CN102178695B - Method for preparing pig-bile-combination-type total bile acid and application thereof - Google Patents
Method for preparing pig-bile-combination-type total bile acid and application thereof Download PDFInfo
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Abstract
The invention relates to a method for preparing pig-bile-combination-type total bile acid. The method comprises the steps of extracting by utilizing ethanol, separating and purifying and drying. Proved by medicinal effectiveness tests for treating colonitis, the pig-bile-combination-type total bile acid prepared by utilizing the method provided by the invention has an inhibitory action on the increase of myeloperoxidase activity and tumor necrosis factor alpha content of model mice and has the advantages of inhibiting the secretion of proinflammatory cytokine, regulating the immune response, effectively blocking the damage of the immunoreactivity to an organism and achieving the purpose of treating colonic inflammation. The pig-bile-combination-type total bile acid prepared by utilizing the method provided by the invention can be adopted as a raw material to be made into various commonly-used preparation types such as tablets, capsules, powders, pills, granules, enemas and the like.
Description
Technical field
The present invention relates to the application of Fel Sus domestica conjunction type total bile acid in medicament, be specifically related to the preparation and the application in the treatment colitis thereof of conjunction type total bile acid.
Background technology
Ulcerative colitis (ulcerative colitis) claim chronic nonspecific colitis again, is a kind of chronic idiopathic inflammatory reaction of mainly involving colon and rectum, belongs to a kind of of inflammatory bowel.Many research confirms that colitis serves as main performance with the serious imbalance of autoimmune system and interior environment factor interaction; Be that a kind of agnogenic big intestinal mucosa chronic inflammatory disease and ulcer sexually transmitted disease (STD) become, pathological change has characteristics such as shallow table property, dispersivity, seriality.The colitis pathological changes mainly is limited to big intestinal mucosa and Submucosa, and clinical is characteristic with diarrhoea, mucus bloody purulent stool, stomachache, and general morbidity is slow, and the course of disease is very long, and how repeatedly state of an illness weight differs, outbreak.After once showing effect, part patient promptly stops; The part patient course of disease is chronic time-continuing process, and its cause of disease maybe be relevant with inheritance susceptible crowd caused Intestinal Mucosal Immunity system exception reaction under multifactor effect.Colitis sees that according to statistics, per 100,000 philtrum sickness rate are respectively: Britain, 79.9, Germany, 109, France, 110, Switzerland, 17.6, New Zealand, 41.3, the U.S., 42 in that American-European countries is more.According to the WMJ latest report, about 250000 people suffer from colitis in the U.S., and its year outpatient service is 250,000 person-times, and number of hospitalized is 20,000 person-times, the annual loss for more than 100 ten thousand working days.Though China does not have accurately statistics, and is in rising trend at China's sickness rate in recent years yet.
The medicine that is used to treat colitis clinically mainly contains aminosallcylic acid class, hormone, immunosuppressant, antibiotic, heparin etc., but is first-selection with aminosallcylic acid class, cortical steroid still at present.The aminosallcylic acid class is that treatment is light, moderate popularity colitis patient's a line medicine.This type preparation mainly contains sulfasalazine, mesalazine, Olsalazine etc.; Its main mechanism is to change the enteric microorganism system; Change the synthetic and electrolyte exchange of prostaglandin in the mucosa; Stop the synthetic of inflammatory mediator and discharge the effect of prevention natural killer cell, mastocyte, neutrophilic granulocyte, mucosa lymphocyte and macrophage, the generation of restricted activity oxygen etc.(Sulfasalazine is to treat colitis at present SASP) to sulfasalazine, and the common drug of especially light, medium-sized colitis because the medicine valency is cheap, determined curative effect, still is a choice drug at home.Sulfasalazine is by 5-aminosalicylic acid and sulfapyridine be combined into, the cracking under the effect of colon bacteria azo reductase of oral back, and sulfapyridine is a carrier part, discharges 5-aminosalicylic acid performance therapeutical effect after making medicine arrive colon.Can make 75% patient's clinical symptoms and sigmoidoscopy improvement and improvement behind the oral sulfasalazine, but the erious adverse reaction of sulfasalazine, at least 20% patient can not take this medicine.Common untoward reaction has headache, gastrointestinal symptom, urticaria, dyspnea, arthralgia, hepatitis etc.Immunosuppressant such as azathioprine, methotrexate, ciclosporin etc. are used for the intractable colitis of serious symptom more, obtained bigger effect in the treatment, but untoward reaction are also apparent in view.
Fel Sus domestica is the Chinese medicine of China, is used for calentura dry thirsty, conjunctival congestion, sore throat, jaundice, pertussis, asthma, has loose bowels, diseases such as dysentery, constipation, carbuncle pyogenic infections from tumour or sore.Comprise cattle sulphur hyocholic acid (Taurohyocholic acid), cattle sulphur Hyodeoxycholic Acid (Taurohyodeoxycholic acid), Taurochenodeoxycholic Acid (Taurochenodeoxycholic acid), sweet ammonia hyocholic acid (Glycohyocholic acid), sweet ammonia Hyodeoxycholic Acid (Glycohyodeoxycholic acid) and sweet ammonia chenodeoxycholic acid (Glycochenodeoxycholic acid) in the Fel Sus domestica conjunction type total bile acid.The application of Fel Sus domestica conjunction type total bile acid in medicament also do not appear in the newspapers at present.
Summary of the invention
Technical problem to be solved by this invention is to overcome the shortcoming of above-mentioned treatment colitis medicine, and a kind of method for preparing of Fel Sus domestica conjunction type total bile acid is provided.
Another technical problem to be solved by this invention is for Fel Sus domestica conjunction type total bile acid a kind of new purposes to be provided.
The method for preparing that solves the problems of the technologies described above the technical scheme that is adopted and be Fel Sus domestica conjunction type total bile acid is made up of following step:
1, ethanol lixiviate
Get fresh pig bile, add 5 times of its volumes amount mass fraction and be 95% ethanol, left standstill 12～14 hours, filter, reclaim solvent, brownish black extractum.
2, separation and purification
The extractum that obtains in the step 1 is added in the 100-160 purpose column chromatography silica gel, and the mass ratio of extractum and column chromatography silica gel is 1: 2, mixes appearance; Volatilize solvent, be ground into 80 order fine powders, fine powder and neutral alumina are loaded in the chromatographic column by mass ratio 1: 10～20 dry method; The use mass fraction is 95% ethanol elution; Collect eluent, on silica gel g thin-layer plate, launch the back with developing solvent and use chromogenic reagent with the capillary tube point sample; Merge eluent when occurring conjunction type Hyodeoxycholic Acid speckle on the lamellae, when speckle disappears, finish.
Above-mentioned developing solvent is that the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1: 0.1 mixed solution, developer are vanillin sulphuric acid test solution or ethanol system sulphuric acid test solution.
3, preparation Fel Sus domestica conjunction type total bile acid
With the eluent decompression and solvent recovery of collecting, 80～100 ℃ of oven dry in baking oven, Fel Sus domestica conjunction type total bile acid, content in cattle sulphur Hyodeoxycholic Acid more than 50%.
In purification procedures 2 of the present invention, the extractum that obtains in the step 1 is added in the 100-160 purpose column chromatography silica gel, the mass ratio of extractum and column chromatography silica gel is 1: 2; Mix appearance, volatilize solvent, be ground into 80 order fine powders; Fine powder and neutral alumina are loaded in the chromatographic column by 1: 15 dry method of optimum quality ratio, and the use mass fraction is 95% ethanol elution, collects eluent; With the capillary tube point sample on silica gel g thin-layer plate; Launch the back with developing solvent and use chromogenic reagent, merge eluent when occurring conjunction type Hyodeoxycholic Acid speckle on the lamellae, when speckle disappears, finish.
The purposes of the Fel Sus domestica conjunction type total bile acid of method for preparing in preparation treatment colitis medicine.
Effective ingredient uses with the form of conventional pharmaceutical formulation with the medicine of Fel Sus domestica conjunction type total bile acid preparation treatment colitis.Described conventional pharmaceutical formulation contains the Fel Sus domestica conjunction type total bile acid as active component, and this active component mixes like solid or the liquid excipient that is suitable for the organic or inorganic of administration in the gastrointestinal with pharmaceutically acceptable carrier in preparation.This pharmaceutical formulation can be solid form such as tablet, granule, capsule; Also can be liquid form such as suspending agent, syrup, enema etc.
Can contain auxiliary substance, stabilizing agent, wetting agent and other additive commonly used in the above-mentioned preparation, like lactose, citric acid, tartaric acid, stearic acid, magnesium stearate, Gypsum Fibrosum powder, sucrose, corn starch, Pulvis Talci, gelatin, agar, pectin, Oleum Arachidis hypogaeae semen, olive oil, cocoa butter, ethylene glycol, ascorbic acid, mannitol etc.
Above-mentioned preparation can be processed according to the conventional preparation technology of various preparations.
The Fel Sus domestica conjunction type total bile acid process that adopts the present invention to obtain is treated the pharmacology test of colitis; Show that Fel Sus domestica conjunction type total bile acid has inhibitory action to model mice activity of myeloperoxidase and the rising of tumor necrosis factor content; Show that Fel Sus domestica conjunction type total bile acid can suppress the secretion of proinflammatory cytokine; Regulate immunoreation, effectively the blocking immunity reactivity reaches therapeutic purposes to the damage that body causes.
The specific embodiment
To further explain of the present invention, but the invention is not restricted to these embodiment below in conjunction with embodiment.
Embodiment 1
With fresh pig bile 100mL is example, and the method for preparing of Fel Sus domestica conjunction type total bile acid is following:
1, ethanol lixiviate
Get fresh pig bile, adding 500mL mass fraction is 95% ethanol, leaves standstill 12～14 hours, filters, and reclaims solvent, gets brownish black extractum;
2, separation and purification
Get above-mentioned extractum 2g, add 100-160 purpose column chromatography silica gel 4g and mix appearance, volatilize solvent, be ground into 80 order fine powders; Gained fine powder and neutral alumina are loaded in the chromatographic column according to 1: 15 ratio dry method of mass ratio; Using mass fraction is 95% ethanol elution, collects eluent, with the capillary tube point sample on silica gel g thin-layer plate; Using the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1: 0.1 blended developing solvent launches; With vanillin sulphuric acid test solution chromogenic reagent,, conjunction type Hyodeoxycholic Acid speckle merges eluent when appearring in lamellae,, finishes speckle when disappearing.
3, preparation Fel Sus domestica conjunction type total bile acid
With the eluent decompression and solvent recovery of collecting, 80～100 ℃ of oven dry in baking oven get Fel Sus domestica conjunction type total bile acid, and content is with cattle sulphur Hyodeoxycholic Acid (C
26H
45O
6NS) meter is more than 50%.
Embodiment 2
With fresh pig bile 100mL is example, and the method for preparing of Fel Sus domestica conjunction type total bile acid is following:
In purification procedures 2, get above-mentioned extractum 2g, add 100-160 purpose column chromatography silica gel 4g and mix appearance, volatilize solvent, be ground into 80 order fine powders; Gained fine powder and neutral alumina are loaded in the chromatographic column according to 1: 10 ratio dry method of mass ratio; Using mass fraction is 95% ethanol elution, collects eluent, with the capillary tube point sample on silica gel g thin-layer plate; Using the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1: 0.1 blended developing solvent launches; With vanillin sulphuric acid test solution chromogenic reagent,, conjunction type Hyodeoxycholic Acid speckle begins to merge eluent when appearring in lamellae,, finishes speckle when disappearing.
Other steps are identical with embodiment 1.Be prepared into Fel Sus domestica conjunction type total bile acid, content is with cattle sulphur Hyodeoxycholic Acid (C
26H
45O
6NS) meter is more than 50%.
Embodiment 3
With fresh pig bile 100mL is example, and the method for preparing of Fel Sus domestica conjunction type total bile acid is following:
In purification procedures 2, get above-mentioned extractum 2g, add 100-160 purpose column chromatography silica gel 4g and mix appearance, volatilize solvent, be ground into 80 order fine powders; Gained fine powder and neutral alumina are loaded in the chromatographic column according to 1: 20 ratio dry method of mass ratio; Using mass fraction is 95% ethanol elution, collects eluent, with the capillary tube point sample on silica gel g thin-layer plate; Using the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1: 0.1 blended developing solvent launches; With vanillin sulphuric acid test solution chromogenic reagent,, conjunction type Hyodeoxycholic Acid speckle begins to merge eluent when appearring in lamellae,, finishes speckle when disappearing.
Other steps are identical with embodiment 1.Be prepared into Fel Sus domestica conjunction type total bile acid, content is with cattle sulphur Hyodeoxycholic Acid (C
26H
45O
6NS) meter is more than 50%.
Embodiment 4
With fresh pig bile 100mL is example, and the method for preparing of Fel Sus domestica conjunction type total bile acid is following:
In the purification procedures 2 of above embodiment 1～3, get above-mentioned extractum 2g, add 100-160 purpose column chromatography silica gel 4g and mix appearance, volatilize solvent, be ground into 80 order fine powders; Gained fine powder and neutral alumina are loaded in the chromatographic column according to 1: 15 ratio dry method of mass ratio; Using mass fraction is 95% ethanol elution, collects eluent, with the capillary tube point sample on silica gel g thin-layer plate; Using the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1: 0.1 blended developing solvent launches; With ethanol system sulphuric acid test solution chromogenic reagent,, conjunction type Hyodeoxycholic Acid speckle begins to merge eluent when appearring in lamellae,, finishes speckle when disappearing.
Other steps are identical with embodiment 1.Be prepared into Fel Sus domestica conjunction type total bile acid, content is with cattle sulphur Hyodeoxycholic Acid (C
26H
45O
6NS) meter is more than 50%.
Embodiment 5
Pharmaceutical formulation Fel Sus domestica conjunction type total bile acid is prepared into tablet, is that example is raw materials used following with adjuvant and quality proportioning thereof to produce 1000 in tablet:
Its preparation method is undertaken by the conventional method of galenic pharmacy tablet, every heavy 0.3g, and every contains Fel Sus domestica conjunction type total bile acid 100mg.
Embodiment 6
Pharmaceutical formulation Fel Sus domestica conjunction type total bile acid is prepared into capsule, is that example is raw materials used following with adjuvant and quality proportioning thereof to produce 1000 of capsules:
Its preparation method is undertaken by the conventional method of galenic pharmacy capsule, every heavy 0.3g, and every contains Fel Sus domestica conjunction type total bile acid 100mg.
Embodiment 7
Pharmaceutical formulation Fel Sus domestica conjunction type total bile acid is prepared into enema, is that example is raw materials used following with adjuvant and quality proportioning thereof to produce enema product 1000mL:
Fel Sus domestica conjunction type total bile acid 3g
Distilled water adds to 1000mL
Its preparation method is undertaken by the conventional method of galenic pharmacy enema, and every bottle of 100mL, every 1ml medicinal liquid contain Fel Sus domestica conjunction type total bile acid 3mg.
Embodiment 8
Pharmaceutical formulation Fel Sus domestica conjunction type total bile acid is prepared into granule, is that example is raw materials used following with adjuvant and quality proportioning thereof to produce granule product 1000g:
Fel Sus domestica conjunction type total bile acid 10g
Sucrose 400g
Dextrin adds to 1000g
Method for preparing is undertaken by the conventional method of galenic pharmacy granule, every bag heavy 10g, and 1g contains Fel Sus domestica conjunction type total bile acid 10mg.
Embodiment 9
Pharmaceutical formulation Fel Sus domestica conjunction type total bile acid is prepared into syrup, is that example is raw materials used following with adjuvant and quality proportioning thereof to produce syrup 1000mL:
Fel Sus domestica conjunction type total bile acid 10g
Sucrose 600g
Distilled water adds to 1000mL
Its preparation method is undertaken by the conventional method of galenic pharmacy syrup, and every bottle of 10mL, 1mL contain Fel Sus domestica conjunction type total bile acid 10mg.
In order to confirm optimum material proportion of the present invention and best process conditions, the inventor tests, and test situation is following:
1, Fel Sus domestica extractum and neutral alumina proportioning are confirmed
Get fresh pig bile 100mL, adding 500mL mass fraction is 95% ethanol, and hold over night is filtered, and reclaims solvent, gets brownish black extractum, gets extractum 2g, adds 100-160 purpose column chromatography silica gel 4g and mixes appearance, volatilizes solvent, is ground into 80 order fine powders; Fine powder and neutral alumina were loaded in the chromatographic column with dry method respectively according to mass ratio in 1: 10,1: 15,1: 20,1: 25; The use mass fraction is 95% ethanol elution; Collect eluent; On silica gel g thin-layer plate, using the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1 with the capillary tube point sample: 0.1 blended developing solvent launches, with ethanol system sulphuric acid test solution chromogenic reagent; When conjunction type Hyodeoxycholic Acid speckle appears in lamellae, merge eluent, when speckle disappears, finish; Measure the absorbance of each eluent with ultraviolet spectroscopy; With the eluent decompression and solvent recovery, 80～100 ℃ of oven dry are weighed in baking oven, calculate the yield of Fel Sus domestica conjunction type total bile acid.Test and result of calculation are seen table 1.
Table 1 Fel Sus domestica extractum and neutral alumina proportioning are to the influence of Fel Sus domestica conjunction type total bile acid decolouring degree and yield
Visible by table 1, the upper prop ratio of Fel Sus domestica extractum and neutral alumina is high more, and the yield of Fel Sus domestica conjunction type total bile acid is low more; Absorbance is more little; Fel Sus domestica conjunction type total bile acid color is shallow more, shows that the amount of used neutral alumina when upper prop separates is big more, and is many more to raw material absorption; But also adsorbed more pigment simultaneously, helped the decolouring of raw material.Take all factors into consideration, it is 1: 10～20 to carry out upper prop and separate that the present invention selects the Fel Sus domestica extractum and the ratio of neutral alumina.
In order to verify the beneficial effect of the Fel Sus domestica conjunction type total bile acid that adopts the present invention's preparation, the inventor has carried out the pharmacology test of treatment colitis to Fel Sus domestica conjunction type total bile acid, and various test situation are following:
1, experiment medicine
Fel Sus domestica conjunction type total bile acid is the pressed powder of yellow to buff, and more than 50%, the secondary redistilled water is prepared, 4 ℃ of airtight preservations in cattle sulphur Hyodeoxycholic Acid for bitter in the mouth, content.
2, laboratory animal
Male Kunming kind healthy mice, body weight (22 ± 2) g, available from Xi'an Jiaotong University Medical College's Experimental Animal Center, the quality certification is numbered 10-004.
3, experiment reagent
2,4,6-TNB (TNBS) is the Sigma Company products, and mouse tumor necrosin & (TNF-α) and myeloperoxidase (MPO) (MPO) enzyme linked immunological kit are available from R&D Systems company.
4, experimental technique
Fel Sus domestica conjunction type total bile acid is made into the aqueous solution of volumetric concentration 20mg/ml, 10mg/ml, 5mg/ml respectively with the secondary redistilled water, and 4 ℃ of refrigerators are preserved subsequent use.
Kunming kind healthy male mice, body weight (22 ± 2) g after 1 week of adaptation, is divided into 5 groups at random under experiment condition, and promptly normal blank control group, modeling group, heavy dose of administration group, middle dosed administration group, low dose of administration group make an experiment.Before the modeling, all mice fasting 24 hours, normal drinking-water; Respectively organize during modeling mice with the slight anesthesia of ether after; Model group and administration group (large, medium and small dosage) are inserted the about 4cm of intestinal with the silica gel hose about diameter 1mm from the mice anus, by 2,4 of every mice 0.5mg; It is 50% alcoholic solution that the 6-TNB injects the 0.1ml mass fraction, and it is 50% alcoholic solution 0.1ml that normal blank control group gives mass fraction from anus.Mice is inverted puts back to conventional raising the in the cage after about 30 seconds.
The colitis modeling began gastric infusion after 12 hours; Normal blank control group and model group mice give normal saline respectively and irritate stomach; Metering among the big metering of administration group Fel Sus domestica conjunction type total bile acid 400mg/kg, the 200mg/kg, the low dose of gastric infusion of 100mg/kg once a day, continue seven days.Tested the 8th day, and put to death mice.Survey the colon index, detect and respectively organize mice tumor necrosis factor activity in the mice serum, and activity of myeloperoxidase in the colon, preparation colon pathological section.
The preparation of the collection of serum and colon's homogenate:
Get blood 1ml from mouse orbit, inject high speed centrifuge to blood, 3000 rev/mins centrifugal 10 minutes, get supernatant and put into-70 ℃ of freezing preservations of refrigerator, be used to detect tumor necrosis factor.
Mice is opened the abdominal cavity and gets total colectomy after putting to death, and surveys total colectomy length.Cut off enteric cavity along mesentery, with ice normal saline flushing intestinal contents, perusal colon outward appearance and variation.Colon divides two parts: a part is put into the liquid nitrogen quick-freezing fast, changes-70 ℃ of cryogenic refrigerators over to and preserves, and is used to detect myeloperoxidase (MPO), and detecting operation is undertaken by the explanation of corresponding reagent box; It is 4% fixing, the section of formalin that the piece of tissue that other gets the about 1cm of far-end colon is put into mass fraction, dyes with the hematoxylin-eosin staining method.
5, experimental result
(1) experiment mice pathology metamorphosis
The normal blank control group mice of perusal intestinal mucosa is not seen pathological changes, and model group mice intestinal wall obviously thickens, intestinal mucosa many places congestion and edema and visible significantly ulcer surface; Fel Sus domestica conjunction type total bile acid small dose group intestinal wall slightly thickens, and congested less, ulcer surface is compared obviously with model group and dwindled; The only local visible small size ulcer of dose groups is congested less in the Fel Sus domestica conjunction type total bile acid; The heavy dose of group of Fel Sus domestica conjunction type total bile acid intestinal wall does not have obviously and thickens, and fold is normal, and only local visible edema does not almost have hyperemia.
Mice colon is observed under optical microscope with the dyeing of hematoxylin-eosin staining method, normal blank group colon each layer clear in structure, and the mucomembranous epithelial cell marshalling, the enteraden rule, goblet cell is abundant, and a matter is not seen congestion and edema; Model group colon mucosal epithelium cell disappearance, the body of gland majority is imperfect, the goblet cell gross distortion, mucosa and Submucosa neutrophil infiltration, vasodilation is congested, the crypts partial destruction, lymph follicle increases, and becomes typical inflammation to change; It is normal that each administration group mice mucosal epithelium begins to recover, and body of gland is arranged comparatively neat, and goblet cell is abundant, and a matter is not almost seen hyperemia, with model group significant difference (P＜0.01) is arranged relatively.
(2) Fel Sus domestica conjunction type total bile acid is to the influence of tumor necrosis factor content in activity of myeloperoxidase and the serum in the mice colon
Fel Sus domestica conjunction type total bile acid treatment group is compared with model group; Activity of myeloperoxidase, tumor necrosis factor content obviously reduce, and significant difference (P＜0.01 or P＜0.05) is arranged, and compare with normal blank group; Difference does not have significance; Show that Fel Sus domestica conjunction type total bile acid can obviously suppress 2,4,6-TNB inducing mouse colon's activity of myeloperoxidase and blood serum tumor necrosin alpha content raise.
Table 2 conjunction type total bile acid is to the influence of TNF-alpha content in MPO activity and the serum in the mice colon
Compare with model group: * P＜0.05 * * P＜0.01.
6, conclusion
Experimental result shows; Fel Sus domestica conjunction type total bile acid has inhibitory action to model mice activity of myeloperoxidase and the rising of tumor necrosis factor content; Fel Sus domestica conjunction type total bile acid can suppress the secretion of proinflammatory cytokine; Regulate immunoreation, the blocking immunity reactivity reaches therapeutic purposes to the damage that body causes effectively.
The function that contains effective composition Fel Sus domestica conjunction type total bile acid medicine: antiinflammatory.
Containing effective composition Fel Sus domestica conjunction type total bile acid medicine cures mainly: colitis.
The specification that contains effective composition Fel Sus domestica conjunction type total bile acid medicine: every heavy 0.3g of tablet, every contains Fel Sus domestica conjunction type total bile acid 100mg; Every heavy 0.3g of capsule, every contains Fel Sus domestica conjunction type total bile acid 100mg; Every bottle of 100mL of enema, every 1ml medicinal liquid contain Fel Sus domestica conjunction type total bile acid 3mg; Every bag heavy 10g of granule, every gram contains Fel Sus domestica conjunction type total bile acid 10mg; Every bottle of 10mL of syrup, every milliliter contains Fel Sus domestica conjunction type total bile acid 10mg.
The usage and dosage that contains effective composition Fel Sus domestica conjunction type total bile acid medicine: oral drugs are obeyed 3 times every day, 1 of 1 of each oral tablet or 1 of capsule or 1 bag of granule or syrup, and children's is cut down according to the circumstance; Enema coloclysis every day 1 time, each 100ml.
The storage that contains effective composition Fel Sus domestica conjunction type total bile acid medicine: the shady and cool dry place of sealing storage.
The effect duration that contains effective composition Fel Sus domestica conjunction type total bile acid medicine: 2 years.
Claims (3)
1. the method for preparing of a Fel Sus domestica conjunction type total bile acid is characterized in that it is made up of following step:
(1) ethanol lixiviate
Get fresh pig bile, add 5 times of its volumes amount mass fraction and be 95% ethanol, left standstill 12～14 hours, filter, reclaim solvent, brownish black extractum;
(2) separation and purification
The extractum that obtains in the step 1 is added in the 100-160 purpose column chromatography silica gel, and the mass ratio of extractum and column chromatography silica gel is 1: 2, mixes appearance; Volatilize solvent, be ground into 80 order fine powders, fine powder and neutral alumina are loaded in the chromatographic column by mass ratio 1: 10～20 dry method; The use mass fraction is 95% ethanol elution; Collect eluent, on silica gel g thin-layer plate, launch the back with developing solvent and use chromogenic reagent with the capillary tube point sample; Merge eluent when occurring conjunction type Hyodeoxycholic Acid speckle on the lamellae, when speckle disappears, finish;
Above-mentioned developing solvent is that the volume ratio of ethyl acetate and methanol, water, glacial acetic acid is 10: 2: 1: 0.1 mixed solution, developer are vanillin sulphuric acid test solution or ethanol system sulphuric acid test solution;
(3) preparation Fel Sus domestica conjunction type total bile acid
With the eluent decompression and solvent recovery of collecting, 80～100 ℃ of oven dry in baking oven, Fel Sus domestica conjunction type total bile acid, content in cattle sulphur Hyodeoxycholic Acid more than 50%.
2. according to the method for preparing of the described Fel Sus domestica conjunction type of claim 1 total bile acid, it is characterized in that: in purification procedures (2), the extractum that obtains in the step (1) is added in the 100-160 purpose column chromatography silica gel; The mass ratio of extractum and column chromatography silica gel is 1: 2, mixes appearance, volatilizes solvent; Be ground into 80 order fine powders, fine powder and neutral alumina are loaded in the chromatographic column by 1: 15 dry method of mass ratio, and the use mass fraction is 95% ethanol elution; Collect eluent, on silica gel g thin-layer plate, launch the back with developing solvent and use chromogenic reagent with the capillary tube point sample; Merge eluent when occurring conjunction type Hyodeoxycholic Acid speckle on the lamellae, when speckle disappears, finish.
3. the Fel Sus domestica conjunction type total bile acid of claim 1 preparation is treated the purposes in the colitis medicine in preparation.
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| CN102775459B (en) * | 2012-07-30 | 2014-12-10 | 南京新百药业有限公司 | Process for preparing hyodeoxycholic acid |
| CN106389471A (en) * | 2016-09-13 | 2017-02-15 | 西北大学 | Application of pulvis fellis suis-combined type cholic acid in preparing drug for treating bronchial asthma |
| CN108717096B (en) * | 2018-06-20 | 2020-10-30 | 南京中医药大学 | Method for identifying fermented arisaema cum bile prepared by pig bile and method for preparing arisaema cum bile by mixed steaming |
| CN111494424A (en) * | 2020-06-16 | 2020-08-07 | 重庆极泽生物科技有限公司 | Preparation method of poultry gall powder |
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