CN100485021C - Natural animal essence muskrat fragrant active component, preparation technology and its use - Google Patents
Natural animal essence muskrat fragrant active component, preparation technology and its use Download PDFInfo
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Abstract
The present invention relates to a natural animal perfume muskrat spice active component which can be used as new natural animal perfume and medicinal material instead of musk, civet, ambergris and castoreum. Said muskrat spice active component contains cycloalkanones with 7C-21C, fatty acid and its esters, cycloalkanols with 3C-33C, aldehydes and alkenes. Its preparation process includes the following steps: adding trypsase to make reaction, then adding 1,3-propylene glycol or acetone, standing still, separation, adding potassium permanganate, adding active carbon, standing still and removing potassium permanganate, centrifugalizing, using 1,3-propylene glycol to make extraction and removing solvent.
Description
Technical field:
The present invention relates to natural animal essence muskrat fragrant active component, preparation technology and uses thereof, more precisely, is to extract the muskrat musk active ingredient by the molecular modification complete processing from muskrat sachet secretory product.
Background technology:
Muskrat musk is the natural animal spices new resources of the another preciousness found by years of researches after four your name's perfume (or spice).The resource of natural animal spices is very deficient, and its quantitative proportion and plant perfume and synthetic perfume are extremely inharmonious, and resource is very rare.Natural musk is that woods musk deer and muskiness glandular sac are secreted, the country of production also seldom, every is produced perfume (or spice) and also only has about 15 grams.Civet is the secretory product of the scent gland capsule of large and small civet, and the output of civet quantity in the world and perfume also seldom.Beaver perfume (or spice) is merely nominal especially.Grane ambra is that the enteron aisle of big marine Physter macrocephalus is discharged, and is a kind of afloat resource.As seen the resource kind of natural animal spices is very few, and quantity is also very limited.Even in this case, these animals of musk deer deer, civet and beaver all watch for animals because of endangered being classified as by domestic and international related organization, and its product Moschus, civet and beaver perfume (or spice) are also forbidden to use.Make that the precious natural animal spices resource that just very lacked originally is few as can not to lack again, formed the special historical stage that many producers with natural animal spices production various product both domestic and external do not have its corresponding product of raw material production.Muskrat musk then is by the scent gland capsule secretion that becomes male muskrat in age, produces fragrant amount per year for every and reaches 2.65 ± 0.76 grams.Muskrat is produced 2-3 tires per year, and about 6 of tire farrowing are the fragrant animals of product that a kind of reproduction speed is fast, feeding cost is low.The utilization of muskrat musk natural animal spices new resources provides competent natural animal spices source for the world undoubtedly.Muskrat musk natural animal spices can be widely used in the blending in daily-use chemical industry, tobacco and wine and fields such as food and dress ornament.Muskrat musk has anti-inflammatory, promotes growth, reduction myocardial consumption of oxygen in that pharmaceutically application is also more extensive, prevents and treats cerebral thrombosis, vasculitis, coronary heart disease and heart many-sided activity such as over-burden.
By the molecular modification processing technology, adopt the muskrat musk natural animal spices of the hot new process of production processing of enzymatic, its product is light yellow oily liquid, contain 31 compositions such as cyclopentadecanone, muskone, 9-alkene dihydrocivetone, 9-alkene dihydrocivetol, the relative density of muskrat musk (20/20 ℃) 0.9080 ~ 0.9060.Refractive index (20 ℃) 1.4800 ~ 1.5000.Acid number (in mgKOH/g)≤4.Have typical animal generative forces of heaven and earth fragrance, breath is clever clearly, lasting is lasting, fragrance is pure.When 1000 times of muskrat musk dilutions, still can smell and debate, at room temperature can place for a long time, steady quality, changed because scent gland secretory product is apt to deteriorate, instability and fragrance deficiency can not be directly as the situation of fragrance applications.Be widely used in industry and fields such as daily use chemicals, perfume industry, medicine, health care of food and tobacco and wine dress ornament.
Muskrat musk is carried out evaluation than system, successively studied the chemical ingredients and the pharmacological action of muskrat musk, the artificial live body of muskrat is got active ingredient and mechanism, the muskrat musk main compound secretion period of perfume (or spice) and product utilization, muskrat musk treatment coronary heart disease and is secreted fragrant rule, the research of muskrat musk spices product innovation, the secretion of muskrat musk cell in vitro and the construction of base of muskrat provenance and breeding supporting technology.Discover that muskrat musk contains compositions such as big cyclic alcohol class, organic aldehydes, alkene class and ketone, fat, ester.The activeconstituents of muskrat musk has anti-inflammatory, promotes growth, reduction myocardial consumption of oxygen, prevents and treats cerebral thrombosis, vasculitis, coronary heart disease and heart many-sided activity such as over-burden.China occupies first place in the world in the research field of muskrat.Gao Yun was engaged in the research of muskrat musk from 1984, successively born multinomial problems such as " the artificial live body of muskrat is got perfume (or spice) and utilization " that assign the Science and Technology Department of Jilin Province, " the muskrat musk main compound is secreted the research in period ", " construction of base of muskrat provenance and the research of breeding supporting technology ", " research of muskrat spices product innovation ", " the activeconstituents research of muskrat musk treatment coronary heart disease ", " research of muskrat musk somatocyte secretion muskrat musk ".In addition, finished " muskrat musk Jilin Province provincial standard " (standard No. is DB22/T 1015-2003) that Jilin Province farming committee assigns, in issue in 2003.These researchs provide than solid basis for the industrialization production of muskrat musk natural animal spices.
Set up the production base of muskrat musk, establish the source mill of muskrat musk spices, the production of development network is for development world natural animal spices resource contributes.Muskrat musk is widely used in the production of blending of industries such as medicine, daily use chemicals, food and tobacco and wine dress ornament and products thereof, set up the new industry of muskrat breed and muskrat musk natural animal spices, with promoting fur manufacturing industry, the feed-processing plant of muskrat, for society provides new employment opportunities.
Summary of the invention:
The object of the present invention is to provide a kind of active ingredient of muskrat musk, described muskrat musk active ingredient contains cycloalkanones, unsaturated fatty acids and ester class thereof, cycloalkanes alcohols, organic aldehydes and vinyl compound, has good biological activity, and is easy to store, toxicity is little, has no stimulation.
Another purpose of the present invention is to provide a kind of preparation technology of muskrat musk active ingredient, and is described simple for process, constant product quality, and the productive rate height is fit to large-scale industrial production.
A further object of the present invention is to provide a kind of purposes of muskrat musk active ingredient.
The invention provides a kind of muskrat musk active ingredient, it is characterized in that: the cycloalkanes alcohols, aldehydes and the alkene constituents that comprise cycloalkanones, lipid acid and the ester class thereof of 11 carbon of seven carbon to two, 13 carbon of three carbon to three.
The content of muskrat musk active ingredient of the present invention is:
C
7-C
21Cycloalkanones composition 0.01%-21%
C
7-C
21Lipid acid and ester class 0.01%-16% thereof
C
3-C
33Cycloalkanol constituents 0.1%-30%
C
3-C
33Aldehydes composition 0.01%-20%
C
3-C
33Alkene constituents 0.01%-20%.
The present invention adopts artificial live body to get the fragrant muskrat musk glandular secretion thing of gathering, and the muskrat musk secretory product of collection obtains natural animal essence muskrat fragrant active component through following complete processing:
1) trypsinase of one times of amount of muskrat musk glandular sac secretory product adding reacted 3-40 hour under-10 ℃ of-120 ℃ of temperature, got mixed solution;
2) 1,3 propylene glycol or the acetone of one times of amount of adding mixed solution, thorough mixing left standstill 1-73 hour;
3) mixed solution separates lower floor and upper solution for 2000 rev/mins, adds potassium permanganate 1-50mg reaction 0.5-100 hour;
4) doubly measure activated carbon with 0.8-10 and left standstill 1-36 hour, remove potassium permanganate;
5) under ℃ of-100 ℃ temperature of centrifugal being placed on-10, be 1-8 time with 1,3 propylene glycol extraction times, add solution amount at every turn and be 3-10 doubly, removing the gained muskrat musk active ingredient of desolvating again is 50-90%.
Analyses such as muskrat musk active ingredient of the present invention thin-layer chromatography, UV spectrum, infrared spectra, gas-chromatography, gas-mass spectrograph, muskrat musk active ingredient main chemical compositions sees Table 1.
The main chemical compositions of table 1 muskrat musk
The present invention also provides the purposes of muskrat musk active ingredient in pharmacy, food, daily-use chemical industry, tobacco and wine dress ornament.
Active ingredient of the present invention is used in preparation treatment myocardial ischemia, anoxic, coronary heart disease, vasculitis and anticoagulant and the thrombotic medicine of inhibition.
Active ingredient of the present invention is the purposes of natural animal spices as lasting agent and fixative.
Muskrat musk active ingredient of the present invention can be made tincture, soft capsule, dripping pill, spray and tablets and other formulations, easy administration as medicine, food.
The present invention finds the result of study of the pharmacologically active of described muskrat musk active ingredient:
1, small white mouse ip muskrat musk 120mg/kg can also promote prostate gland to urge seminal vesicle when promoting body weight gain, and the growth of immune organ thymus gland and spleen illustrates that muskrat musk promotes that growth of animal is many-sided.
2, muskrat musk has the activity of SOD in the tangible promotion liver, improve the ability of removing the superoxide anion that produces because of lipid peroxidation, stop the formation of lipid peroxide mda, intracellular peroxidatic reaction of lipid process slows down, demonstrate stronger activity of fighting against senium, the activity of fighting against senium of muskrat musk presents tangible dose-effect relationship, and promptly oral 120mg/Kg is anti-ageing more obvious than oral 60mg/Kg and 30mg/Kg's.
3, muskrat musk has the very significantly effect of stable interior environment, is mainly reflected in muskrat musk (12mg10ml test tube) and can resists the fine blood haemolysis that goes that oozes NaCl solution at the height of 0.4% hypotonic NaCl and 0.8%.By abdominal injection muskrat musk 12mg 8 days, again can be to anti-erythrocyte antih(a)emolysin in the body of 0.45%NaCl solution.The antagonism erythrocyte membrane breaks, and the effect of stablizing erythrocyte membrane is remarkable.
4, muskrat musk anti-hemolysis activity research shows, muskrat musk can resist the hemolytic action of the hypotonic and hypertonic solution of 0.4%-0.8% gradient, only presents slightly soluble blood extremely hypotonic 0.35% and 0.30%.Muskrat musk has extremely significantly stablizes erythrocytic activity.
5, muskrat musk shows that to the pharmacology activity research result of anesthetized dog muskrat musk can reduce myocardial consumption of oxygen, reduces the utilization of blood oxygen, bring high blood pressure down, and reducing heart rate, coronary blood flow increasing, over-burden that good prospects for application is arranged to the clinical treatment heart.
6, muskrat musk has anti-inflammatory, bacteriostatic pharmacologically active.
7, muskrat musk is by toxicological experiment, and muskrat musk toxicity is little, has no stimulation, and is safe in utilization.
Can draw by above research and the invention has the advantages that:
1, the invention provides the natural animal essence muskrat fragrant active component that can replace Moschus, thereby determine the medicinal use of muskrat musk, opened up new natural animal spices new resources.
2, the muskrat musk safety non-toxic of the present invention preparation is non-stimulated, room temperature storage for a long time, and steady quality, the pharmacologically active aspect is better than muskrat musk glandular secretion thing, is indicating that it has good application prospects.
3, raw material sources of the present invention are abundant, and preparation technology is simple, can make in flakes, formulations such as ball, spray, capsule, and are easy to use.
4, folk prescription of the present invention be mixed with medicine have anti-inflammatory, hypoxia tolerance, reduction myocardial consumption of oxygen, reduce blood oxygen utilization, bring high blood pressure down, reducing heart rate, coronary blood flow increasing and various active such as anti-ageing, anticoagulation and thrombolysis, help preventing and treating coronary heart disease, cardiac hypertrophy, over-burden and prevent and treat critical illness such as cerebral thrombosis, vasculitis for heart.
5, clinical trial shows, the oral instant effect of natural animal spices muskrat musk of the present invention's preparation has no side effect.
6, scent gland secretory product is the oyster white dope, deposits apt to deteriorate and variable color naturally, and fragrance is lighter, and the urine flavor is arranged, can not be directly as the natural animal fragrance applications.The secondary processing of the present invention by scent gland secretory product is that the technical matters of molecular modification is developed the muskrat musk active ingredient.This product is light yellow stream shape liquid.Relative density (25/25 ℃) 0.9084.Refractive index (20 ℃) 1.4851.Acid number (in mgKOH/g) 3.3.Have typical animal generative forces of heaven and earth fragrance, fragrance is clearly clever, soft, lasting is lasting, head is fragrant slightly with the fruital of ester class, and fragrance is purer.Still can smell when muskrat musk spices dilutes 1000 times and debate, fragrance is stronger.And contain on the muskone basis at muskrat musk glandular secretion thing saturated fatty acid is become unsaturated fatty acids, removed adiaphorous protein, increased that it is anti-ageing, remove free radical, reduce myocardial consumption of oxygen, reduce blood oxygen utilization, bring high blood pressure down, reducing heart rate, coronary blood flow increasing, press down because of bolt forms, strengthens anti-inflammatory action, shorten vetanarcol the length of one's sleep, therapeutic action such as promote to grow.
7, utilize muskrat musk glandular secretion thing as medicine and lasting agent, must just can play the fragrance effect of its due effect and assurance cosmetics of everyday use by preparation technology of the present invention, and have only muskrat musk just can reach the muskrat musk standard by preparation technology's preparation of the present invention.
It is of the present invention that preparation is simple, constant product quality, and the productive rate height is fit to large-scale industrial production.Adopt the muskrat musk active ingredient of the present invention's preparation to contain a large amount of unsaturated fatty acidss, increased its biological activity, be suitable for as natural drug medicinal material and spices, through experimental results demonstrate that the muskrat musk active ingredient not only has anti-inflammatory, pharmacologically active such as anti-ageing and antibacterial, and aspect cardiovascular, good pharmacologically active is arranged also, it can significantly reduce myocardial consumption of oxygen, reduce the utilization of blood oxygen, bring high blood pressure down, reducing heart rate, coronary blood flow increasing and anti-ageing, anticoagulation and thrombolysis etc. help preventing and treating coronary heart disease, cardiac hypertrophy, over-burden and prevent and treat cerebral thrombosis for heart, critical illness such as vasculitis.Muskrat musk can be widely used in the blending of daily-use chemical industry, food, tobacco and wine dress ornament as new natural animal spices simultaneously.Especially the muskrat musk active ingredient can be used as lasting agent and fixative in cosmetic industry.And the muskrat musk active ingredient has no side effect, and is non-stimulated, but prolonged preservation under the room temperature, steady quality is easy to store, and alternative Moschus, civet, ambergris and beaver perfume (or spice) become new natural animal spices and medicine medicinal material.
Embodiment
Below by test the example further set forth active ingredient of the present invention beneficial effect.
The pharmacological evaluation of test example 1 natural animal essence muskrat fragrant active component of the present invention.
One, the activity of fighting against senium of muskrat musk
1, muskrat musk is to the influence of male mouse growth childhood
Get 24 of male white mouses, body weight 11.75 ± 2g divides three groups, each 9 of control group and muskrat musk groups, 6 of androlin groups.Experiment periods is fed machine-processed feed block, sunflower seeds, and drinking-water is picked-up arbitrarily.3 groups of small white mouse difference ip muskrat musk 120mg/Kg, androlin 6, Omg/Kg and physiological saline 5.0ml/Kgo successive administration 13 days were weighed after administration on the the 3rd, 6,8,10 and 13 day.The results are shown in Table 2.
Table 2 muskrat musk is to the influence that do not become small white mouse body weight gain in age (x ± SE)
Annotate: compare with control group * p＜0.05**p＜0.01***P＜0.001, down together.
Table 2 as seen, ip muskrat musk the 6th day, the small white mouse body weight increases to some extent than control group, the 8th～13 day, body weight gain was rapid, with control group relatively, significant difference, P＜0.01.Its action intensity is similar to ip androlin 6.0mg/Kg.
2, muskrat musk is to male childhood of small white mouse prostate gland---the influence of seminal vesicle and testicular weight
To test respectively organize mouse in 1 and weigh after, sacrificed by decapitation, win prostate gland---seminal vesicle, testis.Thymus gland and spleen are weighed, and the results are shown in Table 3.
Table 3 muskrat musk is to the influence that do not become each allelotaxis of mouse in age (x ± SE)
3, muskrat musk is to the influence of superoxide dismutase activity and mda content in the hepatic tissue
Get and respectively organize the activity that the small white mouse hepatic tissue is measured superoxide-dismutase.Experimental result sees Table 4.
Table 4 mouse perfume (or spice) is to the influence of superoxide dismutase activity and mda content in the hepatic tissue (x ± SE)
Table 4 as seen, three dosage groups of muskrat musk all have in various degree enhancement to the activity of mouse liver superoxide-dismutase, especially with ip muskrat musk 120mg the strongest (P＜0.01), other two groups also demonstrate enhanced trend, demonstrate tangible dose-effect relationship.
Muskrat musk also has and reduces the MDA content effect more significantly, has also demonstrated dose-effect relationship simultaneously.
Two, muskrat musk is to the stabilization of small white mouse erythrocyte membrane
1, muskrat musk is to stablizing the effect of small white mouse erythrocyte membrane
Abdominal injection muskrat musk 12mg/Kg, successive administration 8 days, control group intravenous injection physiological saline 5.0ml/Kg.After the last administration 1 hour; extract cardiac blood 0.2ml and be prepared into 2% red cell suspension; getting 0.2ml puts to 0.45% sodium chloride solution; be inverted 2 times; the centrifugal 2min of 2000 * g; get supernatant liquor and survey the OD value, estimate the provide protection of muskrat musk, the results are shown in Table 5 the small white mouse erythrocyte membrane with the OD value in 420nm.
Table 5 muskrat musk is to the provide protection group OD value of small white mouse erythrocyte membrane (x ± SE)
Table 5 result as seen, iv muskrat musk 0.4mg can obviously protect the effect of mouse red blood cell film.
2, the anti-hemolysis activity of muskrat musk
Get 10 test tubes, add respectively 0.30%, 0.35%, 0.40%, 0.45%, 0.50%, 0.55%, 0.60%, 0.65%, 0.70% with 0.80% the different hypotonic and high sodium chloride solution that oozes concentration, in vitro put into 0.2% in every and remove fine blood 0.1ml, add the 12mg muskrat musk, observe the outer hemolytic activity of muskrat musk antagonist.
The result shows that muskrat musk has extremely strong anti-hemolysis activity.It can resist the hemolytic action of the hypotonic and hypertonic solution of 0.4% to 0.8% different gradients, only presents slightly soluble blood extremely hypotonic 0.35% and 0.30%.As seen, muskrat musk has the extremely significantly activity of stabilizing cell membrane.
The toxicological experiment of test example 2 muskrat musk active ingredients of the present invention
Sample: muskrat musk glandular secretion thing provides for muskrat musk spices product innovation seminar of Jilin Province's special product institute, and acquisition time is 6-August, adopts artificial live body to get fragrant method (Chen Yushan etc. 1996), gathers the back and places refrigerator (18 ℃) storage.
The muskrat musk spice product is the product of muskrat musk glandular secretion thing through molecular modification (secondary processing) preparation, and institute provides by Jilin Province's produce.
Laboratory animal: rabbit, male and female dual-purpose, body weight (2.5-3.0Kg).Cavy, male and female dual-purpose, body weight 250-300g.Outbreeding system Kunming kind small white mouse, male and female dual-purpose, body weight 20-24g.Provide by Norman Bethune Medical University, Jilin Province Experimental Animal Center.
Method and result
1, scent gland secretory product and muskrat musk spice product are to the toxic influence of mouse skin
Get 70 of small white mouses, male and female half and half, body weight 20 ± 2.0g is divided into 7 groups at random, and 10 every group, the animal stable in two days, first group is the blank group, smears consubstantiality distilled water to small white mouse.2nd, 3,4 groups are produced the high, medium and low concentration group of scent gland secretory product respectively, smear scent gland secretory product and muskrat musk spices to every mouse of each group respectively.Back part of animal defeathering after the grouping, with Japanese trichogen (consonance Industry Co., Ltd) depilation, about 2-2.5cm raised three days, smeared by each group and different concns, saw Table 6.Smeared respectively 30 days, and observed small white mouse and smear place's skin variation and active situation, and record.After the experiment, animal continued to stablize 5 days, each group small white mouse is carried out broken sturdy the testing of skin,, cut off 5mm diameter otch with surgical scissors at center, defeathering place with above-mentioned grouping mouse, cause skin to break the skin model, alcohol (70%) sterilization two days later, was smeared 5 times/three days by dosage described in the table and group, observe skin once more and change active situation, and record.Then whole male and female mouse are dropped into breeding.Observe scent gland secretory product and muskrat musk spices to reproductive performance and embryo's distortion effect.
The result of visual inspection shows that all the mouse diet is normal, and activity is not broken full, smooth, the no abnormal variation of mouse skin of skin freely.Broken skin mouse group mouse contrast and experimental group skin all heal, and any toxic side effect do not occur.
The mouse sexuality that drops into breeding is normal, and female mice is all become pregnant, and litter size, young young mouse weight and control group no significant difference see Table 7.Outward appearance and cut open inspection and distortion all occurs.Scent gland secretory product and muskrat musk spices do not have teratogenesis.
Table 6 scent gland secretory product and muskrat musk spices are to mouse skin toxicity, broken skin skin healing effect
Table 7 muskrat musk and muskrat musk spices are to MOUSE REPRODUCTION and the young teratogenesis of tire
2, scent gland secretory product and muskrat musk spices are to the irritating influence of rabbit skin
Get 50 of rabbit, male and female are regardless of, body weight 2.5-3.0Kg, be divided into seven groups, give consubstantiality ponding (distilled water), the 2nd, 3,4 group scent gland secretory product 1,2,3 respectively for the blank group for first group, 5th, 6,7 groups are respectively muskrat musk spices group, and every experimental rabbit smears scent gland secretory product and muskrat musk spices dosage sees Table 8.Respectively each group rabbit is fixed, respectively with back two 10 * 6cm skin of face defeathering, with alcohol and iodine sterilization defeathering place skin, each is organized rabbit and according to dosage smeared scent gland secretory product and muskrat musk spices respectively 30 days, observation is tried the rabbit agents area no change, and response intensity criterion sees Table 9.
Table 8 scent gland secretory product and muskrat musk spices are to the skin irritant influence of rabbit
The criterion of table 9 skin irritation test response intensity
Observations shows, 48 hours observation rabbit agents area skin after the administration, each organizes that rabbit there is no that hyperemia, bubble form, pleat is mashed, exfoliation, degeneration necrosis, the variant dosage group of scent gland secretory product and muskrat musk spices does not all have any unusual phenomenon to rabbit skin, skin is not had any toxicity to stimulate, with sample after 30 days, its activity, search for food normal.Safe in utilization.After 30 days, rabbit is put to death, take medicine position skin, carry out the on-the-spot cut sections for microscopic examination of disease, smear position skin and do not have any abnormal change.More there is not the canceration of bringing out.High dose group scent gland secretory product and the external application of muskrat spices do not have any stimulation and carcinogenesis.
3, scent gland secretory product and muskrat musk spices are to the anaphylactoid influence of cavy
Get 35 of cavys, male and female are regardless of, and body weight 250-300g is divided into 7 groups at random.Respectively each group guinea pig back is taken off the skin of two 6 * 4cm areas, carry out defeathering, simultaneously with alcohol and iodine disinfection.Treated cavy, the 1st group is that control group is given the distilled water with volume, smears scent gland secretory product 0.2,0.5,0.8 gram for the 2nd, 3,4 group, smears muskrat musk spices 0.2,0.5,0.8 gram respectively for the 5th, 6,7 group, each treated animal was smeared 5 days continuously, and observation is tried the cavy agents area no change.Criterion is as follows:
_ _ no abnormal reaction
± roll up, perpendicular hair, other is no abnormal
+ grab nose, perpendicular hair, uneasiness, breathe urgent, slight cyanosis
++ perpendicular hair, breathing significant difficulties, cyanosis, myasthenia of the limbs, belly are creeped with pasting.
+++death
Observe after medication, scent gland secretory product and muskrat musk spices all do not have any unusual performance.
Stopped to smear behind scent gland secretory product and the muskrat musk spices 7 days, and attacked 3 days (same dose and identical stimulating method) with scent gland secretory product and muskrat musk spices once again, so that observe scent gland secretory product and the muskrat musk spice product does not have any anaphylaxis to cavy yet.Prove that further scent gland secretory product and muskrat musk spices are safe in utilization, untoward reactions such as no allergy.
4, scent gland secretory product and muskrat musk spices are observed the acute toxicity of the disposable injection of small white mouse vein
Get two groups of 20 branches of small white mouse, 10 every group, body weight 31 ± 2.6 grams by only giving small white mouse intravenous injection (tail vein) scent gland secretory product and muskrat musk spices 0.8 gram (maximum injection dosage), are observed the toxic reaction of scent gland secretory product and muskrat musk spices.
The result shows, behind two groups of injected in mice scent gland secretory product and muskrat musk spices 0.8 gram, all occurs deadly, and different is that after the injection back mouse performance uneasiness, all the differing appearance activity reduces.In the future inferior, movable normal, to observe after seven days, none only occurs dead.
The muskrat musk vanishing cream No. 1, No. 2, No. 3, No. 4 of test example 3 muskrat seminars of Gao Yun development and muskrat musk perfume is anti-ageing, the activity research of anti-inflammatory and aspect such as antibacterial.
One, the muskrat musk medicinal cosmetic is to the active influence of mouse skin SOD
132 mouse web portion hairs are cut clean ellipse to 3cm, experiment in two batches.Each 66 of the two batches of mouse are divided equally 6 groups for every batch.1～5 group is respectively muskrat musk perfume, muskrat musk vanishing cream 1,2,3 and No. 4, and 6 groups is control group.First batch of mouse once is coated with cream and perfume by every mornings 8.30 beginning of dosage shown in the table, and control group is coated with water.Continuous two weeks.End batch mouse every morning 9.00 and afternoon 3.00 in 2 weeks is coated with cream and perfume secondary.After last was coated with cream and perfume 1h, the sacrificed by decapitation animal took off and is coated with cream and perfume place skin is cut into fragment, pressed the Geller method and separated SOD, measured by methods such as Lan Kaiwei.The results are shown in Table 10.
Table 10 muskrat musk makeup are to the influence of SOD in Mice (μ g/g skin)
Annotate: compare with control group ※ P＜0.05 ※ ※ P＜0.01 ※ ※ ※ P＜0.001, down together.
No matter No. 1, muskrat musk vanishing cream and No. 3 smear for 1 time or twice every day, all can obviously strengthen the activity of mouse skin SOD, but it is more obvious to smear the activity of SOD every day for twice.Once smear the active not obvious of SOD No. 2 and No. 4, be coated with cream twice every day and demonstrate stronger SOD activity but be increased to, 1～No. 4 cream of muskrat musk all demonstrates tangible dose-effect relationship.Muskrat musk perfume does not have the activity of obvious raising mouse skin SOD.
Two, the muskrat musk medicinal cosmetic is to the influence of mouse lipofuscin
Get the previous experiments animal livers and open luxuriant method methyl alcohol by indigo plant: chloroform (2: 1) extracts liquid 5ml and prepare tissue homogenate in glass homogenizer, behind 40 ℃ of incubation 5min of homogenate, the centrifugal 10min of 1500g, supernatant liquor measure lipofuscin content on the RF-510 spectrophotofluorometer.Rather be the fluorescence primary standard with 0.1 μ g/ml (preparation of 0.05M sulfuric acid) quinoline during mensuration, excitation wavelength 365nm, wavelength of transmitted light 435nm, slit 10nm, gain 2/9.The results are shown in Table 11.
Table 11 muskrat musk makeup are to the influence of mouse lipofuscin (μ g/g liver)
The result shows, except that muskrat musk perfume, no matter once a day or embrocate for twice the content that reduces lipofuscin in the mouse liver is very significantly all arranged, action intensity is also similar for all the other 4 kinds of muskrat balm frosts.It is not obvious that muskrat musk perfume is embrocated the effect that reduces Mouse Liver lipofuscin content once a day, but be increased to twice every day the effect that reduces lipofuscin more significantly arranged.
Three, the muskrat musk medicinal cosmetic is to the influence of mice serum lipid peroxide (LPO) content
Previous experiments animal broken end is got blood and is prepared serum, presses thiobarbituricacid (TBA) determination of color Serum LPO content.Get and add 0.61%TBA 1ml after serum 0.3ml adds 20% Tricholroacetic Acid 2.5ml mixing, boiling water takes out after boiling 30min, flowing water is cooled to room temperature, add the pigment that propyl carbinol 4ml vibration mixing extracting TBA and mda condensation produce, the centrifugal 10min of 1500 * g gets supernatant liquor with 751 1 type spectrophotometric determination content.TEP with same operation during mensuration is a standard, the propyl carbinol zeroing, and wavelength is 535nm.The results are shown in Table 12.
Table 12 muskrat musk makeup are to MDA content influence in the mice serum
The result shows that 5 kinds of muskrat musk makeup are except that perfume, and all the other 4 kinds of cream frosts all have the effect that more significantly reduces LPO content in the mice serum, and twice of every day is more obvious than primary effect, is tangible dose-effect relationship.
Four, the muskrat musk medicinal cosmetic is to (the influence of MAO-B) of Mouse Liver monoamine oxidase
Aforementioned mouse sacrificed by decapitation is got hepatic tissue by five strict homogenate of sample prescription legal system, classified centrifugal preparation monoamine oxidase source, and with the substrate of benzylamine as MAO, colorimetric estimation MAO-B activity.The results are shown in Table 13.
Table 13 muskrat musk makeup are to the influence of Mouse Liver monoamine oxidase
Table 13 as seen, (MAO-B) has differently restraining effect to monoamine oxidase in the murine liver tissue for 5 kinds of medical cosmetic products of muskrat musk.Especially obviously with No. 2,3 and 4, cream frost.No. 1, cream frost and perfume only reach to smear for twice every day wipes the activity that just demonstrates the inhibition monoamine oxidase.
Five, the muskrat musk medicinal cosmetic is to the influence of the scorching effect of last mouse-anti
Get 60 of mouse, divide equally 6 groups, 1～5 group is respectively muskrat musk perfume, muskrat balm frost 1,2,3 and No. 4, and 6 groups is control group.Embrocate muskrat balm frost or perfume (0.1g or 0.05ml/ are only) at mouse web portion (cutting clean belly wool) continuously, once a day, after last is embrocated 1h, cause the scorching mouse both sides ear of cutting afterwards by only being coated in two ears with dimethylbenzene 0.05ml, weigh by the ear circle tissue that the refined cloud method of Xu is laid same position fixed size with rubber plug puncher, with left and right sides ear weight difference evaluation swelling degree.
The result shows, control group swelling degree is 9.59 ± 2.81mg, muskrat musk perfume is 5.51 ± 2.27 (P＜0.01), muskrat balm frost is 5.47 ± 2.89mg (P＜0.05) No. 1, No. 2 is 5.13 ± 1.25mg (P＜0.001), No. 3 is 6.31 ± 1.6 (P＜0.05), and No. 4 is 6.43 ± 2.19 (P＜0.05).Show that 5 kinds of muskrat musk makeup all have stronger anti-inflammatory action.But it is especially more obvious with No. 2, cream frost and perfume antiphlogistic effects.
Six, the muskrat musk medicinal cosmetic is to the influence of mouse part skin dermal tissue morphological structure
Get 36 mouse, male and female are regardless of, and divide six groups, and 6 every group, embrocate the muskrat musk makeup once every day in two weeks, and after last was embrocated 1h, skin was cut in carefulness, and routine is manufactured tissue slice, row HE dyeing.Observe the metamorphosis of respectively organizing the mouse subcutaneous dermal tissue.
The result shows that capillary vessel is abundant in five kinds of muskrat balm frost group mouse subcutaneous (except the perfume) corium, the body of gland prosperity, and the reticulin fiber in the reticular tissue, collegen filament and spandex fiber etc. all increase than contrast.Demonstrate the basis of skin moisten skin care preferably.Muskrat musk perfume does not have this effect.
Seven, the muskrat musk medicinal cosmetic is to the influence of bacteriostatic action
The streptococcus aureus of selection standard, Staphylococcus albus, intestinal bacteria and Pseudomonas aeruginosa are made flat board with nutrient agar medium (chemical reagent work of shanghai Medicine assay office product), with 1.5 * 10
8Ml/ bacterium liquid is applied to respectively in the flat board, burrows on flat board then, and every hole dia is 6mm, and every hole adds muskrat balm creme 0.1g, cultivates 24h for perfume 0.05ml37 ℃, measures antibacterial ring size.The results are shown in Table 14.
Table 14 muskrat musk medicinal cosmetic is to the influence of bacteriostatic action
The result shows that 5 kinds of makeup all have bacteriostatic activity to some extent to golden Portugal, white area and three kinds of bacterial strains of large intestine, and wherein muskrat musk perfume activity is the strongest, and cream has the moderate bacteriostatic action 1, No. 3, No. 2 bacteriostatic action of cream frost a little less than.
Eight, muskrat musk EM frost is observed the treatment of seborrhea skin fire and seborrheic keratosis
Use muskrat musk EM frost treatment seborrheic dermatitis and seborrheic keratosis and obtained curative effect preferably.In 20 routine seborrheic keratosis, produce effects 3 examples, effective 15 examples, total effective rate reaches 90%, and medication can be seen effect in 2 months.In addition, in 30 routine seborrhea facial dermatitis diseases, cure 9 examples, effective 20 examples, total effective rate reaches 96.7%, and effect is obvious, and inflammation promptly begins to disappear in 3～5 days after medication, and gargalesthesia alleviates.
Further set forth the preparation method of active ingredient of the present invention by the following examples
Embodiment 1
Muskrat is provided by Gao Yun, the artificial live body of non-narcotic method is got the fragrant natural muskrat musk of gathering, the trypsinase that adds one times of amount, under-10 ℃ of temperature, reacted 3 hours, get mixed solution, add 1 of one times of amount of mixed solution, 3 propylene glycol, thorough mixing left standstill 1 hour, and mixed solution separates lower floor and upper solution for 2000 rev/mins, add potassium permanganate 1mg reaction 0.5 hour, left standstill 1 hour with 0.8 times of amount activated carbon, remove potassium permanganate, under the centrifugal being placed on-10 ℃ temperature, with 1,3 propylene glycol extraction times are 1 time, and add solution amount is 3 times at every turn, and removing the gained muskrat musk active ingredient of desolvating again is 50%.
Embodiment 2
Muskrat is provided by Gao Yun, and the artificial live body of non-narcotic method is got the fragrant natural muskrat musk of gathering, and adds the trypsinase of one times of amount, reacts 12 hours under 20 ℃ of temperature, gets mixed solution; Add the acetone of one times of amount of mixed solution, fully mix, left standstill 5 hours; Mixed solution is separated lower floor and upper solution for 2000 rev/mins, add potassium permanganate 10mg reaction 5 hours; Left standstill 10 hours with 2 times of amount activated carbon, remove potassium permanganate; Centrifugal being placed under 20 ℃ of temperature is 2 times with 1,3 propylene glycol extraction times, and add solution amount is 5 times at every turn, and removing the gained muskrat musk active ingredient of desolvating again is 54.6%.
Embodiment 3
Muskrat is provided by Gao Yun, and the artificial live body of non-narcotic method is got the fragrant natural muskrat musk of gathering, and adds the trypsinase of one times of amount, reacts 24 hours under 30 ℃ of temperature, gets mixed solution; Add 1,3 propylene glycol of one times of amount of mixed solution, fully mix, left standstill 24 hours; Mixed solution is separated lower floor and upper solution for 2000 rev/mins, add potassium permanganate 20mg reaction 40 hours; Left standstill 24 hours with 6 times of amount activated carbon, remove potassium permanganate; Centrifugal being placed under 30 ℃ of temperature is 4 times with 1,3 propylene glycol extraction times, and add solution amount is 6 times at every turn, and removing the gained muskrat musk active ingredient of desolvating again is 72.9%.
Embodiment 4
Muskrat is provided by Gao Yun, and the artificial live body of non-narcotic method is got the fragrant natural muskrat musk of gathering, and adds the trypsinase of one times of amount, reacts 30 hours under 60 ℃ of temperature, gets mixed solution; The acetone that adds one times of amount of mixed solution, thorough mixing left standstill 60 hours, and mixed solution separates lower floor and upper solution for 2000 rev/mins, add potassium permanganate 40mg reaction 80 hours, left standstill 30 hours with 8 times of amount activated carbon, remove potassium permanganate, centrifugal being placed under 60 ℃ of temperature, with 1,3 propylene glycol extraction times are 6 times, and add solution amount is 8 times at every turn, and removing the gained muskrat musk active ingredient of desolvating again is 86.5%.
Embodiment 5
Muskrat is provided by Gao Yun, and the artificial live body of non-narcotic method is got the fragrant natural muskrat musk of gathering, and adds the trypsinase of one times of amount, reacts 40 hours under 120 ℃ of temperature, gets mixed solution; Add 1,3 propylene glycol of one times of amount of mixed solution, fully mix, left standstill 73 hours; Mixed solution is separated lower floor and upper solution for 2000 rev/mins, add potassium permanganate 50mg reaction 100 hours; Left standstill 36 hours with 10 times of amount activated carbon, remove potassium permanganate; Centrifugal being placed under 100 ℃ of temperature is 8 times with 1,3 propylene glycol extraction times, and add solution amount is 10 times at every turn, and removing the gained muskrat musk active ingredient of desolvating again is 90.0%.
Claims (6)
1, a kind of natural animal essence muskrat fragrant active component, it is characterized in that: comprise cycloalkanes alcohols, aldehydes and the alkene constituents of cycloalkanones, lipid acid and the ester class thereof of 11 carbon of seven carbon to two, 13 carbon of three carbon to three, by following prepared:
1) trypsinase of one times of amount of muskrat musk glandular sac secretory product adding reacted 3-40 hour under-10 ℃ of-120 ℃ of temperature, got mixed solution;
2) 1,3 propylene glycol or the acetone of one times of amount of adding mixed solution, thorough mixing left standstill 1-73 hour;
3) mixed solution separates lower floor and upper solution for 2000 rev/mins, adds potassium permanganate 1-50mg reaction 0.5-100 hour;
4) doubly measure activated carbon with 0.8-10 and left standstill 1-36 hour, remove potassium permanganate;
5) under ℃ of-100 ℃ temperature of centrifugal being placed on-10, be 1-8 time with 1,3 propylene glycol extraction times, add solution amount at every turn and be 3-10 doubly, removing the gained muskrat musk active ingredient of desolvating again is 50-90%.
2, natural animal essence muskrat fragrant active component according to claim 1 is characterized in that: the content of described muskrat musk active ingredient is:
C
7-C
21Cycloalkanones composition 0.01%-21%
C
7-C
21Lipid acid and ester class 0.01%-16% thereof
C
3-C
33Cycloalkanol constituents 0.1%-30%
C
3-C
33Aldehydes composition 0.01%-20%
C
3-C
33Alkene constituents 0.01%-20%.
3, a kind of preparation technology of natural animal essence muskrat fragrant active component is characterized in that:
This technology comprises the following steps:
1) trypsinase of one times of amount of muskrat musk glandular sac secretory product adding reacted 3-40 hour under-10 ℃ of-120 ℃ of temperature, got mixed solution;
2) 1,3 propylene glycol or the acetone of one times of amount of adding mixed solution, thorough mixing left standstill 1-73 hour;
3) mixed solution separates lower floor and upper solution for 2000 rev/mins, adds potassium permanganate 1-50mg reaction 0.5-100 hour;
4) doubly measure activated carbon with 0.8-10 and left standstill 1-36 hour, remove potassium permanganate;
5) under ℃ of-100 ℃ temperature of centrifugal being placed on-10, be 1-8 time with 1,3 propylene glycol extraction times, add solution amount at every turn and be 3-10 doubly, removing the gained muskrat musk active ingredient of desolvating again is 50-90%.
4, the application of natural animal essence muskrat fragrant active component according to claim 1 in pharmacy, food, daily-use chemical industry, tobacco and wine dress ornament.
5, the application of natural animal essence muskrat fragrant active component according to claim 1 in preparation treatment myocardial ischemia, anoxic, coronary heart disease, vasculitis and anticoagulant and the thrombotic medicine of inhibition.
6, natural animal essence muskrat fragrant active component according to claim 1 is as the purposes of lasting agent and fixative.
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| CN104962511B (en) * | 2015-07-07 | 2018-10-09 | 中国农业科学院特产研究所 | A kind of method of Muskrat fragrant secretory cell Isolation and culture and application |
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