CN104592329A - Novel method for preparing high-purity rutin - Google Patents

Abstract

The invention relates to a novel method for preparing high-purity rutin. The method sequentially comprises the following steps: (1) dissolving rutin NF11 as a raw material to obtain a dissolving solution; (2) filtering the dissolving solution to obtain a filtrate; (3) concentrating the filtrate; (4) extracting and removing impurity; (5) centrifuging and rinsing; and (6) drying. The impurity in rutin is only removed by virtue of a special method, the disadvantage that mother liquor always contains partial rutin due to repeated crystallization in the traditional process is avoided and the limitations that methods reported in other literatures are very high in cost, the industrialization cannot be achieved and only small samples can be prepared in a laboratory are overcome. The method has the characteristics of low cost, high content, high yield, simplicity in operation, easiness in industrialization and the like.

Description

A kind of high-purity rutoside novel preparation method

Technical field

The present invention relates to a kind of high-purity rutoside novel preparation method.

Background technology

Rutin is the carbohydrate ligands of the flavonol be extensively present in plant, two glycosides are glucose and rhamnosyl, for faint yellow or light green needle crystal or crystalline powder, under normal circumstances containing three crystal water, in 95 ~ 97 DEG C of situations, drying can lose two crystal water, in 110 DEG C of vacuum (1.33 × 10 ³pa) under condition, dry 12h becomes anhydride, and anhydride becomes brown at 125 DEG C, and 195 ~ 197 DEG C become snappiness plastic, 214 ~ 215 DEG C of decomposition, meet light color and turn dark, have bitter taste, fusing point 190 ~ 192 DEG C, the water of Absorbable rod three molecule becomes three water rutins in an atmosphere.The easy moisture absorption of anhydride, different name violaguercitrin, vitamin P, rutin.1g is dissolved in 7mL and boils methyl alcohol, 8000mL water, 200mL boiling water.Be dissolved in pyridine, methane amide, basic solution, be slightly soluble in ethanol, acetone, ethyl acetate, be dissolved in chloroform, dithiocarbonic anhydride, ether, benzene, ethereal solution hardly.Dilute solution meets iron(ic) chloride in green, in strongly-acid solvent, separate out yellow mercury oxide.Rutin has multiple biological activity, as antioxygenation; Sun-proof, absorb action of ultraviolet radiation; Improve motor capacity; The protective effects on gastric mucosa; Osteoporosis; Anti-aging effects; Reduce blood sugar effect; Pancreatitis effect; To the provide protection of ischemic organ; Inhibiting tumor cell effect; Antivirus action; Anti-inflammatory, V _psample effect; Analgesic activity; For preventing and treating Intracerebral hemorrhage, hypertension, retinal hemorrhage, purple scar and acute hemorrhagic ephritis; Treatment chronic tracheitis, efficient 84.8% and 98%.The derivative of rutin mainly contains: study of rutin complex, Quercetin, rhamnosyl, troxerutin, rutin sulfuric ester and dihydroquercetin etc.The novel form of rutin, mainly contains: rutin cyclodextrin inclusion compound, rutin effervesce particle, rutin solid dispersion system, rutin HPMC controlled release tablet, co-precipitates of rutin, rutin mixture etc.Rutin has a wide range of applications in fields such as medicine, daily use chemicals, food, has become the focus of research in recent years.

Rutin extraction method has following several: (1) hot water extraction method Chen You plum reports sophora bud meal, and with boiling water boiling 2 times, each 10min water liquid lets cool, and separate out precipitation, hot water recrystallization obtains rutin.This method utilizes rutin to dissolve in boiling water, lets cool the feature that postprecipitation goes out, and due to rutin, solubleness is very little in the hot water, but this method water consumption all too is large, has no actual utility value.(2) methanol extraction Sun Wenji etc. report sophora bud meal and extract twice with methanol eddy, and filtrate merges concentrated, filter, obtain crystallization.Mother liquor again thickening filtration is precipitated, and merges precipitation with washed with diethylether, obtains crystallization.Methyl alcohol is comparatively wide in extract by solvents application, the composition of low-pole and stronger polarity can be extracted simultaneously, but pigment content is larger.Rutin is weak is dissolved in methyl alcohol, and methanol eddy method can be adopted to extract, but GAMMA-Carotene, chlorophyll content has regulation in NF11 version, and the rutin look valency that this method is extracted is high, and cost is high too, and treatment capacity is little, cumbersome, and domestic only have few producer to apply at present.(3) alkaline ethanol extraction method Lee family spring etc. report sophora bud meal with 95% ethanol add liming borax adjust pH8.5, backflow 30min, the dregs of a decoction extract according to upper method second time, and extracted twice liquid merges concentrated, concentrated solution adds hydrochloric acid and adjusts pH3 to be precipitated, and drying obtains crude product rutin.Crude product rutin adds water heating, adds liming and adjusts pH8 to dissolve, insulation 15min, adds phosphoric acid and adjusts pH6.5 to be incubated 45min, filter, obtain filtrate and precipitation, precipitate to wash, water liquid and above filtrate merge, add hydrochloric acid and adjust pH3, at 40 DEG C, leave standstill 8h, precipitation washing obtains fine work rutin.The character that this method Extraction parts utilizes rutin to precipitate under easy molten acidity under alkalescence is by Extraction of rutin.(4) thermokalite water extraction method Yang Hong etc. report sophora bud meal with pH8 ~ 9 buck and boil, and alkaline solution adjusts pH2 ~ 3 suction filtration, and precipitation heating water dissolves, suction filtration, and filtrate cools, and filters, precipitates to wash, dry to obtain rutin.This method is extracted with heat alkali liquid, and yield is high, but impurity is also large, the flavones parent nucleus destructible of rutin, refining also improper with hot water.(5) microwave loss mechanisms Li Min etc. report with Extraction of rutin rate for evaluation index, adopt the continuous extraction equipment of pipeline type microwave, with 10 times amount (v/w) for extraction agent, extract 5min at 65 DEG C, methanol concentration 90%, decreases environmental pollution, but cost is high, treatment capacity is little.Bibliographical information is with microwave power 320W, and alcohol concn 65%, solid-liquid ratio 1: 18 (w/v), microwave time 4min, extraction yield, less than 19%, is not suitable for industrial production.In traditional leaching process, the mode of carrying out due to energy accumulation and process of osmosis is irregular, and extract selectivity very poor, microwave extraction can heat different components selectivity, has good selectivity.The alternative extraction solvent of microwave extraction is many, extraction can be got liquid and be heated to more than atmospheric boiling point in the short period, improve the boiling point of solvent, and can not decompose effective constituent, shorten extraction and get the time, improve extraction and get efficiency.(6) extraction with ultrasonic-NaOH gets the heavy method Guo of acid is that girl etc. reports and boils extraction with soda-lime, and the ultrasonic micro-0.5h that boils of pH9 adjusted by milk of lime, is acidified to pH4 at filtrate and 70 ~ 75 DEG C, stir evenly, leave standstill, wash acid, drying, extraction yield 17.83%, this method laboratory can be done.Supersound extraction utilizes solvent and extracts between sample and produces sound wave cavatition, and ultrasonic wave, at the alternating action of positive phase and minus phase, can make bubble formation in solution, increase, explosion, increase the contact area of solvent and solid, thus raising rate of mass transfer, improve extraction efficiency, ultrasonic wave can be utilized clean in reality, sterilization, atomization, nondestructive testing, dry etc., be a kind of widely used mature technology.(7) cable-styled method alkali is carried the heavy Yang Xin river of acid etc. and is reported employing soxhlet extraction, by sophora flower powder to 20 ~ 40 order, and the borax water liquid of 12 times amount (v/w) 3%, with saturated lime water bar pH9, temperature 95 ~ 100 DEG C, extracts 30min, extracts 2 times, extracting solution adjusts pH3 with hydrochloric acid, leave standstill 4h, treatment capacity is large, easy to operate, cost is low, is applicable to industrial production.(8) supercritical CO ₂extraction process Jiang Zhong is beautiful reports employing supercritical CO ₂extraction process, with the preferred processing parameter of orthogonal experiment, Radix Et Rhizoma Fagopyri Tatarici moisture 3%, extracting pressure 30Mpa, temperature 35 DEG C, time 80min, rutin yield 1.3%.CO ₂have the double properties of gas and liquid under overcritical, material has the solvability of height in this fluid, CO ₂middle entrainment agent can the polarity of alter, and by opposed polarity, different boiling, the composition of different molecular weight extracts successively, reaches the object of separation and purification.CO ₂extraction process is a kind of new technology of development in recent years, and have no solvent residue, extraction yield is high, the survivable characteristic of target component, but due to apparatus expensive, treatment capacity is little, cleaning trouble.(9) aqueous two-phase system poplar Qinghai etc. report the double-aqueous phase system extraction rutin formed with hydrophilic ionic-liquid Tetrafluoroboric acid 1-butyl-3-Methylimidazole and ammonium sulfate, result display ionic liquid 1.5 ~ 2mL, the double-aqueous phase system that ammonium sulfate 1.5g is formed is better, add rutin 1.0 ~ 1.5mL, pH controls 4 ~ 6, and percentage extraction reaches 90%.Preparation double water-phase solution system, utilizes each composition solubleness in double water-phase solution system in thing to be extracted to vary in size and obtains, be a kind of novel liquid-liquid separation technology, be used successfully to protein at present, the separation of microbiotic and traditional Chinese medicine ingredients.(10) macroreticular resin absorbing method Wen Puhong etc. report and are separated rutin in Radix Et Rhizoma Fagopyri Tatarici with D16 macroporous resin, pulverized by Radix Et Rhizoma Fagopyri Tatarici, extraction using alcohol, extracting solution concentrates, filter, filtrate is with D16 resin absorption, first with washing, then with 70% ethanol elution, collect elutriant, concentrated, dry, rutin purity is 38.76%.This method is mainly for separating of the rutin in the lower Radix Et Rhizoma Fagopyri Tatarici of content, and its purity is not high yet, also needs further separation and purification.Macroporous resin adsorption isolation technique is a kind of novel process utilizing sorbent material target component absorption in Chinese medicine aqueous extract invalid components to be separated.This method have equipment simple, easy to operate, save the energy, cost is low, product purity is high, the advantage such as nonhygroscopic, therefore macroreticular resin absorbing method traditional Chinese medicine research and the application in producing increasingly extensive.The separation this method being applied to Effective Component of Chinese Medicine achieves quite significant achievement.In recent years, Flavonoids by Macroporous Adsorption Resin has been widely used in the separation of Effective Component of Chinese Medicine with refining.(11) sophora bud is pulverized by the superfine report of enzymolysis process extraction Gong Sheng, and add water mixing, adds enzyme liquid, suction filtration, concentrated.Optimised process is hydrolysis temperature 45 DEG C, pH4.5, and cellulase add-on is 0.03%, enzymolysis time 2h, and solid-liquid ratio is 1: 55.Cellulase can make the main component cellulose degradation of plant cell wall, destroys cell walls, is conducive to composition and leaches, improve yield.(12) tensio-active agent is worked in coordination with super the reporting of extraction method Gong Sheng and is worked in coordination with extraction with tensio-active agent, and yield reaches 17.5%, purity 92.5%, can shorten extraction and sedimentation time.Tensio-active agent is the molecule formed by two kinds of distinct particles, particle has an extremely strong lipophilicity, another kind of then have extremely strong wetting ability, after being dissolved in the water, tensio-active agent can reduce the surface tension of water, and improves the solubility of organic compound.(13) alkaline methanol refluxing extraction Wen Puhong etc. report in the sophora bud meal borax water liquid adding 1.2 times amount (w/ww) 0.4%, place 24h, add the methyl alcohol of 2 ~ 5 times amount (w/w) again, and adjusting pH8 ~ 9 with milk of lime, backflow 20min, extracts 2 times, filtrate adjusts pH3 with hydrochloric acid, crystallization, yield is more than 20%, and content reaches 97%.This method yield is high, and cost is low, is applicable to the refining of industrial production, particularly rutin.(14) sequential extractions Zhang Chaoyan etc. report after sophora bud sodium bicarbonate and the process of alkyl phenol ethylene oxide affixture, and enzyme is killed in boiling, and with limewater extraction, yield can reach 22%, content 98%, and especially residue reduces 77.2% than original technique.The simplest yield of sequential extractions operation is high, and cost is low, is suitable for industrial production.Trochol in the sophora bud, sophoradiol, Flos Sophorae Immaturus A prime, Flos Sophorae Immaturus B prime, Flos Sophorae Immaturus C prime, without phenolic hydroxyl group, is insoluble to alkali lye.Rutin and rue enzyme coexist in the sophora bud, rue enzyme is the protein with specific catalytic effect, high temperature can make its inactivation, thus avoid the hydrolysis of rutin in leaching process, rutin has 4 phenolic hydroxyl groups and shows slightly acidic, the saturated limewater stronger by alkalescence makes solvent, be conducive to rutin salify to dissolve, also can remove a large amount of polysaccharide slime formers matter contained in the sophora bud, but in the stronger situation of alkalescence, 2 phenolic hydroxyl groups at ortho position are easily oxidized, under highly basic, particularly heat calcium can form inner complex with rutin and separate out precipitation, PH is too low simultaneously makes rutin form salt of pretending, thus product yield is reduced.

Rutin process for purification has following several: (1) dextrane gel method Zhao Wen is refined etc. reports dextrane gel method and utilize molecular size to be separated, molecule large due to dextrane gel inside can not be entered, thus first by wash-out, little rear by wash-out of molecule, thus compositions different for molecular size is separated, by rutin with after dissolve with methanol, add dextrane gel capital, with methanol-eluted fractions, collect methanol solution, concentrated, crystallization.(2) alkali extraction-acid precipitation Gong Sheng is superfine reports thick rutin and adds suitable quantity of water, dissolves, filter with liming, and filtrate, with hydrochloric acid acid adjustment, leaves standstill, and filter, obtain refined rutin, purity reaches 99%.The actual slightly acidic of rutin that utilizes of this method is dissolved in alkali lye, the character of Precipitation again after acidifying, but some compositions similar to rutin character still cannot remove.(3) Li Jiayong etc. report crude product rutin with liming heating for dissolving, then heating and thermal insulation, drip acid solution, and insulation is filtered, the acid adjustment of filtrate acid solution, leaves standstill, and separates out the refined rutin of precipitation.This fado one step Acid precipitation, the rutin impurity obtained is few, content is high.(4) the thick congruence of the molecular conformation transformation prince of Dharma reports utilization at room temperature with alcohol dissolving rutin, and filter, filtrate places crystallization, and crystallization again after mother liquor concentrations, obtains high-content rutin.This method utilizes and changes the two-dimension plane structure of rutin into 3-D solid structure, is conducive to solvent molecule and enters, and increases solvability, dissolves in some impurity of alcohol after can removing heating.(5) report such as Ferric Ammonium Citrate method of purification horse mandate is by molten for crude product rutin with Ferric Ammonium Citrate, filters, lets cool, acidifying, filters, water washing and precipitating, dry fine work rutin.This method utilizes adjacent diphenol hydroxyl polymeric in Ferric Ammonium Citrate and rutin molecule, makes the molecular structure of rutin become heterocycle shape from plane, and molecular gap increases, and in water, solubleness improves.

Take a broad view of domestic and foreign literature, the method extracting rutin is a lot, and there is extraction in current China, it extracts principle: because containing phenolic hydroxyl group in the structure of rutin, with soluble in water after alkali salify, in this salts solution, add acid, regulator solution pH value, then rutin is free again separates out.The extraction of current rutin is described above, this invention with NF11 version rutin for raw material, the impurity in rutin is only removed by special methods, there is not traditional technology periodic crystallisation, shortcoming all the time containing part rutin in mother liquor, or the method for other bibliographical informations cannot the problem of industrialization, can only make small sample, but cost is very high in laboratory.It is low that this invention has cost, and content is high, and yield is high, easy and simple to handle, is easy to the features such as industrialization.

Summary of the invention

The dissolving of the one NF11 version rutin.

The filtration of the second lysate.

3rd filtrate concentrates.

4th extracts removal of impurities.

5th centrifugal rinsing.

6th is dry.

Embodiment

Mode one: add dimethyl formamide 700kg in 1000L reactor, drops into the NF11 version rutin 100kg (HPLC content 92.8%) of accurate weighing in reactor, heats while stirring, and is heated to 80 ~ 100 DEG C and dissolves 0.5h.Filtered while hot removing insolubles and mechanicalness impurity, collect filtrate.Concentrating under reduced pressure filtrate, vacuum tightness is-(0.092 ~ 0.094) MPa, and temperature is 60 ~ 80 DEG C, and cryoconcentration after first high temperature is concentrated, is concentrated into proportion 1.20 ~ 1.30, obtains enriched material.In enriched material, add reagent dehydrated alcohol 300kg, be heated to 60 ~ 70 DEG C, stir uniformly scattered paste shape, heating 1h, centrifugal, stir 30 ~ 60min.Scattered paste shape thing is with centrifuge, and centrifugate reddish-brown, again with ethanol rinse after centrifuge dripping, until centrifugate is limpid.Vacuum-drying, loads drip pan by fine work rutin, controls evenly, thickness 1 ~ 2cm, sends in microwave dryer, first dry 0.5h under 50 DEG C of low temperature, then is warming up to 70 DEG C of dry 0.5h, obtains fine work rutin 92.5kg, content 99.2%

Mode two: add dimethyl sulfoxide (DMSO) 350kg in 500L reactor, adds the NF11 version rutin 50kg (HPLC content 92.8%) of precise under stirring, be heated to 60 ~ 100 DEG C and dissolve 0.5h.Filtered while hot removing insolubles and mechanicalness impurity, collect rutin and dissolve filtrate.Vacuum tightness is-(0.090 ~ 0.092) MPa under concentrating under reduced pressure, temperature is 40 ~ 80 DEG C, and cryoconcentration after first high temperature is concentrated, is concentrated into proportion 1.25 ~ 1.3, obtains rutin enriched material.In enriched material, add 200kg ethyl acetate, be heated to 60 ~ 70 DEG C and fully stir 30 ~ 60min, stir uniformly scattered paste shape.Scattered paste shape thing is with centrifuge, and centrifugate is yellow, again with ethanol rinse after centrifuge dripping, until centrifugate is limpid.Obtain jonquilleous rutin, vacuum-drying, fine work rutin is loaded drip pan, control evenly, thickness 1 ~ 2cm, send in Vacuumdrier, first dry 3.0h under 50 DEG C of low temperature, then be warming up to 80 DEG C of dry 2.0h.Obtain fine work rutin 46.3kg, content 98.5%

Mode three: add morpholine 640kg in 1000L reactor, by the NF11 version rutin 80kg (HPLC content 92.8%) of accurate weighing under stirring, heats while stirring, and is heated to 8 ~ 100 DEG C and dissolves 0.5h.Filtered while hot removing insolubles and mechanicalness impurity, collect rutin filtrate.Concentrating under reduced pressure filtrate, vacuum tightness is-(0.093 ~ 0.096) MPa, and temperature is 50 ~ 80 DEG C, and cryoconcentration after first high temperature is concentrated, is concentrated into proportion 1.20 ~ 1.25, obtains rutin enriched material.320kg acetone is added in enriched material, fully stirs, be heated to 60 ~ 70 DEG C, stir uniformly scattered paste shape, stir 30 ~ 60min.Scattered paste shape thing is with centrifuge, and centrifugate reddish-brown, again with ethanol rinse after centrifuge dripping, until centrifugate is limpid.Vacuum-drying, loads drip pan by fine work rutin, controls evenly, thickness 1 ~ 2cm, sends in Vacuumdrier, first dry 2.0h under 60 DEG C of low temperature, then is warming up to 80 DEG C of dry 3.0h, obtains fine work rutin 71.8kg, content 99.6%

Mode four: add 250kg pyridine in 500L reactor, drops into the NF11 version rutin 50kg (HPLC content 92.8%) of accurate weighing in reactor, is heated to 70 ~ 100 DEG C and dissolves 1.0h.Filtered while hot removing insolubles and mechanicalness impurity, collect filtrate.Concentrating under reduced pressure filtrate, vacuum tightness is-(0.090 ~ 0.096) MPa, and temperature is 60 ~ 90 DEG C, and cryoconcentration after first high temperature is concentrated, is concentrated into proportion 1.15 ~ 1.20, obtains rutin enriched material.In enriched material, add 200kg trichloromethane, be heated to 40 ~ 50 DEG C, fully stir, stir uniformly scattered paste shape.Scattered paste shape thing is with centrifuge, and centrifugate is yellow, again with ethanol rinse after centrifuge dripping, until centrifugate is limpid.Vacuum-drying, loads drip pan by fine work rutin, controls evenly, thickness 1 ~ 2cm, sends in Vacuumdrier, first dry 3.0h under 50 DEG C of low temperature, then is warming up to 80 DEG C of dry 2.0h.Obtain fine work rutin 46.8kg, content 98.2%

Mode five: accurate weighing 50kg adds sulfur oxychloride 280kg in 1000L reactor, by the NF11 version rutin 50kg (HPLC content 92.8%) of accurate weighing, is heated to 60 ~ 100 DEG C and dissolves 1h.Filtered while hot removing insolubles and mechanicalness impurity, collect filtrate.Concentrating under reduced pressure filtrate, vacuum tightness is-(0.092 ~ 0.094) MPa, and temperature is 40 ~ 80 DEG C, and cryoconcentration after first high temperature is concentrated, is concentrated into proportion 1.15 ~ 1.20, obtains rutin enriched material.In enriched material, add dithiocarbonic anhydride 200kg, be heated to 60 ~ 70 DEG C, fully stir, stir uniformly scattered paste shape, stir 30 ~ 60min.Scattered paste shape thing is with centrifuge, and centrifugate is yellow, uses ethanol rinse again, until centrifugate is limpid after centrifuge dripping.Vacuum-drying, loads drip pan by fine work rutin, controls evenly, thickness 1 ~ 2cm, sends in Vacuumdrier, first dry 3.0h under 50 DEG C of low temperature, then is warming up to 80 DEG C of dry 2.0h.Obtain fine work rutin 45.8kg, content 98.6%.

Claims (7)

1. a high-purity rutoside novel preparation method, is characterized in that, with NF11 version rutin for raw material, the method is successively through comprising the following steps:

(1) dissolving of NF11 version rutin.

(2) filtration of rutin lysate.

(3) concentrating under reduced pressure of rutin filtrate.

(4) the extraction removal of impurities of rutin enriched material.

(5) the rutin centrifugal rinsing after removal of impurities.

(6) drying of fine work rutin.

2. a kind of high-purity rutoside novel preparation method according to right 1, it is characterized in that: one of in step (1) middle dimethyl formamide, dimethyl sulfoxide (DMSO), morpholine, pyridine, sulfur oxychloride and piperidines or mixed solution, consumption is 5 ~ 10 times (V/W) of rutin amount, is heated to 60 ~ 100 DEG C and dissolves 1h.

3. a kind of high-purity rutoside novel preparation method according to right 1 or 2, is characterized in that: filtered while hot removing insolubles and mechanicalness impurity in step (2), collects filtrate.

4. a kind of high-purity rutoside novel preparation method according to right 1 or 2, it is characterized in that: concentrating under reduced pressure in step (3), vacuum tightness is-(0.092 ~ 0.094) MPa, temperature is 40 ~ 80 DEG C, cryoconcentration after first high temperature is concentrated, be concentrated into proportion 1.15 ~ 1.30, obtain enriched material.

5. a kind of high-purity rutoside novel preparation method according to right 1 or 2, it is characterized in that: one of to add in enriched material in step (4) in reagent alcohol, ethyl acetate, acetone, methylene dichloride, trichloromethane, dithiocarbonic anhydride, ether, benzene, abundant stirring, be heated to 60 ~ 70 DEG C, stir uniformly scattered paste shape, reagent dosage is 1 ~ 4 times amount (W/W) of rutin amount, stirs 30 ~ 60min.

6. a kind of high-purity rutoside novel preparation method according to right 1, it is characterized in that: in step (5), scattered paste shape thing is with centrifuge, after centrifuge dripping, no matter solvent for use is different when dissolving, centrifugal rinsing is all with ethanol rinse, until centrifugate is limpid.

7. a kind of high-purity rutoside novel preparation method according to right 1, it is characterized in that: with microwave drying or vacuum-drying in step (6), fine work rutin is loaded drip pan, control evenly, thickness 1 ~ 2cm, send in Vacuumdrier, first dry 1.0h under 50 DEG C of low temperature, then be warming up to 80 DEG C of dry 0.5h.