CN105434539A - Composition of lotus flavones - Google Patents
Composition of lotus flavones Download PDFInfo
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- CN105434539A CN105434539A CN201510731265.3A CN201510731265A CN105434539A CN 105434539 A CN105434539 A CN 105434539A CN 201510731265 A CN201510731265 A CN 201510731265A CN 105434539 A CN105434539 A CN 105434539A
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Abstract
The invention discloses a composition of lotus flavones. The composition is prepared by the following steps: adding an ethanol solution to crude powder of lotus leaves; dissolving an extract with distilled water and purifying the solution with macroporous resin to obtain flavone powder; and treating the powder by a high-performance liquid chromatograph to obtain the composition of the lotus flavones with the content of 90%-98%. The composition is novel in conception, simple in process and low in production cost; the obtained compound is high in purity; and industrial production is easy to achieve.
Description
Technical field
The present invention relates to a kind of natural medicine field, be specifically related to a kind of compositions and preparation method of lotus flavone.
Background technology
Folium Nelumbinis is the blade of nymphaeaceae plant lotus (Nelumbonucifera6aertn.), main product in Jiangsu, Zhejiang, Hunan, Hubei, the ground such as Anhui, resource is very abundant, and output is huge.Folium Nelumbinis is except containing except the routine chemical componentses such as carbohydrate, lipid, protein, tannin, also main containing alkaloid, polyphenol and flavone three major types compound, and it all has at edible and medicinal applications applies more widely.Because Folium Nelumbinis has good antioxidation, fat-reducing slimming, blood pressure lowering, the effect such as antibacterial, and wide material sources, cheap, it is one of plant having Development volue most.But the utilization rate of Folium Nelumbinis plant resources is very low at present, often be only that about hundred tons are applied to food, medicine is produced, the adjuvant of Chinese herbal medicine decoction or other health product, remaining most of Folium Nelumbinis is all rot lotus Tanaka as fertilizer, and Folium Nelumbinis resource is not effectively utilized.Modern medicine study finds, Folium Nelumbinis has Adjust-blood lipid, scavenging free radicals, the effect such as antibacterial, and toxicity is minimum, and its main pharmacological component is Flavonoid substances.
CN101095724A discloses: a kind of technique extracting lotus flavone.Relate to the preparation technology extracting flavone from Folium Nelumbinis.Precipitation-extraction-ultrafiltration-absorption with macroporous adsorbent resin multiple techniques is adopted to refine lotus flavone; concentrated dealcoholysis postprecipitation eliminates water-insoluble impurity; petroleum ether extraction removes oil-soluble impurities; water-soluble macromolecule impurity removed by ultrafilter membrane; macroporous adsorbent resin selective absorption lotus flavone; stepwise elution collects dissimilar high-purity flavone, for pharmaceuticals industry uses the technique providing the extraction lotus flavone of large-scale production.
CN104844551A discloses: to be a kind ofly separated, the method for purify lotus flavone and polysaccharide simultaneously, after adopting alkaline aqueous solution lixiviate Folium Nelumbinis, obtain extracting solution, by the macroporous resin compositions of DA-201, D101 and DM-301, the lotus flavone in extracting solution and polysaccharide are adsorbed, water is first adopted to rinse macroporous resin column, by separation of polysaccharides, then adopt alcoholic solution to rinse macroporous resin column and obtain lotus flavone; The present invention utilizes the nature difference of lotus flavone and polysaccharide, influencing each other in various component extraction process, active and extraction ratio are taken into full account, adopt the macroporous resin compositions of DA-201, D101 and DM-301 to extract lotus flavone and polysaccharide simultaneously, there is the advantage that extraction ratio is high, purity is high; Adopt macroporous resin compositions to carry out separating-purifying to it, there is the advantage that technique is simple, production cost is low, product quality is high, with short production cycle, be applicable to large-scale production.
CN1923829A discloses: the preparation method of polyamide separation and purification lotus flavone and lotus leaf alkaloid, with the Folium Nelumbinis extract of the Folium Nelumbinis extract of Folium Nelumbinis crude extract, membrane separation for removing impurities or resin separation purification for raw material, add water and stir into suspension, polycaprolactam lotus flavone, non-adsorbed portion lease making decompression and solvent recovery vacuum concentration becomes lotus leaf alkaloid; The lotus flavone be adsorbed on polyamide washes depolarization impurity with water and pigment also uses polar organic solvent aqueous solution eluting, and eluent becomes lotus flavone through decompression and solvent recovery vacuum concentration;
CN101139320A discloses: adopt acidic hydrophilic organic solvent extraction nuciferine and lotus flavone, after extracting solution recovery section solvent, adopt cation exchange resin enriching and purifying nuciferine, residual feed liquid adopts polycaprolactam purification lotus flavone further.
CN104926766A discloses: a kind of method of simultaneous extraction Quercetin and nuciferine from Folium Nelumbinis, and nuciferine is water insoluble, and analogous alkaloid is also insoluble in aqueous alkali, and lotus flavone is dissolved in hot water, is soluble in aqueous alkali, first in alkaline environment, extract lotus leaf alkaloid with high purity methanol again with after potass extraction lotus flavone, alkali extract does not almost have alkaloid, part flavone is still had in alcohol extract, alcohol extract reclaims after alcohol concentrates and uses acid dissolve alkaloid, flavonoid thing is insoluble to acid, remove the Flavonoid substances in alkaloid after filtration, sour water alkalize to obtain alkaloid precipitation, alkali extracting solution filters to obtain acid non-soluble substance (flavonoid) merging treatment of flavone crude product and alcohol extraction part after acid is sunk, the supernatant alkalization aqueous alkali of alcohol extraction part and alkali being put forward part merges upper prop, use D101 resin absorption, part biological alkali residual in Recycling of waste liquid and flavonoid.
" high performance preparative liquid chromatography method is separated and prepares flavone compound from Folium Nelumbinis " (" chromatograph ", in January, 2007) discloses:: adopt high performance preparative liquid chromatography method to be separated from Folium Nelumbinis (NelumbonuciferaGaertn) and prepare Flavonoids in Lotus Leaves.With 60% alcohol reflux Folium Nelumbinis, be separated through D-101 post and polyamide column chromatography after crude extract is concentrated, be separated on SymmetryPrepTMC18 post again, carry out gradient elution (flow velocity is for 5.0mL/min) with water-acetonitrile for mobile phase, obtain 3 kinds of flavone compounds.Through ultraviolet spectra, infrared spectrum, nuclear magnetic resonance, NMR and mass spectral analysis, determine that these 3 kinds of materials are respectively hyperin, isoquercitrin and astragalin.The purity of 3 kinds of prepared compounds is all more than 97%, and wherein astragalin obtains for being separated from Folium Nelumbinis first.
" technical study of AB-8 purification by macroporous resin Folium Nelumbinis total flavones " (" Food Additives Used in China ") discloses: concentration is the adsorption column of lotus flavone extracting solution 100mL by AB-8 macroporous resin of 2.0mg/mL, interval 10mL collects flavone residual liquid: when resin blade diameter length ratio 1: 10; Adsorption flow rate 3BV/h; Sample solution pH value 5.0; Sample solution concentration is at 2.0mg/mL; Use the ethanol of 3BV consumption 90% as eluant; When parsing flow velocity is 1.5BV/h, lotus flavone purity is 53.44%.
" lotus flavone extracts, the research of separation and purification and antioxidant activity thereof " (Anhui Polytechnic University's Master's thesis, 2014) disclose: adopt water extraction to extract crude flavonoid powder from Folium Nelumbinis, with half preparative HPLC, compartment analysis is carried out to the lotus flavone after AB-8 macroporous resin preliminary purification, establish the best chromatographiccondition of lotus flavone high efficiency separation and mensuration: chromatographic column be WatersC18 reversed-phase column (150mmx " 0mmI.D., 5fim), mobile phase A: acetonitrile (chromatographically pure), Mobile phase B: ultra-pure water (containing 0.1% formic acid), sample size is 600pL, flow velocity is 3mL/min, column temperature is 30 DEG C, determined wavelength is 254mn, condition of gradient elution is: 0-20min, 5%A95%B-40%A60%B.Analyzing the five kinds of lotus flavone monomers obtained is Quercetin-3-O-Mulberry cloth bioside, hyperin, Quercetin-3-O-glucuronic acid, isoquercitrin and Quercetin.
Research report at present about Folium Nelumbinis total flavones is more, but concrete composition is indefinite, for this reason, needing to set up a kind of easy, economic method to obtain the lotus flavone powder of definite ingredients, supplying raw materials for preparing highly purified monomeric compound further.Separation purifying technique specially for Quercetin-3-O-β in Folium Nelumbinis-D-Glucose aldehydic acid glycosides has no any Patents and bibliographical information.
Summary of the invention
The present invention relates to:
A compositions for lotus flavone, in compositions, the weight content of flavone is 40-98%.
A compositions for lotus flavone, in compositions, the weight content of flavone is 40-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 30-65%.
A compositions for lotus flavone, in compositions, the weight content of flavone is 65-70%.
A compositions for lotus flavone, in compositions, the weight content of flavone is 65-70%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 60-65%.
5, a compositions for lotus flavone, in compositions, the weight content of flavone is 90-98%.
A compositions for lotus flavone, in compositions, the weight content of flavone is 90-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 81-96%.
A compositions for lotus flavone, in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 90-98%.
A compositions for lotus flavone,
The compositions of lotus flavone is through HPLC liquid phase analysis, and main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides appears in 22-23min, and the condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min,
Mobile phase is: acetonitrile-0.1% formic acid water, methanol-0.1% formic acid water, acetonitrile-water, methanol-water four kinds of mobile phases;
Mobile phase is preferably: acetonitrile-water: 20-80 volume ratio isocratic elution.
A compositions for lotus flavone,
The compositions of lotus flavone is through HPLC liquid phase analysis, and main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides appears in 22-23min, and another chromatographic peak appears in 23-24min; Main peak is 12: 1 with the ratio of the peak area of another chromatographic peak; The condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min,
Mobile phase is: acetonitrile-0.1% formic acid water, methanol-0.1% formic acid water, acetonitrile-water, methanol-water four kinds of mobile phases;
Mobile phase is preferably: acetonitrile-water: 20-80 volume ratio isocratic elution.
An extracting method for the compositions of lotus flavone,
(1) Folium Nelumbinis is dried below 60 DEG C, pulverize after prepare Folium Nelumbinis coarse powder;
(2) alcoholic solution adding 30-80% in Folium Nelumbinis coarse powder carries out circumfluence distillation, Extracting temperature is 60-95 DEG C, extract solid-liquid ratio and count 1: 10-1: 50 with quality and the ratio of volume, extraction time 2-4h, repeat lixiviate 2-4 time, after extracting solution concentrate drying, obtain the extract that flavone weight content is 3-5%; Said extracted thing adopts distilled water to dissolve, and suspension adopts purification with macroreticular resin further;
(3) distilled water and the 15-25% ethanol elution of 8-12 times of bed volume is adopted, water-solubility impurity such as removing polysaccharide, albumen etc.;
(4) adopt 30-40% ethanol to macroporous resin eluting;
(5) by concentrated for 30-40% ethanol elution warp, dry, pulverizing, mix homogeneously, obtain extract, in extract, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is the 60-65% of extract;
(6) above-mentioned powder dissolves through distilled water, centrifugal rear preparative high performance liquid chromatography instrument carries out separation preparation, take acetonitrile-water as mobile phase, flow velocity is 10-30mL/min, volatilize solvent, vacuum drying, obtain compositions, in compositions, the weight content of flavone is 90-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 81-96%.
An extracting method for the compositions of lotus flavone,
(1) Folium Nelumbinis is dried below 60 DEG C, pulverize after prepare Folium Nelumbinis coarse powder;
(2) alcoholic solution adding 30-80% in Folium Nelumbinis coarse powder carries out circumfluence distillation, Extracting temperature is 60-95 DEG C, extract solid-liquid ratio and count 1: 10-1: 50 with quality and the ratio of volume, extraction time 2-4h, repeat lixiviate 2-4 time, after extracting solution concentrate drying, obtain the extract that flavone weight content is 3-5%; Said extracted thing adopts distilled water to dissolve, and suspension adopts purification with macroreticular resin further.
(3) distilled water and the 15-25% ethanol elution of 8-12 times of bed volume is adopted, water-solubility impurity such as removing polysaccharide, albumen etc.;
(4) adopt 30-40% ethanol to macroporous resin eluting;
(5) by concentrated for 30-40% ethanol elution warp, dry, pulverizing, mix homogeneously, obtain extract, in extract, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is the 60-65% of extract;
(6) above-mentioned powder dissolves through distilled water, centrifugal rear preparative high performance liquid chromatography instrument carries out separation preparation, take acetonitrile-water as mobile phase, flow velocity is 10-30mL/min, volatilize solvent, vacuum drying, obtain compositions, in compositions, the weight content of flavone is 90-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 81-96%;
Described in step 2, macroporous adsorbent resin comprises nonpolar macroporous adsorption resin, LX-60 and X-5; Low pole macroporous adsorbent resin, HPD722 and LX-5;
Intermediate-polarity macroporous adsorption resin, HPD750 and AB-8;
Containing Quercetin-3-O-β-D-Glucose aldehydic acid glycosides in lotus flavone compositions;
Extract in step 5, belongs to the compositions of lotus flavone.
In step 5, the compositions of lotus flavone is through HPLC liquid phase analysis, chromatogram there are two characteristic peaks, what 22-23min occurred is main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides, and another chromatographic peak appears in 23-24min, and main peak is 12: 1 with the ratio of the peak area of another chromatographic peak; The condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min, acetonitrile-water: 20-80 volume ratio isocratic elution.
Mobile phase of the present invention comprises:
Acetonitrile-0.05% formic acid water, acetonitrile-0.1% formic acid water, acetonitrile-0.14% formic acid water, acetonitrile-0.2% formic acid water, acetonitrile-0.24% formic acid water, acetonitrile-0.3% formic acid water, methanol-0.1% formic acid water, acetonitrile-water, methanol-water;
Preferred: acetonitrile-0.1% formic acid water (10-90 volume ratio), acetonitrile-water (20-80 volume ratio) isocratic elution.
An extracting method for lotus flavone, is characterized in that: the selection and optimization of the extraction conditions of powder.The extraction effect that it is extractant that the present invention compares with hot water, alkali liquor, ethanol.
The present invention is best with the extraction effect of ethanol.Extraction ratio is: 87%-95%
The present invention with 30%, 40%, 50%, 60%, 70%, the ethanol water of 80%6 concentration extracts the flavone compound in Folium Nelumbinis, and the extraction ratio of the ethanol water of 50% is the highest.
An extracting method for lotus flavone, is characterized in that: herbal extract, 15-25%, 30-40%, 40-50% ethanol elution are analyzed through HPLC respectively, the following Fig. 1-4 of chromatogram.
Elution requirement: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min, 20% acetonitrile isocratic elution.Reference substance is the Quercetin-3-O-β-D-Glucose aldehydic acid glycosides standard substance of purity more than 98%.
The selection and optimization of high performance preparative liquid chromatography condition
The selection and optimization of mobile phase: the present invention compares acetonitrile-0.1% formic acid water, methanol-0.1% formic acid water, acetonitrile-water, methanol-water four kinds of mobile phases to the separating effect of chromatographic peak.
Gradient elution program: the present invention, by changing flow velocity, progressively adjusting organic facies ratio in mobile phase, segments step by step,
Preferred detection wavelength 360nm, column temperature 25 DEG C, flow velocity 10-30mL/min, the chromatographic separation condition of 20% acetonitrile isocratic elution.Quercetin-3-O-β-D-Glucose aldehydic acid glycosides the standard substance of reference substance purity more than 98%.
Different gradient elution program chromatogram is as Fig. 5-7.
In mobile phase aqueous phase, formic acid concn has considerable influence to chromatographic peak separating degree, adds formic acid than not adding formic acid and can obtain good separating degree; Organic facies ratio has considerable influence to retention time, and organic facies proportion is larger, and retention time is shorter, and separating degree is poorer; Column temperature also has considerable influence to separating degree, and too high too low separating degree is all poor;
Therefore select determined wavelength 360nm, column temperature 25 DEG C, flow velocity 10-30mL/min,
Therefore preferably acetonitrile-0.1% formic acid water (15: 85 volume ratio) or acetonitrile-water (20-80 volume ratio) isocratic elution are chromatographic separation condition.
A kind of compositions of lotus flavone is through HPLC liquid phase analysis, what 22-23min occurred is main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides, there is another chromatographic peak in 23-24min, the condition that main peak is 12: 1HPLC liquid phase analysis with the ratio of the peak area of another chromatographic peak is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min, 20% acetonitrile isocratic elution.
A compositions for lotus flavone, the computational methods of flavone weight content:
W=F
1* A
sample* V
sample* (1+1/12) * 100%/m
group, wherein F
1=C
right/ A
right
W, A
sample, V
sample, m
group, C
right, A
rightbe defined as follows:
W is weight percentage composition
A
samplefor the peak area of Quercetin-3-O-β in sample liquid-D-Glucose aldehydic acid glycosides
V
samplefor sample liquid volume
M
groupfor composition quality
C
rightfor reference substance concentration
A
rightfor reference substance peak area
A compositions for lotus flavone,
The computational methods of the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides:
W=F
1* A
sample* V
sample* 100%/m
group, wherein F
1=C
right/ A
right
W, A
sample, V
sample, m
group, C
right, A
rightbe defined as follows:
W is weight percentage composition
A
samplefor the peak area of Quercetin-3-O-β in sample liquid-D-Glucose aldehydic acid glycosides
V
samplefor sample liquid volume
M
groupfor composition quality
C
rightfor reference substance concentration
A
rightfor reference substance peak area
(Fig. 8,9) nuclear magnetic data and (Figure 10) mass spectrometric data are shown in the qualification of Quercetin-3-O-β in Folium Nelumbinis-D-Glucose aldehydic acid glycosides.
Advantage of the present invention and beneficial effect are:
(1) the present invention adopts Folium Nelumbinis that is cheap, wide material sources to be flavone in a kind of Folium Nelumbinis of raw material separation and purification.The compound purity obtained is high, and technique is comparatively simple, and the investment such as equipment is less, easily amplifies, is comparatively suitable for large-scale industrialized production etc.
(2) the present invention significantly can improve the weight content of Quercetin-3-O-β in compositions-D-Glucose aldehydic acid glycosides, and its weight content can be increased to 60-65% by the 3-5% in medicinal substances extract, after preparing the preparation of efficient liquid phase, be increased to 90-98%.
Accompanying drawing explanation
Fig. 1: the HPLC chromatogram of herbal extract
The HPLC chromatogram of Fig. 2: 15-25% eluent
The HPLC chromatogram of Fig. 3: 30-40% eluent
The HPLC chromatogram of Fig. 4: 40-60% ethanol elution
Fig. 5: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 25mL/min, the HPLC chromatogram of 25% acetonitrile-water isocratic elution
Fig. 6: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 25mL/min, the HPLC chromatogram of 23% acetonitrile-water isocratic elution
Fig. 7: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 25mL/min, mobile phase is acetonitrile-0.3% formic acid water, the HPLC chromatogram of 22% isocratic elution
The hydrogen spectrum of Fig. 8: Quercetin-3-O-β-D-Glucose aldehydic acid glycosides
The carbon spectrum of Fig. 9: Quercetin-3-O-β-D-Glucose aldehydic acid glycosides
The mass spectrum of Figure 10: Quercetin-3-O-β-D-Glucose aldehydic acid glycosides
Detailed description of the invention
The present invention is described in further detail by following examples.
Embodiment 1
The method step of a kind of lotus flavone of employing macroporous adsorbent resin separation and purification involved in the present invention is as follows:
(1) Folium Nelumbinis dried, pulverize, obtain Folium Nelumbinis coarse powder;
(2) 300g Folium Nelumbinis coarse powder is taken, the solid-liquid ratio of 1: 30 (W/V) is adopted to add 9 liter of 50% alcoholic solution in this Folium Nelumbinis powder, 90 DEG C of circumfluence distillation time 3h, repeat lixiviate 3 times, after extracting solution merges, concentrate drying obtains extract, and in extract, flavone weight content is 3%; Said extracted thing dynamic loading macroporous adsorbent resin LX-60 after distilled water ultrasonic dissolution carries out purification;
(3) adopt the distilled water of 10 times of bed volumes and 25% ethanol to carry out eluting respectively, fully clean a large amount of water-solubility impurity such as polysaccharide, albumen;
(4) 35% ethanol is adopted to carry out eluting to macroporous resin;
(5) 35% ethanol elution through concentrated, vacuum decompression is dry, pulverize, after mix homogeneously, obtain powder, the weight content of the Quercetin-3-O-β in powder-D-Glucose aldehydic acid glycosides is 60%.
(6) above-mentioned powder adopts preparative high performance liquid chromatography instrument to carry out separation and purification after distilled water ultrasonic dissolution is centrifugal, flow velocity is 16mL/min, determined wavelength 360nm, column temperature 25 DEG C, acetonitrile-water (20-80 volume ratio) isocratic elution, namely obtain yellow crystals, in this crystal, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 98%.
Embodiment 2
(1) Folium Nelumbinis dried, pulverize, obtain Folium Nelumbinis coarse powder;
(2) 200g Folium Nelumbinis coarse powder is taken, the solid-liquid ratio (W/V) of 1: 40 is adopted to add 8 liter of 70% alcoholic solution in this Folium Nelumbinis coarse powder, 80 DEG C of circumfluence distillation time 4h, repeat lixiviate 2 times, after extracting solution merges, concentrate drying obtains extract, and in extract, flavone weight content is 3%.Said extracted thing adopts the dynamic loading of macroporous adsorbent resin LX-5 to carry out purification after distilled water ultrasonic dissolution;
(3) adopt distilled water and 18% ethanol elution of 8 times of bed volumes respectively, fully clean a large amount of water-solubility impurity such as polysaccharide, albumen;
(4) 30% ethanol is adopted to carry out eluting to macroporous resin;
(5) 30% ethanol elution is obtained powder after concentrated, lyophilization, pulverizing, mix homogeneously, in powder, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 62%.
(6) above-mentioned powder is through distilled water ultrasonic dissolution, after suspension is centrifugal, preparative high performance liquid chromatography instrument is adopted to carry out separation and purification, flow velocity is 20mL/min, determined wavelength 360nm, column temperature 25 DEG C, acetonitrile-water (25-75 volume ratio) isocratic elution, namely obtain yellow crystals, in this crystal, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 95%.
Embodiment 3
(1) Folium Nelumbinis dried, pulverize, form Folium Nelumbinis coarse powder;
(2) 400g Folium Nelumbinis coarse powder is taken, the solid-liquid ratio (W/V) of 1: 20 is adopted to add 8 liter of 60% alcoholic solution in this Folium Nelumbinis coarse powder, 70 DEG C of circumfluence distillation time 2h, repeat lixiviate 4 times, after extracting solution merges, concentrate drying obtains extract, and in extract, flavone weight content is 4%.Said extracted thing adopts the dynamic loading of macroporous adsorbent resin AB-8 to carry out purification after distilled water ultrasonic dissolution;
(3) adopt distilled water and 20% ethanol elution of 9 times of bed volumes respectively, fully clean a large amount of water-solubility impurity such as polysaccharide, albumen;
(4) ethanol of 32% is adopted to carry out eluting to macroporous resin;
(5) 32% ethanol elution through concentrated, vacuum decompression is dry, pulverize, obtain powder after mix homogeneously, in powder, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 64%.
(6) above-mentioned powder is through distilled water ultrasonic dissolution, after suspension is centrifugal, preparative high performance liquid chromatography instrument is adopted to carry out separation and purification, flow velocity is 18mL/min, determined wavelength 360nm, column temperature 25 DEG C, acetonitrile-water (20-80 volume ratio) isocratic elution, namely obtain yellow crystals, confirm through structure detection, in this crystal, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 96%.
Embodiment 4
A compositions for lotus flavone is 40-98% flavone containing weight content, and the HPLC analytical method of flavone is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4mL/min, acetonitrile-water (20-80 volume ratio) isocratic elution.
Embodiment 5
A kind of compositions of lotus flavone, be 40-98% flavone containing weight content, the weight content of wherein main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 30-65%, the HPLC analytical method of flavone is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.5mL/min, acetonitrile-water (20-80 volume ratio) isocratic elution.
Embodiment 6
A compositions for lotus flavone is 65-70% flavone containing weight content.The HPLC analytical method of flavone is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.5mL/min, acetonitrile-water (20-80 volume ratio) isocratic elution.
Embodiment 7
A compositions for lotus flavone is 65-70% flavone containing weight content, and the weight content of wherein main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 60-65%.The HPLC analytical method of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.6mL/min, mobile phase acetonitrile-0.3% formic acid water (20-80 volume ratio) isocratic elution.
Embodiment 8
A compositions for lotus flavone is 90-98% flavone containing weight content.The HPLC analytical method of flavone is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.6mL/min, mobile phase acetonitrile-0.05% formic acid water (20-80 volume ratio) isocratic elution.
Embodiment 9
A compositions for lotus flavone is 90-98% flavone containing weight content, and the weight content of wherein main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 90-98%.The HPLC analytical method of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.6mL/min, mobile phase acetonitrile-0.24% formic acid water (20-80 volume ratio) isocratic elution.
Embodiment 10
A compositions for lotus flavone, the compositions of lotus flavone is through HPLC liquid phase analysis, and what 22-23min occurred is main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides;
The condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.6mL/min, mobile phase acetonitrile-0.14% formic acid water (20-80 volume ratio).
Embodiment 11
A compositions for lotus flavone, the compositions of lotus flavone is through HPLC liquid phase analysis, and what 22-23min occurred is main peak Quercetin-3-O-β-D glucuronide, and another chromatographic peak appears in 23-24min; Main peak is 12: 1 with the ratio of the peak area of another chromatographic peak.
The condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.6mL/min, mobile phase acetonitrile-0.14% formic acid water (20-80 volume ratio) isocratic elution.
Embodiment 12
Tested by antioxidation, the dosage that digital proof is identical, the specific activity medicinal substances extract through the flavone composition of extraction purification is good.
Although be described the specific embodiment of the present invention, those skilled in the art will appreciate that and can carry out multiple change and modification to the present invention under the prerequisite not departing from scope of the present invention or spirit.Thus, this invention is intended to contain all these dropping within the scope of claims and coordinate thereof change and modify.
Claims (10)
1. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of flavone is 40-98%.
2. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of flavone is 40-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 30-65%.
3. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of flavone is 65-70%.
4. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of flavone is 65-70%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 60-65%.
5. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of flavone is 90-98%.
6. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of flavone is 90-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 81-96%.
7. a compositions for lotus flavone, is characterized in that: in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 90-98%.
8., according to the compositions of a kind of lotus flavone of claim 1-7, it is characterized in that:
The compositions of lotus flavone is through HPLC liquid phase analysis, and main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides appears in 22-23min, and the condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min,
Mobile phase is: acetonitrile-0.1% formic acid water, methanol-0.1% formic acid water, acetonitrile-water, methanol-water four kinds of mobile phases;
Mobile phase is preferably: acetonitrile-water: 20-80 volume ratio isocratic elution.
9., according to the compositions of a kind of lotus flavone of claim 1-7, it is characterized in that:
The compositions of lotus flavone is through HPLC liquid phase analysis, and main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides appears in 22-23min, and another chromatographic peak appears in 23-24min; Main peak is 12: 1 with the ratio of the peak area of another chromatographic peak; The condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min,
Mobile phase is: acetonitrile-0.1% formic acid water, methanol-0.1% formic acid water, acetonitrile-water, methanol-water four kinds of mobile phases;
Mobile phase is preferably: acetonitrile-water: 20-80 volume ratio isocratic elution.
10., according to the extracting method of the compositions of a kind of lotus flavone of claim 1-7, it is characterized in that:
(1) Folium Nelumbinis is dried below 60 DEG C, pulverize after prepare Folium Nelumbinis coarse powder;
(2) alcoholic solution adding 30-80% in Folium Nelumbinis coarse powder carries out circumfluence distillation, Extracting temperature is 60-95 DEG C, extract solid-liquid ratio and count 1: 10-1: 50 with quality and the ratio of volume, extraction time 2-4h, repeat lixiviate 2-4 time, after extracting solution concentrate drying, obtain the extract that flavone weight content is 3-5%; Said extracted thing adopts distilled water to dissolve, and suspension adopts purification with macroreticular resin further;
(3) distilled water and the 15-25% ethanol elution of 8-12 times of bed volume is adopted, water-solubility impurity such as removing polysaccharide, albumen etc.;
(4) adopt 30-40% ethanol to macroporous resin eluting;
(5) by concentrated for 30-40% ethanol elution warp, dry, pulverizing, mix homogeneously, obtain extract, in extract, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is the 60-65% of extract; ;
(6) above-mentioned powder dissolves through distilled water, centrifugal rear preparative high performance liquid chromatography instrument carries out separation preparation, take acetonitrile-water as mobile phase, flow velocity is 10-30mL/min, volatilize solvent, vacuum drying, obtain compositions, in compositions, the weight content of flavone is 90-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 81-96%;
An extracting method for the compositions of lotus flavone, is characterized in that:
(1) Folium Nelumbinis is dried below 60 DEG C, pulverize after prepare Folium Nelumbinis coarse powder;
(2) alcoholic solution adding 30-80% in Folium Nelumbinis coarse powder carries out circumfluence distillation, Extracting temperature is 60-95 DEG C, extract solid-liquid ratio and count 1: 10-1: 50 with quality and the ratio of volume, extraction time 2-4h, repeat lixiviate 2-4 time, after extracting solution concentrate drying, obtain the extract that flavone weight content is 3-5%; Said extracted thing adopts distilled water to dissolve, and suspension adopts purification with macroreticular resin further.
(3) distilled water and the 15-25% ethanol elution of 8-12 times of bed volume is adopted, water-solubility impurity such as removing polysaccharide, albumen etc.;
(4) adopt 30-40% ethanol to macroporous resin eluting;
(5) by concentrated for 30-40% ethanol elution warp, dry, pulverizing, mix homogeneously, obtain extract, in extract, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is the 60-65% of extract;
(6) above-mentioned powder dissolves through distilled water, centrifugal rear preparative high performance liquid chromatography instrument carries out separation preparation, take acetonitrile-water as mobile phase, flow velocity is 10-30mL/min, volatilize solvent, vacuum drying, obtain compositions, in compositions, the weight content of flavone is 90-98%, and in compositions, the weight content of Quercetin-3-O-β-D-Glucose aldehydic acid glycosides is 81-96%;
Described in step 2, macroporous adsorbent resin comprises nonpolar macroporous adsorption resin, LX-60 and X-5; Low pole macroporous adsorbent resin, HPD722 and LX-5;
Intermediate-polarity macroporous adsorption resin, HPD750 and AB-8;
Containing Quercetin-3-O-β-D-Glucose aldehydic acid glycosides in lotus flavone compositions;
In step 5, the compositions of lotus flavone is through HPLC liquid phase analysis, chromatogram there are two characteristic peaks, what 22-23min occurred is main peak Quercetin-3-O-β-D-Glucose aldehydic acid glycosides, and another chromatographic peak appears in 23-24min, and main peak is 12: 1 with the ratio of the peak area of another chromatographic peak; The condition of HPLC liquid phase analysis is: determined wavelength 360nm, column temperature 25 DEG C, flow velocity 0.4-1.0mL/min, acetonitrile-water: 20-80 volume ratio isocratic elution.
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| CN111387477A (en) * | 2020-05-06 | 2020-07-10 | 江西师范大学 | Microcapsule embedding technology for improving digestion stability of lotus leaf flavone |
| CN114441687A (en) * | 2022-02-17 | 2022-05-06 | 山东福瑞达生物股份有限公司 | Fingerprint spectrum construction method and application of antioxidant lotus leaf extract |
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