CN104958393A - Extract of potentilla anserina rhizome and medical application thereof - Google Patents
Extract of potentilla anserina rhizome and medical application thereof Download PDFInfo
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- CN104958393A CN104958393A CN201510309906.6A CN201510309906A CN104958393A CN 104958393 A CN104958393 A CN 104958393A CN 201510309906 A CN201510309906 A CN 201510309906A CN 104958393 A CN104958393 A CN 104958393A
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Abstract
The invention discloses an extract of potentilla anserina rhizome and medical application thereof. A preparation method for the extract comprises the following steps: (1) crushing dried potentilla anserina rhizome; (b) carrying out heat reflux extraction with 75% ethanol, combining extracting liquid and concentrating the extracting liquid until alcohol smell does not exist so as to obtain concentrate; (c) successively extracting the concentrate obtained in the step (b) with petroleum ether, ethyl acetate and water-saturated n-butanol so as to respectively obtain petroleum ether extract, ethyl acetate extract and n-butanol extract; and (d) enriching the n-butanol extract with macroporous resin, carrying out elution with 10% ethanol weighing 8 times of a column volume so as to remove polysaccharide, then carrying out elution with 65% ethanol weighing 10 times of the column volume, collecting eluate produced by elution with 65% ethanol and subjecting the eluate to pressure-reduced concentration and spray drying. The extract of potentilla anserina rhizome is capable of protecting the liver and can be used for preparing a drug used for protecting the liver.
Description
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to a kind of Radix potentillae anserinae rhizome extract and the pharmaceutical preparation containing this extract, this Radix potentillae anserinae rhizome extract can be used for preparing liver-protective medicine.
Background technology
Plant Radix potentillae anserinae is perennial herb, is mainly distributed in Qinghai, Gansu, Tibet.Root bottom enlarges into spindle or oval tuber, and stem is crawled, and at joint, take root in place, often lands and grows new plant, outer by Fu Sheng or partly carry out thin pubescence or come off near without hair.Basal leaf is for being interrupted winglike compound leaf.Obviously grow thickly when blooming, lobule 6-11 couple, to raw or alternate, stem leaf is little or degenerate; Petiole is by Fu Sheng or partly carry out thin pubescence, sometimes comes off near without hair; Lobule stockless or terminal leaflet have short handle, and little a pair of the top leaf base is downward to be converged with rachis, and base portion lobule is gradually little of Radix Aconiti Lateralis Preparata shape; Basal leaf stipule film quality, brown, and petiole is linked to be constitute sheath-like, outer by thin pubescence or come off near without hair.Stem leaf stipule keratin, light green, many drastic cracks; Vanelets is usually oval, long l-2.5cm, wide 0.5-1cm, and tip circle is blunt, base portion wedge shape or wealthy wedge shape, and there is most serrated irregular at edge or in splintery, above green, by thin pubescence or come off near without hair, below close by silvery white thin,tough silk hair.Single flower axil is raw; Bennet is dredged pubescence; Flower diameter 1.5-2cm; Sepal 5, triangular egg, the anxious point of tip or gradually point, calycule sheet 5, oval or elliptic-lanceolate, full edge or have 2-4 tooth, closely isometric or slightly short with sepal; Petal 5, obovate, tip is circular, than long 1 times of sepal, yellow; Spray side is raw.Achene is avette, and the hollow point of tool, there is grain at back.The florescence 5-7 month.
Potentilla anserina be Rosaceae potentilla plants potentilla anseriana (
potentilla anserinel.) tuber, has effect of promoting the production of body fluid to quench thirst, invigorating spleen and reinforcing stomach, vigorate qi and replenish the blood, astringing to arrest bleeding, antidiarrheal, cough-relieving, sharp expectorant, cures mainly the diseases such as haematemesis, hematochezia, metrorrhagia, malaria carbuncle skin ulcer, insufficiency of the spleen diarrhoea, dysentery.18 seed amino acids of Radix potentillae anserinae rich in starch, fatty acid and needed by human body and multivitamin, have higher medical treatment and nutritive value.The book such as " Chinese medicine voluminous dictionary ", " Chinese herbal medicine is commonly used in Tibet " is all on the books to this medicine.These product are very abundant at Qinghai Province's wild resource, and Medicinal for a long time, is particularly applied more in Tibetan medicine.Yang Na etc. send out the polysaccharide extracting process having inquired into Radix potentillae anserinae by water extraction; and (Yang Na etc. are studied to the antioxidant activity of extract; Chinese food journal; 2nd phase in 2014; 60th ~ 66 pages), there is not yet the research report about potentilla anserina extract and liver-protective medicinal usage thereof.
Summary of the invention
The object of the present invention is to provide a kind of Radix potentillae anserinae rhizome extract, the preparation method of this extract and liquid phase analysis method, the pharmaceutical preparation containing this extract and utilize this extract to prepare the purposes of liver-protective medicine.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
A kind of Radix potentillae anserinae rhizome extract, this extract is prepared by following methods:
A the Radix potentillae anserinae rhizome of drying is pulverized by (); B () extracts with 75% alcohol heat reflux, merge extractive liquid, is concentrated into and obtains concentrated solution without alcohol taste; C () uses petroleum ether, ethyl acetate and water saturated n-butanol extraction successively to step (b) concentrated solution, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; D () n-butyl alcohol extract macroporous resin enrichment, first with 10% alcohol flushing, 8 column volume removing polysaccharide, then uses 65% ethanol elution, 10 column volumes, collects 65% eluent, concentrating under reduced pressure, spraying dry.
Further, macroporous resin described in step (d) is AB-8 type macroporous resin.
In order to be controlled Radix potentillae anserinae rhizome extract differences between batches prepared by Different sources, batch medical material by the method setting up HPLC finger printing, the liquid phase analysis method of described extract is:
Chromatographic column: Diamonsil C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% phosphoric acid solution (triethylamine adjust ph to 6.2);
Gradient elution program: 0.01 ~ 70min, A 20% → 80%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 240nm; Column temperature: 30 DEG C;
Sample size: 10 μ L.
Pharmaceutical preparation, the described Radix potentillae anserinae rhizome extract containing treatment effective dose and pharmaceutically acceptable carrier.
The application of described Radix potentillae anserinae rhizome extract in the liver-protective medicine of preparation.
The application of described pharmaceutical preparation in the liver-protective medicine of preparation.
When extract of the present invention is used as medicine, directly can uses, or use in the form of a pharmaceutical preparation.
Described pharmaceutical preparation contains the Radix potentillae anserinae rhizome extract of the present invention for the treatment of effective dose, and all the other are acceptable on materia medica, nontoxic to humans and animals and pharmaceutically suitable carrier of inertia and/or excipient.
Described pharmaceutically suitable carrier or excipient are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation adjuvant.Pharmaceutical preparation of the present invention is used with the form of per weight dose.Extract of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, drop pill, micropill, suspensoid, Emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into aqueous or oily solution, aseptic powder injection, liposome or the Emulsion etc. of sterilizing.
Advantage of the present invention: the present invention by extracting Radix potentillae anserinae root medicinal material, remove impurity, enrichment process, the extract with the liver protecting effect can be obtained; Liquid phase analysis method provided by the invention may be used for the finger printing setting up this extract, for controlling Different sources, the differences between batches of extract prepared by batch medical material.
Accompanying drawing explanation
Fig. 1 is Radix potentillae anserinae rhizome extract HPLC chromatograms.
Detailed description of the invention
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Main agents and material: Radix potentillae anserinae root medicinal material is purchased from Hui nationality's Chinese Medicinal Materials Markets; Food-grade ethanol is purchased from Shanghai Ling Feng chemical reagent company limited; Pharmaceutical grade AB-8 macroporous resin is purchased from sky tunami letter resin company limited; Acetonitrile is HPLC level, is purchased from TEDIA; 85% phosphoric acid is HPLC level, is purchased from TEDIA; Chromatographic grade pure water is heartily pure water.HL7702 liver cell line is provided by Shanghai Life Sciences Research Institute, Chinese Academy Of Sciences.DMEM in high glucose culture medium, DMEM in high glucose/F12=1:1 culture medium, hyclone are all purchased from HycLone.EDTA is purchased from Shanghai reagent one factory.Trypan blue is purchased from Beijing Chemical Plant.Sugar-free DMEM culture medium is purchased from Gibco.Pancreatin, MTT, DMSO are all purchased from Amresco.LDH method for releasing detection kit is purchased from GENMDE.ALT biochemistry detection test kit, AST biochemistry detection test kit, LDH biochemistry detection test kit are all purchased from Beckman reagent company limited of the U.S..Western and IP cell pyrolysis liquid, PMSF, BCA determination of protein concentration test kit, MDA detection kit are all purchased from green skies biological study institute.
CO
2incubator (SheL-Lab 2300), electric heating constant temperature incubator (Shanghai leap medical apparatus and instruments factory), anaerobic culture box (Changsha Chang Jin Science and Technology Ltd.), microplate reader (U.S. BioTek Epoch), full automatic biochemical apparatus (Beckman LX20), centrifuge (German Eppendorf centrif μ ge 5415D).
Embodiment 1: prepared by Radix potentillae anserinae rhizome extract
Pulverized by the Radix potentillae anserinae rhizome of 10kg drying, extract (25L × 3 time) with 75% alcohol heat reflux, merge extractive liquid, is concentrated into and obtains concentrated solution (6L) without alcohol taste; Petroleum ether (6L × 3 time), ethyl acetate (6L × 3 time) and water saturated n-butyl alcohol (6L × 3 time) is used to extract successively to concentrated solution, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract 305g respectively, with AB-8 macroporous resin (2kg after n-butyl alcohol extract dissolves with 5L distilled water, column volume 1.5L) enrichment, 10% alcohol flushing, 8 column volumes (12L) are first used to remove polysaccharide, use 65% ethanol elution, 10 column volumes (15L) again, collect 65% eluent, concentrating under reduced pressure, spraying dry obtains Radix potentillae anserinae rhizome extract extract powder 185g.
Embodiment 2: liquid-phase chromatographic analysis
Need testing solution is prepared: in the brown volumetric flask of extract extract powder 5mg to 50mL that Example 1 method is obtained, add 30mL 20% acetonitrile solution ultrasonic dissolution, after being cooled to room temperature, continue to add 20% acetonitrile solution standardize solution.
Analytical method:
High performance liquid chromatograph: Agilent 1260, binary pump;
Chromatographic column: Diamonsil C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% phosphoric acid solution (triethylamine adjust ph to 6.2);
Gradient elution program: 0.01 ~ 70min, A 20% → 80%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 240nm; Column temperature: 30 DEG C;
Sample size: 10 μ L.
Analyze with 10 batches of Different sources, a batch extract prepared by Radix potentillae anserinae rhizome, set up finger printing and mate, 1 ~ No. 9 peak all occurs in 10 batch sample chromatograms as a result.Therefore demarcate 9 peaks for total fingerprint peaks, set up the HPLC finger printing of this extract accordingly, the results are shown in Figure 1.
Embodiment 3: Radix potentillae anserinae rhizome extract pharmacological testing
One, test method
1, cell grouping
Cell is divided into 6 groups, often organizes 6 multiple holes: (1) normally cultivation group (N group): complete medium, cultivate under normal condition; (2) ischemia-reperfusion group (IR group): after cultivating 1h under complete medium normal condition, simulation anoxia 8h, Reperfu-sion 4h; (3) potentilla anserina extract group: RE1 group: potentilla anserina extract 0.5ng/mL pretreatment 1h; RE2 group: potentilla anserina extract 5ng/mL pretreatment 1h; RE3 group: potentilla anserina extract 50ng/mL pretreatment 1h; Simulation anoxia 8h, Reperfu-sion 4h; (4) normal saline group (NS group): with the normal saline pretreatment 1h with potentilla anserina extract equal volume, simulation anoxia 8h, Reperfu-sion 4h.
2, the pretreatment of potentilla anserina extract
(1) preparation of potentilla anserina extract: 1mg potentilla anserina extract is dissolved in 500mL normal saline.Liquid-transfering gun draws 2.5mL in first centrifuge tube, is mixed with the potentilla anserina extract solution that concentration is 500ng/mL with normal saline dilution to 10mL.Draw from first centrifuge tube in 1mL to the second centrifuge tube, be mixed with to 10mL the potentilla anserina extract solution that concentration is 50ng/mL with normal saline dilution.From second centrifuge tube, draw 1mL in the 3rd centrifuge tube, be mixed with to 10mL the potentilla anserina extract solution that concentration is 5ng/mL with normal saline dilution.Perform labelling.
(2) potentilla anserina extract pretreatment: normal cultivation group and ischemia-reperfusion group are changed with the complete medium that every hole 100 μ L is fresh.The every hole of RE1 group adds the potentilla anserina extract solution 10 μ L of 5ng/mL and fresh complete medium 90 μ L, the every hole of RE2 group adds the potentilla anserina extract solution 10 μ L of 50ng/mL and fresh complete medium 90 μ L, and the every hole of RE3 group adds the potentilla anserina extract solution 10 μ L of 500ng/mL and fresh complete medium 90 μ L.The every hole of normal saline group adds normal saline 10 μ L and fresh complete medium 90 μ L.Shake 96 orifice plates gently, make medicinal liquid fully dark even, put into CO
21h is hatched in incubator.
3, the foundation of the in-vitro simulated Ischemia-Reperfusion Injury Model of hepatocyte:
(1) in-vitro simulated ischemic period: after pretreatment time terminates, from cell culture incubator, take out first piece of 96 orifice plate, the normal every hole of cultivation group 100 μ L complete mediums are changed, and put into CO
2continue in incubator to cultivate 8h.From CO
2second piece of 96 orifice plate is taken out in incubator, ischemia-reperfusion group, potentilla anserina extract pretreated group and the every hole of normal saline group 100 μ L sugar-free DMEM culture medium displacement complete mediums, put into anaerobic culture box, cultivate in box the sterile vials maintenance saturated humidity put into and fill 50mL aquesterilisa.Anaerobic culture box is put into 37 DEG C of constant-temperature incubation casees, air inlet connects 94%N
2-5%CO
2-1%O
2mist, gas outlet connects oxygen detection instrument.Pass into mist with the gas flow of 2L/min, when oxygen detection instrument demonstrates QI KOU oxygen concentration <1%, regulate mixed gas flow 300mL/min to maintain gas outlet oxygen concentration <1%.With this simulated ischemia process 8h.
(2) in-vitro simulated refilling process: from CO
2take out first piece of 96 orifice plate in incubator, the fresh complete medium of 100 μ L is changed in the normal every hole of cultivation group, puts into CO
2continue in incubator to cultivate 4h.From anaerobic culture box, take out second piece of 96 orifice plate, ischemia-reperfusion group, potentilla anserina extract pretreated group and the every hole of the normal saline group complete medium that 100 μ L are fresh replaces sugar-free DMEM culture medium, puts into CO
2(37 DEG C, 5%CO under normal condition in incubator
2, 95% air, saturated humidity) cultivate 4h, simulation refilling process.
3, Indexs measure
3.1MTT method detects Activity of hepatocytes
(1) MTT solution preparation: take 250mgMTT with electronic micro balance, put into small beaker, adds 50mLPBS and stirs 30min, make it abundant dissolving, be the MTT solution that concentration is 5mg/mL.Degerming with 0.22 μm of micropore filter in superclean bench, be packed as and often prop up 1mL, 4 DEG C keep in Dark Place for subsequent use.
(2) cell prepares: carry out cell grouping as stated above, and separately establishes zeroing hole.And carry out potentilla anserina extract pretreatment and in-vitro simulated Ischemia-Reperfusion Injury as stated above.
(3) colour generation: the every hole of last time point adds the MTT solution 20 μ L of 5mg/mL eventually, puts into 37 DEG C, 5%CO
2, 95% air, saturated humidity CO
2continue in incubator to cultivate 4h.Stop cultivating, careful suction abandons supernatant in hole, and every hole adds 100 μ LDMSO solution, and micro oscillator vibrates 10min, and crystal is fully dissolved.
(4) colorimetric: select 570nm wavelength, microplate reader measures each hole trap (A), record result.Experiment repetition 2 times.
3.2 LDH release cell proliferation and toxicity detection
Operate by GENMED LDH release cell proliferation and toxicity immue quantitative detection reagent box description, experiment repetition 2 times.
(1) prepare cell to be measured as stated above, and separately establish the cell (sample maximum enzyme activity control wells) of acellular culture fluid (background empty map hole) and untreated follow-up cracking, perform labelling.
(2) detection time to regulation puts first 1 hour, from CO
2take out 96 porocyte culture plates to be measured in incubator, add 10 μ LGENMED lysates in the cell hole of untreated follow-up cracking, add 10 μ LGENMED replenishers in all the other all detect aperture simultaneously.Put back to CO
2continue cultivation in incubator and namely specify detection time in 1 hour.
(3) from CO
2take out 96 porocyte culture plates to be measured in incubator, put into orifice plate centrifuge 10min, speed is 250g.
(4) carefully pipette 50 μ L supernatant respectively in the respective aperture of 96 new orifice plates, perform labelling simultaneously.
(5) mix GENMED reactant liquor, every hole adds 50 μ LGENMED reactant liquors respectively, more every hole adds 10 μ LGENMED nitrite ions respectively, shakes 96 orifice plate mixings.
(6) at room temperature hatch 30min, avoid illumination.
(7) every hole adds 10 μ LGENMED stop buffers respectively.
(8) at once put microplate reader into and measure absorbance (A), select wavelength 570nm.
(9) according to following formulae discovery cell mortality:
The actual extinction reading of processing sample=treatment samples sample wells extinction reading-sample controls hole extinction reading-background empty map hole extinction reading
The actual extinction reading=sample maximum enzyme activity control wells extinction reading-sample controls hole extinction reading-background empty map hole extinction reading of sample cell maximum enzyme activity
Cell mortality=(the actual extinction reading of processing sample actual extinction reading ÷ sample cell maximum enzyme activity) × 100%
The mensuration of 3.3 hepatocyte function indexs:
Carry out cell grouping, pretreatment and simulated ischemia reperfusion injury as stated above.Be collected in Ep pipe by each porocyte culture supernatant 100 μ L respectively to whole last time point, often pipe adds 100 μ L complete mediums respectively and is diluted to 200 μ L, centrifugal 5min under room temperature, and speed is 1200r/min.Get supernatant and detect ASL, ALT and LDH level under full automatic biochemical apparatus.Experiment repetition 2 times.
Three, result and conclusion
1, potentilla anserina extract pretreatment is on the impact of ischemical reperfusion injury Activity of hepatocytes
IR group Activity of hepatocytes decline (P<0.05) more obvious than N group, shows that in-vitro simulated Ischemia-Reperfusion Injury successfully causes hepatocyte ischemical reperfusion injury.Give 5ng/ml potentilla anserina extract pretreatment 1 hour, the hepatocellular vigor of ischemical reperfusion injury comparatively ischemia-reperfusion group and normal saline group strengthens (P<0.05), but fail to return to normal group Activity of hepatocytes level (P<0.05), normal saline group then with ischemia-reperfusion group no significant difference (P>0.05).And the potentilla anserina extract pretreatment 1 hour of 0.5ng/ml and 50ng/ml concentration, all fail to strengthen the hepatocellular vigor of ischemical reperfusion injury (P>0.05), difference also not statistically significant (P>0.05) between its Activity of hepatocytes and normal saline group.The results are shown in Table 1(
▲represent and compare with N group, P<0.05, difference has statistical significance;
★represent and compare with IR group and NS group, P<0.05, difference has statistical significance).
2, the cell proliferation of potentilla anserina extract pretreatment on hepatocyte ischemical reperfusion injury and the impact of toxicity
Compared with normal group, in-vitro simulated ischemical reperfusion injury makes cell mortality >90%.Give 5ng/ml potentilla anserina extract pretreatment 1 hour, heptocellular death rate declines about 35%, difference has statistical significance (P<0.05), but its cell mortality is still than normal group height about 50%, and difference has statistical significance (P<0.05); And though normal saline group heptocellular death rate slightly declines (being about 8%) than ischemia-reperfusion group, no significant difference.The potentilla anserina extract pretreatment of 0.5ng/ml and 50ng/ml concentration 1 hour, heptocellular death rate reduces by 6% and 13% respectively than ischemia-reperfusion group, but no significant difference (P>0.05); And close with the heptocellular death rate of normal saline group (P>0.05).The results are shown in Table 2(
▲represent and compare with N group, P<0.05, difference has statistical significance;
★represent and compare with IR group and NS group, P<0.05, difference has statistical significance).
3, potentilla anserina extract pretreatment is on the impact of ischemical reperfusion injury hepatocyte liver function index
Compare with normal group, the ALT level in ischemical reperfusion injury group cell culture supernatant has no obvious rising (P>0.05); And AST, LDH level and AST/ALT ratio significantly raise, difference has statistical significance (P<0.05).Give 5ng/ml potentilla anserina extract pretreatment 1 hour, AST, LDH level in liver cell culture supernatant and AST/ALT ratio comparatively ischemia-reperfusion group and normal saline group reduce, but still compared with normal group high (P<0.05); And ALT level compares with ischemia-reperfusion group and normal saline group, no significant difference (P>0.05).ALT, AST, LDH level in normal saline group cell culture supernatant and no significant difference (P>0.05) between AST/ALT ratio and ischemia-reperfusion group.0.5ng/ml and 50ng/ml potentilla anserina extract pretreatment 1 hour, all fails to make AST, LDH level in liver cell culture supernatant reduce (P>0.05).The results are shown in Table 3(
▲represent and compare with N group, P<0.05, difference has statistical significance;
★represent and compare with IR group and NS group, P<0.05, difference has statistical significance).
This research shows, and potentilla anserina extract (RE2 group) makes hepatocyte strengthen the tolerance of the ischemical reperfusion injury experienced afterwards, and show as Activity of hepatocytes and strengthen, cell mortality reduces.Above result prompting, potentilla anserina extract pretreatment can alleviate human liver cell ischemical reperfusion injury, plays hepatocytoprotection.
Table 1 mtt assay detects potentilla anserina extract pretreatment to the impact of ischemical reperfusion injury Activity of hepatocytes
The breeding of table 2 LDH method for releasing hepatocyte and toxicity detection by quantitative cell mortality
The change (n=5) of ALT, AST, LDH level in table 3 liver cell culture supernatant
Embodiment 4: the preparation of tablet
By embodiment 1 method first obtained extract, add excipient, pelletizing press sheet in itself and excipient weight than the ratio for 1:10.
Embodiment 5: the preparation of oral liquid
By embodiment 1 method first obtained extract, oral liquid method for making makes oral liquid routinely.
Embodiment 6: the preparation of capsule or granule
By embodiment 1 method first obtained extract, add excipient in itself and excipient weight than the ratio for 1:9, make capsule or granule.
Embodiment 7: the preparation of injection
Obtain extract by embodiment 1 method, inject with water, fine straining, injection is made in embedding sterilizing.
Embodiment 8: the preparation of aseptic powder injection
By embodiment 1 method first obtained extract, be dissolved in sterile water for injection, stirring makes molten, filters with aseptic suction funnel, more aseptic fine straining, and be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains injectable powder.
Claims (6)
1. a Radix potentillae anserinae rhizome extract, is characterized in that described extract is prepared by following methods:
A the Radix potentillae anserinae rhizome of drying is pulverized by (); B () extracts with 75% alcohol heat reflux, merge extractive liquid, is concentrated into and obtains concentrated solution without alcohol taste; C () uses petroleum ether, ethyl acetate and water saturated n-butanol extraction successively to step (b) concentrated solution, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; D () n-butyl alcohol extract macroporous resin enrichment, first with 10% alcohol flushing, 8 column volume removing polysaccharide, then uses 65% ethanol elution, 10 column volumes, collects 65% eluent, concentrating under reduced pressure, spraying dry.
2. extract according to claim 1, is characterized in that: macroporous resin described in step (d) is AB-8 type macroporous resin.
3. Radix potentillae anserinae rhizome extract according to claim 1, is characterized in that: the liquid phase analysis method of described extract is:
Chromatographic column: Diamonsil C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% phosphoric acid solution (triethylamine adjust ph to 6.2);
Gradient elution program: 0.01 ~ 70min, A 20% → 80%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 240nm; Column temperature: 30 DEG C;
Sample size: 10 μ L.
4. pharmaceutical preparation, is characterized in that: the Radix potentillae anserinae rhizome extract according to claim 1 containing treatment effective dose and pharmaceutically acceptable carrier.
5. the application of Radix potentillae anserinae rhizome extract according to claim 1 in the liver-protective medicine of preparation.
6. the application of pharmaceutical preparation according to claim 4 in the liver-protective medicine of preparation.
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| CN105111269A (en) * | 2015-10-09 | 2015-12-02 | 杭州启澄科技有限公司 | Novel limonin compound as well as preparation method and medical application thereof |
| CN105125652A (en) * | 2015-10-12 | 2015-12-09 | 刘高志 | Tsinling larkspur root extract and preparing method and medical application thereof |
| CN105125624A (en) * | 2015-10-17 | 2015-12-09 | 淄博夸克医药技术有限公司 | Euphorbia tithymaloides extract and preparing method and medical application thereof |
| CN106581087A (en) * | 2015-10-19 | 2017-04-26 | 淄博夸克医药技术有限公司 | Sabina chinensis leaf extract and preparation method and medical applications thereof |
| CN105168349A (en) * | 2015-10-21 | 2015-12-23 | 淄博夸克医药技术有限公司 | Salvia plebeian extract as well as preparation method and medical application thereof |
| CN105147907A (en) * | 2015-10-22 | 2015-12-16 | 淄博夸克医药技术有限公司 | Dasheen flower extract, as well as preparation method and medicinal application thereof |
| CN105327127A (en) * | 2015-10-29 | 2016-02-17 | 田丽华 | Smallflower epipactis flower extract resisting brain aging and preparing method of smallflower epipactis flower extract |
| CN105193999A (en) * | 2015-10-29 | 2015-12-30 | 王淑英 | Preparation method of extract of all grass of capituligrerae and medical application |
| CN105616424A (en) * | 2015-12-27 | 2016-06-01 | 温州统益生物医药科技有限公司 | Application of triterpenoid in preparation of drug for protecting liver |
| CN105481878A (en) * | 2015-12-31 | 2016-04-13 | 胡文杰 | Novel limonin compound and preparation method and medical application thereof |
| CN105768099A (en) * | 2016-03-04 | 2016-07-20 | 吴正锋 | Blood sugar-reducing healthcare food with addition of rhizoma anemarrhenae extract |
| CN115487232A (en) * | 2022-09-29 | 2022-12-20 | 西藏自治区农牧科学院农产品开发与食品科学研究所 | Extraction and preparation process of potentilla anserina total flavonoids |
| CN115487232B (en) * | 2022-09-29 | 2024-04-02 | 西藏自治区农牧科学院农产品开发与食品科学研究所 | Extraction and preparation process of potentilla anserina total flavonoids |
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