CN1095466C - Soya total flavone, its prepn. method and use thereof - Google Patents
Soya total flavone, its prepn. method and use thereof Download PDFInfo
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- CN1095466C CN1095466C CN99119344A CN99119344A CN1095466C CN 1095466 C CN1095466 C CN 1095466C CN 99119344 A CN99119344 A CN 99119344A CN 99119344 A CN99119344 A CN 99119344A CN 1095466 C CN1095466 C CN 1095466C
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Abstract
The present invention relates to soybean total flavonoid and an extraction technology and a novel medicine purpose thereof. The present invention discloses the extraction technology of soybean total flavonoid, and also discloses the application of soybean total flavonoid for preparing medicine for treating hyperlipidemia, medicine for reducing triglyceride in the liver, medicine for regulating metabolic disturbance and medicine for reducing lipid peroxide.
Description
The invention relates to the invention of soybean total flavones preparation method and new medicine use thereof, specifically is to extract total flavones from the soybean base, and the soybean total flavones is prepared into the protective foods and the medicine of treatment hyperlipidaemia.
Modern medicine study shows that (hyperlipoproteinemia HLP) is a kind of lipid metabolism disease to hyperlipidaemia, it is one of important risk factor that causes cardiovascular and cerebrovascular diseases and fatty liver, have only the scientific and rational strategy of employing, control blood fat effectively, could prevent the generation of relative disease.Natural drug treatment hyperlipidaemia has and is widely used, determined curative effect, and the advantage that toxic side effect is little, thereby be subjected to patient's favorable comment and clinical medicine expert's attention.
Medical book of all times is often classified soybean as and can be made human long-lived healthy primary food.Ming Dynasty's LI Shi-Zhen thinks that " food clothes soybean is made us the longue skin, beneficial color, fills out marrow, adds strength, and qi-restoratives can be eaten."; Energy " beneficial gas and taste in wide, dissipate-swelling is full, following large intestine foul smell ".
About 1,380 ten thousand tons of China's soybean annual production, account for the third place in the world, beans and bean product contain multiple anti-cancer, anticancer, prevention and treatment cardiovascular disorder material, comprising daizeol, soybean saponin, lectin, proteinase inhibitor, Mierocrystalline cellulose, calcium, selenium, phytic acid, folic acid etc., utilize abundant soybean resource production protective foods and medicine its realistic meaning and huge economic benefit to be arranged to improving China people body health quality.
The object of the invention provides soybean total flavone extracting method and new medicine use thereof, and this new purposes is the application of soybean total flavones in preparation control hyperlipidemia.
The present invention uses modern scientific method, extracts the soybean general flavone, and has studied the effect of its anti-lipid rising, find that the soybean general flavone can effectively suppress increasing of blood fat, the triglyceride that reduces in the liver is piled up, and the regulating lipid metabolism disorder reduces the generation of serum and liver lipid peroxidation thing.Its pharmacological action is suitable with import medicine promise weighing apparatus, but cheap, provides new medication foundation for preventing and treating hyperlipidemia, has finished the present invention.
Soybean total flavones of the present invention extracts acquisition from the soybean base, this method is to be raw material with the soybean base, and all flavones conjugates are converted into flavonoid glycoside, and extracts the material that is rich in daizeol from said soybean base raw material.
The method that reclaims daizeol from the soybean base of the present invention comprises: extract total flavones with ethanol from the soybean base, the total flavones here comprises compounds such as Genistein conjugate, Genistein glucoside, Genistein glucoside unit, Daidzin conjugate, Daidzin, daidzein; The flavones conjugate is converted into flavonoid glycoside; Use a kind of solvent recuperation daizeol then, and make higher healthcare products of daizeol content or medicine.
Concrete grammar is as follows:
With 40-90% extraction using alcohol soybean base twice, merge twice extracting solution after, recovered alcohol.The extracting solution of this method gained can extract the 70-95% of daizeol total amount.
Regulate the extracting solution of above-mentioned gained then with NaOH, making pH is 7-11, keep extracting solution 20-80 ℃ 1-12 hour, this method can be converted into flavonoid glycoside with 95% flavones conjugate.
In the extracting solution of above-mentioned gained, add an amount of volume propyl carbinol, extracted 1-12 hour for the first time; Collect the butanols part, add an amount of volume propyl carbinol again, extracted 1-12 hour for the second time; Collect the butanols part, merge secondary butanols extraction liquid, behind the recovery butanols, get the pale brown look solids of exsiccant, be the soybean total flavones.
Soybean total flavones of the present invention can be prepared into any drug form and any healthcare products thereof that are suitable for clinical use according to the preparation method of routine as active constituents of medicine, comprises tablet, capsule, granule, oral liquid, pill, paste etc.
Experimental example soybean total flavones is to the therapeutic action one of experimental rat hyperlipidaemia. experiment material 1 animal
The Wister male rat, 6-8 week, body weight 150-180g/, totally 60.Provide by animal housing of the academy of traditional Chinese medicine.2 medicines
1) soybean extraction: every milliliter contains total flavones 5mg; 2) positive control drug: the promise weighing apparatus, American Pie moral pharmaceutical factory produces, the 900mg/ sheet; 3) Sodium cholic acid: Haidian District microbiological culture media products factory is produced; 4) propylthiouracil: 5) reagent and medicine box: (1) triglyceride level (TG) test kit, produce numbering: 990319 by Beijing Chemical Plant's clinical reagent subsidiary factory; (2) total cholesterol is measured test kit (TC), is produced numbering: 990408 by Beijing Chemical Plant's clinical reagent subsidiary factory; (3) serum high-density LP test kit, clinical reagent subsidiary factory in Beijing Chemical Plant's produces, numbering: 990522.Two. experimental technique 1, preparation high lipid food: 1% cholesterol, 10% lard, 0.2% Thyreostat I, 0.5% cholic acid, 5% sucrose, 83.3% basal feed.Each thing mixing adds water and stirs, and the stripping and slicing of flattening is put into drying baker and dried, about 100 ℃ of temperature.2, animal grouping; feed and administration: get 60 of rats; be divided into normal group at random; model group; permanent group of positive drug promise (each 10) and total flavones group (30) normal group give normal diet and feed every days; all the other are respectively organized and give high fat every day and raise (20g/ only), resupply capacity arm's length basis feed after treating all to eat up.Every of positive controls every day, the treatment component was big to promise weighing apparatus 16.76mg, in, every of little three dosage group every day give respectively general flavone 2ml/100g body weight, 1ml/100g body weight and 0.5ml/100g body weight, more than equal gastric infusions, once a day.Normal group, model group all give physiological saline, every day every 2ml/100g body weight.All animals freely drink water, and raise altogether 15 or 30.3, collection of specimens: all rats stop high lipid food the day before yesterday of drawing materials, and fasting 12 hours.Broken end is got blood and liver, preparation serum and liver homogenate, be stored in-20 ℃ standby.4, observation item and measuring method:
(LDL-C HDL-C), adopts enzyme process, measures with 724 type visible spectrophotometers, and test kit is produced by Beijing Chemical Plant's clinical reagent subsidiary factory for cholesterol (TC), triglyceride level (TG), high and low density lipoprotein cholesterol; Method is adopted in the extraction of TC, TG in the hepatic tissue, measures and uses enzyme process; The determining the protein quantity phenol reagent process; MDA improvement Yagi spark gap state husband method survey mda (malondiadehycle, MDA).5, statistical procedures: adopt t check three. the result:
This experiment causes hyperlipemia model with the high lipid food rat of feeding, and behind the feed high lipid food, high fat rat is compared body weight and rises slowly with normal group, but model group and treatment group body weight and no significant difference.In two weeks after high lipid food is fed, model group TC is existing obviously to raise, but TG is still normal, and around feeding time increased to, serum TG, two of TC all were higher than normally, and two groups of data compare significance (P＜0.005) (table 1).Around the historical facts or anecdotes time of testing is decided to be.After around animal is fed, the big or middle dosage group of soybean total flavones rat blood serum TG.TC obviously reduces than model group, difference significance (P＜0.05); Positive control drug (promise weighing apparatus) group Serum TC also obviously reduces than model group, difference significance (P＜0.05), but not obvious to the TG effect.Soybean total flavones and promise weighing apparatus group compare, and the big or middle dosage of Chinese medicine reduces the TG effect and is better than positive control drug, reduces TC difference meaningless (P＞0.05).For further observing triglyceride level heap in the hepatic tissue, the TG in the hepatic tissue has also been measured in this experiment.The result shows, in the soybean general flavone, small dose group can obviously reduce its level, with model group significance (P＜0.05) (table 2) relatively.
The big-and-middle dosage group of the soybean total flavones serum high-density LP that can obviously raise is with model group data significance (P＜0.05) relatively, permanent group of promise and soybean total flavones small dose group high density lipoprotein increasing, but not statistically significant.The model group serum low-density LP has significantly than normal group and increases, and two groups of data are significance (P＜0.001) relatively, and permanent group of promise and the big or middle dosage group of soybean total flavones reduce low-density lipoprotein extremely significantly meaning (P＜0.001) (table 3).The demonstration of serum MDA determination data, the permanent group of promise, heavy dose of group, middle dosage group all can obviously reduce the MDA level, with the relatively more equal significance (P＜0.005) of model group; The demonstration of hepatic tissue MDA determination data, the permanent group of promise, heavy dose of group all can obviously reduce the MDA level, with the relatively more equal significance (P＜0.05) (table 4) of model group.
Table 1 soybean total flavones is to the influence of rat blood serum triglyceride and cholesterol (the group mouse serum TG inhibiting rate serum TC inhibiting rate of X ± SE)
(mmol/l (%) is (%) (mmol/1) for number
) dosage group 9 2.90 ± 0.3# 34.91 ± 6.7 24.42 ± 3.1# 29.03 ± 9.1 small dose group 10 4.95 ± 0.5 11.27 ± 12.1 29.11 ± 1.7#, 15.41 ± 5.07* compares P＜0.05 with normal group among the permanent group of normal group 8 2.77 ± 0.4-2.59 ± 0.35-model group 8 4.29 ± 0.2*-34.41 ± 3.1*-promise 9 4.63 ± 0.5#-4.06 ± 18.2 20.53 ± 2.9#, 40.34 ± 8.6 heavy dose of group 10 2.86 ± 0.3# 35.78 ± 6.8 19.59 ± 1.1# 43.78 ± 3.2; # and model group be p＜0.05 relatively.
(X ± SE) the group mouse is counted HDL-C rate of increase LDL-C inhibiting rate to table 2 soybean total flavones to the influence of rat blood serum lipoprotein
(mmol/l) (%) (mmol/l) (%) among the permanent group of normal group 8 0.86 ± 0.1-1.75 ± 0.4-model group 8 0.65 ± 0.1*-26.67 ± 3.8*-promise 9 0.97 ± 0.1# 47.71 ± 15.9 16.78 ± 2.4#, 33.52 ± 12.9 heavy dose of group 10 1.48 ± 0.2# 106.1 ± 40.6 0.76 ± 0.8# 32.20 ± 7.6 dosage group 9 1.20 ± 0.3# 63.57 ± 61.5 19.79 ± 3.3# 17.90 ± 8.3 small dose group, 10 1.15 ± 0.1#, 51.38 ± 26.4 23.85 ± 2.6 6.17 ± 17.0* compare P＜0.05 with normal group; # and model group be p＜0.05. relatively
(X ± SE) group N TG nmol/g inhibiting rate % TC nmol/g inhibiting rate % normal group 8 0.04 ± 0.0-7.09 ± 4.3-model group 8 19.66 ± 5.6*-82.65 ± 17.0*-positive drug group 9 8.40 ± 3.1# 57.25 ± 14.7 61.45 ± 7.6# 25.65 ± 9.7 heavy dose of group 10 10.51 ± 2.8# 46.57 ± 14.4 46.09 ± 9.9#, 36.02 ± 12.1 intermediate amounts are organized 9 3.28 ± 1.5# 83.33 ± 7.5 64.36 ± 10.0#, 22.13 ± 12.1 small dose group, 10 6.52 ± 2.43#, 66.86 ± 12.06 48.74 ± 8.0#, 41.03 ± 9.7* and normal group comparison P＜0.05 to table 3 soybean general flavone on the impact of liver tissues of rats triglyceride and cholesterol; # and model group be p＜0.05 relatively.
Table 4 soybean general flavone is on the impact of rat blood serum and hepatic tissue MDA (dosage group 9 48.36 ± 16.7# 46.33 ± 5.7 0.11 ± 0.03 20.24 ± 8.5 small dose group 10 75.18 ± 42.99 17.27 ± 15.5 0.20 ± 0.05-45.4 ± 12.2* and normal group comparison P＜0.05 among group N serum MDA nmol/ml inhibiting rate % hepatic tissue MDAnmol/mg inhibiting rate % normal group 8 30.16 ± 12.39-0.02 ± 0.01-model group 8 91.12 ± 15.65*-0.13 ± 0.05*-positive drug group 9 38.51 ± 18.29# 57.00 ± 6.6 0.09 ± 0.02# 29.11 ± 5.5 heavy dose of group 10 30.89 ± 11.78# 65.26 ± 4.1 0.08 ± 0.02# 40.82 ± 5.2 of X ± SE); # and model group be p＜0.05. four relatively. discuss:
This result of study shows the effect that extract of soybean (general flavone) has obvious antagonism experimental hyperlipidemia, fat to accumulate in liver.
Duplicate the important method that the animal hyperlipemia model is the disease of studying atherosclerotic.Though rat is to tolerance of high fat diet, inexpensive, the source is wide, and it is convenient to get blood, and blood volume is many than mouse, has easily to feed, and easily irritates stomach, and resistibility is strong, and feeding habits and people are close.After adding cholate and Thyreostat I, can form promising result.Therefore making high blood lipid model with rat still is subject to people's attention, bibliographical information is arranged recently, if high lipid food adds Vit D3, all around, except that blood fat and cholesterol be higher than normal, can see the rat aorta intimal thickening, middle film smooth muscle cell proliferation, there is obvious calcified plaque in some middle level, position, tentatively causes Atherosclerosis Model.
This experiment adopts high lipid food to feed rat after 2 weeks, and cholesterolemia obviously raises, but the blood triglyceride rising is not obvious.Feed the rat high lipid food after 4 weeks, compare with normal rat, TC raises nearly 16 times, and TG raises about 2 times, so experimental period was decided to be for 4 weeks.
This experiment shows that the soybean general flavone has obvious adjusting improvement effect to hyperlipidemia rats, can significantly reduce Serum TC, TG, and inhibiting rate is respectively 29.03 ± 9.1%--40.34 ± 8.6%.Significantly reduce rat blood serum LDL-C simultaneously, rising HDL-C level reduces TG level in the hepatic tissue, to reach the effect of regulating lipid metabolism disorders.Think that traditionally HDL is the protection factor of coronary heart disease, has study of anti-atherogenic effect.Rising HDL level can reduce the coronary heart disease incidence separately.HDL has the effect in control agent inner tissue cholesterol storehouse, and it can get off the cholesterol absorption on surrounding tissue and the arterial wall, and it is transported to metabolism in the liver, discharges from bile.Therefore HDL can prevent that lipid from depositing at arterial wall, prevents that fat from piling up in liver, has anti arteriosclerosis and lipotropic effect.
Unrighted acid in the biomembrane (FFA) can with oxygen radical generation peroxidatic reaction of lipid, form MDA (such as MDA etc.) and new oxygen radical, cause cellular damage and dysbolism.Liver is the major organs of lipid and lipoprotein metabolism, and liver cell is impaired, and lipid and lipoprotein metabolism disorder can cause that blood fat raises.Liver MDA has reflected the hepatic tissue level of lipid peroxidation, has indirectly reflected the degree of hepatocellular injury and dysbolism.And blood MDA content has reflected body tissue's level of lipid peroxidation, indirectly the degree of corresponding whole lipidosis.This experiment shows, MDA content increase in model group liver tissues of rats and the blood, and there is the pathogenesis of lipid peroxidation really in the prompting hyperlipemia model.Total flavones group rat MDA content descends and points out the antioxygenation of total flavones, the antagonism peroxidatic reaction of lipid, thus protected hepatocellular eubolism function.
The positive control drug promise weighing apparatus of this experiment is the medicine of commonly using clinically for the clofibrate derivative.This experiment shows that the soybean general flavone truly has good effect for reducing blood fat, and it reduces TC, TG., MDA, and rising HDL and reduction LDL effect and equal promise weighing apparatus are quite.But the promise weighing apparatus has side effect and value costlinesses such as tangible digestive tract reaction.
Embodiment 1
Doubly measure 40-90% extraction using alcohol soybean base 2-4 hour of volume with 4-15, again with 4-15 times of volume 40-90% extraction using alcohol soybean base 2-4 hour.After merging twice extract, recovered alcohol.This method can be extracted the 70-95% of daizeol total amount.
Embodiment 2
With embodiment 1 gained extracting solution, regulate with NaOH that to make pH be 7-11, keep extracting solution 20-80 ℃ 1-12 hour, this method can be converted into flavonoid glycoside with 95% flavones conjugate.
Embodiment 3
With embodiment 2 gained extracts, add the n-butanol that 1-5 doubly measures volume, extract 1-12 hour for the first time; Collect the butanols part, add again the n-butanol that 1-5 doubly measures volume, extract 1-12 hour for the second time; Collect the butanols part, merge secondary butanols extract, behind the recovery butanols, get dry brown color solids, be the soybean general flavone.
Embodiment 4
Get soybean total flavones 1000 grams, add medical starch 500 grams, mix, the ethanol granulation is granulated, and compressing tablet makes tablet.
Embodiment 5
Get soybean general flavone 1000 grams, add medical starch 500 grams, mix, encapsulated, and get final product.
Claims (7)
1, the extracting method of soybean total flavones is characterized in that this method for 40-90% extraction using alcohol soybean base twice, merge twice extracting solution after, recovered alcohol is promptly.
2, according to the extracting method of claim 1, it is characterized in that further regulating in the described extracting method extracting solution of above-mentioned gained with NaOH, making pH is 7-11, keep extracting solution 20-80 ℃ 1-12 hour.
3, according to the extracting method of claim 1, it is characterized in that in the described extracting method that further adding 1-5 in the extract of above-mentioned gained doubly measures n-butanol, extracts 1-12 hour for the first time; Collect the butanols part, add again 1-5 and doubly measure n-butanol, extract 1-12 hour for the second time; Collect the butanols part, merge secondary alcohol extract, behind the recovery butanols, get dry brown color solids.
4, the application of soybean general flavone in the medicine of preparation treatment hyperlipidemia.
5,, it is characterized in that said application is the application of soybean total flavones in the medicine of preparation adjusting lipid metabolism disorders according to the purposes of claim 4.
6,, it is characterized in that said application is the application of soybean total flavones in the medicine of preparation reduction lipid peroxide according to the purposes of claim 4.
7,, it is characterized in that said application is the soybean total flavones and reduces application in the medicine of triglyceride and cholesterol in the blood, in the liver in preparation according to the purposes of claim 4.
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| CN1211583A (en) * | 1997-06-17 | 1999-03-24 | 希尔斯硅股份有限公司 | Polyorganosiloxanes with dialdoxyorganosiloxy end groups |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1211583A (en) * | 1997-06-17 | 1999-03-24 | 希尔斯硅股份有限公司 | Polyorganosiloxanes with dialdoxyorganosiloxy end groups |
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