CN101890048B - Total bile acid extract of bear bile powder and preparation method and application of injection thereof - Google Patents
Total bile acid extract of bear bile powder and preparation method and application of injection thereof Download PDFInfo
- Publication number
- CN101890048B CN101890048B CN2009101432410A CN200910143241A CN101890048B CN 101890048 B CN101890048 B CN 101890048B CN 2009101432410 A CN2009101432410 A CN 2009101432410A CN 200910143241 A CN200910143241 A CN 200910143241A CN 101890048 B CN101890048 B CN 101890048B
- Authority
- CN
- China
- Prior art keywords
- extract
- ethyl acetate
- acid
- powder
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Steroid Compounds (AREA)
Abstract
The invention relates to a total bile acid extract of bear bile powder and a preparation method and application of injection thereof. The total bile acid extract of the bear bile powder is prepared by the following steps of: performing reflux extraction on the bear bile powder by using ethanol; recovering the ethanol from extracting solution, and concentrating the extracting solution; and performing alkaline hydrolysis, neutralization, acidification, ethyl acetate extraction, ethyl acetate crystallization and ethanol crystallization. The invention also provides a method for preparing the injection from the total bile acid extract of the bear bile powder. The prepared total bile acid extract of the bear bile powder and the injection thereof can be used for preparing medicaments for resisting bacteria and viruses, relieving cough and reducing sputum and protecting liver and gallbladder. The invention can improve the extraction yield and purity of ursodesoxycholic acid in the total bile acid extract of the bear bile powder and reduce the chenodeoxycholic acid content simultaneously, so the ursodesoxycholic acid content in the total bile acid extract of the bear bile powder is no less than 70 percent and the chenodeoxycholic acid content is no more than 20 percent. The ursodesoxycholic acid content in the total bile acid extract of the bear bile powder is higher, and the ursodesoxycholic acid yield is high and is over 18 percent generally.
Description
Technical field
The present invention relates to the preparation method of Chinese medicine preparation, be specifically related to preparation method and the purposes of a kind of total bile acid extract of bear bile powder and injection thereof.
Background technology
Chinese medicine more and more receives more concern at present, but, due to complicated component, technique falls behind, the quality standard shortcoming, and clinical use untoward reaction is more, has been acknowledged as the excessive risk product.Tracing it to its cause, is mainly that extracts active ingredients purity is not high, invalid components and principal component is not indefinite.
The dry product that Fel Ursi powder is ursidae animal black bear or brown bear bile, adopt natural Fel Ursi juice or drain-out bear gall's juice.Fel Ursi powder has that heat clearing away is relieving convulsion, the improving eyesight disinsection efficiency.Contain the Multiple components such as bile acids, bilins, amino acids; wherein bile acids is its main effective ingredient; heat-clearing and toxic substances removing, relieving cough and resolving phlegm, choleretic effect are arranged; conjugate form mainly with taurine or glycine exists, and can obtain multiple free bile acid after hydrolysis, as ursodesoxycholic acid, chenodeoxycholic acid, cholic acid, deoxycholic acid etc.; wherein the ursodeoxycholic acid content is the highest; chenodeoxycholic acid takes second place, and the two is isomers, often not easily separated.From pharmacological action, that ursodesoxycholic acid and chenodeoxycholic acid all have is antibiotic, antiinflammatory, analgesic, cough-relieving and the effect of dissolving cholelithiasis, but the effect of ursodesoxycholic acid is better than chenodeoxycholic acid, moreover the side effect of chenodeoxycholic acid is more obvious, there will be stomachache, diarrhoea phenomenon during oral administration, can produce certain hemolytic during intravenously administrable, and ursodesoxycholic acid has no adverse reaction under positive usual amounts substantially.Therefore to reduce as far as possible the content of its contained chenodeoxycholic acid when producing ursodesoxycholic acid.
Fel Ursi powder has been reported and launch for the preparation of injection; but the goal object of its extraction is generally that to take tauroursodeoxycholic acid and Taurochenodeoxycholic Acid be main total binding bile acid; although simple process; but be difficult for purifying; especially can not control the ratio of two kinds of main conjugated bile acid, side effect is larger.
The method of from Fel Ursi powder, extracting at present ursodesoxycholic acid be mainly by the basic hydrolysis saponification, in and the steps such as acidify, acetic acid ethyl ester extract, ethyl acetate crystallization and recrystallization complete, yield approximately 10%, in the gained crystal, the content of ursodesoxycholic acid is lower, generally in 60% left and right, the CDCA acid content is higher, generally more than 25%, and the color of crystal is darker, be generally yellow or faint yellow, illustrate that impurity content is more, be unsuitable for the medicine for some special forms, as injection etc.
Summary of the invention
One of purpose of the present invention is to provide a kind of preparation method of total bile acid extract of bear bile powder, and the method can improve ursodesoxycholic acid extract yield and purity in total bile acid extract of bear bile powder, reduces the content of chenodeoxycholic acid simultaneously.
Another object of the present invention is to provide the method for making injection with above-mentioned total bile acid extract of bear bile powder.
Another purpose of the present invention is to provide total bile acid extract of bear bile powder prepared by said method and the pharmaceutical applications of ejection preparation thereof, is mainly used in the pharmaceutical applications of the aspects such as antibiotic, antiviral, relieving cough and resolving phlegm and hepatic cholagogic.
Total bile acid extract of bear bile powder preparation method of the present invention specifically is achieved through the following technical solutions.
A kind of preparation method of total bile acid extract of bear bile powder, it comprises the steps:
A, ethanol extraction: Fel Ursi powder is mixed to ebuillition of heated reflux, extract, 0.5-2 hour with ethanol and active carbon;
B, saponification: ethanol extract is reclaimed to ethanol to distinguishing the flavor of without alcohol, concentrated solution thin up, then reinforcing body sodium hydroxide, stirring and dissolving is carried out saponification under 90-120 ℃, and the response time is 16-24 hour;
C, neutralization reaction: saponification adds hydrochloric acid to regulate pH value to 1-6 after finishing, separate out precipitation, filters and obtains the Fel Ursi powder extract crude product;
D, crude product drying: the Fel Ursi powder extract crude product adopts 100 ℃ of following temperature dryings;
E, ethyl acetate are extracted: by the crude product crushed after being dried, add active carbon and ethyl acetate backflow to extract 2-4 time, extraction time is 0.5-1.5 hour;
F, ethyl acetate crystallization: acetic acid ethyl acetate extract in step e is merged, reclaim ethyl acetate, 0-30 ℃ of placement, crystallization;
G, alcohol crystal: the ethyl acetate crystal is added to dissolve with ethanol and make solution, add activated carbon decolorizing, filter, pure liquid is concentrated, and crystallization filters, crystallizing and drying and get final product.
Wherein, in step a, described concentration of alcohol is 50%-95%, the 5-15 that the ethanol consumption is Fel Ursi powder weight times, and the 10%-20% that activated carbon dosage is Fel Ursi powder weight.
Wherein, in step b, described concentrated solution is diluted with water to 8-12 times of Fel Ursi powder weight, the 10%-20% that sodium hydroxide weight is dilute liquid medicine weight.
Wherein, in the neutralization reaction of step c, described concentration of hydrochloric acid is 2M-12M.
Wherein, in step e, the 10%-20% that described activated carbon dosage is the crude product consumption, the 5-15 that the ethyl acetate consumption is the crude product consumption is doubly.
Wherein, can also increase the step by re-crystallizing in ethyl acetate in step f.
Wherein, in step f, acetic acid ethyl acetate extract merges 2-8 times that amount of solution is crude product weight.
Wherein, in step g, the 4-8 that the consumption that adds ethanol is ethyl acetate crystal weight doubly, adds the 1%-5% that active carbon is solution weight, and the 2-4 that the concentrated liquid measure of pure liquid is ethyl acetate crystal weight doubly.
Above consumption all calculates according to weight, and ethanol used can substitute with methanol, and hydrochloric acid used can substitute with sulphuric acid, and corresponding process conditions are constant.
The present invention also provides the total bile acid extract of bear bile powder that said method is made to make the method for injection, comprises the steps:
The total bile acid extract of bear bile powder that said method is made injects water, with sodium hydroxide solution, regulates pH value to 7-8, use activated carbon decolorizing after adding water for injection, decarbonization filtering, and fine straining, cool, and fill is sealed, sterilizing and get final product.
Wherein, the total bile acid extract of bear bile powder consumption is according to the ursodesoxycholic acid cubage, and injection loading amount 2-20ml/ props up.
Fel Ursi powder total bile acid injection preferred content specification of the present invention is 20mg/ml (in ursodesoxycholic acid), and using dosage is according to state of an illness adjustment corresponding to individual instances.
The total bile acid extract of bear bile powder that the present invention also provides said method to prepare and ejection preparation thereof the purposes in antibiotic, the antiviral of preparation, relieving cough and resolving phlegm and hepatic cholagogic medicine.
In order to overcome the deficiencies in the prior art, adopt safety non-toxic, simple and practical method, provide a kind of and can either improve ursodesoxycholic acid extract yield and purity in total bile acid extract of bear bile powder, reduce the method for CDCA acid content simultaneously, make the content of ursodesoxycholic acid in total bile acid extract of bear bile powder be not less than 70%, the CDCA acid content is not higher than 20%.For reaching goal of the invention, the present invention is first by the Fel Ursi powder alcohol reflux, extracting solution reclaim ethanol concentrated after, then carry out basic hydrolysis, in and acidify, ethyl acetate extraction, ethyl acetate crystallization, alcohol crystal, obtain total bile acid extract of bear bile powder.In addition, the present invention also provides and take the total urso cholic acid extract and make the method for injection as raw material.
Compared with the prior art, the present invention has following advantage:
The first, the preparation of Fel Ursi powder total bile acid has increased the ethanol extraction step, can effectively remove the impurity such as contained protein, aminoacid, pigment, part fat, inorganic salt, reach the purpose of primary purification, improved hydrolysis saponification effect, more be conducive to further separation and purification;
The second, the preparation of Fel Ursi powder total bile acid has increased the alcohol crystal step, utilizes the difference of ursodesoxycholic acid and chenodeoxycholic acid dissolubility in ethanol, by alcohol crystal, can effectively reduce the content of chenodeoxycholic acid in crystal, improves the content of ursodesoxycholic acid;
Three, the present invention not only in the total urso cholic acid extract ursodeoxycholic acid content higher, and yield is high, generally more than 18%, thereby makes limited resource obtain more scientific and reasonable utilization, has reduced production cost;
Four, this law be take Fel Ursi powder and is prepared ursodesoxycholic acid and chenodeoxycholic acid complex without oxidation and reduction reaction process as raw material, and less demanding to working condition, environmentally safe, be applicable to large-scale industrialization production;
Five, in the injection made from the total urso cholic acid extract, contained to survey the clear and definite component content of composition and structure high for solid content, all reaches more than 80%, of light color, closely colourless, stable and controllable for quality.
Total urso cholic acid extract prepared by the inventive method and the pharmaceutical research of injection thereof are as follows.
1, refrigeration function research
Get some of body weight 120～225kg healthy rabbits, laboratory temperature is controlled at 25 ℃ of left and right.3d before test, survey normal anus temperature every day 2 times, chooses body temperature and change not higher than 50 of the rabbit of 0.3 ℃, is divided at random 5 groups.Measure 3 anus temperature (every 30min 1 time) before administration, take the body temperature meansigma methods as basal body temperature.The administration group is pressed table 1 dosage iv, and Normal group is with method to the isometric(al) normal saline, and positive control adopts arginine aspirin.Each treated animal auricular vein is injected (iv) endotoxin 5EU/kg simultaneously.After endotoxin is attacked 1,2,3,4,5h measures rabbit anus temperature.Experiment shows, each dosage group of total urso cholic acid injection can obviously suppress the rising of rabbit body temperature due to endotoxin, wherein, the action time of high dose group and intensity obviously are better than low dose group, with the matched group comparing difference, significance (P<0.05, P<0.01) is arranged, show that the total urso cholic acid injection has obvious refrigeration function, the results are shown in Table 1.
The refrigeration function (n=10, x ± s) of fever in rabbits due to table 1 total urso cholic acid injection induced by endotoxin
(1)Compare P<0.05 with matched group;
(2)P<0.01
2, antibacterial action research
Get 100 of mices, body weight (18 ± 1) g, male and female half and half, be divided into 5 groups at random, 20 every group, all gives 30 LD of staphylococcus aureus
50Challenging dose, then by table 2 medicine and dosage grouping (total urso cholic acid group dosage is by ursodesoxycholic acid), intravenous administration (iv), matched group iv equal-volume normal saline, positive control adopts the amoxicillin injection.Once a day, successive administration is 5 days.The death toll of 5 interior each treated animals after observed and recorded infects.Experimental result shows, each dosage group of total urso cholic acid injection is to the lethal protective effect all had in various degree of infection of staphylococcus aureus mice, in Table 2.
The protective effect of table 2 total urso cholic acid injection to the death of infection of staphylococcus aureus induced mice
3, antivirus action research
(1), infected by influenza FM
1Cause the impact of mice pneumonia
Get 60 of mices, body weight (15 ± 1) g, be divided into 6 groups at random by table 2,10 every group; Except Normal group, all the other are with 15 times of LD
50Influenza virus FM
1Intravenous injection.The administration group is pressed table 2 dosage, 1d start injection administration before infecting, and every day 1 time, continuous 5d, Normal group is with method to the isometric(al) normal saline, and positive control adopts ribavirin injection.Within the 6th day, weigh (prohibiting water 8h), put to death animal, dissect and get the lung cleaning, remove the tissues such as trachea, hilar lymph node, use the filter paper suck dry moisture, weigh, calculate one by one as follows lung exponential sum suppression ratio: heavy (the g)/body weight (g) * 100 of lung index=lung; Lung index (%)=
(The average lung index of the average lung index-test group of virus control group
)The average lung index of/virus control group * 100%.Experiment shows, each dosage group infected by influenza FM
1Due to strain, pneumonopathy becomes obvious inhibitory action, with virus control group comparing difference, significance (P<0.05, P<0.01) is arranged, and the results are shown in Table 3.
Table 3 infected by influenza FM
1Cause the impact (n=10) of mice pneumonia
(1)Compare P<0.01 with Normal group;
(2)With the virus control group, compare, P<0.05,
(3)P<0.01 (2)
(2), infected by influenza FM
1The dead protective effect of infecting mouse
Get 120 of mices, body weight (15 ± 1) g, be divided into 6 groups at random by table 4, and 20 every group, except Normal group, all the other are with 30 times of LD
50The influenza virus intravenous injection, every 0.05ml.The administration group is pressed table 4 dosage, and before infecting, 1d starts intravenous administration, every day 1 time, and continuous 5d, Normal group is with method to the isometric(al) normal saline, and positive control adopts ribavirin injection.From infecting Continuous Observation 14d from virus, record day by day animal symptom and death toll.Calculate as follows dead protective rate (%), extending life rate (%): dead protective rate (%)=(virus control group mortality rate-test group mortality rate) * 100%; Extending life rate (%)=
(Test group natural law-matched group natural law of on average surviving of on average surviving
)/ matched group natural law * 100% of on average surviving.Experimental result, compare with the virus control group, and the total urso cholic acid injection can obviously extend the average survival natural law of infecting mouse, improves dead protective rate (P<0.05, P<0.01), infected by influenza FM
1Infecting mouse has dead protective effect, shows that the total urso cholic acid injection has obvious interior resisting virus effect to mice.The results are shown in Table 4.
Table 4 couple mice influenza virus FM
1The dead protective effect of virus strain infection
(1)Compare P<0.05 with the virus control group;
(2)P<0.01
4, relieving cough and resolving phlegm effect research
(1), antitussive action
Get 100 of Kunming mouses, be divided at random 5 groups, 20 every group.Before experiment, water 12h is can't help in the mice fasting, then respectively organizes mice by table 5 dosed administration, and 1 time/d, be total to 5d, wherein Normal group gives the equal-volume distilled water, and positive control adopts codeine phosphate inj.1h after the last administration, put into fixed container by mice, and make mice accept constant voltage ammonia to stimulate to the scheduled time and draw and cough, and the logarithmic interval of ammonia mist stimulation time is 0.1, stimulates after stopping and taking out immediately mice, observes cough number of times in 1min.Obtain with sequential method (upper purgation) spray time (EDT that causes the half mouse cough
50), and calculate the R value, if the R value is greater than 130%, illustrate that medicine has antitussive action; If the R value is greater than 150%, explanation has remarkable antitussive action.Experimental result shows: each dosage group of total urso cholic acid injection R value all is greater than 130%, and the R value increases with the increase of dosage.In Table 5.
The antitussive action of table 5 total urso cholic acid injection
(2), resolve phlegm effect
Get 75 of SD rats, be divided at random 5 groups, 15 every group.Before experiment, water 12h is can't help in the rat fasting, with after the urethane intraperitoneal injection of anesthesia, face upward position fixing, cut off neck middle part skin, isolate trachea, between upper two cartilaginous rings of thyroid cartilage lower edge, with entry needle Jianzha County one aperture, then to centripetal direction in trachea, insert one of internal diameter 0.8mm capillary glass tube, make capillary tube just contact the trachea lower surface, to draw the sputum at trachea rear portion.Record the normal secretory volume of 2h before administration, then press table 6 dosage intravenous administration, wherein Normal group is to the equal-volume distilled water, and positive control adopts the Bisolvon injection, then observes after administration sputum secretory volume in 2h, with the resolve phlegm effect of investigation medicine.Result shows: after each dosage group administration of total urso cholic acid injection, before expectoration amount and administration, compare, difference has statistical significance (P<0.05 or P<0.01), and there was no significant difference before and after the Normal group administration.In Table 6.
The resolve phlegm effect of table 6 total urso cholic acid injection
(1)Compare P<0.05 after administration with before administration;
(2)P<0.01
5, hepatic cholagogic effect research
(1), the total urso cholic acid injection is to the protective effect of rat carbon tetrachloride acute hepatic injury model
Get 90 of SD rats, male and female half and half, divide 6 groups at random, 15 every group.Modeling method is: the normal saline of the capacity such as normal group lumbar injection, positive control adopts diammonium glycyrrhizinate injection, all the other each group injection 40%CCl
4Solution (olive oil dissolving), give 2 times weekly, amounts to 15 days.Result shows, the ALT of each dosage groups such as total urso cholic acid injection, AST level all obviously reduce (P<0.05, P<0.01) than model group.In Table 7.
The impact (x ± s, n=15) of table 7 total urso cholic acid injection on rat model Serum ALT and AST
(1)Compare P<0.01 with Normal group;
(2)With model group, compare, P<0.01,
(3)P<0.05.
(2), the total urso cholic acid injection is to rat isothiocyanic acid-1-naphthalene ester acute icteric liver injury model protective effect
Get 90 of SD male rats, divide at random 6 groups, 15 every group.Isothiocyanic acid-1-naphthalene ester dissolves with olive oil.The normal saline of the capacity such as normal group and model group lumbar injection, positive control adopts Yinzhihuang Injection, and total urso cholic acid injection etc. is respectively organized intraperitoneal injection continuous 7 days, every day 1 time.Result shows, with model group, compares, and each dosage group of total urso cholic acid injection all can obviously reduce the level (P<0.05, P<0.01) of TBIL, DBIL and IBIL, and has a certain amount of effect relationship.In Table 8.
The clear impact (x ± s, n=15) on rat model TBIL, DBIL and IBIL of table 8 expectorant heat
(1)Compare P<0.01 with Normal group;
(2)With model group, compare, P<0.01,
(3)P<0.05
(3), the facilitation of total urso cholic acid injection to the rat bile flow
Get 75 of body weight 180～220g male rats, be divided at random 15 groups.Before experiment, water is can't help in the 12h fasting.During experiment every Mus with chloral hydrate 0.60mg/kg intraperitoneal injection of anesthesia after, facing upward position is fixed on fixing head, cut 2cm along the abdomen median line, open abdominal cavity, find the bile duct of white flexible in the descendant duodenum mesentery, wear 2 rhizoid lines under it, the ligation pars papillaris, make " V " shape otch to the liver direction, insert plastic tube, with small beaker, collect bile.First collect 30min bile, then respectively organize rat and give medicine by duodenum by table 9, the capacity normal saline such as Normal group injection, positive control adopts arginine aspirin.Collect bile 1 time every 30min after administration, totally 3 times, record bile flow, after the calculating administration, bile flow increases percentage rate.The result demonstration, with comparison before administration, each dosage group of total urso cholic acid injection all can obviously improve bile flow, and difference has significant P<0.05.Prompting total urso cholic acid injection has the effect that increases bile flow.In Table 9.
The impact (x ± s, n=15) of table 9 total urso cholic acid injection on the rat bile flow
(1)Each group and front relatively P<0.01 of administration,
(2)P<0.05.
The accompanying drawing explanation
Fig. 1 is total urso cholic acid extract ursodesoxycholic acid of the present invention and chenodeoxycholic acid assay chromatogram
Fig. 2 is total urso cholic acid injection ursodesoxycholic acid of the present invention and chenodeoxycholic acid assay chromatogram
Assay adopts HPLC-ELSD: take octadecylsilane chemically bonded silica as filler; Take 0.05% trifluoroacetic acid as mobile phase A (t=0min, c=50%) (t=10min, c=0), and acetonitrile is Mobile phase B (t=0min, c=50%) (t=10min, c=100%), gradient elution.
Wherein, retention time is located as ursodesoxycholic acid in 4.6 minutes, and retention time is located as chenodeoxycholic acid in 6.4 minutes.
The specific embodiment
Describe by the following examples the preparation method of total bile acid extract of bear bile powder of the present invention and injection thereof in detail, but the protection domain be not intended to limit the present invention.
The preparation of embodiment 1. total bile acid extract of bear bile powder
Get Fel Ursi powder 1000g, the active carbon that adds 5 times of amount 50% ethanol and Fel Ursi powder amount 10%, agitating heating reflux, extract, 0.5 hour, filter, filtrate recycling ethanol is to distinguishing the flavor of without alcohol, and concentrated solution is diluted with water to 8 times of medical materials and measures dilute liquid medicine, adds the solid sodium hydroxide of amount of liquid medicine 10%, stirring and dissolving, 90 ℃ of lower heating hydrolysis 16 hours.Hydrolyzed solution adds 2M hydrochloric acid to regulate pH value to 1.0 under stirring, separate out precipitation, filters, and taking precipitate, wash with water to neutrality, and 100 ℃ dry, pulverize, and obtain crude product 496g.To add the active carbon of crude product amount 10% in crude product, mix homogeneously, add 5 times of amount ethyl acetate backflow of crude product and extract 2 times, and each 0.5 hour, filter while hot, merge extractive liquid,, filtrate is reclaimed ethyl acetate to 2 times of amounts of crude product, and 0 ℃ is standing 24 hours; Filter, must precipitate 245g, precipitation adds 4 times of amount dissolve with ethanols and obtains medicinal liquid, adds the active carbon of amount of liquid medicine 1% to stir decolouring, filters, alcohol liquid is concentrated into 490ml, standing, separates out white crystals, filters, crystallizing and drying, pulverize, and obtains Fel Ursi powder extract, and yield is 24.6%.Wherein the content of ursodesoxycholic acid is 78.8%, and the content of chenodeoxycholic acid is 14.5%.
The preparation of embodiment 2. total bile acid extract of bear bile powder
Get Fel Ursi powder 1000g, the active carbon that adds 10 times of amount 75% ethanol and Fel Ursi powder amount 15%, agitating heating reflux, extract, 1.5 hours, filter, filtrate recycling ethanol is to distinguishing the flavor of without alcohol, and concentrated solution is diluted with water to 10 times of medical materials and measures dilute liquid medicine, adds the solid sodium hydroxide of amount of liquid medicine 15%, stirring and dissolving, 105 ℃ of lower heating hydrolysis 20 hours.Hydrolyzed solution adds 8M hydrochloric acid to regulate pH value to 1.5 under stirring, separate out precipitation, filters, and taking precipitate, wash with water to neutrality, and 20 ℃ of vacuum dryings are pulverized, and obtain crude product 514g.To add the active carbon of crude product amount 15% in crude product, mix homogeneously, add 10 times of amount ethyl acetate backflow of crude product and extract 3 times, and each 1 hour, filter while hot, merge extractive liquid,, filtrate is reclaimed ethyl acetate to 3 times of amounts of crude product, and 20 ℃ are standing 24 hours; Filter, must precipitate 256g, precipitation adds 6 times of amount dissolve with ethanols and obtains medicinal liquid, adds the active carbon of amount of liquid medicine 3% to stir decolouring, filters, and pure liquid is concentrated into 768ml, standing, separates out white crystals, filters, and crystallizing and drying, pulverize, and obtains Fel Ursi powder extract.Yield is 22.2%.Wherein the content of ursodesoxycholic acid is 80.9%, and the content of chenodeoxycholic acid is 13.2%.
The preparation of embodiment 3. total bile acid extract of bear bile powder
Get Fel Ursi powder 1000g, the active carbon that adds 15 times of amount 95% ethanol and Fel Ursi powder amount 20%, agitating heating reflux, extract, 2 hours, filter, filtrate recycling ethanol is to distinguishing the flavor of without alcohol, and concentrated solution is diluted with water to 12 times of medical materials and measures dilute liquid medicine, adds the solid sodium hydroxide of amount of liquid medicine 20%, stirring and dissolving, 120 ℃ of lower heating hydrolysis 24 hours.Hydrolyzed solution adds 12M hydrochloric acid to regulate pH value to 2 under stirring, separate out precipitation, filters, and taking precipitate, wash with water to neutrality, and-45 ℃ of lyophilizations are pulverized, and obtain crude product 502g.To add the active carbon of crude product amount 20% in crude product, mix homogeneously, add 15 times of amount ethyl acetate backflow of crude product and extract 4 times, and each 1.5 hours, filter while hot, merge extractive liquid,, filtrate is reclaimed ethyl acetate to 5 times of amounts of crude product, and 30 ℃ are standing 24 hours; Filter, must precipitate 252g, precipitation adds 8 times of amount dissolve with ethanols and obtains medicinal liquid, adds the active carbon of amount of liquid medicine 5% to stir decolouring, filters, and pure liquid is concentrated into 1008ml, standing, separates out white crystals, filters, and crystallizing and drying, pulverize, and obtains Fel Ursi powder extract.Yield is 19.6%.Wherein the content of ursodesoxycholic acid is 82.0%, and the content of chenodeoxycholic acid is 12.8%.
The preparation of embodiment 4. Fel Ursi powder total bile acid injections
Get total bile acid extract of bear bile powder 20g (in ursodesoxycholic acid), inject water 500ml, with 10% sodium hydroxide solution, regulate pH value to 7.5, heating makes dissolve complete, cools, and mends and adds water to 1000ml, add the 3g needle-use activated carbon, 85 ℃ of insulations are decoloured 30 minutes, and the titanium rod filters, filtering with microporous membrane, cool fill (10ml/ props up), sealing, sterilizing, obtain.
The above preparation method to total bile acid extract of bear bile powder provided by the present invention and injection thereof and pharmaceutical applications are described in detail.It is to be noted; the described content of the specific embodiment is for better implementing the present invention preferred embodiment; protection scope of the present invention is not limited to the described technical scheme of above-mentioned embodiment; and should be as the criterion with the described flesh and blood of claims, any possible technologic change is only otherwise the flesh and blood that breaks away from the claims in the present invention all belongs to the scope that the present invention protects.
Claims (10)
1. the preparation method of a total bile acid extract of bear bile powder, is characterized in that comprising the steps:
A, ethanol extraction: Fel Ursi powder is mixed to ebuillition of heated reflux, extract, 0.5-2 hour with ethanol and active carbon;
B, saponification: ethanol extract is reclaimed to ethanol to distinguishing the flavor of without alcohol, concentrated solution thin up, then reinforcing body sodium hydroxide, stirring and dissolving is carried out saponification under 90-120 ℃, and the response time is 16-24 hour;
C, neutralization reaction: saponification adds hydrochloric acid to regulate pH value to 1-6 after finishing, separate out precipitation, filters and obtains the Fel Ursi powder extract crude product;
D, crude product drying: the Fel Ursi powder extract crude product adopts 100 ℃ of following temperature dryings;
E, ethyl acetate are extracted: by the crude product crushed after being dried, add active carbon and ethyl acetate backflow to extract 2-4 time, extraction time is 0.5-1.5 hour;
F, ethyl acetate crystallization: acetic acid ethyl acetate extract in step e is merged, reclaim ethyl acetate, 0-30 ℃ of placement, crystallization;
G, alcohol crystal: the ethyl acetate crystal is added to dissolve with ethanol and make solution, add activated carbon decolorizing, filter, pure liquid is concentrated, and crystallization filters, crystallizing and drying and get final product.
2. preparation method according to claim 1, is characterized in that, in step a, described concentration of alcohol is 50%-95%, the 5-15 that the ethanol consumption is Fel Ursi powder weight times, and the 10%-20% that activated carbon dosage is Fel Ursi powder weight.
3. preparation method according to claim 1, is characterized in that, in step b, described concentrated solution is diluted with water to 8-12 times of Fel Ursi powder weight, the 10%-20% that sodium hydroxide weight is dilute liquid medicine weight.
4. preparation method according to claim 1, is characterized in that, in the neutralization reaction of step c, described concentration of hydrochloric acid is 2M-12M.
5. preparation method according to claim 1, is characterized in that, in step e, and the 10%-20% that described activated carbon dosage is the crude product consumption, the 5-15 that the ethyl acetate consumption is the crude product consumption is doubly.
6. preparation method according to claim 1, is characterized in that, increases the step by re-crystallizing in ethyl acetate in step f.
7. preparation method according to claim 1, is characterized in that, in step f, acetic acid ethyl acetate extract merges 2-8 times that amount of solution is crude product weight.
8. preparation method according to claim 1, it is characterized in that, in step g, the 4-8 that the consumption that adds ethanol is ethyl acetate crystal weight doubly, add the 1%-5% that active carbon is solution weight, the 2-4 that the concentrated liquid measure of pure liquid is ethyl acetate crystal weight doubly.
9. the total bile acid extract of bear bile powder made according to the described method of claim 1-8 any one is made the method for injection, comprises the steps:
The total bile acid extract of bear bile powder made is injected to water, with sodium hydroxide solution, regulate pH value to 7-8, use activated carbon decolorizing after adding water for injection, decarbonization filtering, fine straining, cool, and fill is sealed, sterilizing and get final product.
10. injection prepared by the total bile acid extract of bear bile powder made according to the described method of claim 1-8 any one and the described method of claim 9 purposes in antibiotic, the antiviral of preparation, relieving cough and resolving phlegm and hepatic cholagogic medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101432410A CN101890048B (en) | 2009-05-21 | 2009-05-21 | Total bile acid extract of bear bile powder and preparation method and application of injection thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101432410A CN101890048B (en) | 2009-05-21 | 2009-05-21 | Total bile acid extract of bear bile powder and preparation method and application of injection thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101890048A CN101890048A (en) | 2010-11-24 |
| CN101890048B true CN101890048B (en) | 2013-12-04 |
Family
ID=43099505
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101432410A Active CN101890048B (en) | 2009-05-21 | 2009-05-21 | Total bile acid extract of bear bile powder and preparation method and application of injection thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101890048B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105395616A (en) * | 2013-12-02 | 2016-03-16 | 李兴惠 | Injection for clearing heat, eliminating phlegm and removing toxicity |
| CN103800343B (en) * | 2014-02-27 | 2015-11-04 | 华南农业大学 | The application of chenodeoxycholic acid on preparation control poultry bird flu medicine |
| CN104041678B (en) * | 2014-05-28 | 2016-05-11 | 山东龙昌动物保健品有限公司 | A kind of bile acid composition and preparation method thereof |
| CN106038601B (en) * | 2016-05-26 | 2020-02-21 | 重庆太极实业（集团）股份有限公司 | Preparation method of bear gall powder with high content, high purity and low fishy smell |
| CN110354143A (en) * | 2018-03-26 | 2019-10-22 | 长春长庆药业集团有限公司 | A kind of extracting method of bear bile powder eye drop stoste |
| CN109125352B (en) * | 2018-10-24 | 2022-01-25 | 上海凯宝药业股份有限公司 | Application of biotransformation bear gall powder in preparation of antitussive medicine |
| CN110368403A (en) * | 2019-07-17 | 2019-10-25 | 上海凯宝药业股份有限公司 | The application in preparing anti-inflammatory drugs of bioconversion bear gall powder |
| CN111272940B (en) * | 2020-03-31 | 2023-04-21 | 上海凯宝药业股份有限公司 | Limited detection method for bear gall powder extract in phlegm-heat clearing injection and fingerprint spectrum thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090494A (en) * | 1993-08-28 | 1994-08-10 | 孙文基 | Bear bile powder enteric capsule and preparation technology thereof |
| CN1110150A (en) * | 1994-04-05 | 1995-10-18 | 曹春林 | Jiawei double coptis medicament and preparing method thereof |
| CN1311002A (en) * | 2000-02-26 | 2001-09-05 | 石丽霞 | Refined bear gall powder and its preparing method |
| CN1981781A (en) * | 2005-12-13 | 2007-06-20 | 王静 | Bear-ball total bound cholic acid injection, its production and usage |
-
2009
- 2009-05-21 CN CN2009101432410A patent/CN101890048B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090494A (en) * | 1993-08-28 | 1994-08-10 | 孙文基 | Bear bile powder enteric capsule and preparation technology thereof |
| CN1110150A (en) * | 1994-04-05 | 1995-10-18 | 曹春林 | Jiawei double coptis medicament and preparing method thereof |
| CN1311002A (en) * | 2000-02-26 | 2001-09-05 | 石丽霞 | Refined bear gall powder and its preparing method |
| CN1981781A (en) * | 2005-12-13 | 2007-06-20 | 王静 | Bear-ball total bound cholic acid injection, its production and usage |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101890048A (en) | 2010-11-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101890048B (en) | Total bile acid extract of bear bile powder and preparation method and application of injection thereof | |
| CN102114044B (en) | Artificially processed bear bile powder and preparation method thereof | |
| CN102885858B (en) | Method for preparing American cockroach extract for treating hepatitis B | |
| CN100490877C (en) | Active extractive of Foliumnerviliae, preparation method and application | |
| CN102885856B (en) | Production method for producing medicine for treating hepatitis B by periplaneta americana extract | |
| CN104496816A (en) | Method for extraction and separation of chlorogenic acid, pinoresinol diglucoside, aucubin and gutta-percha from eucommia ulmodies oliv raw material | |
| CN101654471A (en) | Technology of large-scale refining production of glycyrrhetate | |
| CN102512542A (en) | Medicinal composition for treating skin wounds and preparation method for medicinal composition | |
| CN104415130A (en) | Reduning injection and preparation method thereof | |
| CN101033445A (en) | Wine capable of protecting liver and preserving one's health and preparing method thereof | |
| CN103819575A (en) | Extraction method for eupolyphaga polysaccharide, and extractive polysaccharide and application of eupolyphaga polysaccharide | |
| CN102908397B (en) | Chinese herbal compound for treating wind-heat type common cold by dredging surfaces and releasing muscles and preparation thereof | |
| CN1951422A (en) | Pharmaceutical composition for treating disease of liver and gallbladder system and preparation and use thereof | |
| CN101584779A (en) | Traditional Chinese medicine for removing calculus | |
| CN101269058A (en) | Spraying agent containing diclofenac acid and uses thereof | |
| CN103735679B (en) | Chinese medicine composition of a kind of diminishing inflammation and clearing pharynx and preparation method thereof | |
| CN101224235B (en) | Treating eperythrozoonosis medicine, preparing method and uses thereof | |
| CN103229895A (en) | Rheum officinale extract and application thereof | |
| CN102205091B (en) | Pharmaceutical composition for treating pain syndrome or arthralgia syndrome and preparation method and application thereof | |
| CN101716244A (en) | Preparation and process of qingkailing eye drops | |
| CN102626451B (en) | Application of rubus parvifolius saponin in preparing anti-fatigue drug | |
| CN110038071A (en) | A kind of Chinese medicine composition and preparation method and application with anti-herpesvirus | |
| CN103432294A (en) | Traditional Chinese medicine oral liquid for treating prawn eye disease and preparation method thereof | |
| CN103285040A (en) | Method for preparing nano periplaneta americana | |
| CN103845463B (en) | Treat Chinese medicine transdermal agent of horse osteoarthritic swelling and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C57 | Notification of unclear or unknown address | ||
| DD01 | Delivery of document by public notice |
Addressee: Wang Yu Document name: Notification of Passing Preliminary Examination of the Application for Invention |
|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |