CN105147907A - Dasheen flower extract, as well as preparation method and medicinal application thereof - Google Patents
Dasheen flower extract, as well as preparation method and medicinal application thereof Download PDFInfo
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Abstract
The invention discloses a dasheen flower extract, as well as a preparation method and medicinal application thereof. Based on total weight of 100 parts, the dasheen flower extract is prepared from the following medicinal materials in parts by weight: 75-85 parts of dried dasheen flowers and 15-25 parts of licorice root. The preparation method comprises the following steps: (a) mixing and grinding 75-85 parts of dasheen flowers and 15-25 parts of dried licorice root, performing heat reflux extraction with ethanol, mixing the extracted liquids, and concentrating until no alcohol smell exists; (b) diluting the ethanol extracted concentrate prepared in the step (a) with water, and sequentially extracting with petroleum ether, ethyl acetate and water saturated n-butyl alcohol to respectively obtain petroleum ether extract, ethyl acetate extract and n-butyl alcohol extract; and (c) enriching the n-butyl alcohol extract with macroporous resin, rinsing 8-12 column volumes with 5-15% ethanol to remove polysaccharide, rinsing 7-9 column volumes with 65-75% ethanol, collecting 65-75% eluate, concentrating at reduced pressure, and drying by spray. The dasheen flower extract has the effect of improving the immunity of mice, and can be developed into medicines for enhancing immunity.
Description
Technical field
The present invention relates to Chinese medicine extract field, Flos Colocasiae Esculentae extract being specifically related to a kind of immunological enhancement and preparation method thereof, this extract can be applied to the medicine that preparation strengthens immunity.
Background technology
Flos Colocasiae Esculentae is the inflorescence of aroid taro.Have another name called taro Seedling flower.Gather when the flowers are in blossom, using fresh herb or dry.
Former plant taro is humidogene draft.Rhizome is avette, normal raw most bottom set, brown, tool cilium.Phyllopodium is raw, 2-3 piece or more, petiole meat, long 20-90cm, green, base portion is constitute sheath-like; Blade ovum shape oval, long 20-50cm, matter is thick, and peltate raw, and the short and sharp point of tip, base portion ear shape, auricle is blunt nosed, Quan Yuan, in wavy.Common peduncle Chang Dansheng, is shorter than petiole; Spathe is different in size, is generally long about 20cm; Pipe portion is green, is about 4cm, thick 2.2cm, long avette; Eaves portion lanceolar or ellipse, be about 17cm, is launched into boat shape, Zheng in edge, faint yellow to green white; Spadix is about 10cm, is shorter than spathe; Female inflorescence is positioned at bottom, long 3-3.5cm, and neuter flower tagmeme is in middle part, and long 3-3.3cm, staminate inflorescence is positioned at top, long 4-4.5cm, and tip is suddenly narrow, and appendages bores shape, is about 1cm.2-8 month phase phase.All there is cultivation south China and various places, North China.
Summary of the invention
The object of the present invention is to provide a kind of Flos Colocasiae Esculentae 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 strengthening immunity.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
A kind of Flos Colocasiae Esculentae extract, this extract is prepared by following methods: (a) by weight, every 100 parts of medical materials comprise dry Flos Colocasiae Esculentae 75 ~ 85 parts and 15 ~ 25 parts dry, Radix Glycyrrhizae, co-grinding, extract with alcohol heat reflux, merge extractive liquid, is concentrated into 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, is obtained petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; C () n-butyl alcohol extract macroporous resin enrichment, first with 5 ~ 15% alcohol flushing, 8 ~ 12 column volume removing polysaccharide, then uses 65 ~ 75% ethanol elution, 7 ~ 9 column volumes, collects 65 ~ 75% eluents, concentrating under reduced pressure, spraying dry.
Further, extracting with alcohol heat reflux the concentration of alcohol adopted described in step (a) is 85 ~ 95%.
Further, extracting with alcohol heat reflux the concentration of alcohol adopted described in step (a) is 90%.
Further, macroporous resin described in step (c) is AB-8 type macroporous resin.
Further, step (c) is: n-butyl alcohol extract AB-8 type macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, then uses 70% ethanol elution, 8 column volumes, collects 70% eluent, concentrating under reduced pressure, spraying dry.
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: AgilentTCC18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% formic acid solution (triethylamine adjust ph to 6.0);
Gradient elution program: 0 ~ 4min, A35%; 4 ~ 14min, A35% → 70%; 14 ~ 16min, A70% → 35%; 16 ~ 20min, A35%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 267nm; Column temperature: 30 DEG C;
Sample size 10 μ L.
Pharmaceutical preparation, the described extract containing treatment effective dose and pharmaceutically acceptable carrier.
Described extract strengthens the application in the medicine of immunity in preparation.
Described pharmaceutical preparation strengthens the application in the medicine of immunity in 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 Flos Colocasiae Esculentae 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 Flos Colocasiae Esculentae of drying and Radix Glycyrrhizae is extracted, remove impurity, enrichment process, can obtain that there is the extract strengthening immunity; 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 Flos Colocasiae Esculentae 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. Flos Colocasiae Esculentae extract is prepared (80 parts of Flos Colocasiae Esculentaes, 20 portions of Radix Glycyrrhizaes)
Crude drug source: Radix Glycyrrhizae is purchased from Hui nationality's Chinese Medicinal Materials Markets.Flos Colocasiae Esculentae is the inflorescence of aroid taro, and harvesting is dried.
Main agents: food-grade ethanol is purchased from Shanghai Ling Feng chemical reagent company limited; AB-8 type macroporous resin is purchased from sky tunami letter resin company limited; Acetonitrile is HPLC level, is purchased from TEDIA; Formic acid is HPLC level, is purchased from TEDIA; Chromatographic grade pure water is heartily pure water.
Preparation method: by dry for 8.0kg Flos Colocasiae Esculentae and the dry Radix Glycyrrhizae co-grinding of 2.0kg, extract 3 times with 90% alcohol heat reflux, each extraction 2 hours, at every turn with 95% ethanol 25L, merge extractive liquid, is concentrated into without alcohol taste (4L); Concentrated solution uses petroleum ether (4L × 3 time), ethyl acetate (4L × 3 time) and water saturated n-butyl alcohol (4L × 3 time) to extract successively, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; N-butyl alcohol extract AB-8 macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, then uses 70% ethanol elution, 8 column volumes, collects 70% eluent, concentrating under reduced pressure, spraying dry (about 225g).
Embodiment 2: 2. Flos Colocasiae Esculentae extract is prepared (75 parts of Flos Colocasiae Esculentaes, 25 portions of Radix Glycyrrhizaes)
Preparation method: by dry for 7.5kg Flos Colocasiae Esculentae and the dry Radix Glycyrrhizae co-grinding of 2.5kg, extract 3 times with 90% alcohol heat reflux, each extraction 2 hours, at every turn with 95% ethanol 25L, merge extractive liquid, is concentrated into without alcohol taste (4L); Concentrated solution uses petroleum ether (4L × 3 time), ethyl acetate (4L × 3 time) and water saturated n-butyl alcohol (4L × 3 time) to extract successively, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; N-butyl alcohol extract AB-8 macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, then uses 70% ethanol elution, 8 column volumes, collects 70% eluent, concentrating under reduced pressure, spraying dry (about 225g).
Embodiment 3: 3. Flos Colocasiae Esculentae extract is prepared (85 parts of Flos Colocasiae Esculentaes, 15 portions of Radix Glycyrrhizaes)
Preparation method: by dry for 8.5kg Flos Colocasiae Esculentae and the dry Radix Glycyrrhizae co-grinding of 1.5kg, extract 3 times with 90% alcohol heat reflux, each extraction 2 hours, at every turn with 95% ethanol 25L, merge extractive liquid, is concentrated into without alcohol taste (4L); Concentrated solution uses petroleum ether (4L × 3 time), ethyl acetate (4L × 3 time) and water saturated n-butyl alcohol (4L × 3 time) to extract successively, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; N-butyl alcohol extract AB-8 macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, then uses 70% ethanol elution, 8 column volumes, collects 70% eluent, concentrating under reduced pressure, spraying dry (about 225g).
Embodiment 4: 4. Flos Colocasiae Esculentae extract is prepared (90 parts of Flos Colocasiae Esculentaes, 10 portions of Radix Glycyrrhizaes)
Preparation method: by dry for 9.0kg Flos Colocasiae Esculentae and the dry Radix Glycyrrhizae co-grinding of 1.0kg, extract 3 times with 90% alcohol heat reflux, each extraction 2 hours, at every turn with 95% ethanol 25L, merge extractive liquid, is concentrated into without alcohol taste (4L); Concentrated solution uses petroleum ether (4L × 3 time), ethyl acetate (4L × 3 time) and water saturated n-butyl alcohol (4L × 3 time) to extract successively, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; N-butyl alcohol extract AB-8 macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, then uses 70% ethanol elution, 8 column volumes, collects 70% eluent, concentrating under reduced pressure, spraying dry (about 225g).
Embodiment 5: 5. Flos Colocasiae Esculentae extract is prepared (70 parts of Flos Colocasiae Esculentaes, 30 portions of Radix Glycyrrhizaes)
Preparation method: by dry for 7.0kg Flos Colocasiae Esculentae and the dry Radix Glycyrrhizae co-grinding of 3.0kg, extract 3 times with 90% alcohol heat reflux, each extraction 2 hours, at every turn with 95% ethanol 25L, merge extractive liquid, is concentrated into without alcohol taste (4L); Concentrated solution uses petroleum ether (4L × 3 time), ethyl acetate (4L × 3 time) and water saturated n-butyl alcohol (4L × 3 time) to extract successively, obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; N-butyl alcohol extract AB-8 macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, then uses 70% ethanol elution, 8 column volumes, collects 70% eluent, concentrating under reduced pressure, spraying dry (about 225g).
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: AgilentTCC18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% formic acid solution (triethylamine adjust ph to 6.0);
Gradient elution program: 0 ~ 4min, A35%; 4 ~ 14min, A35% → 70%; 14 ~ 16min, A70% → 35%; 16 ~ 20min, A35%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 267nm; Column temperature: 30 DEG C;
Sample size 10 μ L.
Analyze with the Flos Colocasiae Esculentae extract of 10 batches of preparation, carry out chromatographic peak coupling, 1 ~ No. 4 peak all occurs in 10 batch sample chromatograms as a result.Therefore demarcate 4 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 Flos Colocasiae Esculentae extract pharmacologically active of 10 batches is similar.
Embodiment 7: extract pharmacological testing
One, materials and methods
1.1 trial drugs: Flos Colocasiae Esculentae extract 1. ~ 5. respectively according to embodiment 1 ~ 5 prepare, be made into suspension with normal saline, 4 DEG C of preservations.Positive drug mannatide, tablet, Lier Pharmaceutical Co., Ltd., Chengdu City, uses mortar pulverize, and normal saline becomes the suspension of 1mg/mL, 4 DEG C of preservations.Cyclophosphamide (CTX), white freeze dried powder, Shanxi Tai Sheng pharmaceutical Co. Ltd, with physiological saline solution, is mixed with the solution of 4mg/mL, and 4 DEG C keep in Dark Place.
1.2 animals: Kunming mouse, male and female half and half, one-level, body weight 20 ± 2g, is provided by Sichuan University's Experimental Animal Center.
1.3 reagent: india ink (CHROMA-GESELLSCHAFTSCHMDGMBH & CO. produces), DNFB (DNCB, Shanghai San'aisi Reagent Co., Ltd.).
1.4 method
1.4.1 Flos Colocasiae Esculentae extract is on the impact of immunosuppression mouse model macrophages phagocytic capacity caused by CTX
216 mices are divided into 18 groups of (blank group, model group, positive drug groups at random by sex body weight, Flos Colocasiae Esculentae extract 1. ~ 5. low middle high dose group), except blank group, all the other respectively organize continuous 4 days of mice equal lumbar injection CTX40mg/kg, simultaneously gastric infusion 10 days.After last administration 1h, mouse tail vein injection india ink (1:4) normal saline solution 0.5mL/ only, gets blood 20 μ L respectively at 2min and 5min eye socket after injection, adds 2mL0.1%Na
2cO
3in solution, in 590nm colorimetric.After sacrifice of animal, get liver, spleen is weighed, calculate phagocytic index K and correct phagocytic index α.
1.4.2 Flos Colocasiae Esculentae extract is on the impact of mice delayed hypersensitivity (DTH)
170 mices are divided into 17 groups at random by sex body weight, abdominal part unhairing 3cm × 3cm, every Mus is coated with 50 μ L1%DNCB (dissolving of 1:1 acetone Oleum Sesami) sensitization, and in second day strengthening sensitization once, except negative group.10 μ L1%DNCB, after 8 days, are applied to mouse right ear two sides, put to death after 24h by gastric infusion, cut left and right ear and to punch respectively cut-off footpath 8mm auricle, weigh.With left and right auricle weight difference for swelling compares group difference.
1.4.3 Flos Colocasiae Esculentae extract is on the impact of immunodeficiency models mice serum hemolysin content caused by CTX
180 mices are divided into 18 groups at random by sex body weight, and except blank group, all the other respectively organize continuous 5 days of mice equal lumbar injection CTX20mg/kg, simultaneously each administration group gastric infusion (0.2mL/10g) 10 days.All groups all in administration the 3rd day lumbar injection 5% sheep red blood cell (SRBC) 0.2mL/ sensitization.After last administration 1h, eye socket gets blood, centrifugalize serum, with normal saline dilution 100 times.Get dilute serum 1mL, mix with 5% sheep red blood cell (SRBC) 0.5mL, 10% complement (guinea pig serum) 0.5mL, put 30min in 37 DEG C of water baths, 0 DEG C of cessation reaction, centrifuging and taking supernatant is in 540nm place colorimetric.
1.4.4 Flos Colocasiae Esculentae extract is on the impact of CTX induced mice Peritoneal Macrophage Phagocytosis
180 mices are divided into 18 groups at random by sex body weight, and often organize 10, except blank group, all the other respectively organize mice equal lumbar injection CTX20mg/kg (0.1mL/10g) continuous 5 days, simultaneously each administration group gastric infusion 10 days.After last administration 1h, each Mus lumbar injection 5% sheep red blood cell (SRBC) normal saline solution 0.4mL, puts to death after 8h, dew peritoneum of cutting open the belly, inject NS2mL, gently rub abdominal part 1min, draw peritoneal fluid and be applied on microscope slide, hatch 30min for 37 DEG C, take out with on normal saline flushing slide not by the SRBC that engulfs and other cells, dry up, methanol fixes 5min, and Giemsa dyes 20min.Washing examines under a microscope 200 macrophages after drying, and calculates phagocytic percentage and phagocytic index.
Two, result
2.1 Flos Colocasiae Esculentae extracts the results are shown in Table 1 (note: * P<0.05, * * P<0.01 compared with model group) to the impact of immunodeficiency models mouse macrophage phagocytic activity caused by CTX.From experimental result, immunodeficiency models group compared with blank group, its phagocytic index and correct phagocytic index significance lower than blank group; Flos Colocasiae Esculentae extract 1. ~ 3. middle and high dosage group and positive group compared with model group, phagocytic index and correct phagocytic index significance higher than model group; Flos Colocasiae Esculentae extract 4. ~ 5. each dosage group compared with model group without significant difference.Result: Flos Colocasiae Esculentae extract 1. ~ 3. middle and high dosage immunosuppression mouse model caused by CTX is had to the effect of immunostimulant.
Table 1 Flos Colocasiae Esculentae extract is on the impact (x ± s) of CTX induced mice macrophages phagocytic capacity
| Group | Dosage/(mg/kg) | Mus number (only) | K value | α value |
| Normal group | - | 12 | 0.0238±0.0047** | 4.11±0.42** |
| Model group | - | 12 | 0.0113±0.0022 | 3.06±0.296 |
| Mannatide | 20 | 12 | 0.0324±0.0078** | 4.60±0.57** |
| Extract is low dose group 1. | 250 | 12 | 0.0129±0.003 | 3.37±0.58 |
| Extract is middle dosage group 1. | 500 | 12 | 0.0230±0.0075** | 3.74±0.76* |
| Extract is high dose group 1. | 1000 | 12 | 0.0262±0.0134** | 3.71±0.75* |
| Extract is low dose group 2. | 250 | 12 | 0.0127±0.0028 | 3.34±0.631 |
| Extract is middle dosage group 2. | 500 | 12 | 0.0229±0.0083** | 3.71±0.82* |
| Extract is high dose group 2. | 1000 | 12 | 0.0258±0.0146** | 3.72±0.723* |
| Extract is low dose group 3. | 250 | 12 | 0.0129±0.002 | 3.35±0.67 |
| Extract is middle dosage group 3. | 500 | 12 | 0.0225±0.0087** | 3.72±0.634* |
| Extract is high dose group 3. | 1000 | 12 | 0.0260±0.0141** | 3.71±0.771* |
| Extract is low dose group 4. | 20 | 12 | 0.0115±0.0025 | 3.15±0.49 |
| Extract is middle dosage group 4. | 250 | 12 | 0.0118±0.0063 | 3.33±0.52 |
| Extract is high dose group 4. | 500 | 12 | 0.0120±0.0124 | 3.35±0.576 |
| Extract is low dose group 5. | 20 | 12 | 0.0113±0.0031 | 3.16±0.412 |
| Extract is middle dosage group 5. | 250 | 12 | 0.0116±0.0071 | 3.31±0.537 |
| Extract is high dose group 5. | 500 | 12 | 0.0119±0.0132 | 3.36±0.601 |
The impact of 2.2 Flos Colocasiae Esculentae extracts on the serum hemolysis cellulose content of CTX induced mice immunodeficiency models the results are shown in Table 2 (notes: * P<0.05, * * P<0.01 compared with model group).From result, model group serum hemolysis cellulose content highly significant is lower than blank group; Flos Colocasiae Esculentae extract 1. ~ 3. in the serum hemolysis cellulose content of high dose group be significantly higher than model group; Flos Colocasiae Esculentae extract 4. ~ 5. each dosage group and model group be without significant difference.
Table 2 Flos Colocasiae Esculentae extract is on the impact (x ± s) of CTX induced mice serum hemolysis cellulose content
| Group | Dosage/(mg/kg) | Mus number (only) | OD value |
| Normal group | - | 10 | 2.71±0.87** |
| Model group | - | 10 | 0.80±0.52 |
| Mannatide | 20 | 10 | 2.03±1.15** |
| Extract is low dose group 1. | 250 | 10 | 1.25±0.91 |
| Extract is middle dosage group 1. | 500 | 10 | 1.95±1.33* |
| Extract is high dose group 1. | 1000 | 10 | 2.13±1.11** |
| Extract is low dose group 2. | 250 | 10 | 1.21±0.93 |
| Extract is middle dosage group 2. | 500 | 10 | 1.94±1.29* |
| Extract is high dose group 2. | 1000 | 10 | 2.10±1.13** |
| Extract is low dose group 3. | 250 | 10 | 1.24±0.86 |
| Extract is middle dosage group 3. | 500 | 10 | 1.92±1.37* |
| Extract is high dose group 3. | 1000 | 10 | 2.11±1.08** |
| Extract is low dose group 4. | 250 | 10 | 0.92±0.75 |
| Extract is middle dosage group 4. | 500 | 10 | 1.05±0.93 |
| Extract is high dose group 4. | 1000 | 10 | 1.09±1.01 |
| Extract is low dose group 5. | 250 | 10 | 0.89±0.79 |
| Extract is middle dosage group 5. | 500 | 10 | 1.03±0.99 |
| Extract is high dose group 5. | 1000 | 10 | 1.10±1.03 |
The impact of 2.3 Flos Colocasiae Esculentae extracts on CTX induced mice Peritoneal Macrophage Phagocytosis the results are shown in Table 3 (compared with model group * P<0.05, * * P<0.01).From result, the phagocytic percentage of model group peritoneal macrophage and phagocytic index highly significant are lower than blank group; Positive drug and Flos Colocasiae Esculentae extract 1. ~ 3. the phagocytic percentage of each dosage group peritoneal macrophage and the equal significance of phagocytic index higher than model group; Flos Colocasiae Esculentae extract 4. ~ 5. each dosage group compared with model group without significant difference.
Table 3 Flos Colocasiae Esculentae extract is on the impact (x ± s) of CTX induced mice Peritoneal Macrophage Phagocytosis
| Group | Dosage/(mg/kg) | Mus number (only) | Phagocytic percentage (%) | Phagocytic index |
| Normal group | NS | 10 | 0.51±0.09** | 0.73±0.20* |
| Model group | NS | 10 | 0.35±0.12 | 0.48±0.23 |
| Mannatide | 20 | 10 | 0.60±0.13** | 0.88±0.22** |
| Extract is low dose group 1. | 250 | 10 | 0.60±0.17** | 0.85±0.29** |
| Extract is middle dosage group 1. | 500 | 10 | 0.61±0.23** | 0.87±0.39* |
| Extract is high dose group 1. | 1000 | 10 | 0.74±0.19** | 1.05±0.33** |
| Extract is low dose group 2. | 250 | 10 | 0.58±0.19** | 0.81±0.33** |
| Extract is middle dosage group 2. | 500 | 10 | 0.60±0.25** | 0.86±0.37* |
| Extract is high dose group 2. | 1000 | 10 | 0.72±0.21** | 1.01±0.35** |
| Extract is low dose group 3. | 250 | 10 | 0.57±0.13** | 0.83±0.27** |
| Extract is middle dosage group 3. | 500 | 10 | 0.59±0.21** | 0.85±0.35* |
| Extract is high dose group 3. | 1000 | 10 | 0.71±0.18** | 1.03±0.30** |
| Extract is low dose group 4. | 20 | 10 | 0.38±0.09 | 0.52±0.25 |
| Extract is middle dosage group 4. | 250 | 10 | 0.39±0.11 | 0.55±0.27 |
| Extract is high dose group 4. | 500 | 10 | 0.42±0.08 | 0.59±0.21 |
| Extract is low dose group 5. | 20 | 10 | 0.37±0.10 | 0.50±0.23 |
| Extract is middle dosage group 5. | 250 | 10 | 0.38±0.13 | 0.53±0.29 |
| Extract is high dose group 5. | 500 | 10 | 0.43±0.10 | 0.57±0.25 |
As fully visible, Flos Colocasiae Esculentae extract has and improves the effect of immune function of mice, but in this effect and Flos Colocasiae Esculentae extract, the content of Radix Glycyrrhizae has important relationship, and the activity of content to Flos Colocasiae Esculentae extract of Radix Glycyrrhizae plays vital 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 (9)
1. a Flos Colocasiae Esculentae extract, it is characterized in that described extract is prepared by following methods: (a) by weight, every 100 parts of medical materials comprise dry Flos Colocasiae Esculentae 75 ~ 85 parts and 15 ~ 25 parts dry, Radix Glycyrrhizae, co-grinding, extract with alcohol heat reflux, merge extractive liquid, is concentrated into 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, is obtained petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; C () n-butyl alcohol extract macroporous resin enrichment, first with 5 ~ 15% alcohol flushing, 8 ~ 12 column volume removing polysaccharide, then uses 65 ~ 75% ethanol elution, 7 ~ 9 column volumes, collects 65 ~ 75% eluents, concentrating under reduced pressure, spraying dry.
2. extract according to claim 1, is characterized in that: extracting with alcohol heat reflux the concentration of alcohol adopted described in step (a) is 85 ~ 95%.
3. extract according to claim 2, is characterized in that: extracting with alcohol heat reflux the concentration of alcohol adopted described in step (a) is 90%.
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 AB-8 type macroporous resin enrichment, first with 10% alcohol flushing, 10 column volume removing polysaccharide, use 70% ethanol elution, 8 column volumes again, collect 70% eluent, concentrating under reduced pressure, spraying dry.
6. extract according to claim 1, is characterized in that: the liquid phase analysis method of described extract is:
Chromatographic column: AgilentTCC18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% formic acid solution (triethylamine adjust ph to 6.0);
Gradient elution program: 0 ~ 4min, A35%; 4 ~ 14min, A35% → 70%; 14 ~ 16min, A70% → 35%; 16 ~ 20min, A35%;
Flow rate of mobile phase: 1.0mLmin
- 1;
Determined wavelength: 267nm; Column temperature: 30 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. extract according to claim 1 strengthens the application in the medicine of immunity in preparation.
9. pharmaceutical preparation according to claim 7 strengthens the application in the medicine of immunity in preparation.
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|---|---|---|---|
| CN201510688870.7A CN105147907A (en) | 2015-10-22 | 2015-10-22 | Dasheen flower extract, as well as preparation method and medicinal application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105902664A (en) * | 2016-06-26 | 2016-08-31 | 林天样 | Traditional Chinese medicine Chinese soapberry fruit extract for treating diarrhea of yak calves |
| CN105994993A (en) * | 2016-06-02 | 2016-10-12 | 苏州灵垦农业科技有限公司 | Feed for improving immune function of dairy cow in early perinatal period and preparation method thereof |
| CN105995060A (en) * | 2016-06-02 | 2016-10-12 | 苏州灵垦农业科技有限公司 | Feed for improving growth performance of fattening sheep and preparation method thereof |
| CN105994979A (en) * | 2016-05-06 | 2016-10-12 | 江苏盐城源耀生物科技有限公司 | Functional ryegrass extract and application thereof in biological feed |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104958333A (en) * | 2015-06-09 | 2015-10-07 | 徐蒙蒙 | Panax japonicus extract and medical application thereof |
| CN104958393A (en) * | 2015-06-08 | 2015-10-07 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | Extract of potentilla anserina rhizome and medical application thereof |
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2015
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104958393A (en) * | 2015-06-08 | 2015-10-07 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | Extract of potentilla anserina rhizome and medical application thereof |
| CN104958333A (en) * | 2015-06-09 | 2015-10-07 | 徐蒙蒙 | Panax japonicus extract and medical application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105994979A (en) * | 2016-05-06 | 2016-10-12 | 江苏盐城源耀生物科技有限公司 | Functional ryegrass extract and application thereof in biological feed |
| CN105994993A (en) * | 2016-06-02 | 2016-10-12 | 苏州灵垦农业科技有限公司 | Feed for improving immune function of dairy cow in early perinatal period and preparation method thereof |
| CN105995060A (en) * | 2016-06-02 | 2016-10-12 | 苏州灵垦农业科技有限公司 | Feed for improving growth performance of fattening sheep and preparation method thereof |
| CN105902664A (en) * | 2016-06-26 | 2016-08-31 | 林天样 | Traditional Chinese medicine Chinese soapberry fruit extract for treating diarrhea of yak calves |
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