CN101670010A - Method for extracting flavonoid glycoside substance - Google Patents
Method for extracting flavonoid glycoside substance Download PDFInfo
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- CN101670010A CN101670010A CN200910308171A CN200910308171A CN101670010A CN 101670010 A CN101670010 A CN 101670010A CN 200910308171 A CN200910308171 A CN 200910308171A CN 200910308171 A CN200910308171 A CN 200910308171A CN 101670010 A CN101670010 A CN 101670010A
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Abstract
The invention provides a method for extracting flavonoid glycoside substance, comprising the following steps of: taking orange pericarp removed with impurities and dry powder of persimmon leaf as rawmaterials, and extracting with 20-80% alcohol; setting aside, precipitating, taking supernatant, adding diatomite that is 1-5% of supernatant in terms of mass, fully stirring, pressing and filtering by a plate filter, membrane-filtering the filtrate B with PES membrane material, and taking the micro-molecular solution passing through the membrane; putting the obtained micro-molecular solution intoa film evaporator to remove impurities by molecular distillation, collecting the required distillate, performing spray drying, and making into powder to obtain the flavonoid glycoside substance. Themethod for extracting flavonoid glycoside substance has quite simple and convenient operation and low cost, and can save a large amount of solvent, reduce the requirements on the device, also reduce pollution on the environment and well guarantee the natural quality of the flavonoid glycoside substance; the content of flavonoid glycoside component in the produced flavonoid extract is high with a total content of more than 70%, and the transfer rate of general flavone can reach more than 95% with less residual pigment.
Description
Technical field
The invention belongs to the extraction separation purification process of natural medicinal ingredients bioflavonoids, be particularly related to a kind of from plant Ebenaceae Diospyros plant persimmon (Diospyros kaki Thunb) leaf and the peel of the drying and ripening of rutaceae orange (Citrus reticulataBlanco) and variety thereof the new technology of separation and Extraction flavonoid glycoside substance, the present invention not only relates to the water ethanol extraction, also relates to the new process of separating the purification flavonoid glycoside substance with kieselguhr filtering, ultrafiltration, with molecular distillation technique.
Background technology
Bioflavonoids (bioflavonoids) is the aldehydes matter that occurring in nature extensively exists, and also claims Citrin, and normal and vitamin C companion deposits, and is the secondary metabolite of plant.Flavonoid is the derivant of chromanane or chromone, and its basic framework has C
6-C
3-C
6Characteristics, promptly by two aromatic rings A and B, a series of chemical compounds that link mutually by central three carbochains.Whether degree of oxidation, link position and three carbochains of B ring on the C ring according to central three carbochains constitute characteristics such as ring-type, flavonoid is divided into flavone and flavonoid glycoside, flavonol and flavonol glycoside, chalcone derivative, aurone aurones class, anthocyanidin and anthocyanin, flavonol, flavanone, flavanonol, carbon-glycoside flavone, bisflavone and dimeric flavones, isoflavone, isoflavanone, mouthful diphenylene ketone oxide class or claims benzo chromone and neoflavone 14 big classes, and sum has more than 2700 approximately.Flavonoid distributes very general in plant kingdom, seldom find in algae, mushroom.
Flavonoid has the various biological function, be mainly: (1) is to the antioxidative regulating action, reported by antioxidation in vitro experiment as single poplar (2007) five kinds of monomeric anti-oxidation functions of polymethoxyflavone in the peel of Citrus reticulata Blanco have been studied, show that five kinds of flavone monomers all have certain anti-linoleic acid oxidability in the peel of Citrus reticulata Blanco, and all be better than rutin; The flavonoid monomer suppresses the liposome oxidability and all shows certain concentration dependence, removes OH free radical ability and respective concentration and also has positive correlation.(2) to the regulating action of lipid metabolism, as woods Depositum urinae preparatum (2000) report, crataegutt can make the mRNA of rat low density lipoprotein receptor (LDLR) transcribe enhancing, and LDLR expresses also and strengthens simultaneously, thereby increases the degraded of low density lipoprotein, LDL (LDL).Lichtenstein (1998) thinks that the soybean prod prevention of arterial is atherosis, the effect of coronary heart disease is owing to Semen sojae atricolor flavonoid etc.(3) inhibitory anti-virus effect, reported the flavonoid material that has bacteriostasis in the citrus peel residue crude extract as Zhang Lijie (2009), staphylococcus aureus, escherichia coli and bacillus subtilis are had good bacteriostasis, and the diameter of inhibition zone is 5.9,5.7 respectively, 6.0mm.Propose among the Yan Fenghai (application number 98113018.6), extract flavonoid glycoside and make pharmacology and bacteriostatic test from leaf of Diospyros kaki L.f., the result shows it with Armeniaca mume Sieb. element and metronidazole effect basically identical, and it has certain curative effect to hematoblastic minimizing and nerve insomnia simultaneously.(4) to endocrine influence, can promote the recovery of beta Cell of islet as Nielsen (1998), Shen (1993) report flavonoid, blood sugar lowering and serum cholesterol, improve carbohydrate tolerance, to adrenolytic blood glucose increasing effect, it can also suppress aldose reductase simultaneously, can treat diabetes and complication thereof.Rbards and Antolovich (1997) research points out that flavonoid can be used for treating diseases such as osteopathia and osteoporosis, its mechanism of action is that it both can suppress the collagen protein synthesis increase of PGE2 (PGE2), the synthetic minimizing of noncollagen protein that can suppress PGE2 again, and when low concentration PGE2, mainly act on the synthetic of noncollagen protein, mainly act on collagen protein synthesis during high concentration PGE2, can be used for the treatment of osteopathia; And can improve thyroid to estrogenic sensitivity, the effect of parafollicular cells of thyroid gland secretion calcitonin is strengthened, finally suppress bone resorption and treat osteoporosis.(5) to the influence of breeding performonce fo animals, give rat skin lower injection daidzein 16 days as (1995) such as the outstanding persons of kingdom, RNA/DNA ratio improves 18.7%, the short muscle growth effect that shows daidzein mainly is the loose and protein anabolism reinforcement of myofibrillar trophism, rather than quickens the result of satellite cell breeding.(6) antitumaous effect has reported that as M Noroozi (1998) the hexa methoxy flavone in the peel of Citrus reticulata Blanco has the platelet aggregation of inhibition, antithrombotic effect, and Heptamethoxyflavone all has stronger tumor-suppression activity etc. to stomach cancer cell and leukaemia.
Although the flavonoid in the natural plants has the various biological function, also there is potential side effect.In plant and plant eater's common evolution, plant has been set up Chemical Defence System to resist plant eater's infringement.In the chemoresistance factor of plant, secondary metabolites confrontation plant eater's resistance mainly contains the effect aspect antixenosis (even host plant has non preference or hinders feeding effect) and the antibiosis (promptly digest and assimilate the plant eater and then play the endotoxin lethality effect or the effect that physiological activity suppresses its growth promoter such as digest and assimilate by reduction) two.The antixenosis of flavonoid and antibiosis confirm by many research work and practical application, result of study show the intravital low-molecular-weight flavonoid of plant because of with the combination of proteins ability a little less than, do not show tangible anti-oxidant action.The flavonoid that molecular weight is too big (mainly refer to the flavanol compound flavone that the degree of polymerization is higher, its molecular weight is more than 3000) is because poorly soluble, be difficult for degraded in animal body, can the nutritive value of plant not exerted an influence yet, can produce remarkable influence (Bravo etc., 1993).
Based on above these effects, the separation and purification of flavone compound is particularly important.The isolation and purification method of flavone compound is a lot, column chromatography, thin layer chromatography, lead salt precipitation, boric acid complexation, the extraction of pH gradient, solvent extraction and the high performance liquid chromatography of using in recent years (HPLC), droplet countercurrent chromatography (DCCC), gas chromatography etc. are arranged, but all there is shortcoming in various degree and limited its suitability for industrialized production, can be used for industrialized supercritical CO
2Extraction is suitable for the last stage of separation process, and adsorption resin method will reach refining to product, and the step that needs is numerous, is used for the purification of flavonoid, and is all undesirable.Molecular distillation (molecular distillation) technology is the non-equilibrium distillation of carrying out under high vacuum condition, can solve the problem that the wide variety of conventional distillation technique cann't be solved, be suitable for the composition of the high added value in the thick product being separated and purifying, and this separation and purification are that other separation means commonly used are difficult to finish, the extraction that is particularly suitable for natural materials with separate, be successfully applied to industries such as petrochemical industry, food, plastics, medicine at present, but also be only limited to the use of liposoluble substance.
Molecular distillation is compared with conventional distillation technique in the application of actual industrialization has following remarkable advantages: operate under the temperature far below the material boiling point (1), and residence time of material is short, separation for high boiling point, temperature-sensitive and readily oxidizable substance material provides the optimal separation method.(2) but the utmost point removes material such as organic solvent, the stink etc. in the liquid effectively, be unusual effective method for adopting behind the solvent extraction precipitation of liquid; (3) molecular distillation can steam the purpose product selectively, removes other impurity, can separate material more than 2 kinds simultaneously by multiple stage separation; (4) fractional distillation process of molecular distillation is a physical process, thereby can protect the not contaminated and infringement of separated material well.
Separation and purification goes out flavone compound from natural plants, not only will have broad application prospects in medicine, food processing industry, has made certain contribution at aspects such as environmental conservation, and is also significant to its physiologic function of further research.
Summary of the invention
The object of the invention be to seek a kind of more economical, from natural product, extract the method for flavones ingredient efficiently.
The technical solution used in the present invention is:
A kind of extracting method of flavonoid glycoside substance, described method is that the dry fragment of getting peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. is that raw material carries out as follows: (1) removes the peel of Citrus reticulata Blanco of impurity, the dried powder of leaf of Diospyros kaki L.f. is a raw material, (2) add 20～80% ethanol that quality is 10～20 times of peel of Citrus reticulata Blanco, the dry fragment quality of leaf of Diospyros kaki L.f. summation, 2～4h is extracted in 30～70 ℃ of insulations, centrifugal, filtration, get filtrate A, filtering residue repeats to extract 1～5 time again, merges each filtrate A that extracts and gets extracting solution; (3) step (2) gained extracting solution is staticly settled, get supernatant, adding quality is the kieselguhr of described supernatant 1～5%, after fully stirring, with the flame filter press filter pressing, uses the filter pressing of 3～10 layers of DANFU silk filter cloth usually, gets liquor B; DANFU silk filter cloth is called for short filter cloth in the present invention; (4) liquor B is carried out membrane filtration with the poly (ether sulfone) film material, molecular cut off 5kD gets the micromolecule solution of filter membrane; (5) get the micromolecule solution that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 1～3ml/min, knifing rotating speed at different levels is 100-300rpm, preferred knifing rotating speed at different levels is 100-200rpm, and circulating water temperatures at different levels are respectively done for oneself 30～70 ℃; The vacuum of controlling first distillator is 600～900Pa, vapo(u)rizing temperature is 40 ℃～60 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 100～300Pa, vapo(u)rizing temperature is 60 ℃～80 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 10～50Pa, vapo(u)rizing temperature is 80 ℃～110 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.01～2Pa, vapo(u)rizing temperature is 150 ℃～300 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～40 ℃ the condition, collects distillate; (6) distillate of Shou Jiing carries out spray drying and makes powdery, promptly described flavonoid glycoside substance.More than said filtrate A, liquor B be meant the filtrate in different process stage in the technical process, distinguish with A, B, the filtrate of repeating to extract gained in the step (2) also all is defined as filtrate A, though it is inequality to repeat the filtrate A that obtains at every turn, but belong to same processing step, be extracting solution after the merging.Membrane evaporator knifing rotating speed at different levels is roughly the same, preferably is controlled between 100～200rpm, and circulating water temperatures at different levels are respectively done for oneself 30～70 ℃; The preferred first distillator circulating water temperature is 30 ℃, and after-fractionating device circulating water temperature is 30 ℃, and the 3rd distillator circulating water temperature is 40 ℃, and the 4th distillator circulating water temperature is 70 ℃.
The present invention filters out from natural plants by external bacteriostatic experiment that to help extracting the leaf of Diospyros kaki L.f. and the peel of Citrus reticulata Blanco that obtain flavonoid glycoside substance be compound, the total extract that obtains by the water alcohol extraction, after adopting kieselguhr filtering, hyperfiltration technique to remove materials such as float class impurity, starch, half soluble cellulose, pectin, protein, chlorophyll, glycoprotein, lipid, adopt molecular distillation technology to obtain flavonoid glycoside substance again.The main points of invention are to adopt molecular distillation technology, and find suitable technological parameter to obtain to adopt small-molecular weight, purity height, stay-in-grade flavonoid glycoside substance.
The extracting method of further flavonoid glycoside substance of the present invention, the dry fragment of described peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. was preferably the dried powder of 20 mesh sieves.
The mass ratio of described peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. can be an arbitrary proportion, arbitrarily the peel of Citrus reticulata Blanco of mass ratio, leaf of Diospyros kaki L.f. extract with method of the present invention and prepare flavonoid glycoside substance and all reasonable effect can be arranged, but the present invention more preferably the mass ratio of peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. be 2: 3.
The extracting method of flavonoid glycoside substance of the present invention, the described sedimentation time is for precipitating 12～48 hours, preferred 12～24 hours.Described filter press has been adopted 3～10 metafiltration cloth filter pressings.
The vacuum that further step of the present invention (5) is preferably controlled first distillator is that 750～850Pa, vapo(u)rizing temperature are 40 ℃～50 ℃, and the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, and this step separates and removes gas and part ethanol; The vacuum of control after-fractionating device is that 150～250Pa, vapo(u)rizing temperature are 65 ℃～75 ℃, and the condition that the cryosurface temperature is 15～25 ℃ is carried out thin film evaporation, separates and removes ethanol and part water; The vacuum of controlling the 3rd distillator is that 20～40Pa, vapo(u)rizing temperature are 80 ℃～100 ℃, and the condition that the cryosurface temperature is 15～25 ℃ is carried out thin film evaporation, separates and removes most of water; The vacuum of controlling the 4th distillator is 0.01～0.5Pa, more preferably 0.01～0.1Pa, vapo(u)rizing temperature is 200 ℃～300 ℃, the cryosurface temperature is to carry out thin film evaporation under 20～30 ℃ the condition, collect distillate, stay excess this moment is high boiling point and the involatile constituent that is removed.
The distillate that spray drying described in the step of the present invention (6) recommends to get step (5) collection directly is used for carrying out spray drying powder process.
Concrete, the extracting method of flavonoid glycoside substance of the present invention carries out as follows: the mass ratio that (1) removes impurity is that 2: 3 peel of Citrus reticulata Blanco, the dry powder of leaf of Diospyros kaki L.f. are raw material; (2) add 55%～60% the ethanol that quality is 10～15 times of peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. dry powder quality summation, 2.5～3h are extracted in 50 ℃ of insulations, and are centrifugal, filter, filtrate A, filtering residue repeats to extract 2～3 times again, merges each filtrate A and gets extracting solution; (3) step (2) gained extracting solution was left standstill 12～24 hours, precipitation is got supernatant, and adding quality is the kieselguhr of described supernatant 2%, after fully stirring, with 6～8 metafiltration cloth filter pressings, gets liquor B with flame filter press; (4) liquor B is carried out membrane filtration with the poly (ether sulfone) film material, molecular cut off 5kD gets the micromolecule solution of filter membrane; (5) get the micromolecule solution that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 2～3ml/min, the vacuum of controlling first distillator is 750～850Pa, vapo(u)rizing temperature is 40 ℃～50 ℃, the cryosurface temperature is 15～25 ℃, circulating water temperature is 30～70 ℃, carry out thin film evaporation under the condition that the preferred cycle coolant-temperature gage is 30 ℃, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 150～250Pa, vapo(u)rizing temperature is 65 ℃～75 ℃, the cryosurface temperature is 15～25 ℃, circulating water temperature is 30～70 ℃, carry out thin film evaporation under the condition that the preferred cycle coolant-temperature gage is 30 ℃, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 20～40Pa, vapo(u)rizing temperature is 80 ℃～90 ℃, 15～25 ℃ of cryosurface temperature, circulating water temperature is 30～70 ℃, carry out thin film evaporation under the condition that the preferred cycle coolant-temperature gage is 40 ℃, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.01～0.05Pa, vapo(u)rizing temperature is 250 ℃～280 ℃, 30 ℃ of cryosurface temperature, knifing rotating speed 300rpm, circulating water temperature is 30～70 ℃, carry out thin film evaporation under 70 ℃ of conditions of preferred cycle coolant-temperature gage, collect distillate; (6) getting distillate that step (5) collects carries out spray drying and makes powdery, promptly described flavonoid glycoside substance.
Beneficial effect of the present invention is mainly reflected in:
1. the diatomite filtration clarification and the ultrafiltration technology that are adopted among the present invention have been removed materials such as the protein in the extracting solution, half soluble fiber, pectin, starch, chlorophyll, glycoprotein, lipid, have saved organic solvent extraction, and polyamide is refining; And membrane material physical and chemical stability height, mechanical strength is good, can repeatedly use; Technological operation is very easy, and is with low cost.
2. molecular distillation method of the present invention is pure physical separating process, can save a large amount of solvents, reduce requirement, reduce pollution simultaneously environment to equipment, and can not bring new impurity and noxious substance into, can guarantee the natural quality of flavonoid glycoside substance well; Though the initial stage one-time investment is bigger, follow-up production cost (comprising material, water power, labour force etc.) is smaller; Be applicable to large-scale industrial production.
3. flavonoid glycoside component content height in the flavone extract that goes out by explained hereafter of the present invention, total content is more than 70%, and the total flavones rate of transform can reach more than 95%, and color and luster is good.After existing skill is stated common employing alcohol extraction or supersound extraction, the flavonoid glycoside substance that acid-alkali treatment or multiple-stage solvent extraction are extracted from plant, but content is very low usually, generally have only about 40%, also have and adopt adsorption resin method to carry out purification, but will reach refining to product, the step that needs is cumbersome.
The specific embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1
(1). the preparation of raw material: clean Zhejiang Province respectively and produce exsiccant peel of Citrus reticulata Blanco, exsiccant leaf of Diospyros kaki L.f., remove impurity wherein, after the pulverizing, cross 20 mesh sieves, respectively peel of Citrus reticulata Blanco dry powder, leaf of Diospyros kaki L.f. powder are stand-by;
(2). the processing of raw material: in peel of Citrus reticulata Blanco dry powder and leaf of Diospyros kaki L.f. dry powder quality ratio is that 2: 3 ratios were got peel of Citrus reticulata Blanco dried powder 0.2kg, leaf of Diospyros kaki L.f. dried powder 0.3kg, add quality and be 60% ethanol water of 10 times of peel of Citrus reticulata Blanco dry powder and leaf of Diospyros kaki L.f. dry powder quality summations, temperature control is at 50 ℃～60 ℃, extract 3h, centrifugal, filtration get filtrate A, and filtering residue extracts 2 times by above method again, merging filtrate A gets extracting solution 7.2L;
(3). filter: step (2) gained extracting solution is staticly settled, and the kieselguhr 0.14kg with the supernatant 7.0L after staticly settling adds supernatant quality 2% to adsorb most of colloid, chlorophyll etc., uses the flame filter press filter pressing with its filtering then; (3) step (2) gained extracting solution is staticly settled, get supernatant, adding quality is the kieselguhr of described supernatant 2%, after fully stirring, with the flame filter press filter pressing, gets 8 layers of DANFU silk filter cloth during filter pressing, gets liquor B 6.2L;
(4). liquor B is carried out membrane filtration with poly (ether sulfone) film material (molecular weight 5kD, 95%), molecular cut off 5kD, and further remove tannin, peptide class etc., get the micromolecule solution 3.6L of filter membrane;
(5). get the micromolecule solution 3.6L that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 2ml/min, knifing rotating speed 100rpm at different levels, the vacuum of controlling first distillator is 800Pa, vapo(u)rizing temperature is 45 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove gas and part ethanol, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 200Pa, vapo(u)rizing temperature is 70 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove ethanol and part water, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 30Pa, vapo(u)rizing temperature is 90 ℃, 25 ℃ of cryosurface temperature, carry out thin film evaporation under the condition that circulating water temperature is 40 ℃, separate and remove most of water, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.02Pa, vapo(u)rizing temperature is 260 ℃, the cryosurface temperature is 30 ℃, carry out thin film evaporation under the condition that circulating water temperature is 70 ℃, collect distillate 0.26L; Stay excess this moment is high boiling point and the involatile constituent that is removed.
(6). get the distillate 0.26L that step (5) collects and make powdery with spray drying tower, promptly flavonoid glycoside substance 0.00603kg is a standard specimen with the rutin, with determined by ultraviolet spectrophotometry wherein general flavone content be 77.4%.
Embodiment 2
(1). the preparation of raw material: clean Zhejiang Province respectively and produce exsiccant peel of Citrus reticulata Blanco, exsiccant leaf of Diospyros kaki L.f., remove impurity wherein, after the pulverizing, cross 20 mesh sieves, respectively peel of Citrus reticulata Blanco dry powder, leaf of Diospyros kaki L.f. powder are stand-by;
(2). the processing of raw material: in peel of Citrus reticulata Blanco dry powder and leaf of Diospyros kaki L.f. dry powder quality ratio is that 2: 3 ratios were got peel of Citrus reticulata Blanco dried powder 0.3kg, leaf of Diospyros kaki L.f. dried powder 0.45kg, add quality and be 40% ethanol of 10 times of peel of Citrus reticulata Blanco dry powder and leaf of Diospyros kaki L.f. dry powder quality summations, temperature control is at 40 ℃～50 ℃, extract 3h, centrifugal, filtration get filtrate A, and filtering residue extracts 2 times by above method again, merging filtrate A gets extracting solution 10.6L;
(3). step (2) gained extracting solution is staticly settled, and the kieselguhr 0.204kg with the supernatant 10.2L after staticly settling adds supernatant quality 2% to adsorb most of colloid, chlorophyll etc., uses the flame filter press filter pressing with its filtering then; After fully stirring,, get 8 metafiltration cloth during filter pressing, get liquor B 8.7L with the flame filter press filter pressing;
(4). liquor B is carried out membrane filtration with poly (ether sulfone) film material (molecular weight 5kD, 95%), molecular cut off 5kD, and further remove tannin, peptide class etc., get the micromolecule solution 4.9L of filter membrane;
(5). get the micromolecule solution 4.9L that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 2ml/min, knifing rotating speed 200rpm at different levels, the vacuum of controlling first distillator is 750Pa, vapo(u)rizing temperature is 40 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove gas and part ethanol, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 150Pa, vapo(u)rizing temperature is 60 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove ethanol and part water, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 40Pa, vapo(u)rizing temperature is 100 ℃, 25 ℃ of cryosurface temperature, carry out thin film evaporation under the condition that circulating water temperature is 40 ℃, separate and remove most of water, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.05Pa, vapo(u)rizing temperature is 270 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 70 ℃, collect distillate 0.42L; Stay excess this moment is high boiling point and the involatile constituent that is removed.
(6) get the distillate 0.42L that step (5) collects and make powdery with spray drying tower, promptly flavonoid glycoside substance 0.00915kg is a standard specimen with the rutin, with determined by ultraviolet spectrophotometry wherein general flavone content be 74.1%.
Embodiment 3
(1). the preparation of raw material: clean Zhejiang Province respectively and produce exsiccant peel of Citrus reticulata Blanco, exsiccant leaf of Diospyros kaki L.f., remove impurity wherein, after the pulverizing, cross 20 mesh sieves, respectively peel of Citrus reticulata Blanco dry powder, leaf of Diospyros kaki L.f. powder are stand-by;
(2). the processing of raw material: in peel of Citrus reticulata Blanco dry powder and leaf of Diospyros kaki L.f. dry powder quality ratio is that 2: 3 ratios were got peel of Citrus reticulata Blanco dried powder 0.2kg, leaf of Diospyros kaki L.f. dried powder 0.3kg, add quality and be 50% ethanol of 12 times of peel of Citrus reticulata Blanco dry powder and leaf of Diospyros kaki L.f. dry powder quality summations, temperature control is at 35 ℃～45 ℃, extract 2.5h, centrifugal, filtration get filtrate A, and filtering residue extracts 2 times by above method again, merging filtrate A gets extracting solution 8.3L;
(3). step (2) gained extracting solution is staticly settled, and the kieselguhr 0.16kg with the supernatant 8.0L after staticly settling adds supernatant quality 2% to adsorb most of colloid, chlorophyll etc., uses the flame filter press filter pressing with its filtering then; After fully stirring,, get 6 metafiltration cloth during filter pressing, get liquor B 7.1L with the flame filter press filter pressing;
(4). liquor B is carried out membrane filtration with poly (ether sulfone) film material (molecular weight 5kD, 95%), molecular cut off 5kD, and further remove tannin, peptide class etc., get the micromolecule solution 3.9L of filter membrane;
(5). get the micromolecule solution 3.9L that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 2ml/min, knifing rotating speed 150rpm at different levels, the vacuum of controlling first distillator is 850Pa, vapo(u)rizing temperature is 50 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove gas and part ethanol, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 150Pa, vapo(u)rizing temperature is 75 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove ethanol and part water, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 40Pa, vapo(u)rizing temperature is 100 ℃, 25 ℃ of cryosurface temperature, carry out thin film evaporation under the condition that circulating water temperature is 40 ℃, separate and remove most of water, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.08Pa, vapo(u)rizing temperature is 280 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 70 ℃, collect distillate 0.31L; Stay excess this moment is high boiling point and the involatile constituent that is removed.
(6). get the distillate 0.31L that step (5) collects and make powdery with spray drying tower, promptly flavonoid glycoside substance 0.00583kg is a standard specimen with the rutin, with determined by ultraviolet spectrophotometry wherein general flavone content be 72.6%.
Embodiment 4
Step (1)～(4) are with the step among the embodiment 1 (1)～(4), the micromolecule solution 3.5L of final filter membrane;
Step (5): get micromolecule solution 3.5L, with petroleum ether, carbon tetrachloride, chloroform, ethyl acetate extracted the micromolecule solution behind the ultrafilter membrane successively, each water intaking layer of each extraction carries out next step extraction, when using ethyl acetate extraction at last, get ethyl acetate layer, the consumption of every kind of reagent is 1/3 of mother solution at that time at every turn, every kind of reagent all extracts 3 times, after obtaining ethyl acetate layer 3.9L, concentrate the removal ethyl acetate, obtain solution 0.33L with Rotary Evaporators, adopt boulton process to carry out drying, obtaining flavonoid glycoside substance 0.00947kg, is standard specimen with the rutin, with determined by ultraviolet spectrophotometry wherein general flavone content be 38.6%.
Embodiment 5
Step (1)～(4) are with the step among the embodiment 1 (1)～(4) method, the micromolecule solution 4.9L that obtains;
(5). get the micromolecule solution 4.9L that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 2ml/min, the vacuum of controlling first distillator is 850Pa, vapo(u)rizing temperature is 45 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove gas and part ethanol, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 350Pa, vapo(u)rizing temperature is 70 ℃, knifing rotating speed 300rpm, 30 ℃ of cryosurface temperature of circulating water temperature are to carry out thin film evaporation under 25 ℃ the condition, separate and remove ethanol and part water, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 30Pa, vapo(u)rizing temperature is 100 ℃, 25 ℃ of cryosurface temperature, carry out thin film evaporation under the condition that circulating water temperature is 40 ℃, separate and remove most of water, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.5Pa, vapo(u)rizing temperature is 280 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 70 ℃, collect distillate 0.46L; Stay excess this moment is high boiling point and the involatile constituent that is removed;
(6). get the distillate 0.46L that step (5) collects and make powdery with spray drying tower, promptly flavonoid glycoside substance 0.00518kg is a standard specimen with the rutin, with determined by ultraviolet spectrophotometry wherein general flavone content be 52.4%.
Embodiment 6
Step (1)～(4) are with the step among the embodiment 1 (1)～(4) method, the micromolecule solution 3.6L that obtains;
(5). get the micromolecule solution 3.6L that step (4) obtains, feeding membrane evaporator separates, described membrane evaporator comprises the level Four distillator, control material flow 1ml/min, knifing rotating speed 100rpm, the vacuum of controlling first distillator is 800Pa, vapo(u)rizing temperature is 45 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove gas and part ethanol, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 200Pa, vapo(u)rizing temperature is 70 ℃, the cryosurface temperature is 25 ℃, carry out thin film evaporation under the condition that circulating water temperature is 30 ℃, separate and remove ethanol and part water, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 30Pa, vapo(u)rizing temperature is 90 ℃, 25 ℃ of cryosurface temperature, carry out thin film evaporation under the condition that circulating water temperature is 40 ℃, separate and remove most of water, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 2Pa, vapo(u)rizing temperature is 260 ℃, 25 ℃ of cryosurface temperature, carry out thin film evaporation under the condition that circulating water temperature is 70 ℃, collect distillate 0.25L; Stay excess this moment is high boiling point and the involatile constituent that is removed;
(6). get the distillate 0.25L that step (8) collects and make powdery with spray drying tower, promptly flavonoid glycoside substance 0.00203kg is a standard specimen with the rutin, with determined by ultraviolet spectrophotometry wherein general flavone content be 40.6%.
The vacuum that the present invention preferably controls the 4th distillator is 0.01～0.5Pa, vapo(u)rizing temperature is 200 ℃～300 ℃, and the cryosurface temperature is 20～30 ℃, and the knifing rotating speed is 250-350rpm, carry out thin film evaporation under 30～70 ℃ of conditions of circulating water temperature, collect distillate.In the 5th step of embodiment 6 vacuum of the 4th distillator be 2Pa more excessive relatively, though also can isolate flavonoid glycoside substance, the total flavones yield is less relatively.
Claims (10)
1. the extracting method of a flavonoid glycoside substance, it is characterized in that described method is that the dry fragment of getting peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. is that raw material carries out as follows: (1) removes the peel of Citrus reticulata Blanco of impurity, the dry fragment of leaf of Diospyros kaki L.f. is a raw material, (2) add 20～80% ethanol that quality is 10～20 times of peel of Citrus reticulata Blanco, the dry fragment quality of leaf of Diospyros kaki L.f. summation, 2～4h is extracted in 30～70 ℃ of insulations, centrifugal, filtration, get filtrate A, filtering residue repeats to extract 1～5 time again, merges each filtrate A that extracts and gets extracting solution; (3) step (2) gained extracting solution is staticly settled, get supernatant, adding quality is the kieselguhr of described supernatant 1～5%, after fully stirring, with the flame filter press filter pressing, gets liquor B; (4) liquor B is carried out membrane filtration with the poly (ether sulfone) film material, molecular cut off 5kD gets the micromolecule solution of filter membrane; (5) getting micromolecule solution that step (4) obtains feeds membrane evaporator and separates, described membrane evaporator comprises the level Four distillator, control material flow 1～3ml/min, knifing rotating speed at different levels is 100-300rpm, circulating water temperatures at different levels are respectively done for oneself 30～70 ℃, the vacuum of controlling first distillator is 600～900Pa, vapo(u)rizing temperature is 40 ℃～60 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 100～300Pa, vapo(u)rizing temperature is 60 ℃～80 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 10～50Pa, vapo(u)rizing temperature is 80 ℃～110 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.01～2Pa, vapo(u)rizing temperature is 150 ℃～300 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～40 ℃ of conditions, collects distillate; (6) getting distillate that step (5) collects carries out spray drying and makes powdery, promptly described flavonoid glycoside substance.
2. the extracting method of flavonoid glycoside substance as claimed in claim 1, the dry fragment that it is characterized in that described peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. was the dried powder of 20 mesh sieves.
3. the extracting method of flavonoid glycoside substance as claimed in claim 1, it is characterized in that described peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. dry fragment mass ratio be 2: 3.
4. the extracting method of flavonoid glycoside substance as claimed in claim 1 is characterized in that the described sedimentation time is for precipitating 12～48 hours.
5. the extracting method of flavonoid glycoside substance as claimed in claim 1 is characterized in that described filter press is with the filter pressing of 3～10 layers of DANFU silk filter cloth.
6. the extracting method of flavonoid glycoside substance as claimed in claim 1, the vacuum that it is characterized in that described step (5) control first distillator is that 750～850Pa, vapo(u)rizing temperature are 40 ℃～50 ℃, and the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition.
7. the extracting method of flavonoid glycoside substance as claimed in claim 1, the vacuum that it is characterized in that described step (5) control after-fractionating device is that 150～250Pa, vapo(u)rizing temperature are 65 ℃～75 ℃, and the condition that the cryosurface temperature is 15～25 ℃ is carried out thin film evaporation.
8. the extracting method of flavonoid glycoside substance as claimed in claim 1, the vacuum that it is characterized in that described step (5) control the 3rd distillator is that 20～40Pa, vapo(u)rizing temperature are 80 ℃～100 ℃, and the condition that the cryosurface temperature is 15～25 ℃ is carried out thin film evaporation.
9. the extracting method of flavonoid glycoside substance as claimed in claim 1, the vacuum that it is characterized in that described step (5) control the 4th distillator is 0.01～0.5Pa, vapo(u)rizing temperature is 200 ℃～300 ℃, and the cryosurface temperature is to carry out thin film evaporation under 20～30 ℃ the condition, collects distillate.
10. the extracting method of the described flavonoid glycoside substance of claim 1, it is characterized in that described method is as follows: the mass ratio that (1) removes impurity is that 2: 3 peel of Citrus reticulata Blanco, the dried powder of leaf of Diospyros kaki L.f. are raw material; (2) add 55%～60% the ethanol that quality is 10～15 times of peel of Citrus reticulata Blanco, leaf of Diospyros kaki L.f. dried powder quality summation, 2.5～3h are extracted in 50 ℃ of insulations, and are centrifugal, filter, filtrate A, filtering residue repeats to extract 2～3 times again, merges each filtrate A and gets extracting solution; (3) step (2) gained extracting solution was left standstill 12～24 hours, precipitation is got supernatant, and adding quality is the kieselguhr of described supernatant 2%, after fully stirring, with 6～8 metafiltration cloth filter pressings, gets liquor B with flame filter press; (4) liquor B is carried out membrane filtration with the poly (ether sulfone) film material, molecular cut off 5kD gets the micromolecule solution of filter membrane; (5) getting micromolecule solution that step (4) obtains feeds membrane evaporator and separates, described membrane evaporator comprises the level Four distillator, control material flow 1～3ml/min, knifing rotating speed at different levels is 100～200rpm, circulating water temperature is respectively done for oneself 30～70 ℃, the vacuum of controlling first distillator is 750～850Pa, vapo(u)rizing temperature is 40 ℃～50 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess A and directly enter the after-fractionating device, the vacuum of control after-fractionating device is 150～250Pa, vapo(u)rizing temperature is 65 ℃～75 ℃, the cryosurface temperature is to carry out thin film evaporation under 15～25 ℃ the condition, must heat up in a steamer excess B and directly enter the 3rd distillator, the vacuum of controlling the 3rd distillator is 20～40Pa, vapo(u)rizing temperature is 80 ℃～90 ℃, carry out thin film evaporation under the condition that the cryosurface temperature is 15～25 ℃, must heat up in a steamer excess C and directly enter the 4th distillator, the vacuum of controlling the 4th distillator is 0.01～0.05Pa, vapo(u)rizing temperature is 250 ℃～280 ℃, carry out thin film evaporation under the condition that the cryosurface temperature is 30 ℃, collect distillate; (6) getting distillate that step (5) collects carries out spray drying and makes powdery, promptly described flavonoid glycoside substance.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863955A (en) * | 2010-05-10 | 2010-10-20 | 孙志高 | Process for hierarchically extracting limonin, flavonoid and dietary fiber from orange peel residues |
| CN110845319A (en) * | 2019-11-28 | 2020-02-28 | 江西中医药大学 | Preparation method of neoflavonoid compound and application of neoflavonoid compound in preparation of medicine for treating osteoporosis |
| CN117503817A (en) * | 2024-01-04 | 2024-02-06 | 广州泽力医药科技有限公司 | Extraction method for improving total flavone content of licorice extract |
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| DE10006147A1 (en) * | 2000-02-11 | 2001-08-16 | Merck Patent Gmbh | Process for the preparation of monoglycosidated flavonoids |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863955A (en) * | 2010-05-10 | 2010-10-20 | 孙志高 | Process for hierarchically extracting limonin, flavonoid and dietary fiber from orange peel residues |
| CN101863955B (en) * | 2010-05-10 | 2015-03-11 | 孙志高 | Process for hierarchically extracting limonin, flavonoid and dietary fiber from orange peel residues |
| CN110845319A (en) * | 2019-11-28 | 2020-02-28 | 江西中医药大学 | Preparation method of neoflavonoid compound and application of neoflavonoid compound in preparation of medicine for treating osteoporosis |
| CN110845319B (en) * | 2019-11-28 | 2023-06-20 | 江西中医药大学 | Preparation method of novel flavonoid compound and application of novel flavonoid compound in preparation of medicament for treating osteoporosis |
| CN117503817A (en) * | 2024-01-04 | 2024-02-06 | 广州泽力医药科技有限公司 | Extraction method for improving total flavone content of licorice extract |
| CN117503817B (en) * | 2024-01-04 | 2024-04-12 | 广州泽力医药科技有限公司 | Extraction method for improving total flavone content of licorice extract |
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