CN106389471A - Application of pulvis fellis suis-combined type cholic acid in preparing drug for treating bronchial asthma - Google Patents
Application of pulvis fellis suis-combined type cholic acid in preparing drug for treating bronchial asthma Download PDFInfo
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Abstract
The invention discloses the application of a pulvis fellis suis-combined type total cholic acid and taurohyodeoxycholic acid in preparing a drug for treating bronchial asthma. The result of the invention shows that, the pulvis fellis suis-combined type total cholic acid and taurohyodeoxycholic acid has an inhibiting effect on the increased content of immunoglobulin, interleukin 4 and interleukin 13 in a model mouse. Meanwhile, the pulvis fellis suis-combined type total cholic acid and taurohyodeoxycholic acid can inhibit the secretion of proinflammatory cytokine, regulate the immunoreaction and effectively prevent the damage of immunoreactivity to human bodies, so that the bronchial asthma can be treated. The prepared pulvis fellis suis-combined type total cholic acid and taurohyodeoxycholic acid is adopted as a raw material to prepare various types of common preparations, such as tablets, capsules, powders, pills, granules and the like.
Description
Technical field
The present invention relates to Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid are in preparation treatment bronchial asthma medicine
In application.
Background technology
Bronchial astehma（Bronchial asthma, abbreviation asthma）One of common chronic respiratory disease, be by
The chronic airway inflammation disease that inflammation cell, inflammatory mediator participate in.This chronic inflammation can lead to airway hyperreactivity
Generation, generally occur within extensively changeable reversible airflow limitation, and cause the panting of recurrent exerbation, out of breath, uncomfortable in chest or cough
Etc. symptom, and often in night and morning outbreak.In recent years, due to the change of weather, environment etc., the incidence of disease of asthma, illness rate
And the death rate is all in ascendant trend year by year.Asthma has had a strong impact on the quality of life of patient, and brings heavy warp to patient
Ji burden, therefore causes extensive attention in this sick global range.
At present, glucocorticoid, β 2 receptor agonist, white three dilute receptor modulators, anticholinergic drug, IgE antibody etc. are
The key agents for the treatment of asthma.However, because individual susceptibility is different, still having some patientss such as smoker, fat asthma to suffer from
Person and non-Th2 raised type asthmatic patient are poor to the reaction of above-mentioned medicine.
Pulvis Fellis Suis are the traditional Chinese medicines of China, have heat-clearing, moisturize, detoxify, relieving cough and asthma the effects such as, dry for pyreticosis
Yearningly, hot eyes, larynx numbness, jaundice, pertussis, asthma, have loose bowels, dysentery, constipation, the disease such as ulcerative carbuncle pyogenic infections.Pulvis Fellis Suis mating type total cholic acid
Include ox sulphur hyocholic acid（Taurohyocholic acid）, ox sulphur hyodesoxycholic acid（Taurohyodeoxycholic
acid）, Taurochenodeoxycholic Acid（Taurochenodeoxycholic acid）, sweet ammonia hyocholic acid（Glycohyocholic
acid）, sweet ammonia hyodesoxycholic acid（Glycohyodeoxycholic acid）With sweet ammonia chenodeoxycholic acid
（Glycochenodeoxycholic acid）.Wherein, the content of ox sulphur hyodesoxycholic acid reaches more than 4%.
According to the inventors knowledge, Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid are in preparation treatment medicine for treating bronchus asthma
Application in thing, yet there are no report.
Content of the invention
It is an object of the invention to provide Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid are in preparation treatment gas
Application in pipe asthmatic medicament, is related to a kind of new application.
The present invention to realize process as follows：
The preparation of Pulvis Fellis Suis mating type total cholic acid：Take Pulvis Fellis Suis medicinal material, add its volume 10 times amount mass fraction be 95% second
Alcohol, ultrasonic extraction makes Pulvis Fellis Suis be completely dissolved, and stands 12-14 hour, filters, recycling design, obtains brownish black medicinal extract.By column chromatography
Silica gel（100～160 mesh）It is added in medicinal extract, medicinal extract is 1 with the mass ratio of column chromatography silica gel：2, mix sample, volatilize solvent, be ground into
80 mesh fine powders, fine powder and neutral alumina in mass ratio 1：15 dry method are loaded in chromatographic column, are washed with the ethanol that mass fraction is 95%
De-, collect eluent, with capillary point sample on silica gel g thin-layer plate, use chromogenic reagent after being launched with solvent, treat lamellae
On occur during mating type cholic acid spot merge eluent, terminate when disappearing to spot, will be molten for the eluent collected recovered under reduced pressure
Agent, in baking oven 80～100 DEG C drying, obtain Pulvis Fellis Suis mating type total cholic acid, content in terms of ox sulphur hyodesoxycholic acid 50% with
On.
Above-mentioned solvent is ethyl acetate is 10 with the volume ratio of methyl alcohol, water, glacial acetic acid：2：1：0.1 mixed solution,
Developer is vanillin-sulfuric acid test solution or ethanol Sulphuric acid test solution.
2nd, the preparation of ox sulphur hyodesoxycholic acid
By Pulvis Fellis Suis mating type total cholic acid obtained above and column chromatography silica gel（100～160 mesh）In mass ratio 1：2 ratio is mixed
Sample, is ground into 80 mesh fine powders, fine powder and column chromatography silica gel（100～160 mesh）In mass ratio 1:25 dry method are loaded in chromatographic column, warp
Column chromatography carries out separating, with volume ratio for 65 repeatedly：25：10：Under 2 methylene chloride-methanol-water-glacial acetic acid mixed solvent
Mutually elute, collect eluent, with capillary point sample on silica gel g thin-layer plate, use chromogenic reagent after being launched with solvent, treat thin
Occur on laminate merging eluent during ox sulphur hyodesoxycholic acid spot, terminate when disappearing to spot, the eluent collected is returned
Receive solvent, recrystallize in acetone, obtain ox sulphur hyodesoxycholic acid, purity reaches more than 95%.
Above-mentioned solvent is ethyl acetate is 10 with the volume ratio of methyl alcohol, water, glacial acetic acid：2：1：0.1 mixed solution,
Developer is vanillin-sulfuric acid test solution or ethanol Sulphuric acid test solution.
The Pulvis Fellis Suis mating type total cholic acid of said method preparation and ox sulphur hyodesoxycholic acid are in preparation treatment bronchial astehma
Purposes in medicine.
The medicine of active ingredient Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid preparation treatment bronchial astehma is used
The form of conventional galenic preparations is using.Described conventional galenic preparations contain the Pulvis Fellis Suis mating type total cholic acid as active component
Or ox sulphur hyodesoxycholic acid, this active component is such as suitable for having of gastrointestinal administration with pharmaceutically acceptable carrier in the formulation
Machine or inorganic solid or liquid excipient mixing.This pharmaceutical formulation can be solid form such as tablet, granule, capsule；
Can also be liquid form such as suspending agent, syrup, enema etc..
Can be containing auxiliary substance, stabilizer, wetting agent and other conventional additive, such as lactose, lemon in above-mentioned preparation
Acid, tartaric acid, stearic acid, magnesium stearate, land plaster, sucrose, cornstarch, talcum powder, gelatin, agar, pectin, peanut oil,
Olive oil, cocoa butter, ethylene glycol, ascorbic acid, mannitol etc..
Above-mentioned preparation can be made according to the conventional preparation technology of various preparations.
The Pulvis Fellis Suis mating type total cholic acid being obtained using the present invention and ox sulphur hyodesoxycholic acid are through treatment bronchial astehma
Pharmacodynamics test, show Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid to asthma mice immunoglobulin (Ig), white
Interleukin 4 and interleukin-11 3 content raise and have inhibitory action, and display Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid can suppress
The secretion of proinflammatory cytokine, adjusts immune response, the damage that effective blocking immunity reactivity causes to body, reaches therapeutic purposes.
Specific embodiment
With reference to embodiment, the present invention is described in more detail, but the invention is not restricted to these embodiments.
Embodiment 1
, the preparation method of Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid is as follows taking Pulvis Fellis Suis 100 g as a example：
1. the preparation of Pulvis Fellis Suis mating type total cholic acid
Take Pulvis Fellis Suis medicinal material 100 g, add its volume 10 times amount mass fraction be 95% ethanol, ultrasonic extraction makes Pulvis Fellis Suis
It is completely dissolved, stands 12-14 hour, filter, recycling design, obtain brownish black medicinal extract.Take medicinal extract 5 g, add the post of 100-160 mesh
Chromatographic silica gel 10 g mixes sample, volatilizes solvent, is ground into 80 mesh fine powders, fine powder and neutral alumina in mass ratio 1：15 dry method are loaded on layer
In analysis post, the ethanol elution being 95% with mass fraction, collect eluent, with capillary point sample on silica gel g thin-layer plate, use second
Acetoacetic ester is 10 with the volume ratio of methyl alcohol, water, glacial acetic acid：2：1：0.1 mixed solution launches for solvent, uses vanillin-sulfuric acid
Test solution develops the color, and occurs merging eluent during mating type cholic acid spot, terminate when disappearing to spot on lamellae.By collect
Eluent decompression and solvent recovery, in baking oven, 80～100 DEG C of drying, obtain Pulvis Fellis Suis mating type total cholic acid, content is gone with ox sulphur pig
Oxycholic acid（C26H45O6NS）Meter is more than 50%.
2. the preparation of ox sulphur hyodesoxycholic acid
Take Pulvis Fellis Suis mating type total cholic acid 2 g obtaining in step 1, add column chromatography silica gel 4 g of 100-160 mesh to mix sample, grind
Become 80 mesh fine powders, fine powder and column chromatography silica gel（100～160 mesh）In mass ratio 1：25 dry method are loaded in chromatographic column, through post repeatedly
Chromatography carries out separating, with volume ratio for 65：25：10：2 methylene chloride-methanol-water-the lower of glacial acetic acid mixed solvent mutually elutes,
Collect eluent, with capillary point sample on silica gel g thin-layer plate, the volume ratio with ethyl acetate and methyl alcohol, water, glacial acetic acid is
10：2：1：0.1 mixed solution launches for solvent, with the colour developing of vanillin-sulfuric acid test solution, treats that ox sulphur pig on lamellae goes
Merge eluent during oxycholic acid spot, terminate when disappearing to spot.By the eluent collected recycling design, tie in acetone again
Crystalline substance, obtains ox sulphur hyodesoxycholic acid, and purity reaches more than 95%.
Embodiment 2
Pharmaceutical formulation Pulvis Fellis Suis mating type total cholic acid prepares piece agent, taking produce 1000, tablet as a example raw materials used and auxiliary material and
Its quality proportioning is as follows：
Pulvis Fellis Suis mating type total cholic acid 50g
Dextrin 50g
Magnesium stearate 2g
Starch adds to 300g
Its preparation method is carried out by the conventional method of galenic pharmacy tablet, every weight 0.3g, the every total cholic acid 50mg of mating type containing Pulvis Fellis Suis.
Pharmaceutical formulation ox sulphur hyodesoxycholic acid prepares piece agent, taking produce 1000, tablet as a example raw materials used and auxiliary material and
Its quality proportioning is as follows：
Ox sulphur hyodesoxycholic acid 10g
Dextrin 50g
Magnesium stearate 2g
Starch adds to 300g
Its preparation method is carried out by the conventional method of galenic pharmacy tablet, every weight 0.3g, the every hyodesoxycholic acid 10mg of sulphur containing ox.
Embodiment 3
Pharmaceutical formulation Pulvis Fellis Suis mating type total cholic acid is prepared into capsule, taking produce capsule 1000 as a example raw materials used and auxiliary
Material and its quality proportioning are as follows：
Pulvis Fellis Suis mating type total cholic acid 50g
Microcrystalline cellulose 20g
Magnesium stearate 1g
Starch adds to 300g
Its preparation method is carried out by the conventional method of galenic pharmacy capsule, every weight 0.3g, the total courage of every mating type containing Pulvis Fellis Suis
Sour 50mg.
Pharmaceutical formulation ox sulphur hyodesoxycholic acid is prepared into capsule, taking produce capsule 1000 as a example raw materials used and auxiliary
Material and its quality proportioning are as follows：
Ox sulphur hyodesoxycholic acid 10g
Microcrystalline cellulose 20g
Magnesium stearate 1g
Starch adds to 300g
Its preparation method is carried out by the conventional method of galenic pharmacy capsule, every weight 0.3g, every hyodesoxycholic acid of sulphur containing ox 10
mg.
Embodiment 4
Pharmaceutical formulation Pulvis Fellis Suis mating type total cholic acid is prepared into granule, taking produce granule product 1000g as a example raw materials used
As follows with auxiliary material and its quality proportioning：
Pulvis Fellis Suis mating type total cholic acid 5g
Sucrose 400g
Dextrin adds to 1000g
Preparation method is carried out by the conventional method of galenic pharmacy granule, every bag of weight 10g, 1g mating type containing Pulvis Fellis Suis total cholic acid 5
mg.
Pharmaceutical formulation ox sulphur hyodesoxycholic acid is prepared into granule, taking produce granule product 1000g as a example raw materials used
As follows with auxiliary material and its quality proportioning：
Ox sulphur hyodesoxycholic acid 1g
Sucrose 400g
Dextrin adds to 1000g
Preparation method is carried out by the conventional method of galenic pharmacy granule, every bag of weight 10g, 1g sulphur containing ox hyodesoxycholic acid 1 mg.
Embodiment 5
Pharmaceutical formulation Pulvis Fellis Suis mating type total cholic acid is prepared into syrup, taking produce syrup 1000 mL as a example raw materials used and
Auxiliary material and its quality proportioning are as follows：
Pulvis Fellis Suis mating type total cholic acid 5g
Honey element 10g
Distilled water adds to 1000mL
Its preparation method is carried out by the conventional method of galenic pharmacy syrup, every bottle of 10mL, the 1mL total cholic acid of mating type containing Pulvis Fellis Suis
5mg.
Pharmaceutical formulation ox sulphur hyodesoxycholic acid is prepared into syrup, taking produce syrup 1000 mL as a example raw materials used and
Auxiliary material and its quality proportioning are as follows：
Ox sulphur hyodesoxycholic acid 1g
Honey element 10g
Distilled water adds to 1000mL
Its preparation method is carried out by the conventional method of galenic pharmacy syrup, every bottle of 10mL, the 1mL hyodesoxycholic acid of sulphur containing ox 1 mg.
In order to verify the beneficial effect of the Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid prepared using the present invention,
Inventor has carried out to Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid treating the pharmacodynamics test of bronchial astehma, respectively
Plant test situation as follows：
1st, Experimental agents
Pulvis Fellis Suis mating type total cholic acid, is the pressed powder to buff for the yellow, bitter, and secondary redistilled water is prepared, 4 DEG C of airtight guarantors
Deposit.Ox sulphur hyodesoxycholic acid, is white solid powder, mildly bitter flavor, and secondary redistilled water is prepared, 4 DEG C of airtight preservations.
2nd, animal used as test
Female KM kind healthy mice, body weight（22±2）G, purchased from Xi'an Jiaotong University Medical College's Experimental Animal Center, the quality certification
Numbering is 61001700000563.
3rd, experiment reagent
Ovalbumin（OVA）Reagent is purchased from Sigma company, mouse immuning ball protein IgE, IL-4（IL-4）With interleukin-11 3
（IL-13）Enzyme linked immunological kit is purchased from R＆D Systems company.
4th, experimental technique
The secondary redistilled water of Pulvis Fellis Suis mating type total cholic acid is made into volumetric concentration 20 mg/ml, 10 mg/ml, 5 mg/ml respectively
The aqueous solution, ox sulphur hyodesoxycholic acid secondary redistilled water is made into volumetric concentration 3 mg/ml, 2 mg/ml, 1 mg/ml respectively
The aqueous solution, 4 DEG C of Refrigerator stores are standby.
Kunming kind Healthy female mouse, body weight（22±2）G, after adapting to 1 week under experimental conditions, is randomly divided into 8 groups, that is,
Normal group, model group, Pulvis Fellis Suis mating type total cholic acid large, medium and small dosed administration group, ox sulphur hyodesoxycholic acid are large, medium and small
Dosed administration group, is tested.
Mice with asthma model is divided into sensitization and is excited two stages using OVA preparation.
Sensitisation phase：Normal group in mouse peritoneal injection 0.2 mL physiological saline sensitization, remaining each group mouse all in
Test the 0th, 7,14 days lumbar injection 0.2 mL and contain the aseptic antigen liquid sensitization of 20 μ g OVA and 2 mg gel aluminum hydroxides.
Excitation phase：Test the 22nd, 23,24 days and remaining each group mouse in addition to Normal group is placed in airtight organic glass
In glass case, Neulized inhalation 1%（w/v）OVA normal saline solution, once a day, 30 min every time, continuous agitation 3 days.Normally right
Excite according to group physiological saline.
Each group mouse, in testing the 18-24 days gastric infusions, is often inferior to first 1 hour drug treatment of atomization.Normal group
Give physiological saline gavage with model group mouse respectively（20 ml/kg）, administration group Pulvis Fellis Suis mating type total cholic acid is by big metering
400 mg/kg, middle metering 200 mg/kg, low dose of 100 mg/kg gastric infusions, administration group ox sulphur hyodesoxycholic acid presses the matter of fundamental importance
Measure 60 mg/kg, middle metering 40 mg/kg, low dose of 20 mg/kg gastric infusions once a day, continue seven days.Last atomization knot
Bundle, after 24 hours, puts to death mouse.IgE content in detection each group mice serum, measures IL-4 and IL-13 in bronchoalveolar lavage fluid and lives
Property, prepare pathologic section.
The collection of serum and the preparation of bronchoalveolar lavage fluid：
After last atomization excites operation 24 hours, eyeball blood sampling is carried out to mouse.Take blood 1 ml from mouse orbit, blood
Injection supercentrifuge, 5000 revs/min are centrifuged 15 minutes, take supernatant to put into -80 DEG C of refrigerator freezings and preserve, for detecting
IgE.
After sacrifice, open chest and expose tracheae to left and right bifurcation, after ligaturing right principal bronchus, right lung is taken off, puts into
Fixing in 10% formalin, washed after 48 hours, then used variable concentrations alcohol（70%、80%、90%、95%、100%）De-
Water, uses FFPE after dimethylbenzene is transparent, is cut into 5 m slabs with slicer, using hematoxylin eosin staining method
Dyeed.Light Microscopic observation pathologic changes, Taking Pictures recording.
After sacrifice, after exposing and separating tracheae, insert remaining needle through pars cervicalis tracheae, lung at the uniform velocity injects 2 mL lifes to the left
Reason salt solution, indwelling 2 minutes, the chest of massage mouse slowly fully reclaimed bronchoalveolar lavage fluid after 30 seconds, repeated equally operation 3 times,
Ensure the rate of recovery>80%.Fully mix bronchoalveolar lavage fluid, be centrifuged 5 minutes with 2000 revs/min, draw supernatant in centrifuge tube,
For measuring cytokine content.
Experimental data is represented with means standard deviation.Experimental data carries out statistical using SPSS17.0 statistical analysis software
Analysis, multigroup mean compares and adopts one-way analysis of variance（ANOVA）,P<0.05 is that difference is statistically significant.
5th, experimental result
（1）Experiment mice pathology metamorphosis
Observe under an optical microscope after the dyeing of mouse lung tissue hematoxylin eosin staining method, Normal group mouse lung group
Knit section bronchial wall completely, thickness is uniform, mucomembranous epithelial cell marshalling has no the epithelial cell and inflammation coming off in tube chamber
Property exudate, has no eosinophil between tube wall and alveolar.Model group mouse lung tissue section bronchial wall circular muscle is plump, glues
Film epithelial cell arrangement disorder, part epithelial cell shedding, visible inflammatory exudate in tube chamber, tube wall and alveolar septum are visible relatively
Multi-lymphocytes, monocyte and eosinophil.Positive controls mouse lung tissue section tunica mucosa bronchiorum epithelial cell
Partial destruction, bronchus tube wall slightly thickens, visible mild hyperaemia and oedema under mucous membrane, visible a little inflammatory exudate under the visual field.
Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid each dosed administration group mouse lung tissue section bronchial wall structure are basic
Recover, no inflammatory exudate in tube chamber, mucomembranous epithelial cell starts to recover, under mucous membrane, congested and water lung is inconspicuous.
（2）Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid are to IL-4, IL-13 activity in mouse bronchoalveolar lavage fluid
And in serum IgE content impact
Pulvis Fellis Suis mating type total cholic acid and Niu Huang hyodesoxycholic acid treatment group compared with model group, IL-4, IL-13 and IgE content
Substantially reduce, have significant difference（P<0.001 orP0.01 orP0.05）, compared with normal blank control group, difference is no
Conspicuousness, shows that Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid can substantially suppress ovalbumin（OVA）Inducing mouse
Bronchoalveolar lavage fluid IL-4 and IL-13 activity and SERUM IgE content raise.
6th, conclusion
Test result indicate that, the bronchial astehma that Pulvis Fellis Suis mating type total cholic acid and ox sulphur hyodesoxycholic acid induce to ovalbumin
Model mice immunoglobulin (Ig), IL-4 and interleukin-11 3 content raise inhibitory action, pig-bile-combination-type total bile acid and ox
Sulphur hyodesoxycholic acid can suppress the secretion of proinflammatory cytokine, adjusts immune response, and blocking immunity reactivity is to body effectively
The damage causing, reaches therapeutic purposes.
The mating type total cholic acid of Pulvis Fellis Suis containing active ingredient or the function of ox sulphur hyodesoxycholic acid medicine：Anti-inflammatory.
The mating type total cholic acid of Pulvis Fellis Suis containing active ingredient or ox sulphur hyodesoxycholic acid medicine cure mainly：Bronchial astehma.
The mating type total cholic acid of Pulvis Fellis Suis containing active ingredient or the specification of ox sulphur hyodesoxycholic acid medicine：Every weight of tablet
0.3g, the every total cholic acid of mating type containing Pulvis Fellis Suis 50 mg, or the every hyodesoxycholic acid of sulphur containing ox 10 mg；Every weight of capsule
0.3 g, the every total cholic acid of mating type containing Pulvis Fellis Suis 50 mg, or the every hyodesoxycholic acid of sulphur containing ox 10 mg；Every bag of weight of granule
10g, the every gram of total cholic acid of mating type containing Pulvis Fellis Suis 5 mg, or the every gram of hyodesoxycholic acid of sulphur containing ox 1 mg；Every bottle of 10mL of syrup, often
The milliliter total cholic acid of mating type containing Pulvis Fellis Suis 5 mg, or the every milliliter of hyodesoxycholic acid of sulphur containing ox 1 mg.
The mating type total cholic acid of Pulvis Fellis Suis containing active ingredient or the usage and dosage of ox sulphur hyodesoxycholic acid medicine：Oral drugs are every
Day take 3 times, each oral tablet 1 or capsule 1 or 1 bag of granule or syrup 1, children are cut down according to the circumstance.
The mating type total cholic acid of Pulvis Fellis Suis containing active ingredient or the storage of ox sulphur hyodesoxycholic acid medicine：At sealing cool place is dried
Storage.
The mating type total cholic acid of Pulvis Fellis Suis containing active ingredient or the term of validity of ox sulphur hyodesoxycholic acid medicine：2 years.
Claims (5)
1. the Pulvis Fellis Suis mating type total cholic acid and/or ox sulphur hyodesoxycholic acid application in preparation treatment bronchial asthma medicine.
2. according to claim 1 application it is characterised in that：Pulvis Fellis Suis mating type total cholic acid main active is ox sulphur pig
Deoxycholic aicd, in terms of ox sulphur hyodesoxycholic acid more than 50%；Ox sulphur hyodesoxycholic acid purity reaches more than 95%.
3. apply it is characterised in that the preparation method of Pulvis Fellis Suis mating type total cholic acid is as follows according to claim 1：Take pig gall
Powder medicinal material, add its volume 10 times amount mass fraction be 95% ethanol, ultrasonic extraction makes Pulvis Fellis Suis be completely dissolved, and stands 12-
14 hours, filter, recycling design, obtain brownish black medicinal extract；100～160 mesh column chromatography silica gels are added in medicinal extract, medicinal extract and post
The mass ratio of chromatographic silica gel is 1：2, mix sample, volatilize solvent, be ground into 80 mesh fine powders, fine powder and neutral alumina in mass ratio 1：15
Dry method is loaded in chromatographic column, the ethanol elution being 95% with mass fraction, collects eluent, with capillary point sample in silica G thin layer
On plate, after being launched with solvent, use chromogenic reagent, occur merging eluent during mating type cholic acid spot, to spot on lamellae
Point terminates when disappearing；By the eluent collected decompression and solvent recovery, 80～100 DEG C of drying in baking oven, obtain Pulvis Fellis Suis and combine
Type total cholic acid, content is in terms of ox sulphur hyodesoxycholic acid more than 50%.
4. apply it is characterised in that the preparation method of ox sulphur hyodesoxycholic acid is as follows according to claim 3：To prepare
Pulvis Fellis Suis mating type total cholic acid and 100～160 mesh column chromatography silica gels in mass ratio 1：2 ratio mixes sample, is ground into 80 mesh fine powders,
Fine powder and 100～160 mesh column chromatography silica gels in mass ratio 1:25 dry method are loaded in chromatographic column, carry out separating through column chromatography repeatedly,
With volume ratio for 65：25：10：2 methylene chloride-methanol-water-the lower of glacial acetic acid mixed solvent mutually elutes, and collects eluent, uses
Capillary point sample, on silica gel g thin-layer plate, uses chromogenic reagent after being launched with solvent, treats that ox sulphur pig on lamellae deoxygenates
Merge eluent during cholic acid spot, terminate when disappearing to spot；By the eluent collected recycling design, tie in acetone again
Crystalline substance, obtains ox sulphur hyodesoxycholic acid, and purity reaches more than 95%.
5. according to claim 3 or 4 application it is characterised in that：Described solvent is ethyl acetate, methyl alcohol, water and ice
The volume ratio of acetic acid is 10：2：1：0.1 mixed solution, developer is vanillin-sulfuric acid test solution or ethanol Sulphuric acid test solution.
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| CN102178695A (en) * | 2011-02-25 | 2011-09-14 | 孙文基 | Method for preparing pig-bile-combination-type total bile acid and application thereof |
| CN104473967A (en) * | 2015-01-05 | 2015-04-01 | 四川神农药业有限公司 | Pig gall powder preparing process |
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- 2016-09-13 CN CN201610817015.6A patent/CN106389471A/en active Pending
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| CN102178695A (en) * | 2011-02-25 | 2011-09-14 | 孙文基 | Method for preparing pig-bile-combination-type total bile acid and application thereof |
| CN104473967A (en) * | 2015-01-05 | 2015-04-01 | 四川神农药业有限公司 | Pig gall powder preparing process |
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