CN103120706B - A kind of liquid chromatograph mass spectrography prepares the method for high-purity rutoside - Google Patents
A kind of liquid chromatograph mass spectrography prepares the method for high-purity rutoside Download PDFInfo
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Abstract
The present invention relates to medical art, it is a kind of method that two-dimensional liquid chromatography-mass spectrometry prepares high-purity rutoside, adopt two-dimensional liquid chromatography-mass spectrometry technology, with methanol-water or acetonitrile-water for mobile phase, take reverse-phase chromatographic column as one dimension preparative chromatography post, carry out component cutting to ginkgo biloba p.e, collecting target components is rutin crude product.Again with reverse-phase chromatographic column for two-dimentional preparative chromatography post, mass spectrum Selective ion mode peak instruct under, component cutting is carried out to rutin crude product, collect, rotary evaporation concentrate, obtain highly purified rutin, purity can reach more than 80%.The present invention, without the need to heating, avoids the doping of hydrolysate, and preparation process is simple, repeatability is high, and operability is good, and ginkgo leaf aboundresources, easily obtains, reduce cost simultaneously, is applicable to large-scale production.
Description
Technical field
The invention belongs to medical art, specifically, relate to a kind of method that two-dimensional liquid chromatography-mass spectrometry prepares high-purity rutoside.
Background technology
Rutin (Rutin), also known as rutin sophorin, is extensively present in plant kingdom.Existing more than 50 kinds of the plant containing rutin had now found that.Rutin belongs to flavone compound, can reduce capillary permeability and fragility, promotes hyperplasia and prevents haemocyte from condensing, the treatment of cardiovascular and cerebrovascular disease plays the important and pivotal role.Meanwhile, rutin also has the effect of the aspects such as anti-inflammatory, antiallergy, diuresis, spasmolysis, antibechic, reducing blood lipid.Clinically rutin be mainly used in high blood pressure supplemental treatment and for prevent and treat because of rutin lack caused by other haemorrhage, as prevent and treat cerebrovascular hemorrhage, hypertension, retinal hemorrhage, purpura, acute hemorrhagic ephritis, chronic bronchitis, colloidal osmotic pressure abnormal, recover the diseases such as capillary elasticity, simultaneously also for prevention and therapy diabetes and merge hyperlipemia.Make raw material with rutin in domestic and international medical industry and produce the cardiovascular drugses such as rutin tablets, troxerutin (monoxerutin), rutin magnesium chromic salts, multivita-glucose hydroxyl fourth rutin, Quercetin.Particularly HR sheet is suitable for inaccessible type cerebrovascular disease, central retinitis, artery sclerosis, coronary heart disease, preinfarction syndrome, thrombophlebitis etc. because of it, expand range of application and the curative effect of rutin product, the demand of rutin is increased further.
For the extraction of rutin, usually adopt alkali to extract sour intermediate processing, this method utilizes rutin to have acidity, can with alkali salify and water-soluble, then acid adding make its become acid after freely to separate out, then obtain purer rutin through recrystallization.Although this method is regarded as comparatively ideal method for many years always, because heating is extracted, often have certain hydrolysate to exist, and productive rate is lower.In recent years, people start progressively Exploration and application new technology, new method extraction purification rutin, comprise hot water and carry cold separation method, hot water puies forward macroporous resin adsorption method of purification, ultrasonic method, supercritical CO_2 extraction method, continuous circumfluence extraction method, the decocting method, (Meng Xiangying such as percolation.The source of rutin, purposes and method for extraction and purification, Changchun Traditional Chinese Medical College's journal, the 2003,19th volume the 2nd phase: 61-64).But these current methods, treatment step is all more loaded down with trivial details, and most method all exists high-temperature heating and occurs hydrolysate, makes subsequent processing steps increase.In order to make full use of medicinal material; rutin is enable to be produced out with script state from medicinal material, without the need to adding through soda acid, without the need to heating; avoid the doping of hydrolysate, therefore need to develop a kind of easy and simple to handle, preparation method of being easy to large-scale production, high efficiency, high-purity rutoside.
Ginkgo leaf is the dry leaf of Ginkgoaceae plant Ginkgo biloba (Ginkgobiloba), and its extract has effect of activating blood circulation to dissipate blood stasis and dredge the collateral.Present pharmacological research shows, is rich in a large amount of flavones and ginkgolides in ginkgo leaf.China is ancient country and the big country of gingko resource, national You20Duo Ge province, city and region establishing in large scale ginkgo, and gingko resource accounts for global 70%, the total output of ginkgo leaf every year at 2 × more than 107kg, resource extreme enrichment.And the production of China rutin is mainly that raw material extracts with the sophora bud at present.The sophora bud is the bud of sophora flower, and resource is few, fully can not meet the demand in market.Therefore utilize ginkgo leaf to extract rutin, the present situation of rutin raw material supply anxiety can be alleviated, expand the source of cardiovascular drugs, have important practical significance.
Two-dimensional liquid chromatography technology is an emerging technology, is to be separated complex sample by the chromatographic column that two character is identical or different.As a kind of ideal tools of Analysis of Complex sample, it has been widely used in the every field such as medical and health, Food Science, environment and agricultural sciences.In recent years, along with two-dimensional liquid chromatography is theoretical and the development of application, two-dimension preparation liquid chromatographic also grows up gradually.Compared with one-dimensional preparative liquid chromatography, two-dimension preparation liquid chromatographic not only has higher peak capacity, and bidimensional has different selective.The component simplified through the first dimension can according to different selective and realize complementary separation in the second dimension.The similar compound of structure effectively may be separated in the second dimension, thus improves efficiency and the compound purity of preparation.
Summary of the invention
For above-mentioned defect, have hydrolysate doping, complex operation in the extraction preparation of current rutin, medicinal material utilization rate is low, the problem of herb resource deficiency, the invention provides a kind of is raw material with ginkgo biloba p.e, adopts two-dimensional liquid chromatography-mass-spectrometric technique to prepare the method for rutin.
For solving the problem, the technical solution adopted in the present invention is:
Two-dimensional liquid chromatography-mass spectrometry prepares a method for high-purity rutoside, it is characterized in that: step is as follows:
High-purity rutoside is prepared from Chinese medical extract;
Chinese medical extract is dissolved in methanol-water solution, obtained Chinese medical extract solution;
Separating and purifying technology is adopted to prepare high-purity rutoside from Chinese medical extract;
Say further:
Described Chinese medical extract is ginkgo biloba p.e.
Furthermore:
Described ginkgo biloba p.e is dissolved in the methanol-water solution of 25%-75%, obtained ginkgo biloba p.e solution, and ginkgo biloba p.e concentration is 10-500mg/mL;
Described ginkgo biloba p.e is dissolved in the methanol-water solution of 40-60%, and ginkgo biloba p.e concentration the best is 150-250mg/mL;
Furthermore:
Described separating and purifying technology is that two-dimensional liquid chromatography-mass spectrometry carries out ginkgo biloba p.e and prepares rutin;
Adopting liquid chromatogram to carry out chromatographic column prepared by one dimension is anti-phase C18, anti-phase C8, ClickOEG.The chromatographic column adopting liquid chromatogram to carry out two dimension preparation is anti-phase C18, anti-phase C8, ClickOEG;
The mobile phase that the preparation of one dimension liquid chromatogram adopts is that organic phase is mixed with water, and elution requirement is brought up to 70% gradient according to the degree such as organic phase 5-65% or 0-50 minute organic phase by 5% and carried out; Organic phase is methyl alcohol or acetonitrile, and aqueous phase is water;
Collect the component that retention time 5-30 minute principal component is rutin, rotary evaporated to dryness, prepares rutin crude product as one dimension; Methanol-water solution rutin crude product 25%-75% being prepared by one dimension dissolves, and is 10-80mg/mL to concentration;
Prepared by rutin crude product to one dimension, carries out two-dimensional liquid chromatography preparation; The mobile phase adopted is that organic phase is mixed with water, and elution requirement is spent according to organic phase 5-30% etc. or brought up to 40% gradient through 0-30 minute organic phase by 5% and carries out, and two dimension preparation instructs Fraction collection according to mass spectrum Selective ion mode; Organic phase is methyl alcohol or acetonitrile, and aqueous phase is water;
During preparation, flow rate of mobile phase is 60-120mL/min, and during preparation, column temperature is room temperature or 25-40 DEG C, and during preparation, sample size is 200-3000 μ L/ pin, and detector is DAD UV-detector and mass detector; During preparation, mobile phase optimum flow rate is 80-100mL/min, and during preparation, column temperature is room temperature, and during preparation, sample size is 2000-2500 μ L/ pin, and UV-detector is 360nm;
The rutin cut collected prepared by two-dimensional liquid chromatography to carry out rotary evaporation and be concentrated into dry, obtain rutin compound.
Furthermore:
It 10-25 minute is the main component containing rutin that described one dimension preparation is collected, and the methanol-water solution that one dimension prepares rutin crude product 40%-60% makes the rutin crude product solution that concentration is 40-60mg/mL.
It is acetonitrile (or containing 0.1% formic acid) and water (or containing 0.1% formic acid) mutually that described one dimension prepares optimal flow, and adopt Gradient methods, acetonitrile ratio was increased to 15%-25% by 10% through 15-20 minute;
Furthermore:
Described two dimension preparation optimal flow is acetonitrile (or containing 0.1% formic acid) and water (or containing 0.1% formic acid) mutually, adopts isocratic condition, and acetonitrile (or containing 0.1% formic acid) concentration is 10-15%, and it is m/z=303 that two dimension prepares mass spectrum Selective ion mode.
Owing to have employed technique scheme, compared with prior art, the present invention compared with prior art has following characteristics:
1), rutin purity is high.Due to the technical method that the present invention is used, be directly prepared for rutin contained in plant, without the need to the change through heating, soda acid, rutin is produced out with the state that itself exists in medicinal material, do not have hydrolysate to adulterate, the rutin purity therefore obtained is high, reaches more than 80%.
2), method is simple, easy and simple to handle.The involved in the present invention method preparing high-purity rutoside, automation equipment degree is high, convenient and simple for operation, just can carry out under normal temperature and pressure, is applicable to the needs of large-scale production.
3), crude drug source is wide, easily obtains, low price.Ginkgo is the distinctive rare tree of China, accounts for 70% of whole world resource, the total output of ginkgo leaf every year at 2 × more than 107kg, resource extreme enrichment.
Three nine-day periods after the winter solstice board Shu Xuening injection, ginkgo leaf tincture are of a specified duration as Applicative time, the traditional Chinese medicine of clear curative effect, use safety, have a wide range of applications, and the high-purity rutoside that the present invention obtains is a kind of important raw material in three nine-day periods after the winter solstice board Shu Xuening injection, ginkgo leaf tincture.And preparation method provided by the invention, process repeatability is high, and operability is good, simultaneously ginkgo leaf aboundresources, easily obtains, and is applicable to the requirement of large-scale production, can be used for the preparation of Shu Xuening injection, ginkgo leaf tincture raw material high-purity rutoside.
Detailed description of the invention
Embodiment:
The present invention take ginkgo biloba p.e as raw material, and adopt two-dimensional liquid chromatography-mass-spectrometric technique to prepare the method for high-purity rutoside, the purity of rutin can reach more than 80%.
The present invention is the method preparing high-purity rutoside from ginkgo biloba p.e, and step is as follows:
1, ginkgo biloba p.e is dissolved in the methanol-water solution of 25%-75%, obtained ginkgo biloba p.e solution, ginkgo biloba p.e concentration is 10-500mg/mL, and best ginkgo biloba p.e is dissolved in the methanol-water solution of 40-60%, and ginkgo biloba p.e concentration the best is 150-250mg/mL.
2, adopting liquid chromatogram to carry out chromatographic column prepared by one dimension is anti-phase C18, anti-phase C8, ClickOEG.The chromatographic column adopting liquid chromatogram to carry out two dimension preparation is anti-phase C18, anti-phase C8, ClickOEG.Best Two way chromatograms column combination is one dimension is anti-phase C18, and two dimension is anti-phase C18.
3, the mobile phase that the preparation of one dimension liquid chromatogram adopts is that methyl alcohol or acetonitrile mix with water, and elution requirement is brought up to 70% gradient according to the degree such as organic phase 5-65% or 0-50 minute organic phase by 5% and carried out.It is acetonitrile (or containing 0.1% formic acid) and water (or containing 0.1% formic acid) mutually that one dimension prepares optimal flow, and adopt Gradient methods, organic phase ratio was increased to 15-25% by 10% through 15-20 minute.
4, collect the component that retention time 5-30 minute principal component is rutin, rotary evaporated to dryness, prepares rutin crude product as one dimension.Rutin crude product methanol-water being prepared by one dimension to dissolve, is 10-80mg/mL to concentration.It is 10-25 minute that one dimension prepares best acquisition time, and be the component mainly containing rutin, being concentrated into concentration is 40-60mg/mL.
5, the sample after concentrating is collected to one dimension, carry out two-dimensional liquid chromatography preparation.The mobile phase adopted is that methyl alcohol or acetonitrile mix with water, and elution requirement is spent according to organic phase 5-30% etc. or brought up to 40% gradient through 0-30 minute organic phase by 5% and carries out.Two dimension preparation optimal flow is acetonitrile (or containing 0.1% formic acid) and water (or containing 0.1% formic acid) mutually, adopts isocratic condition, and acetonitrile (or containing 0.1% formic acid) concentration is 10-15%; Two dimension preparation instructs Fraction collection according to mass spectrum Selective ion mode, and mass spectrum Selective ion mode is m/z=303.
6, during preparation, flow rate of mobile phase is 60-120mL/min, and during preparation, column temperature is room temperature or 25-40 DEG C, and during preparation, sample size is 200-3000 μ L/ pin, and detector is DAD UV-detector and mass detector, and mass spectrum Selective ion mode instructs Fraction collection; Optimum flow rate is 80-100mL/min, and column temperature is room temperature, and sample size is 2000-2500 μ L/ pin, and UV-detector is 360nm.
7, two-dimensional liquid chromatography being prepared the rutin cut rotary evaporation collected is concentrated into dry, obtains highly purified rutin compound.
Be below according to the specific embodiment of the present invention, in conjunction with actual, the invention provides four kinds of embodiments.
Embodiment 1:
Take ginkgo biloba p.e 10g, be dissolved in the methanol-water solution of 50mL50%, obtained ginkgo biloba p.e solution, concentration is 200mg/mL, crosses 0.45 μm of miillpore filter, carries out the preparation of one dimension liquid chromatogram.One dimension liquid chromatogram adopts XUnionC18, and mobile phase adopts acetonitrile (containing 0.1% formic acid) to be organic phase, and water (containing 0.1% formic acid) is aqueous phase, gradient elution mode: organic phase concentration brought up to 25% by 10% through 20 minutes.Adopt DAD UV-detector 360nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 500 μ L/ pins, and flow rate of mobile phase is 90mL/min, collects the cut of 12-14 minute, carries out rotary evaporation and be concentrated into dry, for one dimension prepares rutin crude product.Methanol-water solution with 40% dissolves rutin crude product, concentration is 50mg/mL, through filtering with microporous membrane, carry out two-dimensional liquid chromatography preparation, chromatographic column is XUnionC18, mobile phase selects acetonitrile (containing 0.1% formic acid) to be organic phase, and water (containing 0.1% formic acid) is aqueous phase, adopts 10% organic phase isocratic elution.Adopt DAD UV-detector 360nm Selective absorber wavelength, mass spectrum Selective ion mode m/z is 303, preparation temperature is room temperature, and sample size is 1000 μ L/ pins, and flow rate of mobile phase is 90mL/min, collecting m/z is the cut of 303, rotary evaporated to dryness, obtains rutin compound, through liquid-phase chromatographic analysis, purity is 99%, and the content that one dimension prepares rutin in rutin crude product is 45%.
Embodiment 2:
Take ginkgo biloba p.e 10g, be dissolved in the methanol-water solution of 100mL25%, obtained ginkgo biloba p.e solution, concentration is 10mg/mL, crosses 0.45 μm of miillpore filter, carries out the preparation of one dimension liquid chromatogram.One dimension liquid chromatogram adopts XTerra, mobile phase adopts methyl alcohol to be organic phase, water is aqueous phase, and organic phase concentration is 65% degree such as grade, adopts DAD UV-detector 360nm Selective absorber wavelength, preparation temperature is 40 DEG C, sample size is 200 μ L/ pins, and flow rate of mobile phase is 60mL/min, collects the cut of 5-7 minute, carrying out rotary evaporation is concentrated into dry, for one dimension prepares rutin crude product.Methanol-water solution with 50% dissolves rutin crude product, concentration is 10mg/mL, through filtering with microporous membrane, carry out two-dimensional liquid chromatography preparation, chromatographic column is XUnionC18, mobile phase selects methyl alcohol (containing 0.1% formic acid) to be organic phase, and water (containing 0.1% formic acid) is aqueous phase, adopts gradient elution mode: organic phase concentration brought up to 40% by 5% through 30 minutes.Adopt DAD UV-detector 360nm Selective absorber wavelength, mass spectrum Selective ion mode m/z is 303, preparation temperature is room temperature, and sample size is 3000 μ L/ pins, and flow rate of mobile phase is 100mL/min, collecting m/z is the cut of 303, rotary evaporated to dryness, obtains rutin compound, through liquid-phase chromatographic analysis, purity is 80%, and the content that one dimension prepares rutin in rutin crude product is 20%.
Embodiment 3:
Take ginkgo biloba p.e 10g, be dissolved in the methanol-water solution of 20mL75%, obtained ginkgo biloba p.e solution, concentration is 500mg/mL, crosses 0.45 μm of miillpore filter, carries out the preparation of one dimension liquid chromatogram.One dimension liquid chromatogram adopts ClickOEG, and mobile phase adopts methyl alcohol (containing 0.1% formic acid) to be organic phase, and water (containing 0.1% formic acid) is aqueous phase, adopts 5% organic equality mode wash-out.Adopt DAD UV-detector 360nm Selective absorber wavelength, preparation temperature is 30 DEG C, and sample size is 3000 μ L/ pins, and flow rate of mobile phase is 120mL/min, collects the cut of 25-30 minute, carries out rotary evaporation and be concentrated into dry, for one dimension prepares rutin crude product.Methanol-water solution with 60% dissolves rutin crude product, concentration is 80mg/mL, through filtering with microporous membrane, carry out two-dimensional liquid chromatography preparation, chromatographic column is ClickOEG, mobile phase selects methyl alcohol (containing 0.1% formic acid) to be organic phase, and water (containing 0.1% formic acid) is aqueous phase, adopts 30% organic phase isocratic elution.Adopt DAD UV-detector 360nm Selective absorber wavelength, mass spectrum Selective ion mode m/z is 303, preparation temperature is 25 DEG C, and sample size is 200 μ L/ pins, and flow rate of mobile phase is 120mL/min, collecting m/z is the cut of 303, rotary evaporated to dryness, obtains rutin compound, through liquid-phase chromatographic analysis, purity is 85%, and the content that one dimension prepares rutin in rutin crude product is 19%.
Embodiment 4:
Take ginkgo biloba p.e 10g, be dissolved in the methanol-water solution of 50mL55%, obtained ginkgo biloba p.e solution, concentration is 200mg/mL, crosses 0.45 μm of miillpore filter, carries out the preparation of one dimension liquid chromatogram.One dimension liquid chromatogram adopts ClickOEG, mobile phase adopts methyl alcohol (containing 0.1% formic acid) to be organic phase, water (containing 0.1% formic acid) is aqueous phase, adopts gradient: organic phase concentration brought up to 70% by 5% through 50 minutes, adopts DAD UV-detector 360nm Selective absorber wavelength, preparation temperature is 25 DEG C, sample size is 2000 μ L/ pins, and flow rate of mobile phase is 80mL/min, collects the cut of 18-22 minute, carrying out rotary evaporation is concentrated into dry, for one dimension prepares rutin crude product.Methanol-water solution with 50% dissolves rutin crude product, concentration is 50mg/mL, through filtering with microporous membrane, carry out two-dimensional liquid chromatography preparation, chromatographic column is XUnionC8, mobile phase selects methyl alcohol (containing 0.1% formic acid) to be organic phase, and water (containing 0.1% formic acid) is aqueous phase, adopts 5% organic phase concentration wash-out.Adopt DAD UV-detector 360nm Selective absorber wavelength, mass spectrum Selective ion mode m/z is 303, preparation temperature is 40 DEG C, and sample size is 200 μ L/ pins, and flow rate of mobile phase is 60mL/min, collecting m/z is the cut of 303, rotary evaporated to dryness, obtains rutin compound, through liquid-phase chromatographic analysis, purity is 90%, and the content that one dimension prepares rutin in rutin crude product is 23%.
Three nine-day periods after the winter solstice board Shu Xuening injection, ginkgo leaf tincture are of a specified duration as Applicative time, the traditional Chinese medicine of clear curative effect, use safety, have a wide range of applications, and the high-purity rutoside that the present invention obtains is a kind of important raw material in three nine-day periods after the winter solstice board Shu Xuening injection, ginkgo leaf tincture.And preparation method provided by the invention, process repeatability is high, and operability is good, simultaneously ginkgo leaf aboundresources, easily obtains, and is applicable to the requirement of large-scale production, can be used for the preparation of Shu Xuening injection, ginkgo leaf tincture raw material high-purity rutoside.
Applicant states, the present invention illustrates detailed process flow of the present invention by above-described embodiment, and single the present invention is not limited to above-mentioned detailed process flow, does not namely mean that the present invention must rely on above-mentioned detailed process flow and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, and to the equivalence replacement of each raw material of product of the present invention and the interpolation of auxiliary element, concrete way choice etc., all drop within protection scope of the present invention and open scope.
Claims (6)
1. liquid chromatograph mass spectrography prepares a method for high-purity rutoside, it is characterized in that: step is as follows:
Ginkgo biloba p.e is dissolved in the methanol-water solution of 25%-75%, obtained ginkgo biloba p.e solution, and ginkgo biloba p.e concentration is 10-500mg/mL;
Separating and purifying technology is adopted to prepare high-purity rutoside from ginkgo biloba p.e;
Separating and purifying technology is that two-dimensional liquid chromatography-mass spectrometry carries out ginkgo biloba p.e and prepares rutin;
Adopting liquid chromatogram to carry out chromatographic column prepared by one dimension is anti-phase C18 or anti-phase C8 or ClickOEG; The chromatographic column adopting liquid chromatogram to carry out two dimension preparation is anti-phase C18 or anti-phase C8 or ClickOEG;
The mobile phase that the preparation of one dimension liquid chromatogram adopts is that organic phase is mixed with water, and elution requirement is brought up to 70% gradient according to the degree such as organic phase 5-65% or 0-50 minute organic phase by 5% and carried out; Organic phase is methyl alcohol or acetonitrile, and aqueous phase is water;
Collect the component that retention time 5-30 minute principal component is rutin, rotary evaporated to dryness, prepares rutin crude product as one dimension; Methanol-water solution rutin crude product 25%-75% being prepared by one dimension dissolves, and is 10-80mg/mL to concentration;
Prepared by rutin crude product to one dimension, carries out two-dimensional liquid chromatography preparation; The mobile phase adopted is that organic phase is mixed with water, and elution requirement is spent according to organic phase 5-30% etc. or brought up to 40% gradient through 0-30 minute organic phase by 5% and carries out, and two dimension preparation instructs Fraction collection according to mass spectrum Selective ion mode; Organic phase is methyl alcohol or acetonitrile, and aqueous phase is water;
During preparation, flow rate of mobile phase is 60-120mL/min, and during preparation, column temperature is room temperature or 25-40 DEG C, and during preparation, sample size is 200-3000 μ L/ pin, and detector is DAD UV-detector and mass detector;
The rutin cut collected prepared by two-dimensional liquid chromatography to carry out rotary evaporation and be concentrated into dry, obtain rutin compound.
2. liquid chromatograph mass spectrography according to claim 1 prepares the method for high-purity rutoside, it is characterized in that: described ginkgo biloba p.e is dissolved in the methanol-water solution of 40-60%, and ginkgo biloba p.e concentration the best is 150-250mg/mL.
3. liquid chromatograph mass spectrography according to claim 1 prepares the method for high-purity rutoside, it is characterized in that: it 10-25 minute is the main component containing rutin that described one dimension preparation is collected, and the methanol-water solution that one dimension prepares rutin crude product 40%-60% makes the rutin crude product solution that concentration is 40-60mg/mL.
4. liquid chromatograph mass spectrography according to claim 3 prepares the method for high-purity rutoside, it is characterized in that: it is acetonitrile and water mutually that described one dimension prepares optimal flow, adopt Gradient methods, acetonitrile ratio was increased to 15%-25% by 10% through 15-20 minute.
5. liquid chromatograph mass spectrography according to claim 3 prepares the method for high-purity rutoside, it is characterized in that: described two dimension preparation optimal flow is acetonitrile and water mutually, adopt isocratic condition, acetonitrile concentration is 10-15%, and it is m/z=303 that two dimension prepares mass spectrum Selective ion mode.
6. liquid chromatograph mass spectrography as claimed in claim 1 prepares the method for high-purity rutoside, it is characterized in that: carry out two-dimensional liquid chromatography when preparing, during preparation, mobile phase optimum flow rate is 80-100mL/min, during preparation, column temperature is room temperature, during preparation, best sample size is 2000-2500 μ L/ pin, and UV detect wavelength is 360nm.
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| CN106279315B (en) * | 2015-05-15 | 2019-05-21 | 北京华润高科天然药物有限公司 | One kind preparing Quercetin -3-O-2 from ginkgo biloba p.e ", the method for 6 "-two rhamnopyranosyl glucosides |
| CN106279317B (en) * | 2015-05-15 | 2019-04-12 | 北京华润高科天然药物有限公司 | A method of preparing Quercetin -3-o- glucoside from ginkgo biloba p.e |
| CN106279359A (en) * | 2016-08-11 | 2017-01-04 | 中国水稻研究所 | A kind of method preparing five kinds of ustilaginoidea virens toxin |
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