CN103483459A - Ultrasound-assisted extraction process of fritillariae thunbergii polysaccharide and health-care product containing fritillariae thunbergii polysaccharide - Google Patents
Ultrasound-assisted extraction process of fritillariae thunbergii polysaccharide and health-care product containing fritillariae thunbergii polysaccharide Download PDFInfo
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- CN103483459A CN103483459A CN201310372605.9A CN201310372605A CN103483459A CN 103483459 A CN103483459 A CN 103483459A CN 201310372605 A CN201310372605 A CN 201310372605A CN 103483459 A CN103483459 A CN 103483459A
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Abstract
The invention discloses an ultrasound-assisted extraction process of fritillariae thunbergii polysaccharide and a health-care product containing the fritillariae thunbergii polysaccharide. The ultrasound-assisted extraction process comprises the following steps: (1) degreasing; (2) performing water extraction, crushing dried fritillariae thunbergii, extracting polysaccharide according to the ultrasound-assisted extraction process, wherein the extraction conditions are that the material-liquid ratio is 1:15, the ultrasonic action power is 120W, the extraction temperature is 80 DEG C, and the extraction time is 1.5 h, repeatedly extracting filter residues for at least three times, and combining filtrates to obtain fritillariae thunbergii polysaccharide water extract; (3) performing alcohol precipitation; (4) removing protein to obtain purified fritillariae thunbergii polysaccharide. The ultrasound-assisted extraction process has the advantages of short extraction time and high efficiency; the health-care product containing the fritillariae thunbergii polysaccharide has the health-care effects of elimination of free radicals and strong oxidation resistance.
Description
Technical field
The invention belongs to compound and extract field, specifically refer to the ultrasonic assisted extraction technique of Thunberg Fritillary Bulb polysaccharide.
Background technology
Polysaccharide is as the natural high moleculer eompound from higher plant, animal and microorganism cells, is the molecule complex structure that formed by the condensation of a plurality of monose molecule, dehydration and huge glucide, is one of four large base substances that form life.Polysaccharide is constantly excavated because its important physiological function reaches to apply widely, and polysaccharide starts from nineteen forty-three as medicine, and people have successfully extracted polysaccharide from many biomaterials at present, and these polysaccharide have extremely important and special physiologically active.
A kind of per nnial herb that Thunberg Fritillary Bulb (Fritillaria thunbergiiMiq.) is the Liliaceae Fritillaria.Underground bulb has the function of heat-clearing dissipating bind, preventing phlegm from forming and stopping coughing, can be used for treating wind-heat invading the lung, phlegm-fire cough, lung carbuncle, acute mastitis, scrofula and sore, is one of " Zhejiang eight flavors ".
Modern pharmacy is to the research of Thunberg Fritillary Bulb activeconstituents mainly for micromolecular compounds such as alkaloids, and remaining waste residue wastes in the process of extracting.Yet in Thunberg Fritillary Bulb, polysaccharide content is high, the exploitation of polysaccharide are studied to rarely seen Study on extraction, there is not yet polysaccharide structures and active research report, ultrasonically auxiliary extracting method research also there is not yet bibliographical information.
Summary of the invention
The objective of the invention is the shortcoming and defect existed in order to overcome prior art, and provide a kind of consuming time few, power consumption is few, the ultrasonic assisted extraction technique of the Thunberg Fritillary Bulb polysaccharide that extraction yield is higher.
Another object of the present invention is to provide a kind of Thunberg Fritillary Bulb polysaccharide and solvent that includes purifying, the concentration >=12mg/mL of the Thunberg Fritillary Bulb polysaccharide of this purifying.
For achieving the above object, technical scheme of the present invention is to provide a kind of healthcare products containing the Thunberg Fritillary Bulb polysaccharide.
(1) grease removal
Thunberg Fritillary Bulb is arranged to impurity elimination, and with 95% ethanol submergence, the water-bath of 80 ℃ refluxes, and at least after 3h, takes out Thunberg Fritillary Bulb, and dry;
(2) water extraction
Get dried Thunberg Fritillary Bulb, pulverize, by ultrasonic assisted extraction technique, extract polysaccharide, extraction conditions is: solid-liquid ratio is 1:15, and ul-trasonic irradiation power is 120W, extracts 80 ℃ of temperature, and extraction time is 1.5h; Filter residue at least repeats to extract three times, and merging filtrate obtains Thunberg Fritillary Bulb polysaccharide water extraction liquid.
(3) alcohol precipitation
Thunberg Fritillary Bulb polysaccharide water extraction liquid is concentrated, 95% the ethanolic soln that adds 3-4 times of volume, the limit edged stirs, cotton-shaped Crude polysaccharides throw out occurs, preserve and be placed in the refrigerator of 4 ℃, at least after 24h, take out, slowly pour out supernatant liquid, centrifugal at least 20min under the rotating speed of lower floor's material more than 4500r/min, centrifugal sediment is Crude polysaccharides;
(4) deproteinated
Crude polysaccharides is dissolved to the sugar soln of making 5%, chloroform-the butanol solution that adds 1/5 volume, fully vibration on velocity-modulated oscillator, at least after 30min, centrifugal at least 10min under rotating speed more than 4500r/min, obtain three layers of liquid, gets the superiors' yellow liquid, repeat 3-5 time, obtain the Thunberg Fritillary Bulb polysaccharide of purifying.
Advantage of the present invention is: the extraction yield of polysaccharide (8.61%) improves 1.18% than the extraction yield of traditional extraction technique (7.43%).And extraction time of the present invention is short, efficiency is high.Specifically referring to the experimental example part in the embodiment of the present application.
For realizing second goal of the invention of the present invention, technical scheme of the present invention is the healthcare products containing the Thunberg Fritillary Bulb polysaccharide, includes Thunberg Fritillary Bulb polysaccharide and the solvent of purifying, the concentration >=12mg/mL of the Thunberg Fritillary Bulb polysaccharide of this purifying.Healthcare products of the present invention have stronger anti-oxidant activity, and the clearance rate of hydroxy radical qiao, up to 98.17%, be can be used as to the health care medicine exploitation.
Below in conjunction with specification drawings and specific embodiments, the present invention is described further.
The accompanying drawing explanation
Fig. 1 Thunberg Fritillary Bulb polysaccharide is removed the graphic representation of hydroxy radical qiao activity;
The graphic representation of Fig. 2 solid-liquid ratio on the impact of polysaccharide extract rate;
The influence curve figure of Fig. 3 temperature to polysaccharide extract rate;
The influence curve figure of Fig. 4 power to polysaccharide extract rate;
Fig. 5 extraction time influence curve figure to polysaccharide extract rate.
Embodiment
Below by embodiment, the present invention is specifically described; only be used to further illustrate the present invention; can not be interpreted as limiting the scope of the present invention, the technician in this field can make some nonessential improvement and adjustment to the present invention according to the content of foregoing invention.
embodiment 1:
1 grease removal
Thunberg Fritillary Bulb is arranged to impurity elimination, accurately take 500g, with 95% ethanol submergence, the water-bath of 80 ℃ refluxes, and takes out Thunberg Fritillary Bulb sample, drying after 3h.
water extraction
Get dried Thunberg Fritillary Bulb, pulverize, by ultrasonic assisted extraction technique, extract polysaccharide, extraction conditions is: solid-liquid ratio is 1:15, and ul-trasonic irradiation power is 120W, extracts 80 ℃ of temperature, and extraction time is 1.5h.Filter residue repeats to extract three times, and merging filtrate obtains Thunberg Fritillary Bulb polysaccharide water extraction liquid.
alcohol precipitation
Thunberg Fritillary Bulb polysaccharide water extraction liquid is condensed into to 500ml, and (bulb of fritillary polysaccharide soln: ethanolic soln=1:4), the limit edged stirs, and cotton-shaped Crude polysaccharides throw out occurs, and be placed in the refrigerator of 4 ℃, preserves to add 95% ethanolic soln of 4 times of volumes.After 24h, take out, slowly pour out supernatant liquid, lower floor's material is centrifugal 20min under the rotating speed of 4500r/min, and centrifugal sediment is Crude polysaccharides.After dry, example weight is 119.7 g.
the method deproteinated
Crude polysaccharides is dissolved to the sugar soln of making 5%, by the Sevag method, except albumen, add chloroform-butanol solution (bulb of fritillary Crude polysaccharides: chloroform-propyl carbinol=5:1), fully vibrate of 1/5 volume on velocity-modulated oscillator.After 30min, centrifugal 10min under the rotating speed of 4500r/min, obtain three layers of liquid.Get the superiors' yellow liquid, repeat 3-5 time, measure protein content with Xylene Brilliant Cyanine G after each deproteinated, until protein is close to nothing.The freeze-drying of polysaccharide sample, obtain purified polysaccharide 38.9g.
embodiment 2:
1 grease removal
Thunberg Fritillary Bulb is arranged to impurity elimination, accurately take 5000g, with 95% ethanol submergence, the water-bath of 80 ℃ refluxes, and takes out Thunberg Fritillary Bulb sample, drying after 3h.
water extraction
Get dried Thunberg Fritillary Bulb, pulverize, by ultrasonic assisted extraction technique, extract polysaccharide, extraction conditions is: solid-liquid ratio is 1:15, and ul-trasonic irradiation power is 120W, extracts 80 ℃ of temperature, and extraction time is 1.5h.Filter residue repeats to extract three times, and merging filtrate obtains Thunberg Fritillary Bulb polysaccharide water extraction liquid.
alcohol precipitation
Thunberg Fritillary Bulb polysaccharide water extraction liquid is condensed into to 1500ml, and (bulb of fritillary polysaccharide soln: ethanolic soln=1:4), the limit edged stirs, and cotton-shaped Crude polysaccharides throw out occurs, and be placed in the refrigerator of 4 ℃, preserves to add 95% ethanolic soln of 4 times of volumes.After 24h, take out, slowly pour out supernatant liquid, lower floor's material is centrifugal 20min under the rotating speed of 4500r/min, and centrifugal sediment is Crude polysaccharides.After dry, example weight is 586.4 g.
the method deproteinated
Crude polysaccharides is dissolved to the sugar soln of making 5%, by the Sevag method, except albumen, add chloroform-butanol solution (bulb of fritillary Crude polysaccharides: chloroform-propyl carbinol=5:1), fully vibrate of 1/5 volume on velocity-modulated oscillator.After 30min, centrifugal 10min under the rotating speed of 4500r/min, obtain three layers of liquid.Get the superiors' yellow liquid, repeat 3-5 time, measure protein content with Xylene Brilliant Cyanine G after each deproteinated, until protein is close to nothing.The freeze-drying of polysaccharide sample, obtain purified polysaccharide 196.2g.
The Thunberg Fritillary Bulb polysaccharide that technique of the present invention is purified is configured to the healthcare products of concentration >=12mg/mL, has excellent anti-oxidant activity.As shown in Figure 1, the Thunberg Fritillary Bulb polysaccharide raises along with the rising of concentration to the clearance rate of hydroxy radical qiao, and obvious dose-effect relationship is arranged between removing ability and concentration.When concentration, in 2 ~ 8mg/mL scope, polysaccharide is remarkable to the scavenging(action) of hydroxy radical qiao, and when concentration increases again, polysaccharide remains unchanged substantially to the scavenging(action) to hydroxy radical qiao.When concentration reaches 12mg/mL, clearance rate is up to 98.17%.
experimental example
the experiment of single factor of 1 ultrasonic-assisted extraction
1.1 the impact of different feed liquid comparison polysaccharide extract rate
Extract temperature 70 C, ul-trasonic irradiation power 120w, under the condition of extraction time 2.5h, change solid-liquid ratio, and the extraction yield of polysaccharide is as Fig. 2.By Fig. 2, can be obtained, the extraction yield of extraction yield polysaccharide improves along with the increase of solid-liquid ratio, and when solid-liquid ratio is brought up to 1:15 from 1:10, extraction yield significantly increases, and from 2.43%, brings up to 6.93%.When solid-liquid ratio is greater than 1:15, along with the raising of solid-liquid ratio, extraction yield has the trend of reduction on the contrary.Therefore, the suitableeest solid-liquid ratio of ultrasonic extraction is near 1:15.
the impact of different extraction temperature on polysaccharide extract rate
Solid-liquid ratio is 1:20, and ul-trasonic irradiation power 120w, under the condition of extraction time 2.5h, changes and extracts temperature, and the extraction yield of polysaccharide is as Fig. 3.By Fig. 3, can be obtained, the extraction yield of extraction yield polysaccharide improves along with the increase of temperature, heats up and is conducive to obtain higher extraction yield, but work as temperature, surpasses 90 ℃, can destroy the glycosidic link of polysaccharide.From figure, show, when extracting temperature and equal 70 ℃, the extraction yield of polysaccharide is 6.78%, and when extracting temperature and be greater than 70 ℃, the extraction yield of polysaccharide tends towards stability, and from many aspects such as energy consumptions, considers, and selects the extraction temperature of 70 ℃ and 80 ℃ comparatively appropriate.
the impact of different capacity on polysaccharide extract rate
Extracting temperature is 70 ℃, and solid-liquid ratio is 1:20, under the condition of extraction time 2.5h, changes hyperacoustic operating power, and the extraction yield of polysaccharide is as Fig. 4.Ultrasonic power improves, and can not only accelerate the circulation of water, accelerates the dissolving of material, can also produce vibration and change the cell interior structure, thereby accelerate the transfer of polysaccharide.By Fig. 3, can be obtained, the extraction yield of extraction yield polysaccharide improves along with the increase of temperature.When power is 160W, extraction yield is 9.02%; When power is greater than 160W, polysaccharide extract rate has a declining tendency.May be to cause the explanation of polysaccharide due to hyperacoustic concuss, cause polysaccharide extract rate to descend.Therefore, ul-trasonic irradiation power the best is 160W ~ 200W.
the impact of different extraction times on polysaccharide extract rate
Extracting temperature is 70 ℃, and solid-liquid ratio is 1:20, under the condition of ul-trasonic irradiation power 120W, changes extraction time, and the extraction yield of polysaccharide is as Fig. 4.Time, the extraction yield of polysaccharide presented ascendant trend in 1.0h-1.5h, and when extraction time is 1.5h, the extraction yield of polysaccharide is 8.49%, when extraction time, is greater than 1.5h, and the extraction yield of polysaccharide obviously descends, between 4%-5%.So the optimum extraction time of polysaccharide is 1.5h.
the orthogonal experiment of ultrasonic extracting process
On the basis of the experiment of single factor of ultrasonic-assisted extraction, on affecting the principal element of Thunberg Fritillary Bulb polysaccharide content, extract temperature, extraction time, solid-liquid ratio and ul-trasonic irradiation power and carry out L
9(3
4) orthogonal experiment.Level of factor analysis in table 1, interpretation of result is in Table 2, and variance analysis is in Table 3.
Table 1 level of factor table
Table 2 L
9(3
4) Orthogonal experiment results
Table 3 orthogonal test analysis of variance table
As known from Table 2, extreme difference is followed successively by extraction time, solid-liquid ratio, extraction temperature and ul-trasonic irradiation power from big to small, explanation is in experimental factor and horizontal extent, the factor primary and secondary that affects the extraction of Thunberg Fritillary Bulb polysaccharide sequentially should be: B>C>A>D, i.e. extraction time>solid-liquid ratio>extraction temperature>ul-trasonic irradiation power.Comprehensive single factor is extracted experiment, finally determines that optimal extract process is: A
2b
2c
3d
1, extraction time is 1.5h, and solid-liquid ratio is 1:15, and extracting temperature is 80 ℃, and ul-trasonic irradiation power is 120W.The extraction yield of polysaccharide is 8.61% with this understanding.
The variance analysis demonstration, the F value of extracting temperature, extraction time, 3 factors of solid-liquid ratio all is greater than F
0.05(2,2)=19.00, this explanation extraction temperature, extraction time, 3 factors of solid-liquid ratio have significant impact to the ultrasonic-assisted extraction of Thunberg Fritillary Bulb polysaccharide, and contribution rate is respectively 26.7%, 36.93%, 32.79%.
This research adopts the method for experiment of single factor and orthogonal test, determined the optimum process condition of the auxiliary Thunberg Fritillary Bulb polysaccharide of ultrasonic wave in conjunction with practical situation: solid-liquid ratio is 1:15, ul-trasonic irradiation power is 120W, extract 80 ℃ of temperature, extraction time is 1.5h, the extraction yield of polysaccharide is the highest, is 8.61%.The ultrasonic-assisted extraction polysaccharide can improve the extraction yield of polysaccharide to a certain extent, with traditional optimum extraction condition, compares, and has consuming time less, and power consumption is few, the advantage that extraction yield is higher.
Claims (2)
1. the ultrasonic assisted extraction technique of a Thunberg Fritillary Bulb polysaccharide is characterized in that comprising the following steps:
(1) grease removal
Thunberg Fritillary Bulb is arranged to impurity elimination, and with 95% ethanol submergence, the water-bath of 80 ℃ refluxes, and at least after 3h, takes out Thunberg Fritillary Bulb, and dry;
(2) water extraction
Get dried Thunberg Fritillary Bulb, pulverize, by ultrasonic assisted extraction technique, extract polysaccharide, extraction conditions is: solid-liquid ratio is 1:15, and ul-trasonic irradiation power is 120W, extracts 80 ℃ of temperature, and extraction time is 1.5h; Filter residue at least repeats to extract three times, and merging filtrate obtains Thunberg Fritillary Bulb polysaccharide water extraction liquid;
(3) alcohol precipitation
Thunberg Fritillary Bulb polysaccharide water extraction liquid is concentrated, 95% the ethanolic soln that adds 3-4 times of volume, the limit edged stirs, cotton-shaped Crude polysaccharides throw out occurs, preserve and be placed in the refrigerator of 4 ℃, at least after 24h, take out, slowly pour out supernatant liquid, centrifugal at least 20min under the rotating speed of lower floor's material more than 4500r/min, centrifugal sediment is Crude polysaccharides;
(4) deproteinated
Crude polysaccharides is dissolved to the sugar soln of making 5%, chloroform-the butanol solution that adds 1/5 volume, fully vibration on velocity-modulated oscillator, at least after 30min, centrifugal at least 10min under rotating speed more than 4500r/min, obtain three layers of liquid, gets the superiors' yellow liquid, repeat 3-5 time, obtain the Thunberg Fritillary Bulb polysaccharide of purifying.
2. the healthcare products containing the Thunberg Fritillary Bulb polysaccharide that technique as claimed in claim 1 is purified, is characterized in that: include Thunberg Fritillary Bulb polysaccharide and the solvent of purifying, the concentration >=12mg/mL of the Thunberg Fritillary Bulb polysaccharide of this purifying.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106397622A (en) * | 2016-10-11 | 2017-02-15 | 重庆三峡医药高等专科学校 | Fritillariae cirrhosae bulbus polysaccharide extraction, separation and purification technology |
| CN108047343A (en) * | 2017-12-06 | 2018-05-18 | 中国科学院新疆理化技术研究所 | The preparation method and applications of Siberian fritillary bulb total starches |
| CN111040040A (en) * | 2019-11-11 | 2020-04-21 | 佳木斯大学 | Preparation method and application of fritillaria ussuriensis polysaccharide zinc complex |
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| CN1765910A (en) * | 2005-11-09 | 2006-05-03 | 天津大学 | Method for extracting simultaneously general alkaloid and starch from fritillary |
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| CN1765910A (en) * | 2005-11-09 | 2006-05-03 | 天津大学 | Method for extracting simultaneously general alkaloid and starch from fritillary |
| CN101601805A (en) * | 2009-06-30 | 2009-12-16 | 长春工程学院 | The detection method of peimine and peiminine in the Chinese medicine Bulbus Fritillariae Cirrhosae extract |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106397622A (en) * | 2016-10-11 | 2017-02-15 | 重庆三峡医药高等专科学校 | Fritillariae cirrhosae bulbus polysaccharide extraction, separation and purification technology |
| CN106397622B (en) * | 2016-10-11 | 2018-10-02 | 重庆三峡医药高等专科学校 | The extraction of bulbus fritillariae cirrhosae polysaccharide and separation purifying technique |
| CN108047343A (en) * | 2017-12-06 | 2018-05-18 | 中国科学院新疆理化技术研究所 | The preparation method and applications of Siberian fritillary bulb total starches |
| CN111040040A (en) * | 2019-11-11 | 2020-04-21 | 佳木斯大学 | Preparation method and application of fritillaria ussuriensis polysaccharide zinc complex |
| CN111040040B (en) * | 2019-11-11 | 2021-10-22 | 佳木斯大学 | Preparation method and application of fritillaria ussuriensis polysaccharide zinc complex |
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