CN105030925A - Plant extract for treating blood stasis disease and pharmaceutic preparation - Google Patents
Plant extract for treating blood stasis disease and pharmaceutic preparation Download PDFInfo
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Abstract
The invention discloses a plant extract for treating the blood stasis disease. A plant extract preparing method includes the steps that a, by weight, 80-90 parts of dried corydalis tuber tuber and 10-20 parts of dried rheum officinale are contained in each 100 parts of medicinal materials to be mixed and smashed, heat reflux extraction is conducted with an ethanol solution with the concentration being 60%, the extracting solutions are combined to be concentrated till no alcohol taste exists, and a concentrated solution obtained through ethanol extraction is obtained; b, the concentrated solution obtained through ethanol extraction in step a is diluted with water, petroleum ether, ethyl acetate and water-saturated n-butyl alcohol are sequentially used for extraction, vacuum concentration is conducted, and a petroleum ether extract, a ethyl acetate extract and a n-butyl alcohol extract are obtained; c, the n-butyl alcohol extract is dissolved with water to be filtered, active ingredients are enriched through macroporous resin, eight column volumes are washed through ethyl alcohol with the concentration being 10% to remove high polar molecules, twelve column volumes are washed through ethyl alcohol with the concentration being 70%, eluant with the concentration being 70% is collected, and the plant extract is obtained after vacuum concentration. The plant extract can be applied to the blood stasis disease and developed into medicine for treating the blood stasis disease.
Description
Technical field
The present invention relates to Chinese medicine extract field, be specifically related to a kind of Rhizoma Corydalis extract and the pharmaceutical preparation containing this extract, this extract can be applied to the medicine of preparation treatment blood stasis disease.
Background technology
Rhizoma Corydalis is the dry tuber of papaveraceae plant corydalis CorydalisyanhusuoW.T.Wang, is pain relieving good medicine ancient simply, and also known as Rhizoma Corydalis, Rhizoma Corydalis, be one of famous eight Zhe's, Dongyang, main product Zhejiang Province, Pan'an one are with.Rhizoma Corydalis has significant analgesia, calmness and syngignoscism, has good clinical effectiveness to various diseases such as coronary heart disease, arrhythmia, gastric ulcers.Just because of Rhizoma Corydalis Be very effective, clinical practice is extensive, in " Chinese Pharmacopoeia " version in 2010, employs Rhizoma Corydalis close in the compound preparation of 30%.
Rhizoma Corydalis main component is alkaloid, is mainly tertiary amine, Amine quarter alkaloid.The amount of tertiary amines alkaloid in crude drug is about 0.65%, and quaternary amines alkaloid (as Corydaline, B prime) is about 0.3%.Up to the present, from Rhizoma Corydalis, be separated the alkaloids composition obtained and about have 30 kinds.From the tuber of Rhizoma Corydalis, propose alkaloid more than 10 altogether plant, wherein there is corydaline (Corydaline) through what identify, dl-tetrahydropalmatine (dl-Tetrahydropalmatine), protopine (Protopine), L-Tetrahydrocoptisine, dl-Tetrahydrocoptisine, L-tetrahydrocolumbamine, corybulbine (Corybulbine), β-homochelidonie (β-Homoche-lidonine), coptisine (Coptisine), 13-Methylpalmatine (De-hydrocorydaline), also has corydalmine (Corydalmine and corybulbine), dehydrogenation corydalmine etc.Except alkaloid, still containing much starch in Rhizoma Corydalis, a small amount of lymphatic temperament, resin, volatile oil, separately containing inorganic microelement.Also containing polysaccharide, hydroxystreptomycin (reticulin), stigmasterol, sitosterol, oleic acid, linoleic acid, linolenic acid, Fumaric acid, 10-29 carbon alcohol etc.
Alkaloid in Rhizoma Corydalis has very strong analgesia, calmness, blood pressure lowering and antiarrhythmic effect.At present, much new research shows, Rhizoma Corydalis also has other physiologically actives widely, as resisted myocardial ischemia, anti-experimental character gastric ulcer, antitumor, antioxidation, to protect the liver.
Summary of the invention
The object of the present invention is to provide and be a kind ofly used for the treatment of the plant extract of blood stasis disease, its preparation method and liquid phase analysis method containing Rhizoma Corydalis, the pharmaceutical preparation containing this extract and utilize this extract to prepare the purposes of the medicine for the treatment of blood stasis disease.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
A kind of plant extract being used for the treatment of blood stasis disease, this extract is prepared by following methods: (a) by weight, every 100 parts of medical materials comprise dry corydalis tuber 80 ~ 90 parts and dry Radix Et Rhizoma Rhei 10 ~ 20 parts, co-grinding, use 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 5 ~ 15% alcohol flushing, 7 ~ 9 column volumes, use 65 ~ 75% ethanol elution, 11 ~ 13 column volumes again, collect 65 ~ 75% eluents, concentrating under reduced pressure and get final product.
Further, in the described alcoholic solution circumfluence distillation of step (a), ethanol solution concentration is 55 ~ 65%.
Further, in the described alcoholic solution circumfluence distillation of step (a), ethanol solution concentration is 60%.
Further, macroporous resin described in step (c) is AB-8 type macroporous resin.
Further, step (c) is: n-butyl alcohol extract water dissolution, filter, with AB-8 type macroporous resin enrichment active component, first remove large polar component with 10% alcohol flushing, 8 column volumes, use 70% ethanol elution, 12 column volumes again, 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 blood stasis disease.
The application of described pharmaceutical preparation in the medicine of preparation treatment blood stasis disease.
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 Rhizoma Corydalis 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 corydalis tuber of drying and Radix Et Rhizoma Rhei extracts, remove impurity, enrichment process, the extract with treatment blood stasis disease 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 Rhizoma Corydalis 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.
Embodiment 1: 1. Rhizoma Corydalis extract is prepared (85 parts of Rhizoma Corydalis, 15 parts of Radix Et Rhizoma Rhei)
Crude drug source: Rhizoma Corydalis and Radix Et Rhizoma Rhei 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 corydalis tuber and the dry Radix Et Rhizoma Rhei co-grinding of 1.5kg, with 60% 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 Rhizoma Corydalis extract and is about 150g.
Embodiment 2: 2. Rhizoma Corydalis extract is prepared (80 parts of Rhizoma Corydalis, 20 parts of Radix Et Rhizoma Rhei)
Preparation method: by dry for 8.0kg corydalis tuber and the dry Radix Et Rhizoma Rhei co-grinding of 2.0kg, with 60% 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 Rhizoma Corydalis extract and is about 150g.
Embodiment 3: 3. Rhizoma Corydalis extract is prepared (90 parts of Rhizoma Corydalis, 10 parts of Radix Et Rhizoma Rhei)
Preparation method: by dry for 9.0kg corydalis tuber and the dry Radix Et Rhizoma Rhei co-grinding of 1.0kg, with 60% 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 Rhizoma Corydalis extract and is about 150g.
Embodiment 4: 4. Rhizoma Corydalis extract is prepared (75 parts of Rhizoma Corydalis, 25 parts of Radix Et Rhizoma Rhei)
Preparation method: by dry for 7.5kg corydalis tuber and the dry Radix Et Rhizoma Rhei co-grinding of 2.5kg, with 60% 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 Rhizoma Corydalis extract and is about 150g.
Embodiment 5: 5. Rhizoma Corydalis extract is prepared (95 parts of Rhizoma Corydalis, 5 parts of Radix Et Rhizoma Rhei)
Preparation method: by dry for 9.5kg corydalis tuber and the dry Radix Et Rhizoma Rhei co-grinding of 0.5kg, with 60% 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 Rhizoma Corydalis 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 Rhizoma Corydalis 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 Rhizoma Corydalis extract pharmacologically active of 10 batches is similar.
Embodiment 7: extract pharmacological testing
One, material and instrument
1.1 laboratory animals: cleaning grade male SD rat, weight (220 ± 10) g, is purchased from Guangxi Medical University's Experimental Animal Center.Production licence number: SCXK (osmanthus) 2009-0002.
1.2 medicines and reagent: respectively according to embodiment 1 ~ 5 prepare Rhizoma Corydalis extract 1. ~ 5..Positive drug FUFANG DANSHEN PIAN (Zhengzhou Ruilong Pharmaceutical Co., Ltd., lot number: 130603); 0.9% sodium chloride injection (A130724H, Guizhou Kelun Pharmaceutical Co., Ltd.); Hemorheology index is surveyed by Yulin City of Guangxi Zhuang Autonomous healthcare hospital for women & children in generation.
The full-automatic lectin from hemolymph analyser (Chongqing Tianhai Medical Equipment Co., Ltd.) of 1.3 key instruments: MVIS-2035; 7180 automatic clinical chemistry analyzers (FDAC Co., Ltd.); XE-5000 blood analyser (Japanese sysmex company); RE-2000A rotary evaporator (Shanghai Yarong Biochemical Instrument Plant); SHZ-D (III) circulating water type vacuum pump (Yuhua Instrument Co., Ltd., Gongyi City); Electronic analytical balance (Beijing Sai Duolisi instrument system company limited) etc.
Two, method
The preparation of 2.1 Rhizoma Corydalis extracts: see embodiment 1 ~ 5, is configured to suspension with normal saline during use.
The pharmacologically active of 2.2 Rhizoma Corydalis extracts
2.2.1 animal grouping and administration: SD rat is divided into 18 groups at random: Normal group and model group (gavage etc. hold normal saline); Radix Salviae Miltiorrhizae Tabellae positive controls (by 100mg/kg gavage FUFANG DANSHEN PIAN solution); Rhizoma Corydalis extract 1. ~ 5. establish respectively basic, normal, high dosage group (respectively gavage give Pterospermi Heterophylli water position 100,200,300mg/kg)
2.2.2 the making of blood stasis model: select swimming power to exhaust legal system for rat blood stasis due to qi deficiency model according to tcm theory " overexertion leading to consumption of QI ".Rat first adaptability swimming instruction 3d, then every day exhausted swimming 1 time, continue 21d.
2.2.3 picture of the tongue is observed: in operation consent and gather tongue figure before drawing materials, and compares colour of the tongue and the Sublingual venation of two groups of rats, and adopts the picture of the tongue comprehensive grading standard from intending to mark.0 point (normally): the micro-green grass or young crops of tongue background color, sublingual vessel beam length are no more than 3/4 of length at the bottom of tongue; 1 point (slight purple dim): the purple dim or sublingual vessel beam length of tongue background color is more than 3/4; 2 points (obviously purple dim): the purple dim and sublingual vessel beam length of tongue background color is more than 3/4.
2.2.4 the mensuration of hemorheology index: in experiment the 21st day, after each group rat presses 0.35mg/kg intraperitoneal injection of anesthesia with 10% chloral hydrate, ventral aorta is taken a blood sample.Detect hemorheological property, erythrocyte rheology and blood coagulation related haematological index.Comprise whole blood viscosity, Plasma Viscosity, erythrocyte aggregation index, platelet count, clotting time (TT), prothrombin time (PT), activated partial thromboplastin time (APPT), Fibrinogen (FIB) content.
2.3 statistical procedures: all data with
represent, data statistics adopts SPSS17.0 statistical software to analyze, and experimental result adopts one factor analysis of variance method to carry out statistical analysis, compares and check with t between group.P < 0.05 is for there being statistical significance.
Three, result
The impact that 3.1 Rhizoma Corydalis extracts are marked on rat's blood stasis model picture of the tongue
Result is as shown in table 1, compares with model group, Rhizoma Corydalis extract 1. ~ 3. the scoring of basic, normal, high dosage group picture of the tongue all significantly reduce (P < 0.05 or P < 0.01), in dose-effect relationship; Rhizoma Corydalis extract 4. ~ the 5. scoring of each dosage group picture of the tongue no significant difference (note: compare with Normal group, * * P < 0.01 compared with model group; Compare with model group, #P < 0.05, ##P < 0.01.)。
The impact (x ± s, n=10) that table 1 Rhizoma Corydalis extract is marked on rat's blood stasis model picture of the tongue
| Group | Dosage/(mg/kg) | Picture of the tongue is marked |
| Normal group | - | 0.00±0.00## |
| Model group | - | 1.40±0.54** |
| Radix Salviae Miltiorrhizae Tabellae group | 100 | 0.40±0.55## |
| Extract is low dose group 1. | 100 | 0.60±0.55# |
| Extract is middle dosage group 1. | 200 | 0.20±0.45## |
| Extract is high dose group 1. | 300 | 0.10±0.32## |
| Extract is low dose group 2. | 100 | 0.65±0.52# |
| Extract is middle dosage group 2. | 200 | 0.23±0.50## |
| Extract is high dose group 2. | 300 | 0.15±0.28## |
| Extract is low dose group 3. | 100 | 0.63±0.57# |
| Extract is middle dosage group 3. | 200 | 0.22±0.42## |
| Extract is high dose group 3. | 300 | 0.11±0.35## |
| Extract is low dose group 4. | 100 | 1.35±0.54 |
| Extract is middle dosage group 4. | 200 | 1.31±0.51 |
| Extract is high dose group 4. | 300 | 1.27±0.37 |
| Extract is low dose group 5. | 100 | 1.33±0.56 |
| Extract is middle dosage group 5. | 200 | 1.29±0.45 |
| Extract is high dose group 5. | 300 | 1.25±0.31 |
3.2 Rhizoma Corydalis extracts are on the impact of rat's blood stasis model whole blood viscosity and Plasma Viscosity
Result is as shown in table 2, compares with model group, Rhizoma Corydalis extract 1. ~ 3. basic, normal, high dosage group whole blood viscosity and Plasma Viscosity all significantly reduce (P < 0.01), its effect presents certain dose-effect relationship; Rhizoma Corydalis extract 4. ~ 5. each dosage group whole blood viscosity and Plasma Viscosity no significant difference (note: compare with Normal group, * * P < 0.01 compared with model group; Compare with model group, ##P < 0.01).
Table 2 Rhizoma Corydalis extract is on the impact (x ± s, n=10) of rat's blood stasis model whole blood viscosity and Plasma Viscosity
3.3 Rhizoma Corydalis extracts are on the impact of rat's blood stasis model erythrocyte aggregation index:
Result is as shown in table 3, compares with model group, Rhizoma Corydalis extract 1. ~ 3. basic, normal, high dosage group erythrocyte aggregation index all significantly reduce (P < 0.01), its effect presents certain dose-effect relationship; Rhizoma Corydalis extract 4. ~ 5. each dosage group erythrocyte aggregation index no significant difference (note: compare with Normal group, * * P < 0.01 compared with model group; Compare with model group, ##P < 0.01).
Table 3 Rhizoma Corydalis extract is on the impact (x ± s, n=10) of rat's blood stasis model erythrocyte aggregation index
| Group | Dosage/(mg/kg) | Erythrocyte aggregation index |
| Normal group | - | 9.28±0.10## |
| Model group | - | 10.55±0.14** |
| Radix Salviae Miltiorrhizae Tabellae group | 100 | 9.63±0.13**## |
| Extract is low dose group 1. | 100 | 8.36±0.06**## |
| Extract is middle dosage group 1. | 200 | 6.75±0.17**## |
| Extract is high dose group 1. | 300 | 6.24±0.19**## |
| Extract is low dose group 2. | 100 | 8.38±0.08**## |
| Extract is middle dosage group 2. | 200 | 6.77±0.19**## |
| Extract is high dose group 2. | 300 | 6.27±0.21**## |
| Extract is low dose group 3. | 100 | 8.39±0.05**## |
| Extract is middle dosage group 3. | 200 | 6.78±0.15**## |
| Extract is high dose group 3. | 300 | 6.24±0.20**## |
| Extract is low dose group 4. | 100 | 10.48±0.10** |
| Extract is middle dosage group 4. | 200 | 10.42±0.16** |
| Extract is high dose group 4. | 300 | 10.36±0.18** |
| Extract is low dose group 5. | 100 | 10.49±0.11** |
| Extract is middle dosage group 5. | 200 | 10.44±0.14** |
| Extract is high dose group 5. | 300 | 10.35±0.17** |
3.4 Rhizoma Corydalis extracts are on the impact of rat's blood stasis model platelet count:
Result is as shown in table 4, compares with model group, Rhizoma Corydalis extract 1. ~ 3. basic, normal, high dosage group platelet count counting all significantly reduce (P < 0.01), its effect is in a certain amount of effect relationship; Rhizoma Corydalis extract 4. ~ 5. each dosage group platelet count counting no significant difference (note: compare with Normal group, * * P < 0.01 compared with model group; Compare with model group, ##P < 0.01).
Table 4 Rhizoma Corydalis extract is on the impact (x ± s, n=10) of rat's blood stasis model platelet count
| Group | Dosage/(mg/kg) | Platelet count (× 10 9/L) |
| Normal group | - | 532.00±65.39## |
| Model group | - | 1130.00±39.75** |
| Radix Salviae Miltiorrhizae Tabellae group | 100 | 770.00±45.76**## |
| Extract is low dose group 1. | 100 | 804.75±42.95**## |
| Extract is middle dosage group 1. | 200 | 664.00±33.97**## |
| Extract is high dose group 1. | 300 | 543.00±41.06## |
| Extract is low dose group 2. | 100 | 807.56±41.79**## |
| Extract is middle dosage group 2. | 200 | 666.14±34.18**## |
| Extract is high dose group 2. | 300 | 544.62±41.72## |
| Extract is low dose group 3. | 100 | 805.13±43.86**## |
| Extract is middle dosage group 3. | 200 | 667.20±33.46**## |
| Extract is high dose group 3. | 300 | 546.97±41.25## |
| Extract is low dose group 4. | 100 | 1100.52±40.95** |
| Extract is middle dosage group 4. | 200 | 1087.34±35.06** |
| Extract is high dose group 4. | 300 | 1061.18±39.85** |
| Extract is low dose group 5. | 100 | 1091.14±39.27** |
| Extract is middle dosage group 5. | 200 | 1083.93±34.57** |
| Extract is high dose group 5. | 300 | 1057.86±40.42** |
3.5 Rhizoma Corydalis extracts are on the impact of rat's blood stasis model blood coagulation four:
Result is as shown in table 5, compares with model group, Rhizoma Corydalis extract 1. ~ 3. basic, normal, high dosage group PT, APTT significantly raise, TT, FIB significantly reduce (P < 0.01), and its effect presents certain dose-effect relationship; Rhizoma Corydalis extract 4. ~ 5. each dosage group PT, APTT, TT, FIB no significant difference (note: compare with Normal group, * * P < 0.01 compared with model group; Compare with model group, ##P < 0.01).
Table 5 Rhizoma Corydalis extract is on the impact (x ± s, n=10) of rat's blood stasis model blood coagulation four
Conclusion: experiment adopts vitality to exhaust swimming legal system and makes rat blood stasis models, and measures the blood circulation promoting and blood stasis dispelling activity of different Rhizoma Corydalis extract, find Rhizoma Corydalis extract 1. ~ 3. all have the effect of blood circulation promoting and blood stasis dispelling, and present certain dose-effect relationship; And Rhizoma Corydalis extract 4. ~ 5. all there is no the effect of blood circulation promoting and blood stasis dispelling.This illustrates that the blood circulation promoting and blood stasis dispelling of Rhizoma Corydalis extract is active has important relationship with the ratio of Rhizoma Corydalis and Radix Et Rhizoma Rhei.
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 with excipient weight than the ratio for 1:9 in it and make.
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 (9)
1. one kind is used for the treatment of the plant extract of blood stasis disease, it is characterized in that described extract is prepared by following methods: (a) by weight, every 100 parts of medical materials comprise dry corydalis tuber 80 ~ 90 parts and dry Radix Et Rhizoma Rhei 10 ~ 20 parts, co-grinding, use 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 5 ~ 15% alcohol flushing, 7 ~ 9 column volumes, use 65 ~ 75% ethanol elution, 11 ~ 13 column volumes again, collect 65 ~ 75% eluents, concentrating under reduced pressure and get final product.
2. extract according to claim 1, is characterized in that: in the described alcoholic solution circumfluence distillation of step (a), ethanol solution concentration is 55 ~ 65%.
3. extract according to claim 2, is characterized in that: in the described alcoholic solution circumfluence distillation of step (a), ethanol solution concentration is 60%.
4. extract according to claim 1, is characterized in that: macroporous resin described in step (c) is AB-8 type macroporous resin.
5. extract according to claim 4, it is characterized in that step (c) is: n-butyl alcohol extract water dissolution, filter, with AB-8 type macroporous resin enrichment active component, first remove large polar component with 10% alcohol flushing, 8 column volumes, use 70% ethanol elution, 12 column volumes again, collect 70% eluent, concentrating under reduced pressure and get final product.
6. 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.
7. pharmaceutical preparation, is characterized in that: the extract according to claim 1 containing treatment effective dose and pharmaceutically acceptable carrier.
8. the application of extract according to claim 1 in the medicine of preparation treatment blood stasis disease.
9. the application of pharmaceutical preparation according to claim 7 in the medicine of preparation treatment blood stasis disease.
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Cited By (5)
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| CN105395815A (en) * | 2015-12-29 | 2016-03-16 | 温州统益生物医药科技有限公司 | Fritillaria yuminensis extract for treating blood stasis 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 |
| CN105963348A (en) * | 2016-06-26 | 2016-09-28 | 林天样 | Traditional Chinese medicine spica prunellae extract for increasing egg laying rate of hen |
| CN105962371A (en) * | 2016-05-16 | 2016-09-28 | 苏州毕诺佳医药技术有限公司 | Application of Chinese wistaria extract to preparation of health food capable of relieving physical fatigue |
| CN107596388A (en) * | 2017-08-28 | 2018-01-19 | 中山大学 | A kind of preparation method of syndrome of blood stasis due to qi deficiency animal model |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105395815A (en) * | 2015-12-29 | 2016-03-16 | 温州统益生物医药科技有限公司 | Fritillaria yuminensis extract for treating blood stasis 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 |
| CN105962371A (en) * | 2016-05-16 | 2016-09-28 | 苏州毕诺佳医药技术有限公司 | Application of Chinese wistaria extract to preparation of health food capable of relieving physical fatigue |
| CN105963348A (en) * | 2016-06-26 | 2016-09-28 | 林天样 | Traditional Chinese medicine spica prunellae extract for increasing egg laying rate of hen |
| CN107596388A (en) * | 2017-08-28 | 2018-01-19 | 中山大学 | A kind of preparation method of syndrome of blood stasis due to qi deficiency animal model |
| CN107596388B (en) * | 2017-08-28 | 2021-03-12 | 中山大学 | Method for making animal model with syndrome of qi deficiency and blood stasis |
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