CN101054421A - Method for ultrasound strengthen, extraction and purification of longan and litchi polysaccharide - Google Patents
Method for ultrasound strengthen, extraction and purification of longan and litchi polysaccharide Download PDFInfo
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- CN101054421A CN101054421A CN 200610011689 CN200610011689A CN101054421A CN 101054421 A CN101054421 A CN 101054421A CN 200610011689 CN200610011689 CN 200610011689 CN 200610011689 A CN200610011689 A CN 200610011689A CN 101054421 A CN101054421 A CN 101054421A
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Abstract
The invention relates to a novel longan and litchi polysaccharide extraction and purification technology. The longan and litchi polysaccharide is extracted by ultrasonic intensified extraction technology, after liquid-solid separation process, through condensing, flocculating or ethanol settling, filtering, freeze drying, crude polysaccharide can be obtained from the extracting liquid; redissolving concentrated polysaccharide crude extracted liquid or crude polysaccharide, then through fractional precipitation or chromatography purifying, drying and smashing to get longan and litchi polysaccharide purified products. For adopting ultrasonic intensified extraction technology can be used to large scale industrialization production, the extracted temperature is reduced from 90-100 DEGC to 30 DEG C, the extracted time is reduced from 4h to 0.5h, the polysaccharide extraction recovery rate is higher than that of traditional extraction method, the extract of the ultrasonic extraction is more favourable to subsequent separation and purification process to reduce extract purification cost of longan and litchi polysaccharide apparently.
Description
Technical field
The invention belongs to the biochemical engineering technology, relate to longan, lichee polysaccharide extraction and new technology for purifying.
Background technology
Polysaccharide is the natural high moleculer eompound that is polymerized by monose, and molecular weight does not wait from several thousand to millions of, is distributed widely in plant, animal and the microorganism.Recent two decades comes, development along with Celluar and Molecular Biology, people recognize that polysaccharide not only participates in constituting the base substance of life, have as functions such as signals between cytoskeleton, transfer cell, the more important thing is that aspects such as finding polysaccharide is regulated at immunity of organism, enhancing cell anti-tumor activity has important biological.
Longan (Dimocarpus Longan Lour, call longan) and lichee (Litchi Chinensis Sonn.), be sapindaceous plant, mainly be distributed in ground such as Guangdong, Guangxi, Fujian, Hainan and Taiwan in China, cultivated area and output all occupy first place in the world, be China's famous subtropical fruit,, be regarded as " treasure in the fruit " always because of having higher nutritive value and multiple nourishing beauty treatment effect.Its fruit of longan, lichee is spherical, and pulp white is transparent, and the juice multi-flavor is sweet, the Ministry of Health with they be decided to be food be again the fruit of medicine.Longan, litchi fruits nutritive value are very high, are regarded as precious tonic since ancient times.But longan, litchi fruits is moisture, sugar degree is high, and the following fruit phase lack, should not stores, corruption easily.Still based on dried food and nuts and pulp, added value of product is lower recently for longan, lichee deep processing.Utilize longan, litchi pulp to extract polysaccharide and not only can realize longan, the deep processing of lichee product, for medicine, protective foods etc. provide unique longan, lichee polysaccharide, increase the plantation added value, this will help increasing the enthusiasm of orchard worker's plantation.
Polysaccharide extracts following method commonly used, add heat extraction, this is the extracting method of at present the most frequently used polysaccharide, after being about to raw material drying, powder essence, use hot water extraction, through separation, concentrated, dry, the refining finished product (Wu Yalin etc. that obtain, the separation of fig polysaccharide, purifying and evaluation, journal of Zhejiang university, 2004,31 (2): 177-179); The Enzymatic Extraction method adds some protease hydrolysis free protein, so that the leaching of polysaccharide (Zhao Suxia, the technology of Enzymatic Extraction mulberry fruit polysaccharide, traditional Chinese medical science research, 2003,16 (6): 19-20) in vat liquor; Microwave extraction method, its principle is to utilize in microwave field, utilize the difference of microwave absorption capacity to make some component of base matter be heated by selectivity, thereby make it obtain separating (Liu Qingmei etc., adopt microwave technology to extract the technical study of laver amylose, Transactions of the Chinese Society of Agricultural Engineering, 2005,21 (2): 153-156).
Chen Zhe is superfine to provide a kind of working method of isolating polysaccharide and transparent beverage from longan.Adopt the ultra-filtration membrane purification technique, select the membrane separation apparatus of 10,000 molecular weight units, monose, VITAMIN, trace element and inorganic salt etc. less than this aperture are drained in the beverage liquid, obtain a kind of transparent nutritional drink of elaboration; Macromolecular polysaccharide greater than this aperture just is separated, and obtains longan polysaccharide (number of patent application CN200410077958.7).Huang Shouqi etc. disclose a kind of longan polysaccharide extracting process, comprise steps (number of patent application CN00107538.1) such as degreasing, double-enzyme hydrolysis, flooding, alcohol precipitation and drying and crushing.Huang Shouqi etc. also disclose a kind of extraction process of lichee polysaccharide, comprise select materials, making beating, enzymolysis, filter and remove residue, batching seasoning, tinning, sterilization, inspection after construction, qualified decals, packing warehouse-in operation (number of patent application CN03100544.6).
Li Xuehua etc. set up hot water extraction-ethanol sedimentation, and enzyme process removes Deproteinization, H in conjunction with the sevag method
2O
2Decoloration process prepares longan polysaccharide and lichee polysaccharide (Li Xuehua etc., the separation and Extraction of longan polysaccharide, lichee polysaccharide and the discussion of antioxygenation thereof, Guangxi Medical University's journal, 2004,21 (3): 342-344).Li Xuehua etc. utilize hot water extraction, ethanol sedimentation extracts lichee polysaccharide, by Deproteinization, the element that discolors polysaccharide is carried out purifying, with gel chromatographic columns sephadex G-25 lichee polysaccharide is carried out grading purification, polysaccharide behind the purifying detects purity (Li Xuehua etc., lichee polysaccharide separation and purification and purity evaluation, Guangxi Medical University's journal of polysaccharide respectively with polyacrylamide gel post P-100, polyacrylamide gel electrophoresis and cellulose acetate membrane electrophoresis method, 2005,22 (3): 366-367).Results of study such as Tang Xiaojun show that the lichee polysaccharide optimum extraction condition is: material-water ratio is 1: 9, and extraction temperature is 100 ℃, and extraction time is 4h; Polysaccharide average quality mark be 4.36% (Tang Xiaojun etc., the extraction conditions of lichee polysaccharide and assay, South China Normal University's journal (natural science edition), 2005, No.2:27-31)
Ultrasonic wave be frequency greater than 20KHz, the elastic mechanical wave of oscillation beyond the human auditory system valve.Ultrasonic wave mainly comes from its cavatition to the strengthening effect of various active ingredients of plants extraction separation.A series of dynamic processes such as micro-bubble when cavitation takes place in liquid nuclear vibrates under the high-strength ultrasonic effect, growth, contraction and collapse.The turbulence effect that produces during cavitation, perturbation effect, interfacial effect and cavity effect have promoted the fragmentation and the mass transfer process of vegetable cell, extract temperature, shorten extraction time, improve the purpose that product extracts yield, reduction product extraction separation cost thereby reach reduction.
The present invention adopts ultrasound-enhanced extraction longan, lichee polysaccharide, make the extraction temperature be reduced to 20-40 ℃ from 90-100 ℃, extraction time is reduced to≤1h from 4h, polysaccharide extract rate is higher than conventional extracting method, adopt technology such as ultrafiltration and concentration, flocculation and flocculation of electric field, chromatography and ethanol precipitation to realize the purifying of polysaccharide, significantly reduced polysaccharide and extracted the purifying cost.
Summary of the invention
The present invention is directed to present longan, lichee polysaccharide and extract problems such as temperature height, extraction time is long, extraction time is long, unique mechanism such as turbulence effect, perturbation effect, interfacial effect and the cavity effect etc. of utilizing the ultrasonic cavitation effect to produce, reach reduce extract temperature, shorten extraction time, improve extraction efficiency, purpose such as reduce production costs.
The present invention adopts the ultrasound assisted extraction technique that can use in scale operation; with water is to extract solvent; utilize orthogonal experimental design and artificial nerve network model; optimized the extraction conditions of longan, lichee polysaccharide; optimum result has been carried out the single factor experiment checking, and yield is extracted in the mass-producing of polysaccharide can reach 15%.Liquid of extracting polysaccharide can get polysaccharide crude after ultrafiltration and concentration, flocculation sediment, drying.Liquid of extracting polysaccharide or polysaccharide crude polysaccharide total yield behind purifying such as ethanol precipitation, chromatography can reach 10%.
The present invention relates to a kind of longan, the ultrasound-enhanced extraction of lichee polysaccharide and purification process thereof, its step is as follows:
(1). the fresh fruit meat that the decortication of fresh longan, lichee, stoning are obtained or exsiccant dry fruit meat be broken to certain fineness with pigment;
(2). the longan after will pulverizing, lichee slurry are regulated suitable liquid-solid ratio, put into ultrasonic extraction device and extract, and get longan, lichee polysaccharide crude extract after the liquid-solid separation;
(3). liquid of extracting polysaccharide adopts ultrafiltration and concentration to finite concentration;
(4). the liquid of extracting polysaccharide after concentrating adds flocculation agent and the electric field-enhanced flocculation sediment that carries out, and perhaps carries out ethanol sedimentation, the centrifugation taking precipitate;
(5). longan, the forced air drying of lichee polysaccharide throw out are got polysaccharide crude;
(6). slightly carry in polysaccharide concentrated solution or the polysaccharide crude lysate at longan, lichee and to add ethanol and carry out fractionation precipitation, precipitation is got in centrifugation, reclaims ethanol; Perhaps carry out column chromatography for separation, the elutriant of getting simple spike also concentrates, and gets the pure product of polysaccharide after the lyophilize.
Embodiment
Embodiment one:
Get the 10g litchi pulp, add water 40 mesh sieves of pulling an oar, Gu regulator solution/, place in the ultrasonic extraction device ultrasonic power 300W, dutycycle 30%, 30 ℃ of temperature, extraction time 20min than being 30: 1.Vat liquor vacuum concentration, ethanol sedimentation, lyophilize get the 1.6g polysaccharide crude.
Embodiment two:
Get the 50g longan pulp, add water 60 mesh sieves of pulling an oar, Gu regulator solution/, place in the 2.5L circulating ultrasonic extraction element ultrasonic power 1200W, dutycycle 30%, 40 ℃ of temperature, extraction time 30min than being 40: 1.Vat liquor vacuum concentration, ethanol sedimentation, precipitation is dissolved again through CM-dextrane gel column purification, according to the elution curve of polysaccharide, collects near the elutriant of absorption peak, and oven dry gets the pure product of 5g polysaccharide.
Embodiment three:
Getting the cleaner liquid 500ml after the longan supersound extraction, is 40 ℃ in temperature, and pH=7 adds the 400ppm chitosan, stirred 5 minutes with 800rpm, and electric field strength E=20V/cm, 100rpm stirs, and electric field action time 30min, polysaccharide extract rate are 70%.After getting the longan supersound extraction,, be 30 ℃ in temperature again through ultrafiltration and concentration liquid 100ml, pH=6, add 200ppm chitosan and 200ppm polystyrene sodium sulfonate, stirred 5 minutes electric field strength E=50V/cm with 1000rpm, 100rpm stirs, and electric field action time 20min, polysaccharide extract rate are 85%.
Embodiment four:
Get the 120g litchi pulp, add water 80 mesh sieves of pulling an oar, Gu regulator solution/, place in the 5.0L circulating ultrasonic extraction element ultrasonic power 1500W, dutycycle 50%, 40 ℃ of temperature, extraction time 40min than being 40: 1.Cleaner liquid ultrafiltration and concentration, ethanol sedimentation, lyophilize get the 16g polysaccharide crude.
Embodiment five:
Getting the cleaner liquid 1000ml after the lichee supersound extraction, is 20 ℃ in temperature, and pH=6.5 adds the 500ppm chitosan, stirred 5 minutes with 900rpm, and electric field strength E=60V/cm, 100rpm stirs, and electric field action time 50min, polysaccharide extract rate are 75%.After getting the lichee supersound extraction,, be 30 ℃ in temperature again through ultrafiltration and concentration liquid 300ml, pH=6, add 300ppm chitosan and 300ppm polystyrene sodium sulfonate, stirred 5 minutes electric field strength E=50V/cm with 1000rpm, 100rpm stirs, and electric field action time 30min, polysaccharide extract rate are 90%.
Embodiment six:
Get the 160g longan pulp, add water 80 mesh sieves of pulling an oar, Gu regulator solution/, place in the 5.0L circulating ultrasonic extraction element ultrasonic power 1800W, dutycycle 40%, 30 ℃ of temperature, extraction time 30min than being 30: 1.Cleaner liquid ultrafiltration and concentration, ethanol sedimentation, precipitation is dissolved again through CM-dextrane gel column purification, according to the elution curve of polysaccharide, collects near the elutriant of absorption peak, and oven dry gets the pure product of 15g polysaccharide.
Claims (6)
1. a longan, the ultrasound-enhanced extraction of lichee polysaccharide and purification process thereof, its step is as follows:
(1). the fresh fruit meat that the decortication of fresh longan, lichee, stoning are obtained or exsiccant dry fruit meat be broken to certain fineness with pigment;
(2). the longan after will pulverizing, lichee slurry are regulated suitable liquid-solid ratio, put into ultrasonic extraction device and extract, and get longan, lichee polysaccharide crude extract after the liquid-solid separation;
(3). liquid of extracting polysaccharide adopts ultrafiltration and concentration to finite concentration;
(4). the liquid of extracting polysaccharide after concentrating adds flocculation agent and the electric field-enhanced flocculation sediment that carries out, and perhaps carries out ethanol sedimentation, the centrifugation taking precipitate;
(5). longan, the forced air drying of lichee polysaccharide throw out are got polysaccharide crude;
(6). slightly carry in polysaccharide concentrated solution or the polysaccharide crude lysate at longan, lichee and to add ethanol and carry out fractionation precipitation, precipitation is got in centrifugation, reclaims ethanol; Perhaps carry out column chromatography for separation, the elutriant of getting simple spike also concentrates, and gets the pure product of polysaccharide after the lyophilize.
2. by the described method of claim 1, it is characterized in that: ultrasound-enhanced extraction conditions is: ultrasonic power is 100w-1800w, dutycycle is 10%-90%, temperature is 10-60 ℃, liquid-solid ratio is 2: 1-50: 1 (v/w, volume/weight), pulp making beating fineness is the 20-200 order, extraction time 5-50min.
3. by the described method of claim 1, it is characterized in that: used retaining molecular weight is 5000 during ultrafiltration and concentration, and the polysaccharide concentration of concentrated solution is 5-50mg/ml.
4. by the described method of claim 1, it is characterized in that: used flocculation agent is chitosan, polystyrene sodium sulfonate, gelatin, tannin etc., flocculation agent consumption 0-1000ppm; Used electric field comprises the AC and DC electric field, and strength of electric field is 0-100V/cm, and 0-60 minute electric field action time, temperature is 20-60 ℃; Stirring velocity is 100-1000rpm, and churning time is 5-60 minute; The pH value is 4-11; Alcohol concn is 30-90% during precipitation, and centrifugal speed is 2000-5000r/min.
5. by the described method of claim 1, it is characterized in that: the forced air drying temperature is 30-80 ℃; The vacuum freezedrying temperature is-50 ℃-30 ℃.
6. by the described method of claim 1, it is characterized in that: the chromatography column material therefor is a gel column, as the CM-dextran, and DEAE-dextran, or absorption resinoid, as AB-8, X-5.
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CN104292354A (en) * | 2014-11-11 | 2015-01-21 | 济南凯因生物科技有限公司 | Extraction method of litchi polysaccharide |
CN105601759A (en) * | 2016-01-07 | 2016-05-25 | 广西医科大学 | Acetylated longan pulp polysaccharide and application of acetylated longan pulp polysaccharide to medicine preparing |
CN110302163A (en) * | 2019-07-23 | 2019-10-08 | 东莞正大康地饲料有限公司 | A kind of florfenicol soluble powder and preparation method thereof |
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CN101838339A (en) * | 2010-04-29 | 2010-09-22 | 黑龙江省科学院大庆分院 | Method for extracting flaxseed gum by utilizing ultrasonic waves |
CN103012615A (en) * | 2013-01-08 | 2013-04-03 | 广东海洋大学 | Method for efficiently extracting sepia acidic polysaccharose |
CN103012615B (en) * | 2013-01-08 | 2015-02-25 | 广东海洋大学 | Method for efficiently extracting sepia acidic polysaccharose |
CN104292354A (en) * | 2014-11-11 | 2015-01-21 | 济南凯因生物科技有限公司 | Extraction method of litchi polysaccharide |
CN105601759A (en) * | 2016-01-07 | 2016-05-25 | 广西医科大学 | Acetylated longan pulp polysaccharide and application of acetylated longan pulp polysaccharide to medicine preparing |
CN105601759B (en) * | 2016-01-07 | 2018-01-19 | 广西医科大学 | A kind of acetylation Arillus longan polysaccharide and its application in pharmacy |
CN110302163A (en) * | 2019-07-23 | 2019-10-08 | 东莞正大康地饲料有限公司 | A kind of florfenicol soluble powder and preparation method thereof |
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