CN103665175A - Fructan extract, and preparation method and application thereof - Google Patents
Fructan extract, and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a fructan extract and a preparation method thereof. The preparation method comprises the following steps: extracting Sichuan ophiopogon root tuber with ethanol to remove impurities, extracting with water, carrying out ethanol fractional precipitation, and drying to obtain a fructan extract (the molecular weight is 2500-8000). The method for preparing the fructan extract has the advantages of short time consumption and low cost, and can perform abundant preparation and adapt to industrial production. Besides, the invention also discloses application of the fructan extract in preparing medicines, foods or health products for losing weight. The inspection on functions of the fructan extract by a diet-induced fat mouse model proves that the fructan extract has an obvious action of losing weight.
Description
Technical field
The present invention relates to a kind of Polylevulosan extract, relate in addition preparation method and the purposes of this Polylevulosan extract.
Background technology
Polylevulosan (fructan) is that the ubiquitous class of occurring in nature mainly be take the carbohydrate general name that fructosyl fructose is formed by connecting as glycosidic link; In each Polylevulosan molecule, all contain 1 molecule glucose, its molecular end does not have reductibility.According to the difference of the polymerization degree between the difference of fructosyl transfer link position and species, Polylevulosan structure is quite complicated.Because Polylevulosan does not substantially decompose in the process of human oral cavity, stomach and small intestine, not digested absorption.Therefore it has good fat-reducing, regulates blood sugar, reducing blood-fat, prevention cardio-cerebral diseases; Improve intestinal function, preventing constipation, diarrhoea; Promote the absorption of mineral substance and synthesize, the cancers such as all right prevent colon cancer.Thereby caused that current medical science, food, biotechnology field are to its interest widely.
The energy metabolism balance of human body is controlled by a set of meticulous regulation mechanism, thereby keeps the stable of body weight.Under the effect of various heredity or environmental factors, human body energy metabolism regulates and breaks down, and causes the picked-up of energy more than consumption, and unnecessary energy is just converted into fat in vivo, forms fat.Obesity is just becoming an epidemic disease more and more attracting people's attention in worldwide, and along with the development of China's economic construction and the raising of living standards of the people, overweight or fat sickness rate certainly has significantly and increases.Be accompanied by increasing of fat morbidity, other Chronic Non-Communicable Diseases relevant to obesity also will be obvious increase trend as the morbidity of diabetes, metabolism syndrome, hypertension, cardiovascular and cerebrovascular diseases, somnopathy, asthma, tumour etc.
Research finds that Polylevulosan can not digestedly absorb in oral cavity, stomach and small intestine, can only be degraded by some probiotics of enteron aisle (bifidus bacillus etc.) fully fermenting, produce short chain fatty acid (acetic acid, propionic acid and butyric acid) and lactic acid, produce caloric value and be less than 1.5Kcal/g.In addition, Polylevulosan also easily under one's belt water-swelling form high viscosity colloid, the emptying time that makes people be difficult for producing hunger sensation and can extend stomach, thereby minimizing food intake, reduce food in the small intestine residence time, can also be in small intestine form mixture with the material such as protein, fat, suppress these material absorbings, reach fat-reducing object.Yet the preparation method who prepares Polylevulosan at present from the tuber of dwarf lilyturf exists cost high, consuming time many more, cannot adapt to the defects such as suitability for industrialized production, cannot prepare in a large number.Meanwhile, the also rarely seen report of research of preparing the Polylevulosan extract activity of certain molecular weight scope in prior art for the river in the specific place of production in the tuber of dwarf lilyturf.
Summary of the invention
Above-mentioned deficiency for prior art, one of technical problem to be solved by this invention be propose a kind of consuming time short, cost is low, can prepare in a large number and adapt to the preparation method who produces extraction Polylevulosan extract the tuber of dwarf lilyturf (the river tuber of dwarf lilyturf) from Sichuan of suitability for industrialized production.
Two of technical problem to be solved by this invention is to provide the Polylevulosan extract that adopts aforesaid method to prepare.
Three of the technical problem to be solved in the present invention is to provide this Polylevulosan extract and has the application in medicine, food or the healthcare products of weight losing function in preparation.
In order to solve the problems of the technologies described above, the process for producing method of a kind of Polylevulosan extract that the present invention proposes, comprises the following steps:
Get dry river Tubers of Ophiopogon japonicus, with 2-8 times of weight 70%-100% alcohol reflux 1-4 time, extracting solution is abandoned it; The dregs of a decoction are carried 1-3 time with 6-14 times of water gaging, merge water extraction liquid, and water extraction liquid is evaporated to 0.8-1.2g/ml; Water extracting liquid is added to 1-5 times of volume 80%-100% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.
Preparation method is preferably: get dry river Tubers of Ophiopogon japonicus, with 4 times of weight, 90% alcohol reflux 2 times, extracting solution is abandoned it; The dregs of a decoction are carried 2 times with 10 times of water gagings, merge water extraction liquid, and water extraction liquid is evaporated to 1.0g/ml; Water extracting liquid adds 3 times of volumes, 95% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.
The bulk drug that the present invention adopts is the river tuber of dwarf lilyturf.River has another name called city, Fu tuber of dwarf lilyturf the tuber of dwarf lilyturf, the continuous tuber of dwarf lilyturf, originates in Santai County, Sichuan Province.Producing region mainly concentrates on nine small towns of basin, Fujiang River, Santai County centered by the town of garden.River tuber of dwarf lilyturf is one of Chinese famous and precious traditional Chinese medicine material, and the plantation of existing more than 500 year is historical, and its feature: piece root is a line and wears pearl is spindle-like, the blunt point in two after dry, middle part grossness, microbend, surperficial oyster white, glossy, matter is harder, micro-perfume (or spice), and taste is sweet denseer, and that chews is clamminess.It has Yin nourishing and lung moistening, clearing heat and relieving fidgetness, reinforcing stomach reg fluid, the effects such as cooling blood for hemostasis.River tuber of dwarf lilyturf is generally planting in annual April, after cultivation in by the end of March, the 2nd to mid-April fine weather gather.River tuber of dwarf lilyturf is planted the compound useful to human body containing amino acid, three kinds of above oligosaccharides and more than one neutral polysaccharides, nine kinds of above homoisoflavone compounds and glycoside, seven kinds of above steroid saponins and vitamin A, B, C, D etc. more than 30.
In addition, the present invention also provides the Polylevulosan extract that adopts aforesaid method to prepare, and the molecular weight ranges of this Polylevulosan extract is 2500 ~ 8000.This Polylevulosan extract is carried out to HPLC-ELSD analysis, obtain HPLC-ELSD collection of illustrative plates as shown in Figure 6, the specific analytical method that this HPLC-ELSD analyzes is as follows:
Chromatographic column: Shodex Sugar-KS802; Column temperature: 25 ℃; Moving phase: distilled water; Flow velocity: 0.5mL/min; Detector: ELSD(gas flow rate: 1.5SLM; Vaporization temperature: 110 ℃; 90 ℃ of atomization temperatures); Sample size: 20 μ l.
Adopt the inventive method to prepare Polylevulosan extract, consuming time short, cost is low, can prepare in a large number and adapt to suitability for industrialized production.
In addition, the present invention also provides this Polylevulosan extract to provide this Polylevulosan extract to have the application in medicine, food or the healthcare products of weight losing function in preparation.
As foodstuff additive, the healthcare products with weight losing function, Polylevulosan extract of the present invention can join in the diet product such as tap water, food, beverage, drinks, seasonings, milk-product, food oils, flour-made food, the effect of performance fat-reducing.
As healthcare products or the medicine with weight losing function, Polylevulosan extract of the present invention can be made into various forms of oral preparations and comprises tablet, capsule, granule or oral liquid etc.The Polylevulosan extract that described oral preparations is preferably 10%-95% by weight percent and pharmaceutical excipient or the additives of 90%-5% form.
The present invention has following effect: pharmacodynamic study shows that Polylevulosan extract of the present invention can be in the situation that not affecting food ration, activity, by increasing body tissue oxygen consumption and suppressing glycolipid composition, absorb, when maintaining normal lean tissue mass, effectively fall lower animal fat content, there is obvious optimum effect of weight reducing.
Accompanying drawing explanation
Fig. 1 respectively organizes the mean body weight growth results schematic diagram of Polylevulosan extract to meals inducing obesity mouse in test example 1 of the present invention, n=12 wherein, Mean ± SD.
Fig. 2 respectively organizes the round the clock OCR result schematic diagram of Polylevulosan extract to meals inducing obesity mouse in test example 1 of the present invention, n=6 wherein, Mean ± SD;
Fig. 3 respectively organizes the breathing entropy result schematic diagram of Polylevulosan extract to meals inducing obesity mouse in test example 1 of the present invention, n=6 wherein, Mean ± SD;
Fig. 4 respectively organizes the round the clock autonomic activities amount result schematic diagram of Polylevulosan extract to meals inducing obesity mouse in test example 1 of the present invention, n=6 wherein, Mean ± SD.
Fig. 5 respectively organizes Polylevulosan extract body composition to meals inducing obesity mouse is formed to result schematic diagram in test example 1 of the present invention, n=6 wherein, Mean ± SD.
Fig. 6 is the HPLC-ELSD collection of illustrative plates of Polylevulosan extract of the present invention.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
The preparation of embodiment 1(Polylevulosan extract)
Get dry Sichuan and produce the piece root 20kg tuber of dwarf lilyturf (the river tuber of dwarf lilyturf), with 4 times of weight, 90% alcohol reflux 2 times, extracting solution is abandoned it; The dregs of a decoction are carried 2 times with 10 times of water gagings, merge water extraction liquid, and water extraction liquid is evaporated to 1.0g/ml; Water extracting liquid adds 3 times of volumes, 95% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.Obtain Polylevulosan extract 4.62kg, yield is: 15.4%.Structural Identification is Polylevulosan.
The preparation of embodiment 2(Polylevulosan extract)
Get dry Sichuan and produce the piece root 30kg tuber of dwarf lilyturf (the river tuber of dwarf lilyturf), with 8 times of weight, 70% alcohol reflux 3 times, extracting solution is abandoned it; The dregs of a decoction are carried 1 time with 14 times of water gagings, merge water extraction liquid, and water extraction liquid is evaporated to 0.8g/ml; Water extracting liquid adds 5 times of volumes, 80% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.Obtain Polylevulosan extract 3.15kg, yield is: 10.5%.Structural Identification is Polylevulosan.
The preparation of embodiment 3(Polylevulosan extract)
Get dry Sichuan and produce the piece root 30kg tuber of dwarf lilyturf (the river tuber of dwarf lilyturf), with 2 times of weight, 100% alcohol reflux 1 time, extracting solution is abandoned it; The dregs of a decoction are carried 4 times with 6 times of water gagings, merge water extraction liquid, and water extraction liquid is evaporated to 1.2g/ml; Water extracting liquid adds 1 times of volume, 100% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.Obtain Polylevulosan extract 3.03kg, yield is: 10.1%.Structural Identification is Polylevulosan.
From embodiment 1-3, the yield of the Polylevulosan extract of embodiment 1 is the highest, so embodiment 1 is preferred embodiment.
Embodiment 4 is containing the preparation of the tablet of Polylevulosan extract
Prescription (1000):
Polylevulosan extract 780g(makes by embodiment 1)
Micropowder silica gel 2g
Magnesium Stearate 2g
By recipe quantity, add Magnesium Stearate to mix Polylevulosan extract, granulation, adds micropowder silica gel, mixes compressing tablet, 1000 (every containing Polylevulosan extract 0.78g).Adult recommends oral consumption 1-4 piece/times, every day 1-3 time.
Embodiment 5 is containing the preparation of the capsule of Polylevulosan extract
Prescription (1000):
Polylevulosan extract 180g(makes by embodiment 1)
Microcrystalline Cellulose 180g
Magnesium Stearate 4g
By recipe quantity, add Microcrystalline Cellulose, Magnesium Stearate to mix Polylevulosan extract, granulation, encapsulated, 1000 (every containing Polylevulosan extract 0.2g).Adult recommends oral consumption 1-6 tablet/time, every day 2-4 time.
Embodiment 6 is containing the preparation of the granule of Polylevulosan extract
Prescription (1000 bags):
By recipe quantity, add Microcrystalline Cellulose, micropowder silica gel, stearic acid, lactose to mix Polylevulosan extract, granulation, pack, 1000 bags (every bag containing Polylevulosan extract 1.8g).The oral consumption 1-2 bag of adult's recommendation/time, every day 1-3 time.
Embodiment 7 is containing the preparation of the beverage of Polylevulosan extract
Formula (1000 bottles):
By formula ratio, Polylevulosan extract is added in 50000mL beverage, make l000 bottle.
Embodiment 8 is containing the preparation of the biscuit of Polylevulosan extract
Formula (100 box):
By formula ratio, saleratus is sprinkling upon in warm water, stirs and to make it to dissolve, add low fine powder and stir together, then add other raw materials (Polylevulosan extract, butter, egg, milk powder) and stir evenly, be kneaded into smooth dough.Thinly slice, put warm place (38 ℃ of left and right) fermentation 10 minutes, roasting training, mounted box.
Below by test example, with pharmacological activity test and the result of Polylevulosan extract of the present invention, further illustrate the present invention.
Test example 1: the impact of Polylevulosan extract on meals inducing obesity (DIO) weight of mice and food ration.
1, animal
Select the C57BL/6J mouse in 4-5 week, male, 100.Raising condition: 22 ± 1 ℃ of temperature, 40%-60% relative humidity, bright 12 hours of controlled light, dark 12 hours, animal freely ingested and drinks water.
2, medicine and reagent
Polylevulosan extract of the present invention (molecular weight 2500 ~ 8000); Normal diet (purchased from Shanghai Slac Experimental Animal Co., Ltd.); High lipid food (60% fatty amount, D12492, purchased from U.S. ResearchDiet company); Sodium chloride injection (purchased from Shijiazhuang Siyao Co., Ltd).
3, experimental technique
3.1 meals inducing obesity mouse modelings
Blank mouse feeds with normal diet.Other C57 mouse all feed with high lipid food, freely enter to take the photograph water food.When mouse is fed to 8 weeks, according to Mouse Weight, by mice group, make respectively to organize body weight consistent, 12 every group.
3.2 laboratory animal echelon design
Table 1
3.3 experiment administrations
The oral distilled water that gives of blank group and model group mouse, the not oral Polylevulosan extract that gives respective concentration of Polylevulosan extract components, once a day, continuous 91 days.
3.4 food ration
Measure and once respectively organize mouse food ration, record weekly.
3.5 body weight increased values
Measure and once respectively organize Mouse Weight weekly, calculate body weight increased value.
3.6 oxygen consumption and the monitoring of autonomic activities amount
Choose each 6 of the solvent control group for the treatment of experiment and Polylevulosan extract high dose group experiment mices, be placed in Oxymas/CLAMS laboratory animal comprehensive monitor system (Columbus Instrument, Columbus, OH), gather 24 hours continuous OCR (VO
2, vol/mas/time) with carbon dioxide generating rate (VCO
2, vol/mas/time) data, further obtain 24 hours and breathe entropy (RER, VCO2/VO
2).RER tolerance interval is (0.707 to 1.0).Meanwhile, apply this system acquisition X/Y/Z axle infrared beam and interrupt amounting to, with the amount of autonomic activities round the clock of monitoring experiment mouse.
3.7 body fat measures of spread
Utilize NMR (Nuclear Magnetic Resonance) spectrum (Nuclear magnetic resonance, NMR) technology, measure the solvent control group for the treatment of experiment and the body composition composition of Polylevulosan extract high dose group experiment mice (each 6), i.e. lipid content, muscle tissue content and moisture.
3.8 statistical method
Application Students ' s test carries out statistical analysis to paired data or many groups data, and p<0.05 represents significant difference, and p<0.01 represents highly significant difference.Use Origin 8.0 softwares (OriginLabCorporation, Northampton, MA, USA), experimental data is carried out to statistical analysis and fitting of a curve.
4, result
4.1 body weight and food ration
Each initial mean body weight of organizing experiment mice comparatively approaches (p>0.05).As shown in Figure 1, after the medicine for treatment of 13 weeks, the mean body weight rate of increase of normal mouse is minimum, is only 56.8% of obese model mouse.And the mean body weight rate of increase of Polylevulosan extract high dose group animal is a little more than Normal group, but significantly lower than obese model mouse group, and be only its 69.7%(p<0.01); In addition, in Polylevulosan extract low dose group and Polylevulosan extract, the mean body weight rate of increase of dosage group is also very low, is respectively 84.3% and 78.2% of obese model mouse.These results show, Polylevulosan extract of the present invention can significantly reduce abdominal obesity Mouse Weight and increase, and has obvious antiobesity action; And this antiobesity action is more obvious along with the increase of Polylevulosan dosage.
Table 2 respectively organize medicine on meals inducing obesity mouse food ration (gram) impact (n=12, Mean ± SD)
Food ration result is as shown in table 2, during whole administration, compares with model control group, and do not have notable difference between food ration the average every day that gives three groups of mouse of Polylevulosan extract; But with respect to the blank group that gives normal diet, the dosage of the Polylevulosan extract mouse of three groups of various dose group but obviously reduces (p < 0.05).This explanation Polylevulosan extract can obviously reduce the food ration of obesity mice.
4.2 oxygen consumption round the clock and autonomic activities amount
Treat after 13 weeks, solvent control group and Polylevulosan extract high dose group are chosen respectively 6 mouse, carry out oxygen consumption in 24 hours and autonomic activities amount monitoring round the clock.As shown in Figure 2, the oxygen consumption round the clock of Polylevulosan extract high dose group mouse is all significantly higher than obese model control group (p<0.01).Compare with obesity mice, the breathing entropy of taking the mouse of Polylevulosan extract high dose group obviously reduces (p<0.05, Fig. 3), and trends towards steatolysis (RER value trend 0.7), and autonomic activities amount is without significantly increasing (Fig. 4).
4.3 body fat distributes
As shown in Figure 5, treat after 13 weeks, the average lipid content of Polylevulosan extract high dose group mouse is lower than obese model mouse; Fatty tissue content in two groups is respectively 15.29 ± 3.13g, 6.52 ± 1.70g, has significant difference (p<0.01); Muscle tissue content in both is respectively 25.15 ± 2.15g, 23.86 ± 0.51g,, there was no significant difference (p>0.05); Compare with obese model control group, the average moisture content of Polylevulosan extract high dose group obviously reduces; Moisture content in two groups is respectively 4.00 ± 0.50g, 2.83 ± 0.16g.
In sum, Polylevulosan extract of the present invention can be substantially in the situation that not affecting mouse food ration, activity, by increasing body tissue oxygen consumption and suppressing glycolipid composition, absorb, when maintaining normal lean tissue mass, effectively fall lower animal fat content, there is obvious optimum effect of weight reducing.Above result shows, Polylevulosan extract of the present invention has significant antiobesity action.
Claims (6)
1. a preparation method for Polylevulosan extract, is characterized in that, the method comprises the following steps:
Get dry river Tubers of Ophiopogon japonicus, with 2-8 times of weight 70%-100% alcohol reflux 1-4 time, extracting solution is abandoned it; The dregs of a decoction are carried 1-3 time with 6-14 times of water gaging, merge water extraction liquid, and water extraction liquid is evaporated to 0.8-1.2g/ml; Water extracting liquid is added to 1-5 times of volume 80%-100% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.
2. preparation method as claimed in claim 1, is characterized in that, the method comprises the following steps: to get dry river Tubers of Ophiopogon japonicus, and with 4 times of weight, 90% alcohol reflux 2 times, extracting solution is abandoned it; The dregs of a decoction are carried 2 times with 10 times of water gagings, merge water extraction liquid, and water extraction liquid is evaporated to 1.0g/ml; Water extracting liquid adds 3 times of volumes, 95% ethanol precipitation, standing, collecting precipitation; The supernatant liquor of gained is repeated to alcohol precipitation 1 time, merge the precipitation of 2 alcohol precipitation gained, lyophilize, is molecular weight at the Polylevulosan extract of 2500 ~ 8000 scopes.
3. adopt the Polylevulosan extract that method prepares described in claim 1 or 2, it is characterized in that, the molecular weight ranges of this Polylevulosan extract is 2500 ~ 8000.
4. a Polylevulosan extract as claimed in claim 3 has the application in medicine, food or the healthcare products of weight losing function in preparation.
5. application as claimed in claim 4, is characterized in that, described medicine or healthcare products are oral preparations.
6. application as claimed in claim 5, is characterized in that, in described oral preparations, the weight percent of Polylevulosan extract is 10%-95%, and surplus is pharmaceutical excipient or additives.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103655604A (en) * | 2012-09-12 | 2014-03-26 | 上海张江中药现代制剂技术工程研究中心 | Application of fructan extract in preparing medicines, foods or health products for treating fatty liver |
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| CN102336931A (en) * | 2011-10-25 | 2012-02-01 | 温州医学院 | Ophiopogonpolysaccharide, and preparation method and application thereof |
| CN102406833A (en) * | 2010-12-01 | 2012-04-11 | 成都中医药大学 | Comprehensive extraction and utilization technology for creeping liriope |
| CN101935359B (en) * | 2010-09-10 | 2012-05-30 | 中国食品药品检定研究院 | Liriope muscari baily polysaccharide and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101935359B (en) * | 2010-09-10 | 2012-05-30 | 中国食品药品检定研究院 | Liriope muscari baily polysaccharide and preparation method thereof |
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| CN102336931A (en) * | 2011-10-25 | 2012-02-01 | 温州医学院 | Ophiopogonpolysaccharide, and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103655604A (en) * | 2012-09-12 | 2014-03-26 | 上海张江中药现代制剂技术工程研究中心 | Application of fructan extract in preparing medicines, foods or health products for treating fatty liver |
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