CN102397373B - Immature bitter orange or bitter orange general flavone extract extracted in decoction way through water and application thereof - Google Patents
Immature bitter orange or bitter orange general flavone extract extracted in decoction way through water and application thereof Download PDFInfo
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Abstract
The invention relates to an immature bitter orange or bitter orange general flavone extract extracted in a decoction way through water, which is characterized in that: immature bitter orange or bitter orange is cut into slices or is pulverized to be decocted for 1-3 times in the water with the volume being 4 to 15 times of that of the raw material, the raw material is decocted and extracted for 1 to 3 hours every time, the decocted liquid is combined and filtered, the filtered liquid passes through a well-processed nonpolar macroporous resin and is sequentially washed by water and 20% ethanol so as to remove the foreign matters, then the filtered liquid is eluted by 25 to 90 percent ethanol and 0. 5 to 2 percent sodium hydroxide, the ethanol eluted liquid is collected, decompressed and concentrated into liquid extract with relative density being about 1.2 to 1.4 (50 DEG C to 70 DEG C), the liquid extract is decompressed and dried at the temperature of 60 to 80 DEG C and pulverized to obtain the immature bitter orange or bitter orange general flavone extract. The extract can be used for preparing medicine for treating motility disturbance type intestine and stomach functional indigestion, epigastric pain, abdominal distension, anorexia, eructation, nausea, vomiting and vomiting caused by chemotherapeutics or other reasons and can be made into a preparation form which is acceptable on the medicine.
Description
Technical field
The present invention relates to a kind of Fructus Aurantii Immaturus of water boiling and extraction or Fructus Aurantii extractive of general flavone and uses thereof.
Background technology
Functional dyspepsia (FD) is a kind of common, frequently-occurring disease.Be called again gastric dynamic dysfunction, or gastric motility disorder.Be a kind of common clinical syndrome, the foreign statistic data shows, 1/3 crowd suffered from FD, accounted for 20%～30% of gop number.Domestic Digestive out-patient 50% left and right that accounts for betides 20%～40% normal population.In addition, can cause vomiting after the Chemotherapy of Tumor Patients more than 80%, preventing or arresting vomiting is mainly take medicines such as paspertin metoclopramide, ondansetrons as main, but the chemical medicine preventing or arresting vomiting nape muscle spasm symptom even might occur with asthenia, dizziness, headache.
Western medical treatment is mainly take motilium, cisapride, mosapride etc. as main at present; But with its a large amount of uses, find to have obvious Cardiotoxity, many patients Q-T interval prolongation and arrhythmia occur after using cisapride, and particularly torsades de pointes type chamber speed waits serious adverse reaction, even causes death.For this reason, FDA Food and Drug Administration is decided by that on July 23rd, 2000 rose and stops using this medicine, and China Drug Administration has also made restriction to the use of cisapride, but still can use under the doctor instructs.
Chinese traditional treatment is with the dispelling the stagnated QI removing food stagnancy, and loose painful abdominal mass class Chinese medicine reduces phlegm; Stop in treatment is stagnant, feeling of fullness distending pain etc., Fructus Aurantii Immaturus is for representing one of medicine.Fructus Aurantii Immaturus, Fructus Aurantii are dry young fruit or the immature fruit of rutaceae Citrus aurantium Linn. Citrus aurantium L. and variety or Fructus Citri sinensis Citrus sinensis Osbeckr.Fructus Aurantii Immaturus, Fructus Aurantii are from Shennong's Herbal, " Mingyi Bielu " record: " except breast side of body accumulation of phlegm in the hypochondrium; by cutting off the water; broken solid; that relieving distension is full ", from Jin Dynasty Lee physician Dong Yuan " differentiation on endogenous " pioneering zhishi daozhi pills to make by hand, clinical formulation application and modern Chinese medicine large-scale production also are used for digestive tract disease up till now, and there is the history of nearly 2,000 years front and back; Fructus Aurantii Immaturus, Fructus Aurantii main component are flavone etc., and its decocting liquid and Hesperidin etc. can significantly strengthen the ahead running of the small intestinal of normal mouse and atropine generation mice inhibition, have the effect of short digestive tract power.But Fructus Aurantii Immaturus, Fructus Aurantii only are used for clinical formulation and Chinese patent medicine preparation on a small quantity, and ample resources is not used widely, and its natural dispelling the stagnated QI removing food stagnancy effect may promote that the effect class same sex of medicine for stomach dynamic is not fully excavated with the modern times.
Summary of the invention
The Fructus Aurantii Immaturus or the Fructus Aurantii extractive of general flavone that the purpose of this invention is to provide a kind of water boiling and extraction are used for gastrointestinal motility obstruction type functional dyspepsia; Upper abdominal pain, abdominal distention, anorexia, belch, feel sick, vomiting that vomiting and chemotherapeutics or other reasons cause.
Fructus Aurantii Immaturus provided by the invention or Fructus Aurantii extractive of general flavone, by Fructus Aurantii Immaturus or Fructus Aurantii are obtained with water boiling and extraction, wherein boiling water extraction method is: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～15 times of water gagings, decoct 1～3 time, the each decoction extracted 1～3 hour, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 25～90% ethanol, 0.5～2% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.20～1.40 (50 ℃～70 ℃) of relative density, drying under reduced pressure, dry temperature is 60～80 ℃, pulverize, and get final product.
preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～13 times of water gagings, decoct 2～3 times, the each decoction extracted 1～2.5 hour, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 30%～85% ethanol, 0.5～1.8% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.25～1.40 (55 ℃～70 ℃) of relative density, drying under reduced pressure, dry temperature is 65～80 ℃, pulverize, and get final product.
preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～11 times of water gagings, decoct 2～3 times, the each decoction extracted 1.5～2.5 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 35%～75% ethanol, 0.5～1.6% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.25～1.35 (55 ℃～65 ℃) of relative density, drying under reduced pressure, dry temperature is 70～80 ℃, pulverize, and get final product.
preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～9 times of water gagings, decoct 3 times, the each decoction extracted 1.5～2 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 40%～65% ethanol, 0.5～1.3% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.30～1.35 (60 ℃～65 ℃) of relative density, drying under reduced pressure, dry temperature is 70～75 ℃, pulverize, and get final product.
Preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～7 times of water gagings, decoct 3 times, the each decoction extracted 2 hours, merged decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water, the ethanol elution below 20% are removed impurity successively, then use 45%～55% ethanol, 0.8～1.2% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure, dry temperature is 75 ℃, pulverize, and get final product.
Preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～8 times of water gagings, decoct 2 times, the each decoction extracted 1.5 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water, the ethanol elution below 20% are removed impurity successively, use again 40%～55% ethanol, 0.7～1.3% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure, dry temperature is 75 ℃, pulverizes, and get final product.
Preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～6 times of water gagings, decoct 2 times, the each decoction extracted 1.5 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water, the ethanol elution below 20% are removed impurity successively, use again 45%～50% ethanol, 0.9～1.1% sodium hydroxide eluting, collect ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure, dry temperature is 75 ℃, pulverizes, and get final product.
Preferably, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, decoct with water 3 times, add for the first time 8 times of water gagings and boiled 2 hours, second and third time adds respectively 6,4 times of amounts and decocted 1 hour, filter merging filtrate, water, 15% ethanol, 45% ethanol, 1% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure, dry temperature is 75 ℃, pulverize, and get final product.
Preferably, wherein 15% ethanol part elution volume is every 1ml resin 3ml eluting, selects every 1ml resin 4ml eluting during 45% ethanol elution.
Preferably, wherein Fructus Aurantii Immaturus or Fructus Aurantii are pulverized by 8 mesh sieves.
Preferably, general flavone content more than 50% wherein.
Preferably, general flavone content more than 80% wherein.
Preferably, wherein also comprise flavonoids and acceptable liposoluble ingredient medically thereof, flavonoids contains one or more in naringin, neohesperidin, Hesperidin, new naringin, Radix seu Folium Tosicodendri Delavayi glucoside and aglycon thereof.
Preferably, wherein naringin content, neohesperidin content reach more than 25% respectively.
Preferably, wherein the macroporous resin applied sample amount by resin volume (ml): crude drug quality (g) is 5: 1.
Preferably, wherein the loading flow velocity of every 1ml resin between 1.25～1.55ml/h.
Preferably, wherein the elution flow rate of every 1ml resin is between 1.25～1.45ml/h, and described non-polar macroporous resin blade diameter length ratio is 1: 10, and after loading, the static adsorption time of medicinal liquid is 1 hour.
Preferably, wherein said non-polar macroporous resin repeats 8 times afterwards with 5% sodium hydroxide solution immersion of 10 times of amounts 2 hours, washes with water to neutrality; 3% hydrochloric acid solution with 10 times of amounts soaked 2 hours again, washed with water to neutrality with the rear use of regenerating.
On the one hand, the invention provides Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone for the preparation of the purposes for the treatment of gastrointestinal motility obstruction type functional dyspepsia disease medicament.
On the one hand, the invention provides the purposes of the vomiting disease medicament that Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone cause for the preparation for the treatment of chemotherapeutics or other reasons.
On the one hand, the invention provides Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone for the preparation of the purposes for the treatment of Upper abdominal pain disease medicament.
On the one hand, the invention provides Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone for the preparation of the purposes for the treatment of abdominal distention, anorexia, belch disease medicament.
On the one hand, the invention provides Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone for the preparation of the purposes of the nauseating disease medicament for the treatment of.
On the other hand, the present invention also provides a kind of Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone preparation, and said preparation is take described Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone as main active.
Experimental data proves, Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone that water boiling and extraction of the present invention obtains can be used for treating gastrointestinal motility obstruction type functional dyspepsia; Upper abdominal pain, abdominal distention, anorexia, belch, feel sick, vomiting that vomiting and chemotherapeutics or other reasons cause.
More importantly, the Fructus Aurantii Immaturus that the invention water boiling and extraction obtains or Fructus Aurantii extractive of general flavone are except having the effect of significant promotion gastric motility, and be also extremely effective to the vomiting of the drug-induceds such as cisplatin, has the unexistent effect of motilium.The Cardiotoxity that there is no again simultaneously cisapride, mosapride, the cardiac safety risk that can thoroughly avoid cisapride, mosapride to cause.
And medical science and study of pharmacy personnel can't in advance under the prerequisite of not doing experiment, learn that in advance Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone that the invention water boiling and extraction obtains have above-mentioned good effect.
In sum, the Fructus Aurantii Immaturus of water boiling and extraction acquisition of the present invention or Fructus Aurantii extractive of general flavone and uses thereof bring significant technique effect.
The specific embodiment
Embodiment 1
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces, add 7 times of water gagings, decoct 3 times, the each decoction extracted 1 hour, merged decoction liquor, filter, the D101 macroporous resin of filtrate by having handled well, water, 18% ethanol elution are removed impurity successively, use again 35% ethanol, 0.5% sodium hydroxide eluting, collect 35% ethanol elution, be evaporated to the approximately fluid extract of 1.25 (55 ℃) of relative density, drying under reduced pressure (60 ℃), pulverize, and get final product.
Embodiment 2
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, by 8 mesh sieves, decoct with water 3 times, add for the first time 8 times of water gagings and boiled 2.5 hours, second and third time adds respectively 6,5 times of amounts and decocted 1 hour, filter merging filtrate, the AB-8 macroporous resin column of filtrate by having handled well, water, 15% ethanol, 45% ethanol, 1% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.31 (65 ℃) of relative density, drying under reduced pressure (70 ℃), pulverize, and get final product.
Embodiment 3
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces, add 8 times of water gagings, decoct 3 times, the each decoction extracted 1.5 hours, merged decoction liquor, filter, the D101 macroporous resin of filtrate by having handled well, water, 16% ethanol elution are removed impurity successively, use again 40% ethanol, 0.6% sodium hydroxide eluting, collect 40% ethanol elution, be evaporated to the approximately fluid extract of 1.20 (50 ℃) of relative density, drying under reduced pressure (65 ℃), pulverize, and get final product.
Embodiment 4
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces, add 9 times of water gagings, decoct 2 times, the each decoction extracted 2 hours, merged decoction liquor, filter, the D101 macroporous resin of filtrate by having handled well, water, 15% ethanol elution are removed impurity successively, use again 45% ethanol, 0.7% sodium hydroxide eluting, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.25 (55 ℃) of relative density, drying under reduced pressure (65 ℃), pulverize, and get final product.
Embodiment 5
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, by 8 mesh sieves, decoct with water 2 times, add for the first time 8 times of water gagings and boiled 2 hours, second and third time adds respectively 7 times of amounts and decocted 1 hour, filter merging filtrate, the D101 macroporous resin column of filtrate by having handled well, water, 14% ethanol, 40% ethanol, 0.9% sodium hydroxide eluting successively, collect 40% ethanol elution, be evaporated to the approximately fluid extract of 1.32 (70 ℃) of relative density, drying under reduced pressure (70 ℃), pulverize, and get final product.
Embodiment 6
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, by 8 mesh sieves, decoct with water 3 times, add for the first time 8 times of water gagings and boiled 2 hours, second and third time adds respectively 7,6 times of amounts and decocted 1.0 hours, filter merging filtrate, the AB-8 macroporous resin column of filtrate by having handled well, water, 16% ethanol, 45% ethanol, 1.1% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.29 (60 ℃) of relative density, drying under reduced pressure (75 ℃), pulverize, and get final product.
Embodiment 7
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, add 10 times of water gagings, decoct 3 times, the each decoction extracted 1 hour, merged decoction liquor, filter, the AB-8 macroporous resin of filtrate by having handled well, water, 14% ethanol elution are removed impurity successively, use again 50% ethanol, 0.8% sodium hydroxide eluting, collect 50% ethanol elution, be evaporated to the approximately fluid extract of 1.30 (55 ℃) of relative density, drying under reduced pressure (65 ℃), pulverize, and get final product.
Embodiment 8
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, by 8 mesh sieves, decoct with water 3 times, add for the first time 8 times of water gagings and boiled 1.5 hours, second and third time adds respectively 6 times of amounts and decocted 1 hour, filter merging filtrate, the D101 macroporous resin column of filtrate by having handled well, water, 12% ethanol, 45% ethanol, 0.8% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure (75 ℃), pulverize, and get final product.
Embodiment 9
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, add 11 times of water gagings, decoct 2 times, the each decoction extracted 1.5 hours, merged decoction liquor, filter, the AB-8 macroporous resin of filtrate by having handled well, water, 13% ethanol elution are removed impurity successively, use again 55% ethanol, 0.9% sodium hydroxide eluting, collect 55% ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure (70 ℃), pulverize, and get final product.
Embodiment 10
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, by 8 mesh sieves, decoct with water 2 times, add for the first time 8 times of water gagings and boiled 2 hours, second and third time adds respectively 6,5 times of amounts and decocted 1.5 hours, filter merging filtrate, the D101 macroporous resin column of filtrate by having handled well, water, 15% ethanol, 45% ethanol, 1% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure (75 ℃), pulverize, and get final product.Embodiment 11
Embodiment 11
Preparation method: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, by 8 mesh sieves, decoct with water 3 times, add for the first time 8 times of water gagings and boiled 2 hours, second and third time adds respectively 6,4 times of amounts and decocted 1 hour, filter merging filtrate, the D101 macroporous resin column of filtrate by having handled well, water, 15% ethanol, 45% ethanol, 1% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to the approximately fluid extract of 1.30 (60 ℃) of relative density, drying under reduced pressure (75 ℃), pulverize, and get final product.
Experimental data 1: extraction process optimization research
Flavonoid glycoside compound generally is soluble in hot water, methanol, ethanol, pyridine and dilute alkaline soln, and indissoluble or be insoluble in the organic solvents such as benzene, ether, chloroform, petroleum ether.Main active in Fructus Aurantii Immaturus is the flavanone constituents take naringin and neohesperidin as representative, and the polarity of these flavanones is all larger than general flavones ingredient, in water soluble and low-concentration ethanol, is soluble in the Diluted Alcohol solution of hot water or heat.
According to the physicochemical property of contained main chemical compositions in medical material, with reference to related documents, simultaneously in conjunction with the trial test result, adopt decocting to boil, the method for purification by macroporous resin on decoction liquor.
Investigate the rate of transform that index is respectively the rate of extract, general flavone content and total flavones
1. the extraction process condition is preferred
1.1 extract the selection of solvent
Fructus Aurantii Immaturus was pulverized 8 mesh sieves, get 12 parts, and be divided into 4 groups, every group average three parts, water, 65% ethanol, 95% ethanol and saturated limewater decoct separately.The results are shown in Table 1.
Table 1 extracts the selection of solvent
Conclusion: as shown in Table 1, the ethanol extraction total flavones rate of transform is lower, and the water extraction total flavones rate of transform reaches more than 90%, but water is lower than 65% ethanol extraction effect, and comprehensive production cost in conjunction with the suitability for industrialized production practical situation, selects water for extracting solvent.
1.2 the selection of medical material granularity
The Fructus Aurantii Immaturus that crushes is sieved, respectively drink three parts of sheets, 8 orders, 20 orders, 40 orders, 60 orders, 120 order, totally 18 parts, decoct with water respectively.The results are shown in Table 2.
The selection of table 2 medical material granularity
Conclusion: by relevant data in table 2 as can be known, when water decocted, the granularity of medical material had very large difference, is not that the medical material granule is more thin better, wherein the rate of transform with the rate of extract of crossing 8 mesh sieves and total flavonoid glycoside is the highest, therefore select 8 orders as the final grinding particle size of medical material.
1.3 the preferred extraction process of orthogonal experiment
According to above experimental result, existing with decocting time in decoction process (A), decoct number of times (B), amount of water (C), decoct before four of soak times (D) as the investigation factor, each factor is established three levels, presses L9 (3
4) orthogonal table carries out orthogonal (table 3, table 4), investigating index is the rate of extract and the total flavonoid glycoside rate of transform.
Table 3 factor level table
Get 27 parts of Fructus Aurantii Immaturus (8 order), every group average three parts, totally nine groups, by table 4L9 (34) orthogonal test table design experiment.The results are shown in Table 4.
Table 4L9 (3
4) orthogonal test table
(1) intuitive analysis:
From the rate of extract Ra optimised process: B
3C
3A
1D
1(B, C are principal element)
Total flavonoid glycoside rate of transform optimised process: B
3C
3A
1D
1(B, C are principal element)
(2) variance analysis
The results are shown in Table 5.
The variance analysis of table 5 take the total flavonoid glycoside rate of transform as index
＊F
1-0.05(2，2)＝19.0
＊＊F
1-0.01(2，2)＝99.0
(3) conclusion: intuitive analysis decocts number of times (B) and solvent for use volume (C) larger to decocting influential effect as can be known, and decocting time (A) and soak time (D) are less to decocting influential effect.Carry out variance analysis take the rate of transform as index, table 5 analysis result has further been verified the intuitive analysis conclusion, and namely set factor C, B have considerable influence to decocting effect successively.Because factor A is less to decocting influential effect, and first liquid of boiling medicine does not also infiltrate the medical material organization internal fully, according to producing practical situation, first fries in shallow oil increase by a hour, determine tentatively that thus decocting condition is: add 18 times of water gagings (with the medical material weight ratio) and carry (8 times, 6 times, 4 times) 3 times, decocting time is respectively 2h, 1h, 1h, and medical material does not soak.
2. demonstration test
Get 3 parts of Fructus Aurantii Immaturus (crossing 8 mesh sieves), the optimal extract process of choosing by orthogonal test carries out demonstration test, that is: (8 times/2h, 6 times/1h, 4 times/1h) decoct respectively 3 times, medical material does not soak 18 times of water gagings.The results are shown in Table 6.
Table 6 demonstration test measurement result
Conclusion: as shown in Table 6, boil 3 times by the selection process decocting, total flavonoid glycoside existing 92% or more by fried, and three sample total flavonoid glycoside rates of transform more consistent (RSD is 1.73%), result shows that last definite optimum extraction process is reasonable, stable.
Experimental data 2: the purification research of citrus aurantium total flavone extract
2.1 separation purifying technique is investigated
Fructus Aurantii Immaturus proposes a large amount of water-solubility impurities after with water extraction, must be further purified processing to water extraction liquid, to improve the content of flavonoids effective constituent, water-solubility impurity mostly is inorganic salt, saccharide, pectin and protein-based, and the active component in Fructus Aurantii Immaturus is flavone compound, according to flavonoid compound characteristic and present China isolation and purification method commonly used, therefore select to come this flavones ingredient of enriching and purifying with macroporous resin.
At first investigate experiment by preliminary applied sample amount, tentatively determine every 1000ml macroporous resin loading 5000ml extracting solution, i.e. resin volume: the crude drug quality is 4: 1.
Again further, investigate by applied sample amount, determine that finally the macroporous resin adsorption 200g crude drug of 1000ml is as final applied sample amount (being 0.2g crude drug/ml resin).
Afterwards, by the investigation of elution volume, determine that 15% ethanol part elution volume is defined as 3000ml (being every 1ml resin 3ml 15% ethanol elution), selects during 45% ethanol elution with 4000ml (1ml resin 4ml 45% ethanol elution).
Again further, by the investigation of loading flow velocity, determine that the loading flow velocity should be chosen in better (being that the loading flow velocity of every 1ml resin is between 1.25～1.55ml/h) between 1250ml/h～1550ml/h.
2.2 orthogonal optimum seeking macroporous resin Optimization Technology
According to above experimental result, existing with elution flow rate, macroporous resin blade diameter length ratio and loading after three of static adsorption times of medicinal liquid as the investigation factor, each factor is established three levels, presses L9 (3
4) orthogonal table carries out orthogonal (table 7, table 8), the investigation index is content and the total flavonoid glycoside rate of transform of the rate of extract, total flavonoid glycoside.
Table 7 factor level table
Get Fructus Aurantii Immaturus (crossing 8 mesh sieves) and extract, extracting solution is divided into 9 parts, and totally nine parts of every part of 4000ml (being equivalent to the 200g crude drug) medicinal liquids are by table 8L9 (3
4) on the macroporous resin (every resin dress 1000ml macroporous resin) of anticipating, use respectively the water of 4000ml, ethanol, 4000ml 45% ethanol and 3000ml 95% ethanol elution of 3000ml 15%.The results are shown in Table 8.
Table 8L9 (3
4) orthogonal test table
(1) intuitive analysis
Analyze the rate of extract optimised process: A according to the R value
1B
3C
1(A is principal element)
Analyze total flavonoid glycoside rate of transform optimised process: A according to the R value
1B
1C
2(A is principal element)
(2) variance analysis
The results are shown in Table 9, table 10.
The variance analysis of table 9 take the rate of extract as index
The variance analysis of table 10 take the total flavonoid glycoside rate of transform as index
ΔF
1-0.19(2，2)＝9.00
＊F
1-0.05(2，2)＝19.0
(3) conclusion: intuitive analysis as can be known, the elution flow rate of resin (A) is principal element, after the blade diameter length ratio of macroporous resin (C) and loading, the static adsorption time (B) of medicinal liquid impact is less.Carry out variance analysis take the rate of transform of the rate of extract and total flavonoid glycoside as index again, in table 9,10, analysis result has further been verified the intuitive analysis conclusion, and namely set factor A has considerable influence to elute effect.Be the leading indicator of this project due to general flavone content, therefore, determine that tentatively optimum condition is A
1B
1C
2That is: elution flow rate is 1250～1450ml/h (elution flow rate that is every 1ml resin is between 1.25～1.45ml/h), and blade diameter length ratio is 1: 10, and after loading, the static adsorption time of medicinal liquid is 1 hour.
2.3 demonstration test and the test of resin access times
Get Fructus Aurantii Immaturus (crossing 8 mesh sieves) and extract, extracting solution is taken as 13 parts, every part of 4000ml (being equivalent to the 200g crude drug), and cold preservation, standby.
A. get 1 part and be splined on 1000ml macroporous resin (the resin column blade diameter length ratio is 1: 10), the loading flow velocity is 1400ml/h, static adsorption 1 hour.Use successively ethanol, 4000ml 45% ethanol and 3000ml 95% ethanol elution of water, 3000ml 15% of 4000ml with the flow velocity eluting of 1400ml/h.Collect 45% ethanol elution and carry out assay.The results are shown in Table 11.
B. with a with after resin to be washed to without alcohol flavor, separately get this resin on the sample of a refrigerator cold-storage, the step reprocessing of pressing a.Resin carries out absorption next time (the same a.b of following steps) successively.
Table 11 demonstration test measurement result
Conclusion: the demonstration test of being undertaken by optimised process as can be seen from Table 11, the sample total flavonoid glycoside content and the total flavonoid glycoside rate of transform that repeat 8 gained all increase.And the indices homogeneous of sample, stable, illustrate that this technique is best, stable.Need later on resin is carried out Regeneration Treatment for 8 times in use simultaneously.
2.4 service test after resin regeneration
5% sodium hydroxide solution of resin with 10 times of amounts soaked 2 hours, wash with water to neutrality; 3% hydrochloric acid solution with 10 times of amounts soaked 2 hours again, washed with water to neutrality.The resin that regeneration is good by demonstration test and a resin access times test lower method absorb-elute successively 3 times, calculates the rate of extract, Liquid Detection total flavonoid glycoside content and the rate of transform.The results are shown in Table 12.
Service test after table 12 resin regeneration
After result shows resin regeneration, operating position is good.
According to above-mentioned separation, purification research, merge extractive liquid, after medicinal material extract filters the D of filtrate by having handled well
101Macroporous resin column, water, 15% ethanol, 45% ethanol, 1% sodium hydroxide eluting, collect 45% ethanol elution successively.
Experimental data 3: the research of citrus aurantium total flavone extract drying means and baking temperature
Get Fructus Aurantii Immaturus 1kg, totally 10 parts, according to above-mentioned definite extraction, purifying process, after extraction, purification, collect 45% ethanol elution, decompression recycling ethanol also is concentrated into the approximately extractum of 1.30 (60 ℃) of relative density, adopts respectively oven drying method, boulton process to compare research to above-mentioned extractum.Oven drying method and boulton process carry out respectively the comparison of 5 kinds of different temperatures (50 ℃, 65 ℃, 75 ℃, 85 ℃, 95 ℃).Investigate the content of total flavonoid glycoside.The results are shown in Table 13 and table 14.
The measurement result of table 13 oven drying method different temperatures
The measurement result of table 14 boulton process different temperatures
From table 13,14 as can be known, oven drying method is long than the dry required time of vacuum on the same temperature level, and effect is not as vacuum drying.The sample color and luster of oven drying method oven dry is relatively poor, be difficult for pulverizing, and along with the rising of temperature, oven drying method easily causes carbonization because of hot-spot, thereby causes the color sample blackout, and effective ingredient is lost.Easy-to-drawly during vacuum drying driedly become cellular, be conducive to pulverize, and color and luster is better, vacuum drying temperature is controlled at 75 ℃ of left and right and is advisable simultaneously, saves time and can not cause the loss of effective ingredient.
Below all select embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts as experimental group
Experimental data 4: effect is told in Fructus Aurantii Immaturus total flavones glucoside extract town
4.1 the Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of pigeon vomiting on copper sulfate
60 of pigeons, body weight 280～350g, male and female half and half.Be divided at random 6 groups: matched group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (20,40,80,160mgkg
-1) 4 dosage group and positive drug group (fourth beautiful jade 40mgkg
-1), 10 every group, ♀
Half and half.Each is organized the pigeon fasting and can't help water 16 hours, next day single administration, matched group gives the distilled water gavage, other group gavage relative medicines, 10mlkg
-1Respectively organize pigeon after 1 hour and give 3.5% copper-bath 10mgkg
-1The gavage modeling.Record is respectively organized pigeon vomiting latent time and is vomitted number of times in 2 hours, and result of the test shows, the Fructus Aurantii Immaturus total flavones glucoside extract (20,40,80,160mgkg
-1) single administration all can not extend the vomiting latent time of pigeon due to copper sulfate, but the Fructus Aurantii Immaturus total flavones glucoside extract (40,80mgkg
-1) single administration can reduce the vomiting number of times of pigeon due to copper sulfate, sees Table 15.
Table 15 Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of pigeon vomiting on copper sulfate
Annotate: compare with matched group,
*P＜0.05,
*P＜0.01
4.2 the Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of pigeon vomiting on cisplatin
80 of pigeons, body weight 280～350g, male and female half and half.Be divided at random 8 groups: matched group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (20,40,80,160mgkg
-1) 4 dosage group and positive drug 3 groups of (fourth beautiful jade 80,160mgkg
-1With Ondansetron Hydrochloride injection 5ugkg
-1) 10 every group, ♀
Half and half.Each is organized the pigeon fasting and can't help water 16 hours, next day single administration, matched group gives the distilled water gavage, other groups give relative medicine, the injection of 1 hour pneumoretroperitoneum gives 13mgkg
-1Cisplatin for inj.Record is respectively organized pigeon vomiting latent time and is vomitted number of times in 4 hours, and result of the test shows, Fructus Aurantii Immaturus total flavones glucoside extract 80mgkg
-1Single administration can extend the vomiting time of pigeon due to cisplatin, but 20,40 and 160mgkg
-1The time all can not extend the vomiting time of pigeon due to cisplatin; The Fructus Aurantii Immaturus total flavones glucoside extract (80,160mgkg
-1) single administration can reduce the vomiting number of times of pigeon due to cisplatin, sees Table 16
Table 16 Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of pigeon vomiting on cisplatin
Annotate: compare with matched group,
*P＜0.05,
*P＜0.01
4.3 the Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of hybrid dog vomiting on apomorphine
56 of hybrid dogs, body weight 8.0～10.0kg, male and female half and half.Be divided at random 7 groups: matched group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (15,30,60,120mgkg
-1) 4 dosage group and 2 groups of (motilium 30mgkg of positive drug
-1With metoclopramide injection 1mgkg
-1), 8 every group, ♀
Half and half.Each is organized the fasting of hybrid dog and can't help water 16 hours, next day single administration, matched group gives the distilled water gavage, other groups give relative medicine gavage, 5mlkg
-1Give 500ugkg after 1 hour
-1The apomorphine subcutaneous injection.Record is respectively organized hybrid dog vomiting latent time and is vomitted number of times in 1 hour.Result of the test shows, the Fructus Aurantii Immaturus total flavones glucoside extract (15,30,60,120mgkg
-1) single administration all can not reduce apomorphine and cause hybrid dog vomiting number of times; The Fructus Aurantii Immaturus total flavones glucoside extract (15,30,120mgkg
-1) also can not extend the vomiting latent time that apomorphine causes the hybrid dog, but Fructus Aurantii Immaturus total flavones glucoside extract 60mgkg
-1The time can extend the vomiting latent time of hybrid dog, see Table 17.
Table 17 Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of hybrid dog vomiting on apomorphine
Annotate: compare with matched group,
*P＜0.05
Experimental data 5: the Fructus Aurantii Immaturus total flavones glucoside extract is to the intestinal progradation
5.1 the Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of the low mice of intestinal function on dopamine
70 of mices, body weight 19～22g is divided into 7 groups at random: matched group, model group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (25,50,100,200mgkg
-1) 4 dosage group and positive drug group (motilium 50mgkg
-1), 10 every group, ♀
Half and half.Each is organized the mice fasting and can't help water 16 hours, next day single administration, matched group gavage distilled water, other groups give the relative medicine gavage.After 30 minutes, except matched group, all the other mices give 1.7mgkg to each mouse stomach
-1The modeling of dopamine hydrochloride inj subcutaneous injection, after 20 minutes, 70 mices gavage respectively 2% sodium carboxymethyl cellulose carbon powder suspension 0.2ml10g
-1, after 20 minutes, mice takes off neck execution, takes out immediately gastrointestinal, and tiling is measured the carbon powder front end to distance and the small intestinal total length of pylorus, calculates ink propulsive rate with ratio between two.Result of the test shows, subcutaneous injection 1.7mgkg
-1The dopamine intestinal that can obviously suppress mice advance, the Fructus Aurantii Immaturus total flavones glucoside extract (25,50,100mgkg
-1) can improve the mice intestinal propelling activity that is suppressed by dopamine, see Table 18.
Table 18 Fructus Aurantii Immaturus total flavones glucoside extract causes on dopamine the impact that the mice intestinal advances
70 of mices, body weight 19～22g is divided into 7 groups at random: matched group, model group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (25,50,100,200mgkg
-1) 4 dosage group and positive drug group (motilium 50mgkg
-1), 10 every group, ♀
Half and half.Each is organized the mice fasting and can't help water 16 hours, next day single administration, matched group gavage distilled water, other groups give the relative medicine gavage.After 30 minutes, except matched group, all the other mices give 2.5mgkg to each mouse stomach
-1The modeling of atropine subcutaneous injection, after 20 minutes, 70 mices gavage respectively 2% sodium carboxymethyl cellulose carbon powder suspension 0.2ml10g
-1, after 20 minutes, mice takes off neck execution, takes out immediately gastrointestinal, and tiling is measured the carbon powder front end to distance and the small intestinal total length of pylorus, calculates ink propulsive rate with ratio between two.Result of the test shows, subcutaneous injection 2.5mgkg
-1The atropine intestinal that can obviously suppress mice advance, Fructus Aurantii Immaturus total flavones glucoside extract 50mgkg-1 can improve the mice intestinal propelling activity by the atropine inhibition, sees Table 19.
Table 19 Fructus Aurantii Immaturus total flavones glucoside extract causes on atropine the impact that the mice intestinal advances
Annotate: compare with matched group,
ΔP＜0.05; Compare with model group,
*P＜0.05
5.3 the Fructus Aurantii Immaturus total flavones glucoside extract causes the impact of the low mice of intestinal function on epinephrine
70 of mices, body weight 19～22g is divided into 7 groups at random: matched group, model group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (25,50,100,200mgkg-1) 4 dosage groups and positive drug groups (motilium 50mgkg-1), 10 every group, ♀
Half and half.Each is organized the mice fasting and can't help water 16 hours, next day single administration, matched group gavage distilled water, other groups give the relative medicine gavage.After 30 minutes, except matched group, all the other mices give 0.5mgkg to each mouse stomach
-1The modeling of adrenalin hydrochloride injection subcutaneous injection, after 20 minutes, 70 mices gavage respectively 2% sodium carboxymethyl cellulose carbon powder suspension 0.2ml10g-1, after 20 minutes, mice takes off neck execution, take out immediately gastrointestinal, tiling, measure the carbon powder front end to distance and the small intestinal total length of pylorus, calculate ink propulsive rate with ratio between two.Result of the test shows, subcutaneous injection 0.5mgkg
-1The epinephrine intestinal that can obviously suppress mice advance, the Fructus Aurantii Immaturus total flavones glucoside extract (25,50,100,200mgkg
-1) all can not improve the mice intestinal propelling activity by epinephrine inhibited, see Table 20.
Table 20 Fructus Aurantii Immaturus total flavones glucoside extract causes on epinephrine the impact that the mice intestinal advances
Annotate: compare with matched group,
The Δ ΔP＜0.01
Experimental data 6: the Fructus Aurantii Immaturus total flavones glucoside extract is on gastric secretion, gastric acid and pepsic impact
6.1 the impact on gastric secretion, gastric acid
48 of rats, body weight 230～270g is divided into 6 groups at random: matched group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (20,40,80,160mgkg
-1) 4 dosage group and positive drug group (motilium 40mgkg
-1), 8 every group, ♀
Half and half.Before test, the rat fasting be can't help water 36 hours, gastric infusion of rat, and matched group gives distilled water, and other groups give relative medicine.The administration while, each rat is opened the abdominal cavity under the ether light anaesthesia, the ligation pylorus, and the rat gastric juice of 5 hours after the collection pylorus ligation, with the centrifugal 5min of 3000r/min, the gastric contents such as place to go food debris are with the accurate weighing gastric juice of graduated cylinder amount.Measure gastric juice 1ml, be placed in conical flask and add the equivalent distilled water, mix homogeneously adds each 2 of 0.5% pair of dimethyl azoaniline alcoholic solution of free acid indicator and total acid indicator 0.5% phenolphthalein alcoholic solution, if mix in gastric juice and contain hydrochloric acid, namely be cherry red.Drip the NaOH of 0.01mol/L with burette,, limit edged shaking flasks disappears to red, till beginning orange colour to occur.Be the terminal point of free hydrochloric acid.Write down the amount that spends till NaOH no longer deepens to redness, be the terminal point of total acidity.Write down twice titration and spend NaOH solution total amount.Calculate concentration and total secretory volume of free hydrochloric acid and total acid.Result of the test shows, the Fructus Aurantii Immaturus total flavones glucoside extract (20,40,80mgkg
-1) gastric secretion of pylorus ligation rat is had no significant effect, but Fructus Aurantii Immaturus total flavones glucoside extract 160mgkg
-1Can reduce the gastric secretion of rat; The Fructus Aurantii Immaturus total flavones glucoside extract (80,160mgkg
-1) concentration of can raise pylorus ligation rat free hydrochloric acid and total acid, but each dosage of Fructus Aurantii Immaturus total flavones glucoside extract (20,40,80,160mgkg
-1) on the secretory volume of free hydrochloric acid and total acid without impact, see Table 21.
Table 21 Fructus Aurantii Immaturus total flavones glucoside extract is to rat gastric secretion, free hydrochloric acid and total acid concentration as influencing factor
Annotate: compare with matched group,
*P＜0.05,
*P＜0.01
The capillary glass-tube that internal diameter 1～2mm thickness is well-balanced is cleaned and is dried, and gets appropriate Ovum Gallus domesticus album and fully beats the even rear filtered through gauze of using, and above-mentioned glass capillary is utilized siphonage, fills Ovum Gallus domesticus album (the pipe planted agent sneaks into without bubble).Then be positioned in 85 ℃ of heat steams and make protein coagulating.After cooling, take out paraffin with protein pipe two ends sealing, standby in the ice storage case.Get the conical flask that gastric juice 1ml puts into 50ml during experiment, add 0.05N hydrochloric acid solution 15ml, shake up, put two of approximately long 2.5cm protein pipe into.Filled in bottleneck, be put in 37 ℃ of calorstats temperature and incubate 24h.Measure the length (mm) of protein pipe two ends transparent part with chi and ask its meansigma methods with the value of four ends, with the pepsic unit=meansigma methods of following formula calculating
2* 16.Result of the test shows, embodiment 11 each dosage of Fructus Aurantii Immaturus total flavones glucoside extract (20,40,80,160mgkg
-1) pepsin activity of pylorus ligation rat is had no significant effect, see Table 22.
The impact of table 22 Fructus Aurantii Immaturus total flavones glucoside extract on rat free hydrochloric acid, total acid secretory volume and pepsin activity
Experimental data 7: the impact of Fructus Aurantii Immaturus total flavones glucoside extract on smooth muscle
7.1 the impact on the gastrointestinal smooth muscle that exsomatizes
Select the healthy adult rat, the head of fiercelying attack causes dusk, takes out rapidly full stomach and duodenum stage casing 1-2cm, and the place cuts off along greater gastric curvature, with containing 95%O
2And 5%CO
2The Krebs liquid (composition: NaCl 117mmol/L, KCl 4.8mmol/L, CaCl of the fully saturated pH 7.35～7.45 of mist
22.5mmol/L, MgCl
21.2mmol/L, NaH
2PO
41.2mmol/L, NaHCO
325mmol/L, Glucose 11mmol/L) repeatedly rinse and exenterate, get respectively at the bottom of the gastric antrum stomach function regulating longitudinal and transverse flesh bar and duodenum stage casing and be about 1cm, wide approximately 0.5cm, the ventilation hook is fixed in the bottom, the upper end is connected with transducer (JH-2 type muscular tension sensor), and it is inserted in the bath pipe that fills oxygen-saturated Krebs liquid (pH 7.35～7.45) 20ml, the adjustment preload is 1g, change liquid once every 20min, by ventilation hook direct oxygen injection in the body lotion, use BL-420E biological function experimental system and record its tension force and frequency.Outer tube through constant current infusion pump [continue, constant temperature (37 ± 0.5 ℃), constant speed (3～4ml/min)] perfusion to keep temperature in bath.After specimen is stable, add medicine ultimate density to be measured to be respectively 0.25mg/L, 0.4mg/L, 0.55mg/L, 0.7mg/L, 0.85mg/L and 1.00mg/L, record and observe medication front and back frequency and tension change.The isolated test result shows, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extract (0.25mg.ml
-1～1.0mg.ml
-1) can reduce longitudinal and transverse shape flesh and duodenum smooth muscle tension force at the bottom of the gastric antrum stomach function regulating, suppress gastrointestinal motility, see Table 23～27.
The impact of table 23 Fructus Aurantii Immaturus total flavones glucoside extract on exsomatize gastric antrum transversal tension force and frequency
Annotate: with comparison before administration,
*P＜0.05,
*P＜0.01
The impact of table 24 Fructus Aurantii Immaturus total flavones glucoside extract on exsomatize gastric antrum longitudinal muscle tension force and frequency
Annotate: with comparison before administration,
*P＜0.05,
*P＜0.01
Table 25 Fructus Aurantii Immaturus total flavones glucoside extract on the stomach that exsomatizes at the bottom of the impact of transversal tension force and frequency
Annotate: with comparison before administration,
*P＜0.05,
*P＜0.01
Table 26 Fructus Aurantii Immaturus total flavones glucoside extract on the stomach that exsomatizes at the bottom of the impact of longitudinal muscle tension force and frequency
Annotate: with comparison before administration,
*P＜0.05,
*P＜0.01
The impact of table 27 Fructus Aurantii Immaturus total flavones glucoside extract on Isolated Duodenum stage casing tension force and frequency
Annotate: with comparison before administration,
*P＜0.05,
*P＜0.01
7.2 on the impact at body duodenum smooth muscle
56 of rats, body weight 180～230g is divided into 7 groups at random: matched group, model group, embodiment 11 Fructus Aurantii Immaturus total flavones glucoside extracts (20,40,80,160mgkg
-1) 4 dosage group and positive drug group (fourth beautiful jade 40mgkg
-1), 8 every group, ♀
Half and half.The rat oral gavage administration, matched group and model group give distilled water, and other groups give relative medicine.Each is organized the rat fasting and can't help water 16 hours, next day single administration, after 30 minutes, except matched group, all the other rats give 1.5mgkg to each rat oral gavage
-1The modeling of dopamine hydrochloride inj subcutaneous injection, chloral hydrate anesthesia, open the abdominal cavity along the abdomen median line, selection duodenum stage casing is 2-3cm approximately, be fixed on intestinal segment two ends on the aperture of intestinal tube stationary pipes both sides with stitching thread, with a stitching thread, one end is sewn on intestinal wall in the middle of intestinal segment, the other end is passed by intestinal tube stationary pipes central authorities and is connected on JH-2 type muscular tension sensor, and records tension force and the frequency of each rat preduodenal by BL-420E biological function experimental system again.Result of the test shows, subcutaneous injection 1.5mgkg
-1Dopamine can obviously be suppressed at tension force and the frequency of body rat preduodenal, the Fructus Aurantii Immaturus total flavones glucoside extract (20,40,80,160mgkg
-1) can improve tension force and frequency by the rat preduodenal of dopamine inhibition, see Table 28.
Table 28 Fructus Aurantii Immaturus total flavones glucoside extract is on the impact in body duodenum stage casing tension force and frequency
Annotate: compare with matched group,
ΔP＜0.05; Compare with model group,
*P＜0.05,
*P＜0.01
Claims (12)
1. Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone are for the preparation for the treatment of abdominal distention, anorexia, the purposes of belch disease medicament, described Fructus Aurantii Immaturus or Fructus Aurantii extractive of general flavone are by obtaining Fructus Aurantii Immaturus or Fructus Aurantii with water boiling and extraction, wherein boiling water extraction method is: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～15 times of water gagings, decoct 1～3 time, the each decoction extracted 1～3 hour, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, wherein the loading flow velocity of every 1ml resin is between 1.25～1.55ml/h, described non-polar macroporous resin blade diameter length ratio is 1: 10, after loading, the static adsorption time of medicinal liquid is 1 hour, wherein the macroporous resin applied sample amount is by resin volume ml: crude drug quality g is 5: 1, water successively, ethanol elution below 20% is removed impurity, use again 25～90% ethanol, 0.5～2% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.20～1.40 fluid extract 50 ℃～70 ℃ mensuration, drying under reduced pressure, dry temperature is 60～80 ℃, pulverize, and get final product, wherein general flavone content is more than 80%, wherein also comprise flavonoids and acceptable liposoluble ingredient medically thereof, flavonoids contains naringin, neohesperidin, Hesperidin, new naringin, Radix seu Folium Tosicodendri Delavayi glucoside and aglycon thereof, naringin content wherein, neohesperidin content reaches more than 25% respectively.
2. purposes as claimed in claim 1, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～13 times of water gagings, decoct 2～3 times, the each decoction extracted 1～2.5 hour, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 30%～85% ethanol, 0.5～1.8% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.25～1.40 fluid extract 55 ℃～70 ℃ mensuration, drying under reduced pressure, dry temperature is 65～80 ℃, pulverize, and get final product.
3. purposes as claimed in claim 2, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～11 times of water gagings, decoct 2～3 times, the each decoction extracted 1.5～2.5 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 35%～75% ethanol, 0.5～1.6% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.25～1.35 fluid extract 55 ℃～65 ℃ mensuration, drying under reduced pressure, dry temperature is 70～80 ℃, pulverize, and get final product.
4. purposes as claimed in claim 3, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～9 times of water gagings, decoct 3 times, the each decoction extracted 1.5～2 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 40%～65% ethanol, 0.5～1.3% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.30～1.35 fluid extract about 60 ℃～65 ℃ mensuration, drying under reduced pressure, dry temperature is 70～75 ℃, pulverize, and get final product.
5. purposes as claimed in claim 4, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～7 times of water gagings, decoct 3 times, the each decoction extracted 2 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 45%～55% ethanol, 0.8～1.2% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.30 fluid extract 60 ℃ of mensuration, drying under reduced pressure, dry temperature is 75 ℃, pulverize, and get final product.
6. purposes as claimed in claim 3, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～8 times of water gagings, decoct 2 times, the each decoction extracted 1.5 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 40%～55% ethanol, 0.7～1.3% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.30 fluid extract 60 ℃ of mensuration, drying under reduced pressure, dry temperature is 75 ℃, pulverize, and get final product.
7. purposes as claimed in claim 6, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, be cut into decoction pieces or pulverizing, add 4～6 times of water gagings, decoct 2 times, the each decoction extracted 1.5 hours, merge decoction liquor, filter, the non-polar macroporous resin of filtrate by having handled well, water successively, ethanol elution below 20% is removed impurity, use again 45%～50% ethanol, 0.9～1.1% sodium hydroxide eluting, collect ethanol elution, be evaporated to relative density and be about 1.30 fluid extract 60 ℃ of mensuration, drying under reduced pressure, dry temperature is 75 ℃, pulverize, and get final product.
8. purposes as claimed in claim 1, concrete boiling water extraction method is as follows: get Fructus Aurantii Immaturus or Fructus Aurantii, pulverize, decoct with water 3 times, add for the first time 8 times of water gagings and boiled 2 hours, second and third time adds respectively 6,4 times of amounts and decocted 1 hour, filter merging filtrate, water, 15% ethanol, 45% ethanol, 1% sodium hydroxide eluting successively, collect 45% ethanol elution, be evaporated to relative density and be about 1.30 fluid extract 60 ℃ of mensuration, drying under reduced pressure, dry temperature is 75 ℃, pulverize, and get final product.
9. purposes as claimed in claim 8, wherein 15% ethanol part elution volume is every 1ml resin 3ml eluting, selects every 1ml resin 4ml eluting during 45% ethanol elution.
10. as the described purposes of claim 1-9 any one, wherein Fructus Aurantii Immaturus or Fructus Aurantii are pulverized by 8 mesh sieves.
11. purposes as described in claim 1～9 any one, wherein the elution flow rate of every 1ml resin is between 1.25～1.45ml/h.
12. purposes as described in claim 1～9 any one, wherein said non-polar macroporous resin repeat 8 times afterwards with 5% sodium hydroxide solution immersion of 10 times of amounts 2 hours, wash with water to neutrality; 3% hydrochloric acid solution with 10 times of amounts soaked 2 hours again, washed with water to neutrality with the rear use of regenerating.
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Effective date of registration: 20190116 Address after: 341000 East Avenue, Shahe Industrial Park Station, Zhanggong District, Ganzhou City, Jiangxi Province Patentee after: Jiangxi Qingfeng Pharmaceutical Co., Ltd. Address before: 341000 No. 8 East Avenue, Shahe Industrial Park Station, Ganzhou City, Jiangxi Province Co-patentee before: Jiangxi Qingfeng Drugs Research Co., Ltd. Patentee before: Jiangxi Qingfeng Pharmaceutical Co., Ltd. |