/V) of 2-4 times of raw material weight again, alcohol extract is taken out in reflux lixiviate 2 hours; Three times so repeatedly, merge three times alcohol extract;
(2) with the alcohol extract that merges for three times steam slip reclaim ethanol and concentrate after, ethanol total extract extractum, ethanol total extract extractum is used water dissolution, petroleum ether, the ethyl acetate with 10-12 times of raw material weight extracts respectively 5 times successively.
(3) divide 5 extractions with ethanol total extract aqueous solution behind petroleum ether, the ethyl acetate extraction with the n-butyl alcohol of 10-12 times of raw material weight, combining extraction liquid, reclaim behind the extractant n-butyl alcohol extract;
The application of a kind of manyflower tickclover herbal extract of the present invention is meant the ethanol total extract of Herba desmodii multifloi and the application that n-butyl alcohol extract is used to prepare the medicine aspect of preventing hepar damnification.
A kind of manyflower tickclover herbal extract of the present invention is used, and is meant the ethanol total extract of Herba desmodii multifloi and the application that n-butyl alcohol extract is used to prepare the liver protecting medicine aspect.
Ethanol total extract and n-butyl alcohol extract to starve Hirudo rhizome or leaf are irritated stomach; With the tetrachloro-methane induction hepatic injury; Detect ALT and AST value in the mice serum, and SOD in the liver tissue homogenate and MDA value, experiment shows: the ethanol total extract and the n-butyl alcohol extract of starving Hirudo rhizome or leaf can effectively reduce the ALT value of carbon tetrachloride hepatic injury mice; AST value and MDA value, and the SOD energy value of the mouse liver cell that can raise significantly.Proof is starved the ethanol total extract of Hirudo rhizome or leaf and the effect that n-butyl alcohol extract has good the liver protecting and control hepatic injury.
Description of drawings: the ethanol total extract ultraviolet spectrometry spectrogram of Fig. 1 Herba desmodii multifloi rhizome; The ethanol total extract ultraviolet spectrometry spectrogram of Fig. 2 Herba desmodii multifloi leaf; The n-butyl alcohol extract ultraviolet spectrometry spectrogram of Fig. 3 Herba desmodii multifloi rhizome; The n-butyl alcohol extract ultraviolet spectrometry spectrogram of Fig. 4 Herba desmodii multifloi leaf;
Abscissa is represented wavelength (nm) among the figure, and ordinate is represented absorbance.Ultraviolet spectrophotometer instrument model: Agilent8453; Quartz colorimetric utensil thickness 1Cm; Wavelength scanning range 200～400nm; Solution medium: ethanol.Concentration: Fig. 1, total ethanol are carried substrate concentration 116.2 μ g/ml; Fig. 2, total ethanol are carried substrate concentration 62.08 μ g/ml; Fig. 3, n-butanol extraction substrate concentration 55.30 μ g/ml; Fig. 4, n-butanol extraction substrate concentration 58.46 μ g/ml can know between wavelength 240-280nm absworption peak is arranged from Fig. 1-4.
The specific embodiment
Take by weighing dry Hirudo leaf 1000 grams of starving and pulverize the back and add 75% (V/V) ethanol waters, reflux, extract, three times (3 * 3000 milliliters), each 2 hours, merge ethanol extract, reclaim solvent, and concentrate ethanol total extract extractum.Ethanol total extract extractum (being called for short the ethanol total extract) is used aqueous dispersion, uses petroleum ether (5 * 2000 milliliters) successively respectively, ethyl acetate (5 * 2000 milliliters), and n-butyl alcohol (5 * 2000 milliliters) extraction merges butanol extraction liquid, reclaims solvent and gets n-butyl alcohol extract.Starving Hirudo leaf ethanol total extract extractum is the brown colloid substance, and its alcoholic solution has absorption maximum through UV scanning at the 271nm place; N-butyl alcohol extract is the brown colloid substance, and its alcoholic solution has absorption maximum through UV scanning at the 274nm place.
Method: intracorporal method-carbon tetrachloride acute liver damage model
Principle: CCl
4Be to use the earliest, hepatic fibrosis derivant the most widely, in the acute liver damage experiment, also be common drug, about CCl
4Liver toxicity mechanism, study morely, have multiple suggestion, but all consistently generally acknowledge that the chain type peroxidization of the formation of free radical and initiation is that it is mainly machine-processed.For CCl
4In the stability study of mouse liver injury model, mainly be CCl through various dose
4Oleum Arachidis hypogaeae semen; Hepatic injury due to after injection detects the superoxide dismutase SOD in glutamate pyruvate transaminase (ALT/GPT), glutamic oxaloacetic transaminase, GOT (AST/GOT) and the hepatic tissue in the mice serum in the certain hour), the content of malonaldehyde (MDA) judges the liver damage situation of mice.ALT and AST mainly are distributed in the hepatocyte of liver, if hepatic necrosis ALT and AST will raise.But these two kinds of enzyme distributions in hepatocyte are different.ALT is distributed in liver cytoplasm, and AST is distributed in liver cytoplasm and the mitochondrion.The chronic hepatitis of acute hepatitis and light disease mainly shows as the rising of ALT.Therefore, AST/ALT＜1; The later stage of chronic hepatitis, liver cirrhosis and liver cancer patient, hepatocellular destructiveness is serious, mitochondrion has also received serious destruction, therefore, AST raises obviously, AST/ALT＞1 even＞2.ALT and AST mainly are distributed in the hepatocyte of liver, and normal value is 0～40 iu.If hepatic necrosis, ALT and AST will raise, and are present the most frequently used liver function indexs.
Instrument: UV-2000 type ultraviolet-uisible spectrophotometer (but You Ni Shanghai Instr Ltd.); CS-Hl type blender (Beijing rich encourage positive scientific & technical corporation), the biochemical incubator (Shanghai one permanent Science and Technology Ltd.) of LRH-150, Multiskan MK3 ELIASA (thermoelectric Shanghai Instr Ltd.); TGL-16 high speed centrifuge (big instrument plant in Jintan City, Jiangsu); The tissue refiner of 985370-395 type (BIOSREC, Mexico), all size pipettor etc.
Reagent: bifendate drop pill (Zhejiang Prov WanBang Pharmaceutical Co., Ltd; Lot number: 080210); ALT, AST, SOD, MDA detection kit (be Nanjing and build up bio-engineering research institute); (packing factory of Shanghai chemical reagents corporation, lot number: 20050115), other reagent are analytical pure to Coomassie brilliant blue G250.
Method:
(20 ± 2g) are divided into 7 groups to mice at random, and 10 every group, blank group, model group are irritated stomach (ig) and give 0.5% CMC-Na solution 20mL/kg every day; The bifendate drop pill positive controls is irritated stomach 100mg/kg every day, starves Hirudo rhizome n-butyl alcohol high dose extract and irritates stomach 500mg/kg, starves Hirudo rhizome n-butyl alcohol low dosage extract and irritates stomach 125mg/kg; Starve Hirudo leaf total extract high dose and irritate stomach 500mg/kg, starve Hirudo leaf total extract low dosage and irritate stomach 250mg/kg, starve Hirudo leaf n-butyl alcohol extract high dose 500mg/kg; Starve Hirudo leaf n-butyl alcohol extract low dosage 125mg/kg administration 8d; Behind last administration 2h, except that blank control group all according to 0.1mL/10g lumbar injection 0.5% carbon tetrachloride oil solution, after water 16h is can't help in fasting; Plucking eyeball gets blood and isolates serum; Detect ALT and AST, isolate liver and weigh, and the liver of the about 300mg of weighing and about 30mg becomes the liver homogenate liquid of 10% (surveying MDA) and 1% (surveying SOD) with 0-3 ℃ of normal saline homogenate; Measure MDA and SOD, Coomassie brilliant blue removes albumen.
The mensuration of ALT, AST, SOD and MDA is all carried out according to the requirement of test kit, and experimental data x ± s representes that experiment statistics adopts the spss17.0 statistical software to carry out one factor analysis of variance.
(1) hungry Hirudo extract is seen table 1 to the result that influences of carbon tetrachloride acute liver damage mice ALT and AST, and carbon tetrachloride model group and blank group compare, and serum alt, AST value all obviously raise, and the modeling success is described.Starve Hirudo rhizome n-butanol extract, leaf total extract and leaf n-butanol extract all can significantly reduce carbon tetrachloride acute liver damage mice ALT and AST energy value, data difference has the utmost point
Table 1 is starved the protective effect of Hirudo extract to the carbon tetrachloride acute liver damage (x ± s)
Each group (except that the sample blank group) and model are compared:
*P＜0.05,
*P＜0.01,
* *P＜0.001;
Model group and blank group compare:
#P＜0.05,
##P＜0.01,
###P＜0.001
The significance meaning.The reduction degree of three ALT and AST value is under the same dose (500mg/kg): leaf n-butanol extract＞leaf total extract＞stem n-butyl alcohol extract, wherein, the above two are higher than positive control drug bifendate (100mg/kg) to the reduction degree of enzyme activity value.The result also shows, the reduction degree of three's various dose group is different, reduces with the degree of its reduction of reduction of dosage, explains that there is dose dependent in the three to the influence of carbon tetrachloride hepatic injury mice ALT and AST enzyme.Can find out from the result: three kinds of manyflower tickclover herbal extracts have fabulous protective effect to the carbon tetrachloride acute liver damage, and have dose dependent.
(2) starve the Hirudo extract to carbon tetrachloride acute liver damage anti-oxidative ability of mice influence carbon tetrachloride model group and blank control group matched group relatively, MDA value is obviously rising all, the reduction of SOD value.Hirudo rhizome n-butanol extract, leaf total extract and leaf n-butanol extract all can significantly reduce the MDA content of carbon tetrachloride acute liver damage mice; Data difference has significance; Wherein the reduction effect of Herba desmodii multifloi rhizome n-butanol extract MDA is the most obvious; Secondly be the leaf n-butanol extract, and the three have dose dependent to the influence of MDA content.The result also shows: three kinds of extracts of Herba desmodii multifloi all can improve the SOD activity value of carbon tetrachloride acute liver damage, and wherein the action effect of rhizome n-butanol extract is best, are the leaf n-butanol extract secondly; The three also has dose dependent to the influence of SOD value.Above result shows that three kinds of hungry Hirudoes get thing and possibly come the liver protecting tissue through oxidative pathway (increased SOD, reduce MDA).
Experimental result shows that carbon tetrachloride model group and blank control group matched group compare, and the MDA value all obviously raises in serum alt, AST value and the hepatocyte, and the SOD value reduces, and the modeling success is described.From total result; Herba desmodii multifloi leaf n-butanol extract has the value of the ALT, AST and the MDA that reduce carbon tetrachloride acute liver damage mice preferably simultaneously and promotes the effect of the SOD activity level of mice; Be the main position of Herba desmodii multifloi hepatoprotective effect, can develop in liver-protecting preparation.
The preparation of embodiment 4, hungry Hirudo extract tablet:
Method for preparing: get above-mentioned hungry Hirudo leaf n-butyl alcohol extract 100g, the n-butyl alcohol of adding an amount of 80% is made the wetting agent mixing and is processed soft material behind adding starch 250g, the arabic gum 15g, under 60 ℃ of temperature, is dried to water content below 3%; Then the thing of oven dry was pulverized 14 mesh sieves; After adding magnesium stearate 5g and Pulvis Talci 20g mixing at last, cross 12 mesh sieve granulate, tabletting; Quality inspection, coating are processed 1000.
It should be noted last that: above embodiment is only in order to explanation; And unrestricted technical scheme of the present invention; Although the present invention is specified with reference to the foregoing description; Those of ordinary skill in the art is to be understood that: still can make amendment or be equal to replacement the present invention, and not break away from any modification or the local replacement of the spirit and scope of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (3)
1. manyflower tickclover herbal extract; It is characterized in that extract is is raw material with the Chinese medicine Herba desmodii multifloi, through the ethanol lixiviate and a kind of Herba desmodii multifloi ethanol extract, reclaim behind the solvent the ethanol total extract; After ethanol total extract aqueous solution is used petroleum ether, ethyl acetate extraction removing impurities successively respectively; The reuse n-butanol extraction behind the recovery n-butyl alcohol extract solvent, gets n-butyl alcohol extract; Ethanol total extract and n-butyl alcohol extract are all the gluey extractum of brown; The alcoholic solution of ethanol total extract and n-butyl alcohol extract scans with uv-spectrophotometric, and absworption peak is arranged between 240-280nm, and Herba desmodii multifloi ethanol total extract, n-butyl alcohol extract all show the characteristic of hepatoprotective and control hepatic injury;
The preparation method for distilling of manyflower tickclover herbal extract, preparation according to the following steps:
(1) ethanol water of exsiccant Herba desmodii multifloi root, stem or leaf being pulverized 60～80% (V/V) that add 2-4 times of raw material weight soaked heating and refluxing extraction 2 hours; Take out alcohol extract; The ethanol water that adds 60～80% (V/V) of 2-4 times of raw material weight again, alcohol extract is taken out in reflux lixiviate 2 hours; Three times so repeatedly, merge three times alcohol extract;
(2) with the alcohol extract that merges for three times steam slip reclaim ethanol and concentrate after, ethanol total extract extractum, ethanol total extract extractum is used water dissolution, petroleum ether, the ethyl acetate with 10-12 times of raw material weight extracts respectively 5 times successively;
(3) divide 5 extractions with ethanol total extract aqueous solution behind petroleum ether, the ethyl acetate extraction with the n-butyl alcohol of 10-12 times of raw material weight, combining extraction liquid, reclaim behind the extractant n-butyl alcohol extract.
2. the application of a kind of manyflower tickclover herbal extract according to claim 1 aspect the medicine of the anti-hepar damnification of preparation, described manyflower tickclover herbal extract is ethanol total extract and n-butyl alcohol extract.
3. the application of a kind of manyflower tickclover herbal extract according to claim 1 aspect preparation the liver protecting medicine, described manyflower tickclover herbal extract is ethanol total extract and n-butyl alcohol extract.
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| CN102743424A (en) * | 2012-07-10 | 2012-10-24 | 郑州大学 | Flos chrysanthemi indici effective ingredient and application thereof |
| CN103342730B (en) * | 2013-06-16 | 2015-06-17 | 浙江大学 | Preparation method of extract of traditional Chinese medicine herb of manyflower ticklover and use of the extract in anti-aging |
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Granted publication date: 20120418 Termination date: 20171228 |