CN105796619A - Application of microspheric silica gel with weak polarity to condensation of Panax notoginsenosides extract - Google Patents
Application of microspheric silica gel with weak polarity to condensation of Panax notoginsenosides extract Download PDFInfo
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- CN105796619A CN105796619A CN201410839621.9A CN201410839621A CN105796619A CN 105796619 A CN105796619 A CN 105796619A CN 201410839621 A CN201410839621 A CN 201410839621A CN 105796619 A CN105796619 A CN 105796619A
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Abstract
The invention relates to an application of microspheric silica gel with weak polarity to condensation of a Panax notoginsenosides extract. The method is mainly carried out by using the microspheric silica gel with weak polarity in a condensation process of the Panax notoginsenosides extract, a highly concentrated solution is obtained, and the hydrolysis rate of Panax notoginsenosides in a common condensation process is reduced.
Description
Technical field
The present invention relates to field of medicine invention, be specifically related to a kind of low pole microsphere silica gel purposes in panax notoginsenoside extract.
Background technology
Radix Notoginseng total arasaponins derives from Radix Notoginseng main root medical material, is usually used in blood circulation promoting and blood stasis dispelling, promotes blood circulation active, has and suppresses platelet aggregation and increase effect of heart and brain blood flow, cardiovascular and cerebrovascular disease is had prevention and treatment.It is mainly composed of arasaponin R1, ginsenosides Rg1, Re and Rb1 and Rd, Radix Notoginseng total arasaponins purge process includes heating extraction, concentration, chromatography and again concentrates, due to mentioned component facile hydrolysis in water, temperature is too high, pH value deviation all can increase its hydrolytic degradation rate too greatly, hydrolyzes to form sapogenin and other impurity and reduces medicinal effects.
The purifying process of Radix Notoginseng total arasaponins all with nonpolar or low pole macroporous adsorbent resin carry out adsorbing-eluting to be to reach the purpose of purification.The adsorbance of macroporous resin is high, but eluting power is poor, after using a large amount of solvent elution, the Radix Notoginseng total arasaponins concentration of gained eluent is low, and adopt the method for concentration of routine, all there is the shortcomings such as big, the cycle length of energy consumption such as vacuum drying, spray drying etc., and in dry run, need heating, easily increase Radix Notoginseng total arasaponins percent hydrolysis.
Also there is patent disclosure membrance concentration technology, such as number of patent application 201010265373.3 (publication number is CN101947250A), denomination of invention is Radix Notoginseng total arasaponins extraction purification new energy-saving process, this disclosure of the invention use film condensing device includes nanofiltration device and attached pretreatment micro-filtration carries out Radix Notoginseng total arasaponins eluent pre-concentration, it is concentrated into volume when being eluent 10% volume, is sent directly into evaporator evaporation and dries to obtain Radix Notoginseng total arasaponins extract.Membrance concentration rate all reaches 90%, and while membrance concentration, partial salts and heavy metal permeable membrane, have the effect of certain desalination and removing heavy metals.This technology actually uses nanofiltration.Nanofiltration is a kind of pressure-driven membrane separating process between reverse osmosis and ultrafiltration, and the pore diameter range of NF membrane is at several ran.Owing to the aperture of nanofiltration is more than reverse osmosis aperture, therefore desirable pressure is less, also results in through material more simultaneously, and Radix Notoginseng total arasaponins loss is bigger.
Polystyrene type low pole adsorbent resin, applicable absorption is all kinds of has certain hydrophobic Chinese medicine ingredients.But owing to such is resin ball-shaped, particle diameter insufficient height is homogeneous, aperture structure is less good, have that resolution is not high, the imperfect shortcoming of selectivity.Utilizing and need more eluent during such resin elution Radix Notoginseng total arasaponins, concentrated effect is not good.
For Problems existing in existing Radix Notoginseng total arasaponins concentration, inventor, through substantial amounts of research, finally proposes the present invention.
Summary of the invention
It is an object of the invention to provide low pole microsphere silica gel purposes in panax notoginsenoside extract concentrates.
Preferably, described low pole microsphere silica gel is the acrylate-divinylbenzene low pole microsphere of tolylation.
Low pole microsphere silica gel using method of the present invention is:
Panax notoginsenoside extract, passes in the chromatographic column loading low pole microsphere silica gel, is that 70-90% ethanol carries out eluting by concentration, collects eluent, obtain Radix Notoginseng total arasaponins concentrated solution.
Preferably, the concentration of alcohol of described eluting is 85%.
Preferably, the blade diameter length ratio of described chromatographic column is 1:3-8.
Preferably, the adsorbance of every milliliter of low pole microsphere silica gel is 0.8-1.4mg Radix Notoginseng total arasaponins.
Low pole microsphere silica gel using method of the present invention is: with in the ratio of blade diameter length ratio 1:3-8 loading low pole microsphere silica gel to chromatographic column, pass into the panax notoginsenoside extract of column volume 8-30 times, adsorption process HPLC method detects chromatographic column effluent liquid, adsorb saturated after, eluting is carried out with the ethanol that concentration is 70-90%, ethanol consumption is 0.5-3 times of chromatographic column volume, eluent Fractional Collections, and detect content of the total saponins in radix notoginseng, after the complete eluting of Radix Notoginseng total arasaponins, merge the eluent that testing result contains Radix Notoginseng total arasaponins, obtain Radix Notoginseng total arasaponins concentrated solution.
Preferably, described low pole microsphere silica gel using method is: take pseudo-ginseng, add 40-80% ethanol extraction, it is 1.10 that extracting solution vacuum is concentrated into relative density, saltout defat, pass through macroporous resin adsorption, after eluting, by panax notoginsenoside extract, passing into blade diameter length ratio is 1:5, load in the chromatographic column of acrylate-divinylbenzene low pole microsphere silica gel of tolylation, 10 times that amount is column volume of panax notoginsenoside extract, adsorption process HPLC method detects chromatographic column effluent liquid, adsorb saturated after, it is the ethanol elution of 85% by concentration, ethanol consumption is 1 times of chromatographic column volume, eluent Fractional Collections, and detect content of the total saponins in radix notoginseng, after the complete eluting of Radix Notoginseng total arasaponins, merge the eluent that testing result contains Radix Notoginseng total arasaponins, obtain Radix Notoginseng total arasaponins concentrated solution.
Radix Notoginseng total arasaponins: PANAXNOTOGINSENOSIDES, molecular formula C47H80O17, derive from the active active ingredient that Radix Notoginseng is extracted.Panax notoginsenoside extract is to be carried by water from Radix Notoginseng or ethanol extraction gained solution.
Microsphere silica gel is a kind of high activity adsorbing material, belongs to amorphous substance, and main component is silicon dioxide, and its chemical molecular formula is mSiO2·nH2O.Itself there is no toxicity, stable chemical nature, do not burn.Water insoluble and any solvent, does not react with any material except highly basic, Fluohydric acid..Mainly it is used as desiccant, moistureproof pearl, eliminating smell agent and various adsorbent, and purifies kerosene, absorption aromatic hydrocarbons etc., in the essential industry product such as tripolycyanamide, phthalic anhydride, maleic anhydride, butadiene rubber, acrylonitrile produce, be used as catalyst and catalyst carrier.
Microsphere silica gel because spherical, aperture height is homogeneous, have high selectivity, high-resolution, high-recovery, eluting concentrate, high repeatability and other advantages.Low pole microsphere silica gel of the present invention, the particularly acrylate of tolylation-divinylbenzene low pole microsphere silica gel, there is high adsorption capacity, high elution property,-Gao eluting characteristics is adsorbed by this silica gel height, the solution system of low concentration dispersed system is adsorbed completely, can Radix Notoginseng total arasaponins eluting is complete with minute quantity eluent, residual quantity is low, and elution efficiency is high.
The low pole microsphere silica gel provided by the invention purposes in panax notoginsenoside extract concentrates has the advantage that
1, low pole microsphere silica gel concentration method of the present invention is without heating, it is not necessary to cold preservation, has that energy consumption is low, easy and simple to handle, main constituent loss is few, collects the feature of being easy to, and high degree decreases the hydrolysis of Radix Notoginseng total arasaponins, improves extraction recovery.
2, adsorbance is big, every milliliter of adsorbable solution containing 0.8-1.4mg Radix Notoginseng total arasaponins of low pole microsphere silica gel.
3, easy eluting Radix Notoginseng total arasaponins.After the solution that adsorption column volume is 10 times, getting final product whole eluting with the ethanol of 1 times amount, can concentrate 10% into original solution, used effluent volume is few, and later stage recovery processing technique is simple.
Detailed description of the invention
Further illustrate the present invention by the examples below.It should be understood that embodiments of the invention are an illustration for the present invention rather than limitation of the present invention.The simple modifications that the present invention is carried out by the essence according to the present invention broadly falls into the scope of protection of present invention.Except as otherwise noted, the percent of the amount of alcohol in the present invention is percentage by volume, and v/v represents the volume ratio of solution, and BV is the multiple of column volume, if 1BV is 1 times of column volume.
Embodiment 1: the preparation method of panax notoginsenoside extract
Take pseudo-ginseng 1000g, add 60 DEG C of heating and refluxing extraction of 60% ethanol 2 times of three seven weight 5 times, 3 hours first times, second time 2 hours, united extraction liquid, filter, it is 1.20 that filtrate is concentrated into relative density, and after concentrated solution defat, D101 absorption with macroporous adsorbent resin eluting crossed by gained extractum, concentration after the oxidized aluminum of eluent, activated carbon adsorption decolouring, obtains panax notoginsenoside extract.
Embodiment 2: the preparation method of panax notoginsenoside extract
Take Radix Notoginseng powder and be broken into coarse powder 1000g, add 80% ethanol 8000ml to extract 7 hours, collect extracting solution, filter, take filtrate recycling ethanol to without alcohol taste, adding water and make every 1ml solution containing 0.5g crude drug, cross macroporous resin column and alumina adsorption, with the ethanol elution of 65%, eluent concentrates, and obtains panax notoginsenoside extract.
Embodiment 3: the preparation method of panax notoginsenoside extract
Take Radix Notoginseng, make 20 order coarse powder, with ethanol extraction 2 times, add the ethanol of 10 times 70% every time, extract 3 hours, united extraction liquid every time, filter, take filtrate recycling ethanol to without alcohol taste, cross the macroporous adsorptive resins absorption of 8 times of three seven weight, collect eluent, adsorption bleaching, recycling design, concentration, obtain panax notoginsenoside extract.
Embodiment 4: the preparation method of panax notoginsenoside extract
Take Radix Notoginseng, be ground into coarse powder, extracting in water 3 times, add the water of 10 times of three seven weight every time, extract 3 hours every time, united extraction liquid, filters, filtrate, crosses the macroporous adsorptive resins absorption of 10 times of three seven weight, collects eluent, adsorption bleaching, recycling design, concentration, obtain panax notoginsenoside extract.
Embodiment 5:
The acrylate of tolylation-divinylbenzene low pole microsphere silica gel is loaded in chromatographic column, blade diameter length ratio is 1:5, pass into the total soap extracting solution of Radix Notoginseng of column volume 10BV, adsorption process HPLC method detects content of the total saponins in radix notoginseng in chromatographic column effluent liquid, adsorb saturated after, use 85% ethanol elution, ethanol consumption is 2 times amount of chromatographic column volume, eluent segmentation takes detection content of the total saponins in radix notoginseng, when amount of alcohol reaches 1 times of chromatographic column volume, start Radix Notoginseng total arasaponins to be detected, during to 1.8 times of volumes, the basic eluting of Radix Notoginseng total arasaponins is complete, collect the eluent containing Radix Notoginseng total arasaponins, obtain Radix Notoginseng total arasaponins concentrated solution, Radix Notoginseng total arasaponins concentrated solution volume is only the 8% of upper prop volume, recovered liquid ult rec reaches 99.9%.Concentrated solution vacuum drying, obtains Radix Notoginseng total arasaponins dry product.
Embodiment 6:
The acrylate of tolylation-divinylbenzene low pole microsphere silica gel is loaded in chromatographic column, blade diameter length ratio is 1:5, pass into the total soap extracting solution of Radix Notoginseng of column volume 10 times, adsorption process HPLC method detects content of the total saponins in radix notoginseng in chromatographic column effluent liquid, adsorb saturated after, use 90% ethanol elution, ethanol consumption is 1 times amount of chromatographic column volume, eluent segmentation takes detection content of the total saponins in radix notoginseng, when amount of alcohol reaches 0.4 times of chromatographic column volume, start Radix Notoginseng total arasaponins to be detected, during to 0.9 times of volume, the basic eluting of Radix Notoginseng total arasaponins is complete, collect the eluent containing Radix Notoginseng total arasaponins, obtain Radix Notoginseng total arasaponins concentrated solution, Radix Notoginseng total arasaponins concentrated solution volume is only the 5% of upper prop volume, recovered liquid ult rec reaches 99.5%.Concentrated solution vacuum drying, obtains Radix Notoginseng total arasaponins dry product.
Embodiment 7:
The acrylate of tolylation-divinylbenzene low pole microsphere silica gel is loaded in chromatographic column, blade diameter length ratio is 1:8, pass into the total soap extracting solution of Radix Notoginseng of column volume 30 times, adsorption process HPLC method detects content of the total saponins in radix notoginseng in chromatographic column effluent liquid, adsorb saturated after, use 70% ethanol elution, ethanol consumption is 3 times amount of chromatographic column volume, eluent segmentation takes detection content of the total saponins in radix notoginseng, when amount of alcohol reaches 1.6 times of chromatographic column volumes, start Radix Notoginseng total arasaponins to be detected, during to 2.6 times of volumes, the basic eluting of Radix Notoginseng total arasaponins is complete, collect the eluent containing Radix Notoginseng total arasaponins, obtain Radix Notoginseng total arasaponins concentrated solution, Radix Notoginseng total arasaponins concentrated solution volume is only the 10% of upper prop volume, recovered liquid ult rec reaches 99.9%.Concentrated solution vacuum drying, obtains Radix Notoginseng total arasaponins dry product.
Experimental example 1: concentrated effect
Radix Notoginseng total arasaponins detection method: measure according to high performance liquid chromatography (annex VI D).
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica for filler；With mobile phase A: acetonitrile, Mobile phase B: water, carry out gradient elution by table 1；Detection wavelength is 203nm.Number of theoretical plate presses ginsenoside Rg1Peak calculates should be not less than 6000, ginsenoside Rg11.5 should be not less than with the separating degree of Re.
Table 1: gradient elution
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0-25 | 19 | 81 |
| 25-35 | 19→20 | 81→80 |
| 35-70 | 20→35 | 80→65 |
| 70-75 | 35→45 | 65→55 |
| 75-76 | 45→90 | 55→10 |
| 76-80 | 90 | 10 |
| 80-81 | 90→19 | 10→81 |
| 81-90 | 19 | 81 |
The preparation of reference substance solution (or reference extract solution): take the drying under reduced pressure Panax Notoginseng saponin R of 12 hours respectively1Reference substance, ginsenoside Rg1Reference substance, ginsenoside Re's reference substance and ginsenoside Rb1Reference substance is appropriate, accurately weighed, adds methanol and makes every 1ml containing Panax Notoginseng saponin R10.3mg, ginsenoside Rg11.2mg, ginsenoside Re 0.2mg, ginsenoside Rb11.0mg, ginsenoside Rd 0.1mg mixed solution, shake up, obtain reference substance solution；Or it is appropriate to take the drying under reduced pressure Radix Notoginseng total arasaponins reference substance of 12 hours, accurately weighed, add methanol and make every 1ml solution containing 2.5mg, shake up, obtain reference extract solution.
The preparation of need testing solution: take this product under content uniformity item appropriate, accurately weighed, add methanol and make every 1ml solution containing 4mg, to obtain final product.
Algoscopy: precision draws reference substance solution (or reference extract solution) and each 5-10 μ l of need testing solution respectively, injects chromatograph of liquid, measures, to obtain final product.
The operating process of embodiment 5 is analyzed, and when passing into 10 times of volumes that the amount of panax notoginsenoside extract reaches chromatographic column, chromatographic column outlet effluent does not detect Radix Notoginseng total arasaponins yet, it was shown that absorption does not reach most saturation.When passing into ethanol elution, when amount of alcohol reaches 1 times of chromatographic column volume, beginning to flow out Radix Notoginseng total arasaponins, during to 1.8 times of volumes, the basic eluting of Radix Notoginseng total arasaponins is complete, and Radix Notoginseng total arasaponins concentrated solution volume is only the 8% of upper prop volume, and recovered liquid ult rec reaches 99.9%.Concrete outcome is in Table 2.
Table 2: specifically detect data
Experimental example result shows: low pole microsphere silica gel of the present invention is big to the adsorbance of Radix Notoginseng total arasaponins, and is extremely easy to eluting, it is possible to reach concentrated effect significantly.
Although, above use generality explanation, detailed description of the invention and test, the present invention is described in detail, but on basis of the present invention, it is possible to it being made some amendments or improves, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (8)
1. low pole microsphere silica gel purposes in panax notoginsenoside extract concentrates.
2. purposes as claimed in claim 1, it is characterised in that described low pole microsphere silica gel is the acrylate-divinylbenzene low pole microsphere of tolylation.
3. purposes as claimed in claim 1 or 2, it is characterised in that described low pole microsphere silica gel using method is: panax notoginsenoside extract, pass in the chromatographic column loading low pole microsphere silica gel, it is that 70-90% ethanol carries out eluting by concentration, collects eluent, obtain Radix Notoginseng total arasaponins concentrated solution.
4. purposes as claimed in claim 3, it is characterised in that described low pole microsphere silica gel using method is: the concentration of alcohol of eluting is 85%.
5. purposes as claimed in claim 3, it is characterised in that described low pole microsphere silica gel using method is: the blade diameter length ratio of chromatographic column is 1:3-8.
6. purposes as claimed in claim 3, it is characterised in that described low pole microsphere silica gel using method is: the adsorbance of every milliliter of low pole microsphere silica gel is 0.8-1.4mg Radix Notoginseng total arasaponins.
7. purposes as claimed in claim 3, it is characterized in that, described low pole microsphere silica gel using method is: with in the ratio of blade diameter length ratio 1:3-8 loading low pole microsphere silica gel to chromatographic column, pass into the panax notoginsenoside extract of column volume 8-30 times, adsorption process HPLC method detects chromatographic column effluent liquid, adsorb saturated after, eluting is carried out with the ethanol that concentration is 70-90%, ethanol consumption is 0.5-3 times of chromatographic column volume, eluent Fractional Collections, and detect content of the total saponins in radix notoginseng, after the complete eluting of Radix Notoginseng total arasaponins, merge the eluent that testing result contains Radix Notoginseng total arasaponins, obtain Radix Notoginseng total arasaponins concentrated solution.
null8. purposes as claimed in claim 3，It is characterized in that，Described low pole microsphere silica gel using method is: take pseudo-ginseng，Add 40-80% ethanol extraction，It is 1.10 that extracting solution vacuum is concentrated into relative density，Saltout defat，Pass through macroporous resin adsorption、After eluting，By panax notoginsenoside extract，Passing into blade diameter length ratio is 1:5、Load in the chromatographic column of acrylate-divinylbenzene low pole microsphere silica gel of tolylation，10 times that amount is column volume of panax notoginsenoside extract，Adsorption process HPLC method detects chromatographic column effluent liquid，Adsorb saturated after，It is the ethanol elution of 85% by concentration，Ethanol consumption is 1 times of chromatographic column volume，Eluent Fractional Collections，And detect content of the total saponins in radix notoginseng，After the complete eluting of Radix Notoginseng total arasaponins，Merge the eluent that testing result contains Radix Notoginseng total arasaponins，Obtain Radix Notoginseng total arasaponins concentrated solution.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109384749A (en) * | 2018-12-26 | 2019-02-26 | 重庆市碚圣医药科技股份有限公司 | A kind of purification process of taxol |
| CN111487349A (en) * | 2020-05-15 | 2020-08-04 | 易达斯生物科技(苏州)有限公司 | Chromatographic column for selective enrichment and purification of saponin and application method thereof |
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| CN102423329A (en) * | 2011-12-13 | 2012-04-25 | 广西梧州制药（集团）股份有限公司 | Decolorization method of panax notoginsenoside extract |
| CN102441016A (en) * | 2011-12-13 | 2012-05-09 | 广西梧州制药（集团）股份有限公司 | Concentration method of pseudo-ginseng total saponin extracting solution |
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| CN102423329A (en) * | 2011-12-13 | 2012-04-25 | 广西梧州制药（集团）股份有限公司 | Decolorization method of panax notoginsenoside extract |
| CN102441016A (en) * | 2011-12-13 | 2012-05-09 | 广西梧州制药（集团）股份有限公司 | Concentration method of pseudo-ginseng total saponin extracting solution |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109384749A (en) * | 2018-12-26 | 2019-02-26 | 重庆市碚圣医药科技股份有限公司 | A kind of purification process of taxol |
| CN111487349A (en) * | 2020-05-15 | 2020-08-04 | 易达斯生物科技(苏州)有限公司 | Chromatographic column for selective enrichment and purification of saponin and application method thereof |
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Application publication date: 20160727 |