CN109021046A - A method of extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf - Google Patents

A method of extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf Download PDF

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CN109021046A
CN109021046A CN201811142856.7A CN201811142856A CN109021046A CN 109021046 A CN109021046 A CN 109021046A CN 201811142856 A CN201811142856 A CN 201811142856A CN 109021046 A CN109021046 A CN 109021046A
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quercitin
siraitia grosvenorii
cauline leaf
extracting
mountain naphthalene
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CN109021046B (en
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陈钱
龙伟岸
黄华学
贺进军
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Hunan Huacheng Biotech Inc
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Hunan Huacheng Biotech Inc
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07HSUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04Heterocyclic radicals containing only oxygen as ring hetero atoms
    • C07H17/06Benzopyran radicals
    • C07H17/065Benzo[b]pyrans
    • C07H17/07Benzo[b]pyran-4-ones
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07HSUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00Processes for the preparation of sugar derivatives
    • C07H1/06Separation; Purification
    • C07H1/08Separation; Purification from natural products

Abstract

A method of extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf, comprising the following steps: (1) the fresh cauline leaf of Siraitia grosvenorii is dried, be crushed, add water, continuous flow upstream homogenate extraction, centrifugal filtration;(2) ultrafiltration, upper large pore resin absorption column absorption, washing, water lotion discard, and eluent is collected in organic solvent elution, and water lotion is collected in washing, is concentrated, dry;(3) it is added in alcoholic solution, dissolves by heating, it is cooling, it stands, filters, be concentrated, it is dry;(4) with phased soln is flowed, filtering, system is collected target phase eluent for liquid-phase chromatographic column, isocratic elution respectively, is concentrated, dry, obtains quercitin and mountain naphthalene glycosides respectively.According to purity >=98.2% of quercitrin glycoside product obtained by the method for the present invention, yield >=90.4%, purity >=98.8% of kaempferia galamga glycoside product, yield >=91.3%;The method of the present invention operating procedure is simple, stablizes, energy consumption, at low cost, it can be achieved that continuous large-scale production.

Description

A method of extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf
Technical field
The present invention relates to a kind of methods for extracting flavonoid glycoside substance, and in particular to one kind mentions simultaneously from Siraitia grosvenorii cauline leaf The method for taking quercitin and mountain naphthalene glycosides.
Background technique
Siraitia grosvenorii is Curcurbitaceae perennial plant, and main product is in Guilin area.Siraitia grosvenorii whole body is all precious: Siraitia grosvenorii is fresh The ingredients such as sweet tea glycosides, protein and polysaccharide rich in fruit, especially mogroside have huge city as natural sweetener Field and economic value;Contain much starch and Siraitia grosvenorii tartaric acid in Siraitia grosvenorii root tuber, Siraitia grosvenorii starch has very high nutriture value Value;Containing abundant grease in arhat fruit seed, main active is squalene, the work with anticancer, anti-cancer and moisturizing beauty treatment With;Contain more flavonoid glycoside substance, predominantly quercitin and kaempferol in Siraitia grosvenorii cauline leaf, flavonoid glycoside substance has aobvious The pharmacology of work, physiological activity, can expand blood vessel increase blood circulation, inhibition thrombosis, at the same have anti-inflammatory, anticancer, it is antibacterial, Hypoglycemic and reducing blood lipid and other effects.
Currently, exploitation is relatively early more mature or extracts mogroside from Lo Han Guo fruit, and Siraitia grosvenorii fresh fruit is adopted After having plucked, cauline leaf, rattan are typically directly thrown away, and not only pollute environment, but also cause the huge wasting of resources.
CN106668234A discloses a kind of extraction and purification process of rose general flavone, process flow are as follows: solvent extraction → macroporous resin purification → polyamide purifying → C18 preparation purifying.Although the technique can get general flavone purity >=90% Product, but entire technical process is more complex, it is more to expend organic solvent amount, it is more difficult to recycle, the rate of recovery of flavones is low, entire technique Realize large-scale industrial production higher cost.
CN102138958A discloses a kind of Momordica grosvenori plant flavones ingredient extraction and purification process, process flow are as follows: second Alcohol extracting → recycling design → macroreticular resin removal of impurities → molten removal of impurities → concentrate drying of elution → concentrate drying → alcohol.Although the technique It can get the product of flavonoid glycoside purity >=50%, and simple process, stabilization, still, the flavones purity after purification is not high, and a step Although the molten removal of impurities of alcohol is simple and effective, the rate of recovery is relatively low.
CN107698638A discloses extraction and purification process, detection method and its utilization of a kind of fructus lycii seed total flavonoid, work Skill process are as follows: drying → broken → extraction → extraction → macroporous resin purification → polyamide purifying.Although obtained by the technique Fructus lycii seed total flavonoid >=40%, still, using the extraction of 80% ethyl alcohol and petroleum ether extraction in extraction process, solvent consumption is big, To the more demanding of production equipment, higher cost.
CN102399252A discloses a kind of preparation method of Vaccarin monomer, process flow are as follows: raw material proposes Take that → → efficiently preparing liquid phase separation, → macroreticular resin is enriched with again → to be eluted → concentration → to dry macroporous resin enrichment.Although the work Vaccarin monomer >=99% obtained by skill, still, the processing step are complicated, use macroporous resin enrichment 2 times, not only molten Agent consumption is big, and the rate of recovery is also low.
CN102050845A discloses method that is a kind of while preparing Calycosin-7-O-BETA-D-glucoside, ononin chemical reference substance, Process flow are as follows: silica gel column chromatography enrichment → resin column removal of impurities → reversed high performance liquid preparative chromatography refines → is dried under reduced pressure.Though The Calycosin-7-O-BETA-D-glucoside and ononin of purity >=98% can be prepared in the right technique simultaneously, still, be enriched with silica gel chromatograph Method, treating capacity is smaller, and organic solvent expends larger and more demanding to production environment, large-scale production higher cost.
CN102503998A discloses a kind of method of quick separating quercitin from Flos Albiziae, process flow are as follows: raw material Extraction → macroporous resin enrichment → concentration crystallization → efficiently prepares liquid phase separation → cooling crystallization → drying.Although the technique can be with Quercitin monomer is obtained, still, since by 2 crystallizations, the rate of recovery is low.
CN104592327A discloses rutin in a kind of hops, the preparation method of isoquercitrin and to its antiallergy, anti- The detection of oxidation activity, process flow are as follows: raw material extraction → abstraction impurity removal → macroporous resin enrichment → half preparation efficient liquid phase point From.Although the technique can obtain rutin and isoquercitrin monomer, due to needing successively to use petroleum ether, ethyl acetate And extracting n-butyl alcohol removal of impurities, complex technical process, solvent consumption is big, higher cost.
Summary of the invention
The technical problem to be solved by the present invention is to overcome drawbacks described above of the existing technology, provide a kind of gained production Product purity is high, high income, operating procedure is simple, stablizes, energy consumption, it is at low cost, it can be achieved that continuous large-scale production slave arhat The method for extracting quercitin and mountain naphthalene glycosides simultaneously in stem end leaf.
The technical solution adopted by the present invention to solve the technical problems is as follows: one kind extracting Mongolian oak simultaneously from Siraitia grosvenorii cauline leaf The method of skin glycosides and mountain naphthalene glycosides, comprising the following steps:
(1) it is crushed, extracts: the fresh cauline leaf of Siraitia grosvenorii is dried, be crushed, add water, continuous flow upstream homogenate extraction, centrifugal filtration obtains Extracting solution;
(2) extracting solution obtained by step (1) ultrafiltration, macroporous resin adsorption: is subjected to ultrafiltration, upper large pore resin absorption column absorption, water Be washed till efflux it is colourless, clarification, it is bright, water lotion discards, organic solvent elution, collect eluent, be washed to organic solvent-free Until, water lotion is collected, is concentrated under reduced pressure, vacuum drying obtains flavonoid glycoside crude product A;
(3) the molten removal of impurities of alcohol: flavonoid glycoside crude product A obtained by step (2) being added in alcoholic solution, is dissolved by heating, cooling, is stood, Filtering is concentrated under reduced pressure, and vacuum drying obtains flavonoid glycoside crude product B;
(4) preparative liquid chromatography separates: by the flowing phased soln of flavonoid glycoside crude product B obtained by step (3), filtering, system is for liquid phase Chromatographic column is separated, then carries out isocratic elution with mobile phase, collects target phase eluent respectively, is concentrated under reduced pressure, vacuum drying, Quercitin and mountain naphthalene glycosides are obtained respectively.
Preferably, in step (1), in the fresh cauline leaf of Siraitia grosvenorii, the mass content of quercitin is 0.04~0.08%, The mass content of mountain naphthalene glycosides is 0.5~1.0%.
Preferably, in step (1), the fresh cauline leaf of Siraitia grosvenorii is crushed to 20~40 mesh.
Preferably, in step (1), the dosage of the water be the fresh cauline leaf quality of Siraitia grosvenorii 4~20 times (more preferable 5~ 15 times).Since grosvenor momordica flavonoid glycosides is highly soluble in water, especially hot water, so it is not only at low cost to adopt water as solvent, Er Qiehuan It ensures safety.
Preferably, in step (1), the temperature of the continuous flow upstream homogenate extraction is 50~90 DEG C, number >=2 of extraction Secondary, the time extracted every time is the more preferable 100~180s of 60~200s().Due to the oxidizable special nature of Flavonoid substances, If Extracting temperature is excessively high or overlong time, extraction recovery can reduce, therefore, can using continuous flow upstream homogenate extraction It realizes serialization, large-scale production, so as to shorten extraction time, while the rate of recovery can also be improved.
Preferably, in step (1), the revolving speed of the centrifugation be 10000~80000r/min(more preferable 40000~ 60000r/min).
Preferably, in step (1), the mode of the centrifugal filtration is first horizontal screw centrifugal filtering, then butterfly centrifugal mistake Filter.
Preferably, in step (2), the ultrafiltration membrane for ultrafiltration is ceramic membrane, and the aperture of the ceramic membrane is 0.2~2.0 μ More preferable 0.3~1.0 μm of m().
Preferably, in step (2), the flow velocity of upper prop is 1.0~4.0BV/h.Since flavonoid glycoside is the biggish substance of polarity, It is easily adsorbed by macroporous absorbent resin, to achieve the purpose that purification & isolation.
Preferably, in step (2), model D-101 type, AB-8 type, the DM-130 type, X-7 of the macroporous absorbent resin One or more of type or XDA-7 type etc..
In the method for the present invention, macroporous absorbent resin is before use, first with 1~3BV, the ethyl alcohol leaching of volume fraction 90~99% 20~30h is steeped, then is washed till with the ethyl alcohol of volume fraction 90~99% that efflux is colourless, free from extraneous odour, is washed to no alcohol taste, then with 3~ The sodium hydroxide solution alkali cleaning of 5BV mass concentration 4~6%, then it is washed to neutrality, finally with the salt of 3~5BV mass concentration 4~6% Acid solution pickling, then it is washed to neutrality, it is spare.
Preferably, in step (2), the volume mass ratio (L/kg) of the macroporous absorbent resin and the fresh cauline leaf of Siraitia grosvenorii is More preferable 1:5~15 1:1~20().
Preferably, in step (2), the diameter height of the large pore resin absorption column is than for more preferable 1:4~6 1:2~8().
Preferably, in step (2), the flow velocity of the washing is 1.0~4.0BV/h.The purpose of washing mainly goes to remove water The stronger impurity of dissolubility, such as monosaccharide, polysaccharide and pectin.
Preferably, in step (2), the dosage of the organic solvent is 1~4BV.
Preferably, in step (2), the flow velocity of organic solvent elution be 0.25~2.00BV/h(more preferable 0.5~ 1.0BV/h).The purpose of organic solvent elution is to elute the flavonoid glycoside of resin adsorption.
Preferably, in step (2), the volume fraction of the organic solvent is 50~90%.
Preferably, in step (2), the organic solvent is food-grade ethanol.
Preferably, in step (2), the temperature of the reduced pressure is 40~70 DEG C, and pressure is -0.1~-0.07MPa, dense Being reduced to solid content is 30~60%.
Preferably, in step (2), the vacuum drying temperature is 40~70 DEG C, and pressure is -0.1~-0.07MPa, when Between be 4~12h.
Preferably, in step (3), the mass volume ratio (kg/L) of the flavonoid glycoside crude product A and alcoholic solution is 1:8~20 (more preferable 1:9~15).Alcohol is molten clean mainly remove be it is a kind of water-soluble preferably, but the impurity that alcohol-soluble is poor, such as albumen Matter etc..
Preferably, in step (3), the temperature of the heating is 50~80 DEG C.
Preferably, in step (3), the alcoholic solution is food-grade anhydrous methanol or volume fraction is 90~99% food Grade ethyl alcohol.
Preferably, described to be cooled to room temperature in step (3).It is preferred that using board-like cooling.
Preferably, in step (3), the time of the standing is 2~4h.The purpose of standing is so that abundant, the impurity that cleans Subsequent filter is convenient in easily layering.
Preferably, in step (3), the temperature of the reduced pressure is 40~70 DEG C, and pressure is -0.1~-0.07MPa, dense Being reduced to solid content is 30~60%.
Preferably, in step (3), the vacuum drying temperature is 40~70 DEG C, and pressure is -0.1~-0.07MPa, when Between be 4~12h.
Preferably, in step (4), the flavonoid glycoside crude product B and mobile phase mass volume ratio (kg/L) are 0.08~0.30: 1.The effect isolated and purified under the ratio is more preferable.
Preferably, in step (4), the mobile phase is organic solvent-aqueous solution that volume ratio is 15~40:95~60. The effect isolated and purified under the ratio is more preferable, and the rate of recovery is higher.
Preferably, in step (4), the organic solvent is one or more of ethyl alcohol, methanol or acetonitrile etc..It is more excellent Choosing, ethyl alcohol.Preferable using the solvent separating effect, convenient for recycling, and cost is lower.
Preferably, in step (4), the filter membrane for filtering is organic phase filter membrane, and aperture is 0.20~0.45 μm.Filtering Main purpose is to remove a small amount of suspended impurity, prevents preparation liquid phase from blocking up column, influences separating effect.
Preferably, in step (4), the flow velocity of sample introduction is 0.25~1.00BV/h.
Preferably, in step (4), the preparative liquid chromatography column is ODS column, and filler is C-18 reversed-phase bonded silica.This Inventor isolates and purifies effect and becomes apparent from the study found that using the column type and filler, and separating degree is higher, and the rate of recovery is also higher.
Preferably, in step (4), the flow velocity of the elution is 0.5~2.0BV/h.The present inventor is the study found that described Separating effect is preferable under flow velocity, and the rate of recovery of target component is higher.
Preferably, in step (4), flowing elution after, according to preparation liquid phase UV detector signal display screen on, the two of appearance The residence time at a independent peak collects the eluent of quercitin and mountain naphthalene glycosides target product respectively.
Preferably, in step (4), the temperature of the reduced pressure is 40~70 DEG C, and pressure is -0.1~-0.07MPa, dense Being reduced to solid content is 30~60%.
Preferably, in step (4), the vacuum drying temperature is 40~70 DEG C, and pressure is -0.1~-0.07MPa, when Between be 4~12h.
It is described to filter preferred plate-frame filtering or plate compression in step (3), (4).
The method of the present invention has the beneficial effect that:
It (1) is in yellow according to quercitrin glycoside product obtained by the method for the present invention and mountain naphthalene glycoside product, quercitin in quercitrin glycoside product Purity >=98.2%, yield >=90.4%, purity >=98.8% of kaempferia galamga glycosides, yield >=91.3% in kaempferia galamga glycoside product;
(2) it since the solvent that the method for the present invention is extracted is water, is extracted compared to conventional organic solvents, more environmentally-friendly, safety, cost It is low;Using continuous flow upstream homogenate extraction, is extracted compared to conventional backflow, both can guarantee serialization, large-scale production, and extraction time Short, the rate of recovery is high;
(3) the preparation liquid phase of the method for the present invention uses organic solvent-water flowing phase, and isocratic elution, compares other organic mixing Solvent, cost is lower, easy to operate, mobile phase energy recycling and reusing, is suitable for large-scale industrial production;
(4) the method for the present invention separates the side combined using the molten removal of impurities of macroporous resin adsorption purifying → alcohol → preparative liquid chromatography Method, not only simple process, stabilization, and ensure that the high-purity of product.
Specific embodiment
Below with reference to embodiment, the invention will be further described.
Siraitia grosvenorii cauline leaf used in the embodiment of the present invention is purchased from Guilin, detects through HPLC, and the quality of quercitin contains Amount is 0.06%, and the mass content of mountain naphthalene glycosides is 0.73%;0.3 μm of aperture, 0.5 μm of ceramics used in the embodiment of the present invention Film is purchased from the long Anyuan Environmental Protection Technology Co., Ltd in Jiangsu;The used D-101 type of the embodiment of the present invention, AB-8 type macroporous absorption Resin is purchased from Xi'an Sunresin New Materials Co., Ltd.;X-5 type macroporous absorption tree used in the embodiment of the present invention Rouge is purchased from Tianjin Nankai Hecheng S&T Co., Ltd.;0.22 μm of aperture, 0.45 μm of organic phase used in the embodiment of the present invention Filter membrane is purchased from Shanghai new Asia purification device factory;ODS column and C-18 reversed-phase bonded silica used in the embodiment of the present invention are purchased In Hanbon Sci. & Tech. Co., Ltd.;Raw material and chemical reagent used in the embodiment of the present invention pass through unless otherwise specified Routine business approach obtains.
Reference example 1
Before use, first using 2BV, the ethyl alcohol of volume fraction 95% impregnates for 24 hours macroporous absorbent resin of the embodiment of the present invention, then uses body The ethyl alcohol of fraction 95% is washed till that efflux is colourless, free from extraneous odour, is washed to no alcohol taste, then the sodium hydroxide with 4BV mass concentration 5% Solution alkali cleaning, then it is washed to neutrality, the hydrochloric acid solution pickling of 4BV mass concentration 5% is finally used, then be washed to neutrality, it is spare.
Embodiment 1
(1) it is crushed, extracts: the fresh cauline leaf of 10t Siraitia grosvenorii being dried, 20 mesh are crushed to, adds 100t water, it is continuous inverse at 60 DEG C Stream homogenate extraction 2 times, each 150s, then at 60000r/min, first horizontal spiral centrifuge centrifugal filtration, then use butterfly centrifugal Machine centrifugal filtration obtains extracting solution 95t;
(2) ultrafiltration, macroporous resin adsorption: 95t extracting solution obtained by step (1) is subjected to ultrafiltration with 0.5 μm of ceramic membrane, ultrafiltration is saturating Liquid is crossed with the flow velocity of 3BV/h, (volume of AB-8 type macroporous absorbent resin is 1000L to upper AB-8 type large pore resin absorption column, and diameter is high Than 1:4) absorption is washed to colourless efflux, clarification, bright, water lotion discards, with 2BV, volume point then with the flow velocity of 2BV/h The food-grade ethanol of number 80% is eluted with the flow velocity of 0.5BV/h, collects eluent, until being washed to no ethyl alcohol, collects water lotion, At 50 DEG C, under -0.1MPa, it is 45%, at 40 DEG C, -0.1MPa that the eluent of collection and water lotion, which are concentrated under reduced pressure into solid content, Under, it is dried in vacuo 6h, obtains flavonoid glycoside crude product A 131.29kg;
(3) 131.29kg flavonoid glycoside crude product A obtained by step (2) the molten removal of impurities of alcohol: is added to the food-grade of 1500L volume fraction 95% In ethanol solution, at 60 DEG C, dissolved by heating, it is board-like to be cooled to room temperature, stand 2h, plate-frame filtering, filtrate at 40 DEG C ,- Under 0.1MPa, being concentrated under reduced pressure into solid content is 50%, at 40 DEG C, under -0.1MPa, is dried in vacuo 8h, obtains flavonoid glycoside crude product B 93.23kg;
(4) preparative liquid chromatography separates: 93.23kg flavonoid glycoside crude product B obtained by step (3) is molten with 1100L mobile phase alcohol-water Liquid (volume ratio 30:70) dissolution, the organic phase filter membrane plate compression in 0.22 μm of aperture of lysate, then, with 0.25BV/h Flow velocity, system separated for liquid-phase chromatographic column (ODS column, filler be C-18 reversed-phase bonded silica), then with mobile phase ethyl alcohol- Aqueous solution (volume ratio 30:70) carries out isocratic elution with the flow velocity of 1.0BV/h, aobvious according to preparation liquid phase UV detector signal In display screen, the residence time at two independent peaks of appearance collects 10~25BV quercitin target phase and the mountain 30~40BV naphthalene respectively Glycosides target phase eluent, at 40 DEG C, under -0.1MPa, being concentrated under reduced pressure into solid content is 55%, and at 40 DEG C, under -0.1MPa, vacuum is dry Dry 6h obtains quercitin 5.58kg and mountain naphthalene glycosides 67.89kg respectively.
Quercitrin glycoside product obtained by the embodiment of the present invention and mountain naphthalene glycoside product are in yellow, are detected through HPLC, gained quercitin The purity of quercitin is 98.2% in product, yield 91.3%, and the purity of mountain naphthalene glycosides is 99.1%, yield 92.2%.
Embodiment 2
(1) it is crushed, extracts: the fresh cauline leaf of 10t Siraitia grosvenorii being dried, 40 mesh are crushed to, adds 150t water, it is continuous inverse at 80 DEG C Stream homogenate extraction 3 times, each 100s, then at 40000r/min, first horizontal spiral centrifuge centrifugal filtration, then use butterfly centrifugal Machine centrifugal filtration obtains extracting solution 145t;
(2) 145t extracting solution obtained by step (1) ultrafiltration, macroporous resin adsorption: is subjected to ultrafiltration, ultrafiltration with 0.3 μm of ceramic membrane Permeate is with the flow velocity of 4BV/h, and (volume of X-5 type macroporous absorbent resin is 1500L to upper X-5 type large pore resin absorption column, and diameter is high Than 1:6) absorption is washed to colourless efflux, clarification, bright, water lotion discards, with 4BV, volume point then with the flow velocity of 4BV/h The food-grade ethanol of number 60% is eluted with the flow velocity of 1.0BV/h, collects eluent, until being washed to no ethyl alcohol, collects water lotion, At 70 DEG C, under -0.08MPa, it is 50%, at 60 DEG C, -0.08MPa that the eluent of collection and water lotion, which are concentrated under reduced pressure into solid content, Under, it is dried in vacuo 8h, obtains flavonoid glycoside crude product A 125.6kg;
(3) the molten removal of impurities of alcohol: 125.6kg flavonoid glycoside crude product A obtained by step (2) is added in the food-grade anhydrous methanol solution of 1200L, At 80 DEG C, dissolved by heating, it is board-like to be cooled to room temperature, 4h is stood, plate-frame filtering, filtrate is at 60 DEG C, under -0.08MPa, subtracts It is 40% that pressure, which is concentrated into solid content, at 60 DEG C, under -0.08MPa, is dried in vacuo 6h, obtains flavonoid glycoside crude product B 91.8kg;
(4) preparative liquid chromatography separates: by 91.8kg flavonoid glycoside crude product B 800L mobile phase methanol-aqueous solution obtained by step (3) (volume ratio 20:80) dissolution, the organic phase filter membrane plate compression in 0.45 μm of aperture of lysate, then, with 0.75BV/h's Flow velocity, system are separated for liquid-phase chromatographic column (ODS column, filler be C-18 reversed-phase bonded silica), then with mobile phase methanol-water Solution (volume ratio 20:80) carries out isocratic elution with the flow velocity of 2.0BV/h, is shown according to preparation liquid phase UV detector signal On screen, the residence time at two independent peaks of appearance collects 10~25BV quercitin target phase and the mountain 30~40BV naphthalene glycosides respectively Target phase eluent, at 60 DEG C, under -0.08MPa, being concentrated under reduced pressure into solid content is 60%, and at 60 DEG C, under -0.08MPa, vacuum is dry Dry 10h obtains quercitin 5.5kg and mountain naphthalene glycosides 66.8kg respectively.
Quercitrin glycoside product obtained by the embodiment of the present invention and mountain naphthalene glycoside product are in yellow, are detected through HPLC, gained quercitin The purity of quercitin is 98.6% in product, yield 90.4%, and the purity of mountain naphthalene glycosides is 99.8%, yield 91.3%.
Embodiment 3
(1) it is crushed, extracts: the fresh cauline leaf of 10t Siraitia grosvenorii being dried, 30 mesh are crushed to, adds 120t water, it is continuous inverse at 70 DEG C Stream homogenate extraction 2 times, each 120s, then at 50000r/min, first horizontal spiral centrifuge centrifugal filtration, then use butterfly centrifugal Machine centrifugal filtration obtains extracting solution 95t;
(2) ultrafiltration, macroporous resin adsorption: 95t extracting solution obtained by step (1) is subjected to ultrafiltration with 0.5 μm of ceramic membrane, ultrafiltration is saturating Liquid is crossed with the flow velocity of 2BV/h, (volume of D-101 type macroporous absorbent resin is 1200L, diameter to upper D-101 type large pore resin absorption column Height ratio 1:5) absorption is washed to colourless efflux, clarification, bright, water lotion discards, with 3BV, volume then with the flow velocity of 3BV/h The food-grade ethanol of score 70% is eluted with the flow velocity of 0.75BV/h, collects eluent, until being washed to no ethyl alcohol, collects washing Liquid, at 60 DEG C, under -0.09MPa, it is 48% that the eluent of collection and water lotion, which are concentrated under reduced pressure into solid content, at 50 DEG C, - Under 0.09MPa, it is dried in vacuo 11h, obtains flavonoid glycoside crude product A 140.3kg;
(3) 140.3kg flavonoid glycoside crude product A obtained by step (2) the molten removal of impurities of alcohol: is added to the food-grade second of 1500L volume fraction 92% In alcoholic solution, at 70 DEG C, dissolved by heating, it is board-like to be cooled to room temperature, stand 3h, plate-frame filtering, filtrate at 50 DEG C ,- Under 0.09MPa, being concentrated under reduced pressure into solid content is 47%, at 50 DEG C, under -0.09MPa, is dried in vacuo 10h, obtains flavonoid glycoside crude product B 89.5kg;
(4) preparative liquid chromatography separates: 89.5kg flavonoid glycoside crude product B obtained by step (3) is molten with 1000L mobile phase acetonitrile-water Liquid (volume ratio 26:74) dissolution, the organic phase filter membrane plate compression in 0.22 μm of aperture of lysate, then, with 0.5BV/h's Flow velocity system is separated for liquid-phase chromatographic column (ODS column, filler be C-18 reversed-phase bonded silica), then with mobile phase acetonitrile-water Solution (volume ratio 26:74) carries out isocratic elution with the flow velocity of 1.5BV/h, is shown according to preparation liquid phase UV detector signal On screen, the residence time at two independent peaks of appearance collects 5~15BV quercitin target phase and the mountain 20~30BV naphthalene glycosides mesh respectively Bid section eluent, at 50 DEG C, under -0.09MPa, being concentrated under reduced pressure into solid content is 56%, at 50 DEG C, under -0.09MPa, and vacuum drying 8h obtains quercitin 5.75kg and mountain naphthalene glycosides 70.5kg respectively.
Quercitrin glycoside product obtained by the embodiment of the present invention and mountain naphthalene glycoside product are in yellow, are detected through HPLC, gained quercitin The purity of quercitin is 98.4% in product, yield 94.3%, and the purity of mountain naphthalene glycosides is 98.8%, yield 95.4%.

Claims (8)

1. a kind of method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf, which comprises the following steps:
(1) it is crushed, extracts: the fresh cauline leaf of Siraitia grosvenorii is dried, be crushed, add water, continuous flow upstream homogenate extraction, centrifugal filtration obtains Extracting solution;
(2) extracting solution obtained by step (1) ultrafiltration, macroporous resin adsorption: is subjected to ultrafiltration, upper large pore resin absorption column absorption, water Be washed till efflux it is colourless, clarification, it is bright, water lotion discards, organic solvent elution, collect eluent, be washed to organic solvent-free Until, water lotion is collected, is concentrated under reduced pressure, vacuum drying obtains flavonoid glycoside crude product A;
(3) the molten removal of impurities of alcohol: flavonoid glycoside crude product A obtained by step (2) being added in alcoholic solution, is dissolved by heating, cooling, is stood, Filtering is concentrated under reduced pressure, and vacuum drying obtains flavonoid glycoside crude product B;
(4) preparative liquid chromatography separates: by the flowing phased soln of flavonoid glycoside crude product B obtained by step (3), filtering, system is for liquid phase Chromatographic column is separated, then carries out isocratic elution with mobile phase, collects target phase eluent respectively, is concentrated under reduced pressure, vacuum drying, Quercitin and mountain naphthalene glycosides are obtained respectively.
2. the method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf according to claim 1, it is characterised in that: In step (1), the fresh cauline leaf of Siraitia grosvenorii is crushed to 20~40 mesh;The dosage of the water is the 4 of the fresh cauline leaf quality of Siraitia grosvenorii ~20 times;The temperature of the continuous flow upstream homogenate extraction is 50~90 DEG C, and number >=2 time of extraction, the time extracted every time is 60~200s;The revolving speed of the centrifugation is 10000~80000r/min;The mode of the centrifugal filtration is first horizontal screw centrifugal Filtering, then butterfly centrifugal filtering.
3. the method according to claim 1 or claim 2 for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf, feature exist In: in step (2), the ultrafiltration membrane for ultrafiltration is ceramic membrane, and the aperture of the ceramic membrane is 0.2~2.0 μm.
4. the method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf described according to claim 1~one of 3, special Sign is: in step (2), the flow velocity of upper prop is 1.0~4.0BV/h;The model D-101 type of the macroporous absorbent resin, AB- One or more of 8 types, DM-130 type, X-7 type or XDA-7 type;The macroporous absorbent resin and the fresh cauline leaf of Siraitia grosvenorii Volume mass ratio is 1:1~20;The diameter height of the large pore resin absorption column compares for 1:2~8.
5. the method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf described according to claim 1~one of 4, special Sign is: in step (2), the flow velocity of the washing is 1.0~4.0BV/h;The dosage of the organic solvent is 1~4BV;It is described The flow velocity of organic solvent elution is 0.25~2.00BV/h;The volume fraction of the organic solvent is 50~90%;It is described organic molten Agent is food-grade ethanol;The temperature of the reduced pressure is 40~70 DEG C, and pressure is -0.1~-0.07MPa, is concentrated into solid content It is 30~60%;The vacuum drying temperature is 40~70 DEG C, and pressure is -0.1~-0.07MPa, and the time is 4~12h.
6. the method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf described according to claim 1~one of 5, special Sign is: in step (3), the mass volume ratio of the flavonoid glycoside crude product A and alcoholic solution is 1:8~20;The temperature of the heating It is 50~80 DEG C;The food-grade ethanol that the alcoholic solution is food-grade anhydrous methanol or volume fraction is 90~99%;The standing Time be 2~4h;The temperature of the reduced pressure is 40~70 DEG C, and pressure is -0.1~-0.07MPa, is concentrated into solid content It is 30~60%;The vacuum drying temperature is 40~70 DEG C, and pressure is -0.1~-0.07MPa, and the time is 4~12h.
7. the method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf described according to claim 1~one of 6, special Sign is: in step (4), the flavonoid glycoside crude product B and mobile phase mass volume ratio are 0.08~0.30:1;The mobile phase is Volume ratio is organic solvent-aqueous solution of 15~40:95~60;The organic solvent is one of ethyl alcohol, methanol or acetonitrile Or it is several;Filter membrane for filtering is organic phase filter membrane, and aperture is 0.20~0.45 μm;The flow velocity of sample introduction be 0.25~ 1.00BV/h;The preparative liquid chromatography column is ODS column, and filler is C-18 reversed-phase bonded silica.
8. the method for extracting quercitin and mountain naphthalene glycosides simultaneously from Siraitia grosvenorii cauline leaf described according to claim 1~one of 7, special Sign is: in step (4), the flow velocity of the elution is 0.5~2.0BV/h;The temperature of the reduced pressure is 40~70 DEG C, pressure Power is -0.1~-0.07MPa, and being concentrated into solid content is 30~60%;The vacuum drying temperature be 40~70 DEG C, pressure be- 0.1~-0.07MPa, time are 4~12h.
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