CN103848920A - Method for extracting isatis root polysaccharide under ultrasonic assistance - Google Patents
Method for extracting isatis root polysaccharide under ultrasonic assistance Download PDFInfo
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- CN103848920A CN103848920A CN201210510656.9A CN201210510656A CN103848920A CN 103848920 A CN103848920 A CN 103848920A CN 201210510656 A CN201210510656 A CN 201210510656A CN 103848920 A CN103848920 A CN 103848920A
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- isatis root
- drying
- banlangen polysaccharide
- degreasing
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Abstract
The invention relates to a method for extracting isatis root polysaccharide under ultrasonic assistance. The method comprises the following steps: drying, smashing and degreasing an isatis root medicinal material; extracting by using an ultrasonic cell crusher; preparing a supernatant obtained by centrifuging into isatis root polysaccharide by using the technologies of concentration, alcohol precipitation, filtration and vacuum drying. Compared with the prior art, a process adopted in the method has the characteristics of rapidness, easiness, convenience, energy conservation and high yield, and a plurality of active substances in the isatis root polysaccharide can be protected. Proven by experiments, the isatis root polysaccharide extracted under ultrasonic assistance has high in-vitro antioxidant activity.
Description
Technical field
The present invention relates to the extracting method of technical field of traditional Chinese medicines, especially relate to a kind of method of ultrasonic-assisted extraction Banlangen Polysaccharide.
Background technology
Root of Indigowoad (Isatidis Radix), call indigo-blue, indigo root, indigo root, it is the dry root of cruciferae isatis (Isatisin digotica Fort.), have effect of clearing heat and detoxicating, cool blood relieve sore throat, poison during clinically for warm epidemic disease, heating pharyngalgia, maculae caused by violent heat pathogen, mumps, scarlet fever, major part pestilence, erysipelas, carbuncle are swollen etc.As the Typical Representative of China's anti virus herb, Root of Indigowoad is the first traditional Chinese medicine research project that NIH subsidizes that obtains.
Polysaccharide is the biomacromolecule of the class formation complexity that is formed by connecting by glycosidic bond by multiple monose molecules, distributes very extensive in natural drug.Polysaccharide plays an important role in many basic vital processes such as infection, tumour cell transfer of biosynthesizing and intercellular identification, fertilization, embryo's formation, neurocyte growth, hormone activation, cell proliferation, virus and bacterium, has therefore become the study hotspot in medical science, trophology and Food science field.It is reported, Banlangen Polysaccharide all has certain promoter action to specific immunity and non-specific immunity, is a kind of more satisfactory natural immunity toughener, has DEVELOPMENT PROSPECT widely as protective foods and medicine.
Application development in recent years comparatively extensive of ultrasonic technology in Effective Component of Chinese Medicine extracts.Utilize hyperacoustic cavatition, mechanical effect and heat effect etc., accelerate release, diffusion and the dissolving of active substance in cell, can significantly shorten extraction time, improve extraction efficiency.At present, use ultrasonic cell disruption instrument to extract Banlangen Polysaccharide and there is not yet bibliographical information.
Summary of the invention
Object of the present invention be exactly provide in order to overcome the defect that above-mentioned prior art exists a kind of fast and convenient, save the energy, the method extracting method that utilizes ultrasonic-assisted extraction Banlangen Polysaccharide that yield is high, and be conducive to protect the many active substances that contain in Banlangen Polysaccharide.
Object of the present invention can be achieved through the following technical solutions:
A method for ultrasonic-assisted extraction Banlangen Polysaccharide, comprises the following steps:
(1) Root of Indigowoad is dried and is ground into Isatis Root;
(2) carry out suction filtration add finite concentration alcohol degreasing in Isatis Root after, then filter residue is carried out to vacuum-drying;
(3) in the Isatis Root after degreasing, add water, use ultrasonic cell disruption instrument to extract Banlangen Polysaccharide, cooling, centrifugation, obtains the supernatant liquor that contains Banlangen Polysaccharide;
(4), by supernatant liquor concentrating under reduced pressure, add equivalent or several times amount dehydrated alcohol, 4 ℃ of hold over night, centrifugal removal mother liquor, collecting precipitation;
(5) successively by ethanol, washing with acetone precipitation, vacuum-drying, obtains the polysaccharide that contains anti-oxidant activity Banlangen Polysaccharide.
In step (1), drying the temperature adopting is 50～60 ℃, and the granularity of pulverizing the Isatis Root obtaining is 60～80 orders.
Degreasing in step (2) adopts 5～10 times (w/v) amount, the 95% alcohol reflux degreasing twice of Isatis Root, each 2h, and described vacuum drying temperature is 20～30 ℃, the time is 24～48h.
In the Isatis Root of step (3) after degreasing, add water, making solid-liquid ratio is 1: 10～1: 100, and the processing parameter that ultrasonic cell disruption instrument extracts is ultrasonic power 400～800W, 30～60 ℃ of temperature, time 10～30min.
Step (4) adopts Rotary Evaporators in the time of concentrating under reduced pressure, 55～60 ℃ of thickening temperatures, and concentrated volume is original volume 1/5～1/10, dehydrated alcohol add-on is 1～5 times of concentrated solution volume; Rotating speed when centrifugal treating is 3500～4500rpm, time 10～15min.
Step (5) adds successively the washing with alcohol of step (4) mother liquor same concentrations and filters in the time of washing, progressively improve alcohol concn, finally use dehydrated alcohol, washing with acetone, it is topped in case moisture is brought in the evaporation of throw out surface solvent into when suction filtration is closely dry, the temperature that vacuum-drying adopts is 40～45 ℃, and be 24～72h time of drying.
Compared with prior art; the technology such as the present invention adopts that pulverizing, degreasing, ultrasonic cell disruption instrument are extracted, centrifugal, concentrated, alcohol precipitation and vacuum-drying are prepared Banlangen Polysaccharide; that the technique proposing has is quick, easy, save the energy, yield high, and is conducive to protect the many active substances that contain in Banlangen Polysaccharide.The extracting method of taking is that ultrasonic cell disruption instrument extracts, compare conventional water bath reflux method, extraction time can be foreshortened to 10min by a few hours, and extraction yield improves nearly one times, the Banlangen Polysaccharide obtaining has good antioxidation activity in vitro, has good ABTS
+free radical scavenging effect, is widely used in anti-oxidant function product.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
Embodiment 1
The extraction of Banlangen Polysaccharide
Root of Indigowoad, in 60 ℃ of oven dry, was pulverized to 80 mesh sieves, added 5 times of amounts (w/v), 95% alcohol reflux twice, each 2h, suction filtration is removed filtrate, and filter residue is in 25 ℃ of vacuum-drying 24h.
Root of Indigowoad 5g after extracting degreasing, the 200mL that adds water (solid-liquid ratio 1: 40), uses ultrasonic cell disruption instrument to extract Banlangen Polysaccharide, ultrasonic power 787w, temperature 60 C, extraction time 10min.By centrifugal the extracting solution that is cooled to room temperature (4500rpm, 15min), collect supernatant liquor, filter residue distilled water wash, and merge with supernatant liquor.
Said extracted liquid is concentrated into 40mL with 60 ℃ of Rotary Evaporators, under agitation slowly adds 4 times of amount dehydrated alcohols, 4 ℃ of hold over night, the centrifugal 15min of 4500rpm/min, collecting precipitation product; Use successively the each 10mL washing precipitation of 80% ethanol, dehydrated alcohol and acetone, topped when suction filtration is closely dry; 45 ℃ of vacuum-drying 72h, weigh.Repeat above-mentioned experiment three times, obtain respectively polysaccharide 1.25g, 1.24g, 1.26g, calculate polysaccharide extract rate, results averaged is 25.01%.
Embodiment 2-18
The optimization of process conditions of ultrasonic-assisted extraction Banlangen Polysaccharide
Adopt the center combination experimental design principle of Box-Benhnken, the scheme of the response surface analysis test of design Three factors-levels, take ultrasonic power, ultrasonic time, 3 processing parameters of ultrasonic temperature as independent variable(s), take polysaccharide extract rate as response value, the processing condition of Optimization for Ultrasonic Wave assisted extraction Banlangen Polysaccharide, test design and the results are shown in Table 1.
Table 1
The Banlangen Polysaccharide optimum extraction condition obtaining by Design-Expert software (Version 7.0.0) analysis is: 58.99 ℃ of ultrasonic power 786.95w, ultrasonic time 10.14min, ultrasonic temperature, the theory extraction value under this condition is 25.14%.Take ultrasonic power 787w, ultrasonic time 10min, ultrasonic temperature 60 C to extract Banlangen Polysaccharide, carry out three parallel laboratory tests, calculate extraction yield and be respectively 24.95%, 24.88% and 25.19%, approach with predictor.
Embodiment 19
The in vitro anti-oxidation of polysaccharide is measured
The Banlangen Polysaccharide obtaining with embodiment 1 carries out ABTS
+the experiment of free radical scavenging effect.Configuration ABTS free radical working fluid, is dissolved in distilled water by Banlangen Polysaccharide sample and is made into a series of concentration.Get 0.1mL sample solution and add 2.9mL ABTS
+free radical working fluid, mixes, and after placement 6min, measures absorbancy in 734nm.Calculate Banlangen Polysaccharide to ABTS according to formula
+the clearance rate of free radical, the results are shown in Table 2.
Clearance rate (%)=[(A
0-A
t)/A0] × 100%
In formula
a0 is 2.9mL ABTS
+solution and the mixed absorbancy of 0.1mL distilled water; A
tfor 2.9mLABTS
+solution and the mixed absorbancy of 0.1mL sample solution.
Table 2
Embodiment 20
A method for ultrasonic-assisted extraction Banlangen Polysaccharide, comprises the following steps:
(1) Root of Indigowoad being dried and is ground at 50 ℃ granularity is 60 object Isatis Roots;
(2) in Isatis Root, add 5 times (w/v), the alcohol reflux degreasing twice that concentration is 95%, each 2h, after carry out suction filtration, then filter residue is carried out to vacuum-drying 48h at 20 ℃;
(3) in the Isatis Root after degreasing, adding water to make solid-liquid ratio is 1: 10, uses ultrasonic cell disruption instrument to extract Banlangen Polysaccharide, ultrasonic power 400W, 30 ℃ of temperature, the reaction times is 30min, then cooling, centrifugation, obtains the supernatant liquor that contains Banlangen Polysaccharide;
(4) supernatant liquor is adopted Rotary Evaporators carry out concentrating under reduced pressure, 55 ℃ of thickening temperatures, are concentrated into 1/5 of original volume by volume, add the dehydrated alcohol of equivalent, 4 ℃ of hold over night, centrifugal removal mother liquor, rotating speed when centrifugal treating is 3500rpm, and the treatment time is 15min, regathers precipitation;
(5) successively by ethanol, washing with acetone precipitation, when washing, add successively the washing with alcohol of step (4) mother liquor same concentrations and filter, progressively improve alcohol concn, finally use dehydrated alcohol, washing with acetone, it is topped in case moisture is brought in the evaporation of throw out surface solvent into when suction filtration is closely dry, finally controlling temperature is 40 ℃, and vacuum-drying 72h, obtains the polysaccharide that contains anti-oxidant activity Banlangen Polysaccharide.
Embodiment 21
A method for ultrasonic-assisted extraction Banlangen Polysaccharide, comprises the following steps:
(1) Root of Indigowoad being dried and is ground at 60 ℃ granularity is 80 object Isatis Roots;
(2) in Isatis Root, add 10 times (w/v), the alcohol reflux degreasing twice that concentration is 95%, each 2h, after carry out suction filtration, then filter residue is carried out to vacuum-drying 24h at 30 ℃;
(3) in the Isatis Root after degreasing, adding water to make solid-liquid ratio is 1: 100, uses ultrasonic cell disruption instrument to extract Banlangen Polysaccharide, ultrasonic power 800W, temperature 60 C, the reaction times is 10min, then cooling, centrifugation, obtains the supernatant liquor that contains Banlangen Polysaccharide;
(4) supernatant liquor is adopted Rotary Evaporators carry out concentrating under reduced pressure, 55 ℃ of thickening temperatures, volume is concentrated into 1/10 of original volume, add the dehydrated alcohol of 5 times of volumes, 4 ℃ of hold over night, centrifugal removal mother liquor, rotating speed when centrifugal treating is 4500rpm, treatment time is 10min, regathers precipitation;
(5) successively by ethanol, washing with acetone precipitation, when washing, add successively the washing with alcohol of step (4) mother liquor same concentrations and filter, progressively improve alcohol concn, finally use dehydrated alcohol, washing with acetone, it is topped in case moisture is brought in the evaporation of throw out surface solvent into when suction filtration is closely dry, finally controlling temperature is 45 ℃, and vacuum-drying 24h, obtains the polysaccharide that contains anti-oxidant activity Banlangen Polysaccharide.
Claims (6)
1. a method for ultrasonic-assisted extraction Banlangen Polysaccharide, is characterized in that, the method comprises the following steps:
(1) Root of Indigowoad is dried and is ground into Isatis Root;
(2) carry out suction filtration add finite concentration alcohol degreasing in Isatis Root after, then filter residue is carried out to vacuum-drying;
(3) in the Isatis Root after degreasing, add water, use ultrasonic cell disruption instrument to extract Banlangen Polysaccharide, cooling, centrifugation, obtains the supernatant liquor that contains Banlangen Polysaccharide;
(4), by supernatant liquor concentrating under reduced pressure, add equivalent or several times amount dehydrated alcohol, 4 ℃ of hold over night, centrifugal removal mother liquor, collecting precipitation;
(5) successively by ethanol, washing with acetone precipitation, vacuum-drying, obtains the polysaccharide that contains anti-oxidant activity Banlangen Polysaccharide.
2. the method for a kind of ultrasonic-assisted extraction Banlangen Polysaccharide according to claim 1, is characterized in that, in step (1), drying the temperature adopting is 50～60 ℃, and the granularity of pulverizing the Isatis Root obtaining is 60～80 orders.
3. the method for a kind of ultrasonic-assisted extraction Banlangen Polysaccharide according to claim 1, it is characterized in that, degreasing in step (2) adopts 5～10 times (w/v) of Isatis Root to measure twice of 95% alcohol reflux degreasing, each 2h, described vacuum drying temperature is 20～30 ℃, and the time is 24～48h.
4. the method for a kind of ultrasonic-assisted extraction Banlangen Polysaccharide according to claim 1, it is characterized in that, in the Isatis Root of step (3) after degreasing, add water, making solid-liquid ratio is 1: 10～1: 100, the processing parameter that ultrasonic cell disruption instrument extracts is ultrasonic power 400～800W, 30～60 ℃ of temperature, time 10～30min.
5. the method for a kind of ultrasonic-assisted extraction Banlangen Polysaccharide according to claim 1, it is characterized in that, step (4) adopts Rotary Evaporators in the time of concentrating under reduced pressure, 55～60 ℃ of thickening temperatures, concentrated volume is 1/5～1/10 of original volume, and dehydrated alcohol add-on is 1～5 times of concentrated solution volume; Rotating speed when centrifugal treating is 3500～4500rpm, time 10～15min.
6. the method for a kind of ultrasonic-assisted extraction Banlangen Polysaccharide according to claim 1, it is characterized in that, step (5) adds successively the washing with alcohol of step (4) mother liquor same concentrations and filters in the time of washing, progressively improve alcohol concn, finally use dehydrated alcohol, washing with acetone, topped when suction filtration is closely dry in case moisture is brought in the evaporation of throw out surface solvent into, the temperature that vacuum-drying adopts is 40～45 ℃, and be 24～72h time of drying.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101250234A (en) * | 2008-04-10 | 2008-08-27 | 北京联合大学生物化学工程学院 | Method for extracting ginseng polysaccharide |
| CN101250232A (en) * | 2008-03-27 | 2008-08-27 | 钱国英 | Extraction technique of sargassum fusiform active polysaccharides |
| CN101367881A (en) * | 2008-09-19 | 2009-02-18 | 南京农业大学 | High-efficiency preparation method for glossy ganoderma polysaccharide |
| CN102002112A (en) * | 2010-12-02 | 2011-04-06 | 蒋熠霞 | Process for extracting ginkgo leaf polysaccharide with aid of ultrasonic |
-
2012
- 2012-12-03 CN CN201210510656.9A patent/CN103848920B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101250232A (en) * | 2008-03-27 | 2008-08-27 | 钱国英 | Extraction technique of sargassum fusiform active polysaccharides |
| CN101250234A (en) * | 2008-04-10 | 2008-08-27 | 北京联合大学生物化学工程学院 | Method for extracting ginseng polysaccharide |
| CN101367881A (en) * | 2008-09-19 | 2009-02-18 | 南京农业大学 | High-efficiency preparation method for glossy ganoderma polysaccharide |
| CN102002112A (en) * | 2010-12-02 | 2011-04-06 | 蒋熠霞 | Process for extracting ginkgo leaf polysaccharide with aid of ultrasonic |
Non-Patent Citations (5)
| Title |
|---|
| 刘依 等: "微波技术在板蓝根多糖提取中的应用", 《中国农业大学学报》 * |
| 唐志华: "微波辅助提取板蓝根多糖的工艺条件研究", 《食品工程》 * |
| 张萍 等: "板蓝根多糖的提取工艺及其清除自由基作用的初步研究", 《河南工业大学学报(自然科学版)》 * |
| 李勇: "《碳水化合物加工技术》", 29 February 2004, 北京：化学工业出版社 * |
| 车荣珍: "超声波辅助提取板蓝根多糖的工艺优化", 《时珍国医国药》 * |
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