A kind of method for distilling of main active components of mulberry leaf and extract purposes thereof
Technical field
The present invention relates to the method that a kind of main active components of mulberry leaf extracts, and extract belongs to the phytochemistry and the modernization of Chinese medicine, diabetes control field preventing and treating diabetes and complication thereof, obesity and treating the application aspect the viral influenza.
Background technology
Diabetes be a kind of be cardinal symptom crowd's gradual endocrinopathy with the carbohydrate metabolism disturbance; Basic pathology physiology is reason such as insulin secretion function relative deficiency or absolute deficiency or Insulin receptor INSR dysfunction in the body; Cause metabolism disorder, comprise sugar, protein, fat, water and electrolyte etc.Generally show as hyperglycemia, glycosuria and hyperlipemia, ketone property acidemia can take place in serious case, even loop attenuation and stupor and threat to life.Diabetes are divided into four types, and wherein maximum type ii diabetes accounts for the 90-95% of diabetes sum.
Activity less, obesity and aging cause type ii diabetes to increase fast.It is reported that China's diabetics was 2.5%, 1996 year 3.21%, 1998 year 3.8% in 1993, rise to now more than 5% that growth rate is accelerated year by year.It is estimated that China's diabetics has reached 5,000 ten thousand people at present, and increase with the speed that increases 1,000,000 people every year newly.
Folium Mori are the Chinese medicine of the medicine-food two-purpose of Ministry of Public Health announcement, in morning among the people application are arranged, and being usually used in treating diabetes with other Chinese prescriptions is diabetes.Modern study has been illustrated the part mechanism of Folium Mori treatment diabetes.Polyhydroxylated alkaloid in the Folium Mori of at first finding by Japanese scholar have with saccharide like structure, but the activity of the disaccharide-hydrolysing enzymes-alpha-glucosidase in can the retroactive inhibition small intestinal slow down glucose and get into blood, reduce post-prandial glycemia.This Alkaloid has and the similar structure of the acarbose of present clinical use and the mechanism of action.Simultaneously, the research proof, alpha-glucosidase inhibitor also has obesity, antiviral therapeutical effect.In addition, there are lot of documents report flavone, polysaccharide and glycoprotein also to have hypoglycemic activity.Comprehensive existing achievement in research, the main active that proves Folium Mori is alkaloid, flavone, polysaccharide.
Publication number is that the Chinese patent of CN1338275A discloses " Chinese medicine extract, Preparation Method And The Use with alpha-glucosidase inhibitor activity ", and this patent has related to and from Chinese medicine Folium Mori, Ramulus Mori, Cortex Mori and Fructus Mori, prepared total alkaloids and be used for diabetes.Publication number is that the patent of CN1250246C discloses " medical composition and its use with alpha-glucoside inhibiting activity ", and this patent has related to the application of compositions in treatment diabetes, hyperlipidemia, antioxidation and antiaging agent of total alkaloids for preparing in Chinese medicine Folium Mori, Ramulus Mori, Cortex Mori and the Fructus Mori and flavone compound Quercetin, catechin and the tea polyphenols that adds.Publication number is that the patent of CN1850166A discloses Folium Mori effective ingredient polysaccharides of Folium Mori and Folium Mori alkaloid preparation of compositions method, and application number is that the patent of CN1981814A discloses " a kind of Folium Mori extract and application and method for preparing in the preparation diabetes medicament " and related to and from Folium Mori, extract total flavones and total alkaloids and mix the back with certain proportion and prepare to become and can prevent and treat the preparation of controlling diabetes and complication thereof.Publication number is that CN1742803A discloses a kind of mode that adopts macroporous adsorbent resin and cation exchange resin to use continuously and extracts flavone and alkaloid.Said method is the representational method for distilling of Folium Mori anti-diabetic composition in recent years.But there is certain deficiency in these methods: perhaps the utilization of Folium Mori extracts active ingredients is incomplete, and perhaps technical process is more loaded down with trivial details, can not separate all main effective ingredient through primary sorption.
To above-mentioned deficiency, we have invented a kind of efficient, economic method for distilling of Folium Mori, can pass through a leaching process; Being about to main active component alkaloid, flavone and the polysaccharide of Folium Mori all extracts; And only need adopt a kind of resin, and technology is simple, and cost is low.
Summary of the invention
The method that the present invention taked is to utilize the large hole cation exchanger resin with ion exchange and adsorption function to adsorb effective ingredient alkaloid and flavone as adsorbent; Can these two kinds of active component of disposable absorption, obtain polysaccharide through methanol or ethanol precipitation simultaneously.The present invention only need adopt a kind of resin, through a leaching process, uses the effect of two kinds of resins before just can reaching, and three types of main active alkaloids, flavone and polysaccharide of Folium Mori are all extracted.Regeneration of resin is also more time saving and energy saving.More need not adopt other complicacies, the higher technology of cost.Whole extraction and separation process does not use other inflammable and explosive organic solvent except methanol or ethanol, non-environmental-pollution, and safety non-toxic is beneficial to scale and extracts production.
For realizing above-mentioned purpose, the present invention has adopted following technical scheme:
1) extract: with Folium Mori is raw material, water or aqueous ethanol or methanol heating extraction;
2) recovery of alcohol: during with methanol or ethanol-water extraction, reclaim under reduced pressure methanol or ethanol obtain aqueous solution;
3) absorption: aqueous extract is through large porous strong acid type or weak-type absorption and exchanger resin, and active component is attracted on the resin.
4) eluting: make eluant with methanol or ethanol/water solution earlier, eluent concentrates and obtains flavones ingredient; Use the ammonia eluting then, concentrate and obtain the alkaloids composition;
5) resin of step 3) concentrates the back with methanol or ethanol precipitation through liquid, and precipitate is polysaccharide.
6), promptly obtain the effective ingredient product of Folium Mori with above-mentioned flavone, alkaloid and polysaccharide concentrated solution mixing after drying.
Extract the ethanol of usefulness or the concentration of methanol, its scope is between 5-95%, and consumption is 5-15 times.Heating-up temperature is between 30-100 ℃.
Used macroporous absorption and exchanger resin, the main strongly acidic cation-exchange with absorption and function of exchange that adopts like the D72 resin, also can use the weak-type macroporous adsorbent resin.Consumption is 0.5-5 a times of raw material weight.
The ethanol that eluting is used or the concentration of methanol, its scope are between 10-95%, and consumption is a 1-10 column volume.The concentration of ammonia is between 0.5%-15%, and consumption is a 1-10 column volume.
Drying means is vacuum drying, spray drying or lyophilization.
By the Folium Mori extract that method for preparing forms, main active is formed for the alkaloid, flavone and the polysaccharide that directly extract from Folium Mori, but not the analogous components that adds in addition.This Folium Mori extract does not add adjuvant promptly can directly be used as electuary; Perhaps add edible adjuvant; Like in starch, microcrystalline Cellulose, dextrin, sucrose, lactose, vitamin C, citric acid and the sodium sulfite any one or a few; Be prepared into dosage forms such as tablet, capsule or oral liquid, be used to prevent and treat diabetes and complication thereof, prevent and treat obesity and treatment viral influenza as medicine or functional food.
The invention has the advantages that extraction and preparation technique is simple, main active can once all obtain, safety, and easy operating does not need special installations such as HTHP, and cost is low.
The specific embodiment
Specify technical scheme of the present invention through embodiment below, but be not limited to cited example.
Embodiment 1
Folium Mori 10kg shreds, and extracts 2 times each 50L in 30 minutes with water boil.Extracting solution leaves standstill cool to room temperature, gets the supernatant through D72 resin column (column volume 10L), and the 20L ethanol elution is used in 10L washing back, uses ammonia/ethanol water (containing ethanol 70%) solution 30L eluting of 1% then.After the ammonia ethanol elution was recovered to no ammonia, the 20L ethanol elution before adding was recycled to the 3L volume, obtains fluid extract.
Resin boils off most of water to 3L through liquid, adds the 7L ethanol precipitation.The centrifugal supernatant of removing mixes with top fluid extract, and vacuum drying is pulverized, and obtains yellow powder 300g, yield 3%.
Get Folium Mori extract 100g, add starch 3000g, microcrystalline Cellulose 30g, with 50% alcohol granulation, tabletting gets tablet, and every 0.6g wherein contains extract 20mg, is equivalent to Folium Mori 0.7g.
Embodiment 2
Folium Mori 10kg shreds, and boils with 50% ethanol/water and extracts 2 times in 30 minutes, each 50L.After extracting solution concentrates and reclaims ethanol, add water 100L dilution, then through D72 resin column (column volume 10L).Handle according to embodiment 1 described mode, obtain yellow powder 400g, yield 4% is the Folium Mori extract of indication of the present invention.
The alpha-glucosidase of above-mentioned Folium Mori extract suppresses active test:
The alpha-glucosaccharase enzymatic solution is measured protein concentration then with the jejunum preparation of Wister rat.The Folium Mori extract water is mixed with the solution for standby of 0.05g/ml.Sample sets with jejunum homogenate 50 μ l, sucrose 50 μ l, be equipped with sample article 50 μ l (final concentration is 5 μ g/ μ l), PBS 50 μ l.Positive control is used acarbose, and final concentration is 0.5 μ g/ μ l.A blank control group is set in addition, and glycosidase wherein is by high-temperature inactivation.Other compositions of criterion group are identical, just do not add sample or glucobay (acarbose).Above-mentioned every group of parallel work 7 pipes were hatched 25 minutes for ready back 37 ℃.Hatch and finish the Na that the back adds 0.1mol/L
2CO
3200 μ l stopped reactions.Every pipe 50 μ l that take a sample measure glucogenic amount by the method that the blood sugar test test kit provides.
Suppression ratio=(1-(glucobay (acarbose) group concentration of glucose-blank control group concentration of glucose)/(criterion group concentration of glucose-blank control group concentration of glucose)) * 100%
According to top formula, the alpha-glucosaccharase enzyme inhibition rate that calculates embodiment 1 and 2 extracts is respectively 90% and 88%.