CN105535068A - Ulmus pumila bark extract, preparation method of ulmus pumila bark extract and application of ulmus pumila bark extract in preparing immunity enhancing medicine - Google Patents
Ulmus pumila bark extract, preparation method of ulmus pumila bark extract and application of ulmus pumila bark extract in preparing immunity enhancing medicine Download PDFInfo
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Abstract
The invention discloses an ulmus pumila bark extract and a preparation method thereof. The preparation method includes the steps that firstly, each 100 parts of medicinal materials include, by weight, 75-85 parts of dried ulmus pumila bark and 15-25 parts of dried radix codonopsis, the medicinal materials are mixed, smashed and subjected to heat reflux extraction with ethyl alcohol, and the extraction solution is fused and then concentrated till the alcohol smell is removed; secondly, the ethyl alcohol extraction concentrated solution obtained in the first step is diluted with water and then extracted with petroleum ether, ethyl acetate and water-saturated n-butyl alcohol in sequence to obtain a petroleum ether extract, an ethyl acetate extract and an n-butyl alcohol extract respectively; thirdly, the n-butyl alcohol extract is enriched with macroporous resin, the enriched n-butyl alcohol extract is washed with 5-15% ethyl alcohol by 8-12 column volumes to remove polysaccharide and then eluted with 65-75% ethyl alcohol by 7-9 column volumes, and a 65-75% elution solution is collected and subjected to vacuum concentration and spray drying. The ulmus pumila bark extract has the effect of improving the immune function of mice and can be developed into the immunity enhancing medicine.
Description
Technical field
The present invention relates to Chinese medicine extract field, Cortex ulmi pumilae 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
The bark of Cortex ulmi pumilae for Ulmaceae elm or the phloem of root bark.Cut off old branch between spring or 8 ~ JIUYUE, strip endothelium immediately and dry.
Former plant elm is deciduous tree, up to 90 meters.Bark taupe, coarse, there is longitudinal furrow to split; Sprig is soft, hairiness, talks lark.Single leaf alternate; Avette or the ellipticity lanceolar of obovate, ellipticity, long 2 ~ 8 centimetres, wide 2 ~ 2.5 centimetres.The sharp point of tip or gradually point, basal circular or wedge shape, the usual single sawtooth in edge, above dirty-green, without hair, below young time have pubescence, only arteries and veins axil adularescent fine hair time old; Long 2 ~ 8 millimeters of petiole, hairiness; Stipule lanceolar, long 1 centimetre, hairiness.Hua Xianye is open, clusters, has short stalk; Calyx 4 ~ 5 splits; Stamen 4 ~ 5, flower pesticide purple; Ovary is flat, style 2.Myriopteron extensum (Wight) K. Schum obovate or subcircular, smooth, tip is jagged.Seed is positioned at central authorities, connects with breach, long 1 ~ 1.5 centimetre.3 ~ April of florescence.Really 4 ~ June of phase.Be born in river levee, ridge and roadside; The foot of a mountain, sand ground also there is growth.China's most area has distribution.Containing cupreol, plant sterol, stigmasterol, multiple sterols and tannin, natural gum, fatty wet goods chemical composition.
Summary of the invention
The object of the present invention is to provide a kind of Cortex ulmi pumilae extract, and provide its preparation method a kind of, and pharmaceutical preparation containing this extract and utilize this extract to prepare the purposes of the medicine strengthening immunity.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
A kind of Cortex ulmi pumilae extract, this extract is prepared by following methods: (a) by weight, every 100 parts of medical materials comprise dry Cortex ulmi pumilae 75 ~ 85 parts and dry Radix Codonopsis 15 ~ 25 parts, 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 Cortex ulmi pumilae 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 ~ 5min, A35%; 5 ~ 10min, A35% → 70%; 10 ~ 15min, A70% → 35%;
15～20min，A35％；
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 240nm;
Column temperature: 30 DEG C;
Sample size 10 μ L.
Pharmaceutical preparation, the described 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 Cortex ulmi pumilae 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 Cortex ulmi pumilae of drying and Radix Codonopsis extracts, 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 Cortex ulmi pumilae 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. Cortex ulmi pumilae extract is prepared (80 parts of Cortex ulmi pumilae, 20 parts of Radix Codonopsis)
Crude drug source: Cortex ulmi pumilae and Radix Codonopsis 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; 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 Cortex ulmi pumilae and the dry Radix Codonopsis 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. Cortex ulmi pumilae extract is prepared (75 parts of Cortex ulmi pumilae, 25 parts of Radix Codonopsis)
Preparation method: by dry for 7.5kg Cortex ulmi pumilae and the dry Radix Codonopsis 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. Cortex ulmi pumilae extract is prepared (85 parts of Cortex ulmi pumilae, 15 parts of Radix Codonopsis)
Preparation method: by dry for 8.5kg Cortex ulmi pumilae and the dry Radix Codonopsis 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. Cortex ulmi pumilae extract is prepared (90 parts of Cortex ulmi pumilae, 10 parts of Radix Codonopsis)
Preparation method: by dry for 9.0kg Cortex ulmi pumilae and the dry Radix Codonopsis 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. Cortex ulmi pumilae extract is prepared (70 parts of Cortex ulmi pumilae, 30 parts of Radix Codonopsis)
Preparation method: by dry for 7.0kg Cortex ulmi pumilae and the dry Radix Codonopsis 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 ~ 5min, A35%; 5 ~ 10min, A35% → 70%; 10 ~ 15min, A70% → 35%; 15 ~ 20min, A35%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 240nm;
Column temperature: 30 DEG C;
Sample size 10 μ L.
Analyze with the Cortex ulmi pumilae 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 Cortex ulmi pumilae extract pharmacologically active of 10 batches is similar.
Embodiment 7: extract pharmacological testing
One, materials and methods
1.1 trial drugs: Cortex ulmi pumilae 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 Cortex ulmi pumilae 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, Cortex ulmi pumilae 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 Cortex ulmi pumilae 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 Cortex ulmi pumilae 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 Cortex ulmi pumilae 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 Cortex ulmi pumilae 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; Cortex ulmi pumilae 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; Cortex ulmi pumilae extract 4. ~ 5. each dosage group compared with model group without significant difference.Result: Cortex ulmi pumilae extract 1. ~ 3. middle and high dosage immunosuppression mouse model caused by CTX is had to the effect of immunostimulant.
Table 1 Cortex ulmi pumilae 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 Cortex ulmi pumilae 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; Cortex ulmi pumilae extract 1. ~ 3. in the serum hemolysis cellulose content of high dose group be significantly higher than model group; Cortex ulmi pumilae extract 4. ~ 5. each dosage group and model group be without significant difference.
Table 2 Cortex ulmi pumilae 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 Cortex ulmi pumilae 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 Cortex ulmi pumilae extract 1. ~ 3. the phagocytic percentage of each dosage group peritoneal macrophage and the equal significance of phagocytic index higher than model group; Cortex ulmi pumilae extract 4. ~ 5. each dosage group compared with model group without significant difference.
Table 3 Cortex ulmi pumilae 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, Cortex ulmi pumilae extract has and improves the effect of immune function of mice, but in this effect and Cortex ulmi pumilae extract, the content of Radix Codonopsis has important relationship, and the activity of content to Cortex ulmi pumilae extract of Radix Codonopsis 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.
The effect of above-described embodiment is essentiality content of the present invention is described, but does not limit protection scope of the present invention with this.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 protection domain of technical solution of the present invention.
Claims (9)
1. a Cortex ulmi pumilae extract, it is characterized in that described extract is prepared by following methods: (a) by weight, every 100 parts of medical materials comprise dry Cortex ulmi pumilae 75 ~ 85 parts and dry Radix Codonopsis 15 ~ 25 parts, 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. Cortex ulmi pumilae 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. Cortex ulmi pumilae 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. Cortex ulmi pumilae extract according to claim 1, is characterized in that: macroporous resin described in step (c) is AB-8 type macroporous resin.
5. Cortex ulmi pumilae 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. Cortex ulmi pumilae 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 ~ 5min, A35%; 5 ~ 10min, A35% → 70%; 10 ~ 15min, A70% → 35%; 15 ~ 20min, A35%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 240nm; Column temperature: 30 DEG C;
Sample size 10 μ L.
7. pharmaceutical preparation, is characterized in that: the Cortex ulmi pumilae 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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| CN201610008881.0A CN105535068A (en) | 2016-01-05 | 2016-01-05 | Ulmus pumila bark extract, preparation method of ulmus pumila bark extract and application of ulmus pumila bark extract in preparing immunity enhancing medicine |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105079182A (en) * | 2015-09-12 | 2015-11-25 | 徐建立 | Gardenia extract and medical application thereof to immuno-enhancement |
| CN105106275A (en) * | 2015-09-16 | 2015-12-02 | 潘光贤 | Melastoma sanguineum extract, preparation method thereof and medical application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105079182A (en) * | 2015-09-12 | 2015-11-25 | 徐建立 | Gardenia extract and medical application thereof to immuno-enhancement |
| CN105106275A (en) * | 2015-09-16 | 2015-12-02 | 潘光贤 | Melastoma sanguineum extract, preparation method thereof and medical application |
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