CN105168305A - Hungaian morningglory root extract for treating gouty arthritis - Google Patents
Hungaian morningglory root extract for treating gouty arthritis Download PDFInfo
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Abstract
The invention discloses a hungaian morningglory root extract for treating gouty arthritis. A preparation method of the extract comprises the following steps: (a) according to parts by weight, mixing and grinding 80-90 parts of dried hungaian morningglory root and 10-20 parts of dried chrysanthemum in terms of 100 parts of medicinal materials, hot-refluxing and extracting by virtue of an ethanol solution of 55%, combining extracting solutions, and concentrating until no alcohol smell exists so as to obtain a concentrated ethanol extracting solution; (b) diluting the concentrated ethanol extracting solution obtained from the step (a) with water, sequentially extracting by virtue of petroleum ether, ethyl acetate and water-saturated normal butanol, decompressing and concentrating so as to respectively obtain a petroleum ether extract, an ethyl acetate extract and a normal butanol extract; and (c) dissolving the normal butanol extract in water, filtering and enriching active ingredients by virtue of macroporous resin; firstly, flushing 8 column volumes with ethanol of 10% to remove high-polarity ingredients, and then eluting 12 column volumes with ethanol of 70%; collecting an eluent of ethanol of 70%, decompressing and concentrating so as to obtain the extract. The extract can be used for treating gouty arthritis and the extract can be developed as medicines for treating the gouty arthritis.
Description
Technical field
The present invention relates to Chinese medicine extract field, be specifically related to a kind of Ipomoea hungaiensis Lingelsh. Et Borza. extract and the pharmaceutical preparation containing this extract, this extract can be applied to the medicine of preparation treatment gouty arthritis.
Background technology
Medical material Ipomoea hungaiensis Lingelsh. Et Borza. is the tuber of Convolvulaceae Merremia plant Ipomoea hungaiensis Lingelsh. Et Borza..Have another name called native egg (" mansion, Zun Yi will "), Yunnan Ipomoea hungaiensis Lingelsh. Et Borza. (" Zhiwu Mingshi Tukao "), Ipomoea hungaiensis Lingelsh. Et Borza. (" Chinese medicine shape sex experience differential method "), laterite melon (" the southern regions of the Yunnan Province book on Chinese herbal medicine " arranges this), Odontobutis obscura (T. Et S.), red natural pond, mountain etc.
The plant that former plant Ipomoea hungaiensis Lingelsh. Et Borza. (formal name used at school: Merremiahungaiensis) is Convolvulaceae Merremia, is the endemic plant of China, is distributed in the ground such as the Guizhou of China's Mainland, Sichuan, Yunnan.This plant is perennial voluble herb, underground tool tuber, spherical or ovum shape, and 2-3 concatenates sometimes, epidermis bronzing, crineous or meat white, starch-containing and have emulsus mucus.Stem is elongated, cylindrical, has thin rib, and most turn-knob, without hair.Elliptic leaf shape, avette or Long Circle, long 2.5-11.5 centimetre, wide 1.2-5 centimetre, top is blunt, nick, gradually point or sharp point, the little mucro of tool, the blunt circle of base portion or wedge shape or micro-in heart-shaped, edge is micro-erodes shape or nearly full edge, and two sides is without hair, and only blade base is by minority echinid, have lateral vein 5-6(-7) right, arteries and veins is with purple sometimes; The long 0.8-3.5 centimetre of petiole, by pubescence.Cyme axil is raw, the long 2-6 centimetre of peduncle, without hair, raw 2-3 or several flowers, or single Semen arachidis hypogaeae axil; Bract is little, flakey, is about 1 millimeter; The long 0.5-2 centimetre of bennet, more sturdy than peduncle, without hair; Sepal is isometric, or foreign side 2 is slightly short, and oval, top is blunt, edge dry film matter, the long 0.7-1.4 centimetre of outer sepal, and the long 1.2-1.5 centimetre of interior sepal, all without hair; Corolla is yellow, funnel-form, long 3.5-6 centimetre, and be with top by faint yellow pubescence in lobe, all the other are without hair; Stamen is Length discrepancy slightly, and filigree base portion expands, by hair; Floral disc ring-type; Ovary is coniform, Room 2, and without hair, stigma 2 is spherical.Capsule Long Circle, high 1-1.3 centimetre, 4 lobes split.The long 5.5-7 millimeter of seed, extremely close by pitchy fine hair.
Summary of the invention
The object of the present invention is to provide a kind of Ipomoea hungaiensis Lingelsh. Et Borza. extract, its preparation method and liquid phase analysis method, the pharmaceutical preparation containing this extract and utilize this extract to prepare the purposes of medicine for the treatment of gouty arthritis.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
A kind of Ipomoea hungaiensis Lingelsh. Et Borza. extract, this extract is prepared by following methods:
A () by weight, every 100 parts of medical materials comprise dry Ipomoea hungaiensis Lingelsh. Et Borza. 80 ~ 90 parts and dry Flos Chrysanthemi 10 ~ 20 parts, co-grinding, and with 55% alcoholic solution circumfluence distillation, merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution without alcohol taste; B (), by step (a) gained ethanol extraction concentrated solution dilute with water, is used petroleum ether, ethyl acetate and water saturated n-butanol extraction successively, concentrating under reduced pressure, is obtained petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; C () n-butyl alcohol extract water dissolution, filters, uses macroporous resin enrichment active component, first remove large polar component with 10% alcohol flushing, 8 column volumes, then use 70% ethanol elution, 12 column volumes, collect 70% eluent, concentrating under reduced pressure and get final product.
In order to be controlled the differences between batches of extract prepared by different batches by the method setting up HPLC finger printing, the liquid phase analysis method of described extract is:
Chromatographic column: AgilentZorbaxExtent-C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.5% phosphoric acid solution;
Gradient elution program: 0.01 ~ 5min, A10% → 15%; 5 ~ 10min, A15% → 48%; 10 ~ 25min, A48% → 78%; 25 ~ 30min, A78% → 10%;
Flow rate of mobile phase: 1.0mLmin
-1; Determined wavelength: 270nm; Column temperature: 35 DEG C; Sample size: 10 μ L.
Pharmaceutical preparation, the described extract containing treatment effective dose and pharmaceutically acceptable carrier.
The application of described extract in the medicine of preparation treatment gouty arthritis.
The application of described pharmaceutical preparation in the medicine of preparation treatment gouty arthritis.
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 Ipomoea hungaiensis Lingelsh. Et Borza. 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 the Ipomoea hungaiensis Lingelsh. Et Borza. of drying and Flos Chrysanthemi extracts, remove impurity, enrichment process, the extract with treatment gouty arthritis can be obtained; Liquid phase analysis method provided by the invention may be used for the finger printing setting up this extract, for controlling the differences between batches of extract prepared by different batches.
Accompanying drawing explanation
Fig. 1 is Ipomoea hungaiensis Lingelsh. Et Borza. extractive 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.
Embodiment 1: 1. Ipomoea hungaiensis Lingelsh. Et Borza. extract is prepared (85 parts of Ipomoea hungaiensis Lingelsh. Et Borza., 15 parts of Flos Chrysanthemis)
Crude drug source: Ipomoea hungaiensis Lingelsh. Et Borza. and Flos Chrysanthemi are purchased from Hui nationality's Chinese Medicinal Materials Markets.
Main agents: 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.
Preparation method: by dry for 8.5kg Ipomoea hungaiensis Lingelsh. Et Borza. rhizome and the dry Flos Chrysanthemi co-grinding of 1.5kg, with 55% alcoholic solution circumfluence distillation (25L × 3 time), merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution (2L) without alcohol taste; Gained ethanol extraction concentrated solution is diluted with water to 3L, petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated n-butyl alcohol (3L × 3 time) is used to extract successively, concentrating under reduced pressure, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract 232g respectively; (c) n-butyl alcohol extract 1.5L water dissolution, medical absorbent cotton filters, with AB-8 macroporous resin (2kg, column volume 1.5L) deposition activity composition, first use 10% alcohol flushing, 8 column volumes (12L) to remove large polar component, then use 70% ethanol elution, 12 column volumes (18L), collect 70% eluent, concentrating under reduced pressure, obtains Ipomoea hungaiensis Lingelsh. Et Borza. extract and is about 150g.
Embodiment 2: 2. Ipomoea hungaiensis Lingelsh. Et Borza. extract is prepared (80 parts of Ipomoea hungaiensis Lingelsh. Et Borza., 20 parts of Flos Chrysanthemis)
Preparation method: by dry for 8.0kg Ipomoea hungaiensis Lingelsh. Et Borza. rhizome and the dry Flos Chrysanthemi co-grinding of 2.0kg, with 55% alcoholic solution circumfluence distillation (25L × 3 time), merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution (2L) without alcohol taste; Gained ethanol extraction concentrated solution is diluted with water to 3L, petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated n-butyl alcohol (3L × 3 time) is used to extract successively, concentrating under reduced pressure, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract 232g respectively; (c) n-butyl alcohol extract 1.5L water dissolution, medical absorbent cotton filters, with AB-8 macroporous resin (2kg, column volume 1.5L) deposition activity composition, first use 10% alcohol flushing, 8 column volumes (12L) to remove large polar component, then use 70% ethanol elution, 12 column volumes (18L), collect 70% eluent, concentrating under reduced pressure, obtains Ipomoea hungaiensis Lingelsh. Et Borza. extract and is about 150g.
Embodiment 3: 3. Ipomoea hungaiensis Lingelsh. Et Borza. extract is prepared (90 parts of Ipomoea hungaiensis Lingelsh. Et Borza., 10 parts of Flos Chrysanthemis)
Preparation method: by dry for 9.0kg Ipomoea hungaiensis Lingelsh. Et Borza. rhizome and the dry Flos Chrysanthemi co-grinding of 1.0kg, with 55% alcoholic solution circumfluence distillation (25L × 3 time), merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution (2L) without alcohol taste; Gained ethanol extraction concentrated solution is diluted with water to 3L, petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated n-butyl alcohol (3L × 3 time) is used to extract successively, concentrating under reduced pressure, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract 232g respectively; (c) n-butyl alcohol extract 1.5L water dissolution, medical absorbent cotton filters, with AB-8 macroporous resin (2kg, column volume 1.5L) deposition activity composition, first use 10% alcohol flushing, 8 column volumes (12L) to remove large polar component, then use 70% ethanol elution, 12 column volumes (18L), collect 70% eluent, concentrating under reduced pressure, obtains Ipomoea hungaiensis Lingelsh. Et Borza. extract and is about 150g.
Embodiment 4: 4. Ipomoea hungaiensis Lingelsh. Et Borza. extract is prepared (75 parts of Ipomoea hungaiensis Lingelsh. Et Borza., 25 parts of Flos Chrysanthemis)
Preparation method: by dry for 7.5kg Ipomoea hungaiensis Lingelsh. Et Borza. rhizome and the dry Flos Chrysanthemi co-grinding of 2.5kg, with 55% alcoholic solution circumfluence distillation (25L × 3 time), merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution (2L) without alcohol taste; Gained ethanol extraction concentrated solution is diluted with water to 3L, petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated n-butyl alcohol (3L × 3 time) is used to extract successively, concentrating under reduced pressure, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract 232g respectively; (c) n-butyl alcohol extract 1.5L water dissolution, medical absorbent cotton filters, with AB-8 macroporous resin (2kg, column volume 1.5L) deposition activity composition, first use 10% alcohol flushing, 8 column volumes (12L) to remove large polar component, then use 70% ethanol elution, 12 column volumes (18L), collect 70% eluent, concentrating under reduced pressure, obtains Ipomoea hungaiensis Lingelsh. Et Borza. extract and is about 150g.
Embodiment 5: 5. Ipomoea hungaiensis Lingelsh. Et Borza. extract is prepared (95 parts of Ipomoea hungaiensis Lingelsh. Et Borza., 5 parts of Flos Chrysanthemis)
Preparation method: by dry for 9.5kg Ipomoea hungaiensis Lingelsh. Et Borza. rhizome and the dry Flos Chrysanthemi co-grinding of 0.5kg, with 55% alcoholic solution circumfluence distillation (25L × 3 time), merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution (2L) without alcohol taste; Gained ethanol extraction concentrated solution is diluted with water to 3L, petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated n-butyl alcohol (3L × 3 time) is used to extract successively, concentrating under reduced pressure, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract 232g respectively; (c) n-butyl alcohol extract 1.5L water dissolution, medical absorbent cotton filters, with AB-8 macroporous resin (2kg, column volume 1.5L) deposition activity composition, first use 10% alcohol flushing, 8 column volumes (12L) to remove large polar component, then use 70% ethanol elution, 12 column volumes (18L), collect 70% eluent, concentrating under reduced pressure, obtains Ipomoea hungaiensis Lingelsh. Et Borza. extract and is about 150g.
Embodiment 6: liquid-phase chromatographic analysis
Need testing solution is prepared: in the brown volumetric flask of extract 5mg to 50mL that Example 1 method is obtained, add 30mL10% acetonitrile solution ultrasonic dissolution, after being cooled to room temperature, continue to add 10% acetonitrile solution standardize solution.Analytical method is as follows:
High performance liquid chromatograph: Agilent1260, binary pump;
Chromatographic column: AgilentZorbaxExtent-C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.5% phosphoric acid solution;
Gradient elution program: 0.01 ~ 5min, A10% → 15%; 5 ~ 10min, A15% → 48%; 10 ~ 25min, A48% → 78%; 25 ~ 30min, A78% → 10%;
Flow rate of mobile phase: 1.0mLmin
-1; Determined wavelength: 270nm; Column temperature: 35 DEG C; Sample size: 10 μ L.
Analyze with the Ipomoea hungaiensis Lingelsh. Et Borza. extract of 10 batches of preparation, carry out chromatographic peak coupling, 1 ~ No. 12 peak all occurs in 10 batch sample chromatograms as a result.Therefore demarcate 12 peaks for total chromatographic peak, set up the HPLC quality control collection of illustrative plates of this extract accordingly, the results are shown in Figure 1.This Ipomoea hungaiensis Lingelsh. Et Borza. extract pharmacologically active of 10 batches is similar.
Embodiment 7: extract pharmacological testing
One, materials and methods
1.1 medicines: Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 5. obtain according to embodiment 1 ~ 5 respectively.Colchicines tablets, lot number 100206, specification 0.5mg/s, Banna Pharmacy Industry Co., Ltd., Xishuangbanna; Get 6 colchicine, grinding, adding distil water is to 200mL, and it is 0.15mg/mL colchicine solution that mixing is mixed with concentration.Crystallite Monosodium urate (MSU), with reference to Coderre method, 194mL distilled water adds 6mL1mol/LNaOH, boil, add 1g uric acid, adjust pH to 7.2 with 1mL1mol/LHCl, stir cooling, 4 DEG C of Refrigerator store 24h, remove supernatant, with filter paper, precipitate moisture is blotted, put into 70 DEG C of drying baker and dry 2h, take out, scrape powder, put into mortar pulverization, sieve with the metallic screen in 250 μm, aperture, make MSU powder for subsequent use.
1.2 reagent: uric acid (UA) test kit, lot number: P100121, Shanghai Foxing Changzheng medical science Co., Ltd; Nitric oxide (NO) test kit, lot number: 20110119, Bioengineering Research Institute is built up in Nanjing; Total protein (TP biuret method) test kit, lot number: 110601, thing development in science and technology company limited of Beijing Rui Ji Hang Seng.
1.3 animals: cleaning grade SD rat, male, body weight 200 ~ 220g, is provided by Zhejiang Academy of Medical Sciences, the quality certification number: SCXK(Zhejiang) 20080033.
1.4 instruments: LP123 electronic balance, weighing apparatus factory of Changshu City, made in CCCP 00000153; YLS-7A rat toes capacity measurer, Shandong Academy of Medical Sciences's equipment station; HEMAVET950 animal blood analyser, ADrewScientificGroupCo; Prestige figure SELECTRA-E full automatic biochemical apparatus, Dutch Rittal GmbH.
1.5 experimental techniques and Testing index
1.5.1 animal grouping and administration: rat is divided into 18 groups at random: Normal group, model control group, positive controls (colchicine 0.0015gkg
-1), Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 5. respectively establish 3 dosage group (100mgkg
-1, 200mgkg
-1, 300mgkg
-1), often organize 10.Administration 5d before modeling, 1 time/d, continues administration after modeling.By reagent group respectively gavage give the Ipomoea hungaiensis Lingelsh. Et Borza. extract of various dose, Normal group gavage gives the water of respective volume, and positive control drug gavage gives the colchicine medicinal liquid of corresponding dosage.
1.5.2 modeling: get 1000mgMSU and add 9mL normal saline, then add lmL tween 80, heated and stirred, (concentration is 100mgmL to be mixed with 10mLMSU suspension
-1), ankle joint flexing after Rat Right after administration 30min, touch with hands and can lay one's hand on and two apophysis portions, after 75% medical alcohol partly sterilised, insert tibialis inboard leg with No. 6 entry needles ankle joint dorsal part on rear side of left and right from 45 ° of directions, 0.1mLMSU suspension is injected into ankle joint chamber, prepares acute gouty arthritis model, stretch song, rotary motion 2min, rats in normal control group physiological saline solution replaces MSU suspension.
1.5.3 Articular swelling: mark with a circle around ankle joint marking pen in outside, Rat Right metapedes ankle joint trident place before modeling, measures Rat Right foot to mark Volume of Displacement twice, averages as arthritic volume before administration; After modeling, 1h, 4h, 6h, 24h measure Rat Right foot to mark Volume of Displacement, by formulae discovery swelling.
Ankle joint Volume of Displacement (mL) before ankle joint Volume of Displacement (mL) after swelling (mL)=cause is scorching-cause is scorching.
1.5.4 hemanalysis: modeling 24h, after last administration 30min, after having surveyed the Volume of Displacement of appointed part, from the blood sampling of rat retroorbital venous clump 5mL, wherein natural blood coagulation 2h, 3500rmin under 4mL room temperature
-1centrifugal 10min, draw serum, reference reagent box description measures serum UA, NO content in accordance with the law; Another 1mL adds 10 μ LEDTA anticoagulants, uses animal blood analyser to measure leukocyte, neutrophilic granulocyte, lymphocyte quantity in accordance with the law.
1.5.5 biochemical and histological indices: on cellophane with sharp keen blade from Rat Right ankle joint dorsal part fast shut-off fascia, cut whole soft tissue and cartilage that ankle joint trident place removes bone, to weigh and the ice normal saline adding 9 times amount makes 10% tissue homogenate, 3500rmin
-1centrifugal 10min, draw supernatant, reference reagent box description measures total protein, NO content in accordance with the law; Spleen, kidney are got in execution, weigh, and calculate spleen system number and kidney system number.
1.6 statistical analysiss: measurement data data result represents with x ± s, compare between group and adopt t inspection.
Two, result
2.1 Ipomoea hungaiensis Lingelsh. Et Borza. extract is on the impact of gouty arthritis rat articular swelling
Compared with Normal group, model control group ankle swelling in rat degree all significantly raises (P<0.01) in 1 ~ 6h after modeling; 1h after modeling, compared with model control group, colchicine group, Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. each dosage group ankle swelling in rat degree all significantly decline (P<0.01); 4h after modeling, compared with model control group, colchicine group, Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. each dosage group ankle swelling in rat degree all significantly decline (P<0.01); 6h after modeling, compared with model control group, colchicine group, Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. high dose group ankle swelling in rat degree all significantly decline (P<0.05); Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. in, low dose group ankle swelling in rat degree also has the trend of reduction; Ipomoea hungaiensis Lingelsh. Et Borza. extract 4. ~ 5. each dosage group in modeling 24h compared with model group all without significant difference.The results are shown in Table 1(note: compare with model group, * P<0.05, * * P<0.01).
Table 1 Ipomoea hungaiensis Lingelsh. Et Borza. extract in modeling 24h on the impact (x ± s, n=10) of gouty arthritis ankle swelling in rat degree
2.2 impacts on gouty arthritis rat spleen body index, kidney body index
Compared with Normal group, model group kidney body index obviously reduces, and spleen body index obviously raises (P<0.05,0.01); Compared with model control group, colchicine group kidney of rats body index significantly raises, and spleen body index significantly reduces (P<0.05); Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. high dose group kidney body index obviously rise (P<0.01), Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. each dosage group spleen body index all have decline (P<0.05,0.01) in various degree; Ipomoea hungaiensis Lingelsh. Et Borza. extract 4. ~ 5. each dosage group on the impact of rat spleen body index, kidney body index and model group without marked difference.In Table 2(note: compare with model control group, * P<0.05, * * P<0.01).
Table 2 Ipomoea hungaiensis Lingelsh. Et Borza. extract is on the impact (x ± s, n=10) of gouty arthritis rat spleen index, renal index
2.3 impacts on leukocyte, neutrophilic granulocyte and lymphocyte level in gouty arthritis rat whole blood
Compared with Normal group, the leukocyte of model control group, neutrophilic granulocyte and lymphocyte quantity have the trend of rising; Compared with model group colchicine and Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. the leukocyte of high dose group rat, neutrophilic granulocyte close lymphocyte quantity and obviously reduce (P<0.05,0.01), Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. all the other each dosage groups all have reduction trend; Ipomoea hungaiensis Lingelsh. Et Borza. extract 4. ~ 5. each dosage group and model control group be than without significant difference.In Table 3(note: compare with model control group, * P<0.05, * * P<0.01).
Table 3 Ipomoea hungaiensis Lingelsh. Et Borza. extract is on gouty arthritis rat leukocyte, neutrophilic granulocyte and lymphocyte impact (x ± s, n=10)
2.4 impacts on gouty arthritis rat blood serum UA level
Each group of rat serum uric acid level result result display, compared with Normal group, model group Level of Serum Uric Acid obviously raises (P<0.05); Compared with model group, Ipomoea hungaiensis Lingelsh. Et Borza. extract 1. ~ 3. in high dose group rat serum uric acid obviously decline (P<0.05); Ipomoea hungaiensis Lingelsh. Et Borza. extract 4. ~ 5. each dosage group and model group be than there are no significant difference.In Table 4(note: compare with model control group, * P<0.05, * * P<0.01).
Table 4 Ipomoea hungaiensis Lingelsh. Et Borza. extract is on the impact (x ± s, n=10) of gouty arthritis rat blood serum UA
Above-mentioned result of the test shows, Ipomoea hungaiensis Lingelsh. Et Borza. extract has therapeutical effect to rat gouty arthritis.The content of Ipomoea hungaiensis Lingelsh. Et Borza. extract ZHONGJUHUA is most important to its performance therapeutical effect.
Embodiment 8: the preparation of tablet
Obtain extract by embodiment 1 method, add excipient, pelletizing press sheet in itself and excipient weight than the ratio for 1:10.
Embodiment 9: the preparation of oral liquid
By embodiment 1 method first obtained extract, oral liquid method for making makes oral liquid routinely.
Embodiment 10: 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 11: the preparation of injection
Obtain extract by embodiment 1 method, inject with water, fine straining, injection is made in embedding sterilizing.
Embodiment 12: 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. an Ipomoea hungaiensis Lingelsh. Et Borza. extract, is characterized in that described extract is prepared by following methods:
A () by weight, every 100 parts of medical materials comprise dry Ipomoea hungaiensis Lingelsh. Et Borza. 80 ~ 90 parts and dry Flos Chrysanthemi 10 ~ 20 parts, co-grinding, and with 55% alcoholic solution circumfluence distillation, merge extractive liquid, is concentrated into and obtains ethanol extraction concentrated solution without alcohol taste; B (), by step (a) gained ethanol extraction concentrated solution dilute with water, is used petroleum ether, ethyl acetate and water saturated n-butanol extraction successively, concentrating under reduced pressure, is obtained petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; C () n-butyl alcohol extract water dissolution, filters, uses macroporous resin enrichment active component, first remove large polar component with 10% alcohol flushing, 8 column volumes, then use 70% ethanol elution, 12 column volumes, collect 70% eluent, concentrating under reduced pressure and get final product.
2. extract according to claim 1, is characterized in that: macroporous resin described in step (c) is AB-8 type macroporous resin.
3. extract according to claim 1, is characterized in that: the liquid phase analysis method of described extract is:
Chromatographic column: AgilentZorbaxExtent-C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.5% phosphoric acid solution;
Gradient elution program: 0.01 ~ 5min, A10% → 15%; 5 ~ 10min, A15% → 48%; 10 ~ 25min, A48% → 78%; 25 ~ 30min, A78% → 10%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 270nm;
Column temperature: 35 DEG C;
Sample size: 10 μ L.
4. pharmaceutical preparation, is characterized in that: the extract according to claim 1 containing treatment effective dose and pharmaceutically acceptable carrier.
5. the application of extract according to claim 1 in the medicine of preparation treatment gouty arthritis.
6. the application of pharmaceutical preparation according to claim 4 in the medicine of preparation treatment gouty arthritis.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105535039A (en) * | 2015-12-28 | 2016-05-04 | 吴芊葭 | Japanese nanocnide herb extract and medical application thereof in treatment of gouty arthritis |
| CN107343893A (en) * | 2017-07-31 | 2017-11-14 | 海南医学院 | Application of Hainan Stephania epigaea extract on treatment urarthritis preparation is prepared |
| CN113237972A (en) * | 2021-04-30 | 2021-08-10 | 江苏卫生健康职业学院 | Traditional Chinese medicine composition with gout resisting effect and fingerprint detection method thereof |
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| CN105535039A (en) * | 2015-12-28 | 2016-05-04 | 吴芊葭 | Japanese nanocnide herb extract and medical application thereof in treatment of gouty arthritis |
| CN107343893A (en) * | 2017-07-31 | 2017-11-14 | 海南医学院 | Application of Hainan Stephania epigaea extract on treatment urarthritis preparation is prepared |
| CN113237972A (en) * | 2021-04-30 | 2021-08-10 | 江苏卫生健康职业学院 | Traditional Chinese medicine composition with gout resisting effect and fingerprint detection method thereof |
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