CN100453085C - Forest frog's egg fat injection and its preparing process - Google Patents
Forest frog's egg fat injection and its preparing process Download PDFInfo
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- CN100453085C CN100453085C CNB011140704A CN01114070A CN100453085C CN 100453085 C CN100453085 C CN 100453085C CN B011140704 A CNB011140704 A CN B011140704A CN 01114070 A CN01114070 A CN 01114070A CN 100453085 C CN100453085 C CN 100453085C
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Abstract
The present invention relates to an oviductus ranaeovum oil injection and preparation technology thereof. The oviductus ranae ovum oil is prepared into an injection which can be taken in an injection medication path. The oviductus ranae ovum oil injection is prepared from Chinese oviductus ranae ovum and auxiliary materials comprising a diluting agent, an emulsifying agent, a cosolvent, an antioxidant, etc. The oviductus ranae ovum oil injection is prepared from 5 to 70% of oviductus ranae ovum oil, 0 to 80% of soybean oil, 0 to 90% of phospholipid, 0 to 1% of Span 80, 0 to 1% of tween 80, 0 to 30% of glycerol, 0 to 5% of vitamin E and 40 to 70% of water for injection, wherein the raw materials are homogenized after being emulsified. The injection has the action of central inhibition and convulsion resistance.
Description
The present invention relates to medical technical field, exactly it is a kind of forest frog's egg fat injection and preparation technology thereof.
Wood frog (Rana Temporaria Chensinensis David) claim that also Oviductus Ranae is a Chordata, two sacrificial guiding principles, and Anura, Ranidae, frog subfamily, wood frog belongs to, tens kinds such as Rana temporaria chensinensis David kind.Wood frog is China's famous and precious tonic of tradition and nourishing Chinese crude drug, and historical data see " decoction pieces is newly joined " the earliest, and record wood frog and fallopian tube (Oviductus Ranae) thereof are mended and not dry, and the lung moistening and production of body fluid promoting is gone into kidney, and lung heat clearing wets one's whistle.The successive dynasties medical book is all recorded it as tonic, and its product is that Oviductus Ranae (wood frog oviduct) and Oviductus Ranae are the dry all of wood frog.
In recent years, Chinese scholar Lee becomes justice wood frog ovum to be carried out containing rich in amino acid in solvent extraction and the analytical proof wood frog ovum 30 various trace elements, multiple unsaturated fatty acid, vitamin D, E, multiple bioactive substances such as estrogen and progestogen (Chinese medicine and natural drug 1994; 29:712).Chinese patent CN1066590A discloses the method for living and getting Oviductus Ranae and ovum; CN1117352A discloses Chinese snow and has breathed out the method that smart (Oviductus Ranae and the ovum of living and getting) and compositions thereof are made health food.Chinese patent CN1212140 discloses the wood frog ovum composition of calcium absorption improving function, and these methods disclose: wood frog ovum contains rich nutrient substances, and body is had many-sided regulating action.
The present invention has adopted CO
2Supercritical extraction technique extracts forest frog's egg fat (number of patent application 0012306.8), and is made into soft capsule, finds that it has remarkable effect for reducing blood fat.Discover that further the forest frog's egg fat injection gives to have powerful anticonvulsant action behind the animal.Therefore, the present invention makes injection with the forest frog's egg fat that different modes extracts, and brings into play its central inhibitory action with the injecting drug use approach.Through the multiple channel retrieval, do not find the evidence that forest frog's egg fat injection is known.
It is raw material that forest frog's egg fat injection adopts wood frog, and the forest frog's egg fat that extracts is made injection.Wherein forest frog's egg fat adopts CO
2Supercritical extraction, solvent extraction or physical squeezing are extracted.Injection contains forest frog's egg fat as constituent, also has other compositions and pharmaceutical adjunct, and as diluent, emulsifying agent, cosolvent, antioxidant etc., other help preparations shaping and stable injection adjuvant also can use.
Diluent can comprise olive oil, Oleum Arachidis hypogaeae semen, Oleum Glycines, Semen Maydis oil, oleic acid, water for injection etc. in adjuvant.Emulsifying agent can comprise phospholipid, spans, Tweens etc.; High PC value soybean phospholipid, sorbester p17, Tween 80 are preferred solvent.Cosolvent can comprise phospholipid, glycerol etc.Antioxidant can comprise vitamin E, vitamin C etc.
The proportioning of each composition is in the forest frog's egg fat injection prescription: forest frog's egg fat 5～70%, Oleum Glycines 0～80%, phosphatidase 10～90%, sorbester p17 0～1%, Tween 80 0～1%, glycerol 0～30%, vitamin E 0～5%, water for injection 40～70%.
Invention is in the past compared, and the present invention has following improvement:
One, the wood frog ovum extract is made injection.
Two, forest frog's egg fat has remarkable anticonvulsant action to the animal injection.
The present invention is described in further detail below in conjunction with embodiment.
1, the supercritical extraction of wood frog ovum
1.1 tried thing: wood frog ovum is provided by trouble ditch wood frog management field, north, Da Beicha village, cities and towns, Qing Yuanxian South Mountain, Liaoning Province, and the place of production is the source of Liuhe, about north latitude 42 degree 3 minutes, east longitude 125 degree 23 minutes.Being tried thing is the dry ovum of wood frog, is the black particle shape, the fishlike smell solid.
1.2 instrument and equipment: 250 liters of supercritical extraction units are developed by the Dalian Guangming Chemical Research And Design Institute.
1.3 experimental technique: remove the wood frog behind the fallopian tube, make it fully dry in the shade, exsiccant ovum is carefully stripped out, be collected in together, choose clean impurity, ovum is ground into coarse powder, take by weighing 200 kilograms, 250 liters of CO pack into
2In the supercritical extraction jar, add 10 kilograms of ethanol, use CO as entrainer
2Supercritical extraction unit pressure is elevated to gradually under 35MPa, 33 ℃ of conditions of temperature and extracted 180 minutes, CO
2Flow 39.8, the pressure of knockout drum 1 are 8MPa, and the pressure of knockout drum 2 is 4Mpa, and separating oil in knockout drum leaves standstill after-filtration and decompression recycling ethanol with ovum oil then, get extract 25kg.Yield is 12.5%.
2, the preparation of forest frog's egg fat injection
2.1 the screening of optimum formula
After making forest frog's egg fat make preparation, stablize, investigate the situation of dissolving each other of forest frog's egg fat and some other adjuvant.
2.1.1 single factor experiment
2.1.1.1 the intersolubility of forest frog's egg fat and spans surfactant relatively
Forest frog's egg fat is pressed 10: 1 and 10: 0.5 two mixed with span (Span) 60,65,80,85 respectively, the gained prescription is designated as A, B, C, D, E, F, G, H, and is cofabrication two parts, a in 30 ℃ of heating mixing, another part mixes in 45 ℃ of heating, examines or check miscible speed and degree.C, D two samples are that Span80 is better as a result, and the miscible time only needs 1 minute in the time of 45 ℃, and wherein D only needs 40 seconds.Miscible slower in the time of 30 ℃.
2.1.1.2 the miscible comparison of forest frog's egg fat and vegetable oil
Forest frog's egg fat is pressed 10: 1,10: 2,10: 3 three mixed with olive oil, Oleum Arachidis hypogaeae semen, Oleum Glycines, Semen Maydis oil and oleic acid respectively, the gained prescription is designated as a, b, c, d, e, f, g, h, I, j, k, l, m, n, o, in 45 ℃ of mixing, the result all can become the liquid of clear, but put to room temperature after 24 hours, olive oil, Oleum Arachidis hypogaeae semen, Oleum Glycines group all have small amount of precipitate, and Semen Maydis oil and oleic acid group are better.
2.1.1.3 forest frog's egg fat and Semen Maydis oil, oleic acid and the screening of Span60 mixed proportion
By forest frog's egg fat: Semen Maydis oil: Span60 and forest frog's egg fat: oleic acid: Span60 is 10: 1: 0.5 and 10: 2: 0 mixed, and gained solution has lamination and precipitate is arranged after placing for 1 week as a result.Adding 10% vitamin E herein in the side fails to solve.
2.1.1.4 forest frog's egg fat mixes with Semen Maydis oil, oleic acid and phospholipid
Span in above-mentioned " 2.1.1.2 " prescription is changed into the phospholipid (comprising German import and home-made injection stage and oral phospholipid) of different size, constant rate, be designated as a1, a2, b1, b2, c1, c2, d1, d2, e1, e2, f1, f2 respectively, experiment is found, oral phospholipid and forest frog's egg fat can not be miscible, the phospholipid of injection stage can be miscible with forest frog's egg fat, Semen Maydis oil, oleic acid, considers the easy oxidation of oleic acid, do not adopt in the prescription in the future; Homemade phospholipid is inexpensive, adopts in the prescription in the future.
Forest frog's egg fat mixes with Semen Maydis oil, Oleum Glycines and homemade phospholipid
With forest frog's egg fat and Semen Maydis oil, Oleum Glycines and phospholipid by being mixed with 8 prescriptions in 10: 2: 2,10: 2: 4,10: 4: 2,10: 4: 4, be designated as 1,2,3,4,5,6,7,8 respectively, be that 10: 4: 2 and 10: 4: 4 are more excellent with the ratio of forest frog's egg fat, Oleum Glycines, phospholipid in all prescriptions as a result, room temperature is placed and is seen variation two weekends, outward appearance is clear still, but when putting, a little layering is arranged to the 3rd week.
2.1.1.6 forest frog's egg fat and Oleum Glycines, phospholipid, ethanol; Forest frog's egg fat and Oleum Glycines, phospholipid, propylene glycol; Forest frog's egg fat and Oleum Glycines, phospholipid, Polyethylene Glycol are respectively by 10: 4: 4: 1,10: 4: 4: 6 prescriptions of 2 two kinds of ratio preparations, be designated as 11,22,33,44,55,66, result's 11,22 the bests of filling a prescription, surplus person has a small amount of layering after placing a week, but prescription before all being better than from mobile aspect.
2.1.1.7 the influence of examination tween80,20 pairs of prescriptions
In above-mentioned prescription, add 5% respectively, 10%tween80 and tween20, the result is in 24 prescriptions of preparation, it is excellent that all solution appearance all do not add the solution of tween.Consider that tween80 is commonly used, so selection tween80 is a recipe ingredient subsequently.
2.1.1.8 associating Span80 and tween80
Tween80 in the above-mentioned prescription is replaced with the mixture (1: 1) of Span80 and tween80, prepare 12 prescriptions, it is relatively poor that the result contains the prescription of Polyethylene Glycol, and other prescription is better.Adding antioxidant vitamin E and BHT (addition is 0.5%) in 6 prescriptions preferably, not influential to the outward appearance of solution, after all prescriptions are placed January, still better.
2.1.2 orthogonal test
On above-mentioned over one hundred prescription basis, filtering out 5 has the factor of considerable influence to the prescription outward appearance, and promptly forest frog's egg fat, Oleum Glycines, phospholipid, Span80 and tween80 divide 4 levels with each factor, adopt L16 (45) orthogonal table arrangement test, carry out formulation optimization.Four levels of wood frog ovum are respectively: 40g, 50g, 60g, 70g; Four levels of Oleum Glycines are: 60g, 50g, 40g, 30g; Four levels of phospholipid are: 40g, 30g, 20g, 10g; Four levels of Span80 are: 1g, 0.75g, 0.5g, 0.25g; Four levels of tween80 are: 0.25g, 0.5g, 0.75g, 1g.By this orthogonal table experiment arrangement.
Result's 1,2,7,10,13 outward appearance optimums of filling a prescription are clear liquid, and long-term placement does not change.Determine that thus the optimal proportion scope is: forest frog's egg fat: Oleum Glycines: phospholipid: span80: tween80=40～60: 30～50: 20～40: 0.5～1: 0.25～0.75.In order to reduce cost, be necessary to reduce the phospholipid consumption, simultaneously, should adopt the prescription of high concentration medicine, so select forest frog's egg fat at last: Oleum Glycines: phospholipid: span80: tween80=60: 30: 20: 0.5: 0.5.Repeat to prepare three times the equal clear of the outward appearance of gained solution with this prescription.
2.2 forest frog's egg fat injection prescription
2.2.1 emulsifying agent consumption:
Forest frog's egg fat and phospholipid are prepared Emulsion by 10: 1～10: 5, when the result is 10: 1 when the weight ratio of being tried thing and phospholipid, be difficult to make Emulsion, and weight ratio is 10: 2～10: 5 o'clock, can be made into Emulsion, after made Emulsion was homogenized with homogenizer, gained Emulsion was stable in January.2.2.2 oil for injection consumption:
With forest frog's egg fat with after injection Oleum Glycines mixed by 10: 5～10: 50, the phospholipid that adds above-mentioned formula ratio, preparation Emulsion, forest frog's egg fat and injection Oleum Glycines ratio are that 10: 10 above prescriptions can be made into Emulsion as a result, homogenize with homogenizer, gained Emulsion epigranular is placed not stratified for a long time.
2.2.3 preparation method
Get and obtain oil phase after the frog oil 50g, injection Oleum Glycines 100g mix, standby; In addition phosphatidase 12 0g and glycerol 50g are dispersed in the 500ml water for injection, make water.Oil phase is added aqueous phase, carry out emulsifying, and homogenize, add to 1000ml, promptly get forest frog's egg fat injection with water for injection with homogenizer.
The pharmacological research of 3 forest frog's egg fat injections
3.1 animal: Kunming mouse, Mus 6 weeks of age, body weight 18～21g are provided by animal housing of Shenyang Pharmaceutical University, random packet is placed on 20～24 ℃ of temperature, constant temperature (55 ± 5%), illumination (08:00-20:00) in 12 hours, 12 hours dark, in the sound damping animal housing, freely ingest, drinking-water adapts to above-mentioned environment and is used for experiment after one week.
3.2 reagent: strychnine nitrate is provided by Shanghai Hefeng Pharmaceutical Co., Ltd., lot number: 000601.
3.3 method:
Get mice and be divided into 5 groups at random, first group is model control group, the blank injection 10ml/kg of lumbar injection (ip); The the 2nd to 5 group of injection forest frog's egg fat 0.11,0.42,1.68mg/kg; The 5th group of ip pentobarbital sodium 2.5mg/kg.Injection wood frog ovum wet goods is after 5 minutes, and mouse peritoneal injection strychnine nitrate 1.5mg/kg counts the number of animals that convulsions takes place thereafter.
3.4 statistics: each is organized between the group of data and relatively uses X
2Inspection statistics.
3.5 result: mice is accepted very fast convulsions after the strychnine injection, and control group mice convulsions incidence rate reaches 84%, and three dosage groups of forest frog's egg fat convulsions incidence rate is respectively 69%, 43% and 33%.Wherein, low dose group does not have statistical significance with the matched group ratio, and middle and high dosage group then can significantly reduce the generation number of animals of fainting from fear, and pentobarbital sodium also significantly reduces convulsions incidence rate (60.4%).
The anticonvulsant action of table 1 forest frog's egg fat (X ± SD)
With model control group than * p＜0.01
Embodiment:
Following embodiment is in order better to set forth the present invention, is not to be used for limiting scope of invention.
Prescription:
Forest frog's egg fat 50 kg
Injection Oleum Glycines 100 kg
Phosphatidase 12 0 kg
Glycerol 50
Water for injection 500 1
Be made into 1,000,000 of forest frog's egg fat injections by certain technology
Method for making: measure by prescription and to obtain oil phase after forest frog's egg fat, injection Oleum Glycines mix, standby; In addition phospholipid and glycerol are dispersed in the water for injection, make water.Oil phase is added aqueous phase, carry out emulsifying, and homogenize with homogenizer, be supplemented to 1000 liters with water for injection at last, filter, with 1ml peace Bottle ﹠ Can dress, sterilization is checked, lettering, packing.Every forest frog's egg fat injection contains forest frog's egg fat 50mg.
Claims (1)
1, convulsion injection is characterized in that: be made up of following component: forest frog's egg fat 5-70%, Oleum Glycines 0-80%, phosphatidase 10-90%, sorbester p17: 0-1%, tween: 0-1%, glycerol 0-30%, vitamin E 0-5%, water for injection 40-70%.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011140704A CN100453085C (en) | 2001-06-15 | 2001-06-15 | Forest frog's egg fat injection and its preparing process |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011140704A CN100453085C (en) | 2001-06-15 | 2001-06-15 | Forest frog's egg fat injection and its preparing process |
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| Publication Number | Publication Date |
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| CN1343496A CN1343496A (en) | 2002-04-10 |
| CN100453085C true CN100453085C (en) | 2009-01-21 |
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| CNB011140704A Expired - Fee Related CN100453085C (en) | 2001-06-15 | 2001-06-15 | Forest frog's egg fat injection and its preparing process |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2101724B1 (en) | 2006-05-11 | 2020-12-02 | Regenics AS | Administration of cellular extracts for rejuvenation |
| EP2296676B1 (en) | 2008-05-09 | 2016-11-02 | Regenics AS | Cellular extracts |
| CA2798137C (en) | 2010-05-06 | 2016-06-28 | Regenics As | Use of fish egg extracts for cellular rejuvenation |
| WO2014091312A2 (en) | 2012-12-10 | 2014-06-19 | Regenics As | Use of cellular extracts for skin rejuvenation |
| CN103436357A (en) * | 2013-07-29 | 2013-12-11 | 长春三德天晟科技有限公司 | Preparation method of rana temporaria chensinensis egg oil without fishy smell |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1117352A (en) * | 1994-04-19 | 1996-02-28 | 高文材 | Chinese forest frog extract |
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- 2001-06-15 CN CNB011140704A patent/CN100453085C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1117352A (en) * | 1994-04-19 | 1996-02-28 | 高文材 | Chinese forest frog extract |
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Granted publication date: 20090121 Termination date: 20110615 |