CN105999765A - Natural pharmaceutical ingredient system separation device based on separation-enriching mode and working method thereof - Google Patents
Natural pharmaceutical ingredient system separation device based on separation-enriching mode and working method thereof Download PDFInfo
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- CN105999765A CN105999765A CN201610494265.0A CN201610494265A CN105999765A CN 105999765 A CN105999765 A CN 105999765A CN 201610494265 A CN201610494265 A CN 201610494265A CN 105999765 A CN105999765 A CN 105999765A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
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Abstract
The invention relates to a natural pharmaceutical ingredient system separation device based on a separation-enriching mode and a working method thereof, which belongs to the technical field of pharmaceutical ingredient separation equipment, and is applied to the fields of research, development and production of the traditional Chinese medicine, plants and marine drugs. The natural pharmaceutical ingredient system separation device consists of three high pressure pumps, a main separation column, 10 to 20 level-1 enriching columns, three level-2 separation columns and 10 to 20 level-2 enriching columns. The chromatographic columns separated in two levels are connected by virtue of a two-position four-way valve to realize the separation, enriching, desorption, absorption and flow path switching of the ingredients; and the main separation column and the level-2 separation columns are connected with a detector and a mixer, and the natural pharmaceutical components and pure compounds are enriched by virtue of solid extraction. The system is controlled by a PLC, a single chip microprocessor and principal computer software, so that the automation of the chromatographic separation process is realized, and the separation preparation efficiency of complicated systems such as the traditional Chinese medicine can be greatly improved.
Description
Technical field
The present invention relates to a kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode and method of work thereof, it belongs to and is continuously separated equipment technical field, is applied to research and development, the production fields such as Chinese medicine, plant amedica, marine drug and food.
Background technology
Natural product from plant, marine organisms, Chinese medicine and biological engineering etc. has become as original new drug, functional food, the source of novel fine chemicals innovation.Natural Medicine Chemistry composition separation and preparation technology including polypeptide, Plant Secondary Metabolites is the key of original new drug research and development; also being emphasis and the difficult point of initiative modern Chinese medicine, it is key based on natural drug Development of New Drugs that the active component in natural product is prepared in scale, separation repeatable, systematic.
Chromatographic technique has prominent advantage to the preparation that separates of the target compound that the physicochemical properties such as molecular weight, structure approximate, become separation and preparation plant amedica, the important method of marine drug isoreactivity composition and means, be also the key technology of R&D of modern TCM.It is particularly suited for separation preparation and the large-scale production of effective ingredient in natural product and Chinese medicine.
Natural drug composition complexity, single chromatographic isolation effect is limited, needs through separating for several times, uses different chromatographic isolation pattern can be only achieved and be satisfied with separating effect in separation process.During conventional Natural products research, although have accumulated rich experience, but the natural medicinal ingredients such as systematic Study Chinese medicine have still been had to the suspicion lacking systematicness.In recent years, the development of Two way chromatograms technology, use several modes filler complementary to separate and pave road for solving this problem undoubtedly.And realized automatization, after realizing position separation, being directly purified compound preparation after carrying out on-line preconcentration can simplify separation preparation process, improves work efficiency to greatest extent.Shao Ping (CN101791491A) devises the two-dimentional preparing chromatography system that a set of online basic, normal, high pressure combines, by six-way valve and software, two preparing chromatography systems are coupled together, during first dimension preparative hplc separates, sample overload loading is run, intercept target fraction entrance the second dimension preparative hplc to separate further, it is suitable for separating principal component and for solving to transship the contradiction between sample introduction and separating power, improves chromatogram separating capacity.Zhang Weibing devises a kind of Quasi dynamic two dimension preparing chromatography system and material separation method (CN102914610A, CN202870047U), by three-way valve, two preparative hplc posts are connected, by Vavle switching, the component that the first dimension preparative hplc post separates is transferred to the second dimension preparative hplc post separate, be equally used for solving the contradiction between overload sample introduction and separating power.Li Heran, Xiao Hongbin disclose the method that (CN102961892A, CN104713973A) uses 2 10 the on-line preconcentration chromatograph separating natural ingredient leading to valve and multiple 2 six-way valves.To some extent solve problem, but still suffer from deficiency on the Automated condtrol of repeatable acquisition effective site, the systematicness separation of effective site and separation process.
Summary of the invention
For solving problems of the prior art; the present invention provides a kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode and method of work thereof; this device is the complex material systems such as plant amedica, marine drug and food to carry out systematicness, scale and seriality separate the automated system of preparation; not only improve shortening and separate the time used; improve work efficiency; simultaneously; the effective site of repeatable acquisition, effective ingredient monomer; developing for such as new Chinese medicine, pharmacology activity research is significant.
The technical solution used in the present invention is: a kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode, it includes liquid-way system, detached dowel, enriching column, detector and automation control system.Described automation control system includes PLC system, HMI interface and PC terminal and running control software；Described PLC system includes Boolean quantity input module, Boolean quantity output module and Analog input mModule, this device includes flash trapping stage and enrichment system and the second-order separation and enrichment system, and described flash trapping stage uses the first solvent tank to be connected to the first blender composition gradient through the second solvent pump separate fluid feed system through the first solvent pump, the second solvent tank with enrichment system；First blender is connected to loading post through the first blender outlet tube, loading column valve；Loading post through loading post outlet tube to flash trapping stage post, flash trapping stage post is connected to one-level enriching column group through flash trapping stage post selector valve, the first flash trapping stage post arm, detector, the second blender valve, blender the first arm, being in series with 10-20 one-level enriching column in one-level enriching column group, each one-level enriching column is provided with 1 two-position four-way valve；Dilution flow container is connected to the second blender through diluting pump, it is provided that diluent is by the enrichment on one-level enriching column of the flash trapping stage component；The waste liquid port that waste liquid enters fraction collectors through the second-order separation post selector valve of the second-order separation post, the second-order separation valve, two grades of enriching column input pipes, two grades of enrichment column valves that flow out after example enrichment are discharged.nullWith enrichment liquid-way system, described the second-order separation includes that the first solvent tank is through the first solvent pump、Second solvent tank is connected to the first blender composition gradient separation fluid feed system through the first blender outlet tube through the second solvent pump、Loading post selector valve、Blender the first arm、Any one Vavle switching in two-position four-way valve，Using the sample in one-level enriching column successively as loading post，Selection through one-level enriching column selector valve，Any one of connection one-level enriching column is switched on suitable 1 the second-order separation post separate，Liquid after separation is through the second-order separation valve、Second flash trapping stage post arm、Flash trapping stage post selector valve、First flash trapping stage post arm、Detector、Second blender valve、Blender the second arm、Two grades of enriching column input pipes are connected to two grades of enriching column groups，Two grades of enriching column groups are in series with 10-20 enriching column，Each two grades of enriching columns are provided with 1 two grades enrichment column valves，Simultaneously、Dilution flow container is connected to the second blender through diluting pump，There is provided diluent liquid to realize the enrichment on two grades of enriching columns of the second-order separation component；The waste liquid port that waste liquid after two grades of enrichments enters fraction collector through two grades of enriching column Vavle switching is discharged；In one-level enriching column, the collection stream of component is connected to first blender composition gradient through the second solvent pump by the first solvent tank collects fluid feed system offer through the first solvent pump, the second solvent tank, collect stream be through the first blender liquid through loading column valve enter one-level enriching column selector valve switching, by the sample eluting successively in 20 enriching columns, through one-level enriching column outlet tube, the second-order separation post selector valve, the second-order separation valve, two grades of enriching column input pipes, two grades of enrichment column valves switch into fraction collector；Sample entrance fraction collector is discharged in the selection being connected to the second blender, the second blender valve, blender the second arm, two grades of enriching column input pipes and two grades of enrichment column valves through diluting pump by dilution flow container of collecting of two grades of enriching column samples.Described PLC system, HMI interface and PC terminal and the first solvent pump, second solvent pump, 3rd solvent pump, loading column valve, flash trapping stage column valve, the second-order separation valve, the second-order separation post selector valve, second blender valve, one-level enrichment column valve, two grades of enrichment column valves are electrically connected, PLC system, HMI interface and PC terminal and software control the first solvent pump, second solvent pump, 3rd solvent pump, loading column valve, flash trapping stage column valve, the second-order separation column valve, the second-order separation post selector valve, second blender valve, one-level enrichment column valve, the break-make conversion of two grades of enrichment column valves, lock out operation and stopping.
The method of work of a kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode, comprises the following steps:
(1) natural medicinal ingredients flash trapping stage process
Started the first solvent pump by PLC system, gradient liquid chromatography solvent system that the second solvent pump, the first blender form, and open loading column valve and the blend sample being in loading post is brought into flash trapping stage post carries out chromatographic isolation, flow into the second blender after detector detection chromatographic isolation；Simultaneously, start diluting pump injection diluent in the second blender, change the characteristic of effluent, by the way of one-level enrichment column valve is according to time-switching, mixture in one-level enriching column, is obtained effective ingredient position according to polarity or compounds category characteristic solid phase extraction concentration respectively；The waste liquid port that the waste liquid of the outflow after example enrichment enters fraction collectors through the second-order separation post selector valve of the second-order separation post, the second-order separation valve, two grades of enriching column input pipes, two grades of enrichment column valves are discharged；；
The target of flash trapping stage process is complicated natural medicinal ingredients to be separated into the effective ingredient position of 20 parts of repeatable acquisitions according to compound polarity or category Properties, for pharmaceutically active systematic study；
(2) natural medicinal ingredients the second-order separation process
PLC system is by controlling one-level enrichment column valve with an one-level enriching column for sample introduction post, first solvent tank is connected to, through the first solvent pump, second solvent tank the second solvent pump, the gradient liquid chromatography solvent system that the first blender forms, through the first blender outlet tube, loading post selector valve, blender the first arm, control the switching of one-level enrichment column valve with an one-level enriching column for sample introduction post；Separating on the second-order separation post through the second-order separation post selector valve, the liquid after separation is connected to two grades of enriching column groups through the second-order separation valve, the second-order separation post arm, flash trapping stage column valve, flash trapping stage post arm, detector, the second blender, the second blender valve, blender the second arm, two grades of enriching column input pipes；Simultaneously, start diluting pump injection diluent in the second blender, change the characteristic of effluent, by the way of two grades of enrichment column valves are according to chromatographic peak collection, Solid-Phase Extraction, mixture is enriched in two grades of enriching columns respectively according to polarity or compounds category characteristic, obtaining the effective ingredient that natural drug is pure, waste liquid is discharged by two grades of enrichment column valves；
The target of the second-order separation is that the effective ingredient position of flash trapping stage is individually separated into monomeric compound；
(3) effective site and the desorption of pure compound
After the flash trapping stage of each natural drug or the second-order separation at position complete, automatically carry out the desorption of position or pure compound with strong solution analysis solvent.
Described flash trapping stage post, the second-order separation post use dynamic axial compression chromatographic column or static axial compression column；Described loading post, one-level enriching column, two grades of enriching columns use static axial compression columns or common performance liquid chromatographic column；Described flash trapping stage post, one-level enriching column, the second-order separation post, the filler of two grades of enrichment chromatographic columns are selected from silica gel, aluminium oxide, bonded-phase silica, macroporous adsorbent resin, ion exchange resin or polyamide.
Described detector is selected from preparative UV-detector, evaporative light scattering detector or differential refraction detector.
Described loading column valve, flash trapping stage post selector valve, the second-order separation valve, the second blender valve, one-level enrichment column valve, two grades of enrichment column valves use two-position four-way valve.
This segregation apparatus is made up of 3 efficient liquid-phase chromatographic pumps, 1 loading post or injector, 1 main detached dowel, 10-20 one-level enriching column, 3 alternative the second-order separation posts, 10-20 2 grades of enriching column, UV-detector and automation control system.
This device realizes separation process in the way of Automated condtrol runs system and automatically runs.2 efficient liquid-phase chromatographic pumps of segregation apparatus provide chromatographic isolation flowing phases, 1 enrichment diluting pump, UV-detector detection chromatographic isolation, Vavle switching to realize chromatographic isolation, enrichment and stream switching etc. and realize by carrying out Automated condtrol with PLC, HMI and upper PCs software.
This device realizes the systematicness of natural medicinal ingredients, the method for work of repeatable acquisition mainly comprises the steps that
(1) on analytical type chromatograph of liquid, use the chromatograph packing material identical with main detached dowel to set up and optimize flow visualizing and the method program of natural medicinal ingredients Gradient chromatography separation method.
(2), in the separable programming embedded device that will set up, under the control of the computer, natural medicinal ingredients is carried out 1,2 grades of automatization's chromatographic isolation.
Natural drug mixture is separated into 10-20 effective site of repeatable acquisition by (3) 1 grades of chromatographs, can be used for the active ingredient system research of Chinese medicine after parsing.
(5) changing solvent orange 2 A and the ratio of B, effective site or the monomeric compound mixture of 20 repeatable acquisitions are passed through 24 switchings leading to valve, automatic eluting is placed in fraction collector, obtains effective site or monomeric compound.
(6) repeat loading, carry out 1 grade separate after, then carry out 2 grades of separation and can obtain the monomeric compound at each position.
The main detached dowel of this device, 2 grades of detached dowels can use dynamic axial compression chromatographic column, static axial compression column；Loading post, 1 grade of enriching column, 2 grades of enriching columns use static axial compression columns or common performance liquid chromatographic column；Main detached dowel, 1 grade of enriching column, 2 grades of detached dowels, fillers of 2 grades of enrichment chromatographic columns are selected from silica gel, aluminium oxide, bonded-phase silica, C18, C8, macroporous adsorbent resin, ion exchange resin or polyamide.
The sample enrichment method that separates of device is online solid phase extraction method, flash trapping stage post, one-level enriching column, the second-order separation post, two grades of enriching columns can use the filler of identical or different type to coordinate, it is achieved the Two way chromatograms systematicness of natural drug effective ingredient separates preparation.
This segregation apparatus realizes in the collecting processes of 1 grade, 2 grades of separation, fraction and position that natural drug effective ingredient systematicness separates and monomeric compound, it separates and the switching of sample collection pattern all has and is automatically obtained with these 24 switchings leading to valve of computer-controlled loading column valve, flash trapping stage post selector valve, the second-order separation valve and the second blender valve, meanwhile, the switching of loading column valve, flash trapping stage post selector valve, the second-order separation valve and the second blender valve also realizes two-stage pillar layer separation and shares the target of 1 UV-detector detection.
The control system of this equipment is carried out Automated condtrol by PLC, single-chip microcomputer and upper PCs software, shows, arranges and be made up of PLC+HMI+PC.Modularity PLC system, meet in, small-scale control requirement；The module of various performances can meet and adapt to Automated condtrol task；Simple and practical distributed frame and general network capabilities so that apply the most flexible；Boolean quantity input module (I): acquisition system dispersed placement, detects external signal and collects process；Boolean quantity output module (O): for control system external drive vigour part, performs to control requirement accordingly, it is achieved system unattended；Analog input mModule (AN): to chromatograph work system monitoring state, be responsible for effective collecting work of process variable.Different working times, working method, the flow of liquid phase pump, maximum working pressure (MWP) can be set according to technological requirement；Chromatogram can be shown the most intuitively, and effectively can operate according to chromatogram.
The invention has the beneficial effects as follows: this device uses two-stage separation-Enrichment Mode to realize the systematicness separation of natural medicinal ingredients, sample passes through solid loading post loading, separate through flash trapping stage post, sample is become according to polarity size separation the fraction of 20 groups of repeatable acquisitions, it is enriched in one-level enriching column, completing flash trapping stage, the fraction of enrichment can carry out bioactivity research or further separating monomer compound as required；One-level enriching column during the second-order separation is as the loading post of the second-order separation, according to compound property difference, the filler being suitable for is selected to carry out stepwise gradient eluting separation on the second-order separation post, detector tracing detection, the monomer of isolated or simple component are enriched on two grades of enriching columns, with in strong eluting liquid wash to fraction collector, it is achieved the systematicness separation of compound.
Accompanying drawing explanation
The process chart of a kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode of Fig. 1.
nullIn figure: 1、First solvent tank，2、Second solvent tank，3、Dilution flow container，4、First solvent pump，5、Second solvent pump，6、Diluting pump，7、First blender，7a、First blender outlet tube，8、Loading post，8a、Loading column valve、9、Flash trapping stage post，9a、Flash trapping stage post selector valve，9b、First flash trapping stage post arm，9c、Second flash trapping stage post arm，10、Loading post outlet tube，11、Detector，12、Second blender，12a、Blender the first arm，12b、Blender the second arm，13、One-level enriching column，13a、One-level enriching column selector valve，13b、One-level enriching column outlet tube，14、Second blender valve，15、The second-order separation post、15a、The second-order separation valve，15b、The second-order separation post selector valve，16、Two grades of enriching columns，16a、Two grades of enriching column input pipes，16b、Two grades of enrichment column valves，17、Fraction collector.
Specific embodiments
Below in conjunction with specific embodiment, the present invention is further elaborated:
A kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode, it includes liquid-way system, detached dowel, enriching column, detector and automation control system.Automation control system includes PLC system, HMI interface and PC terminal and running control software；PLC system includes Boolean quantity input module, Boolean quantity output module and Analog input mModule, this device includes flash trapping stage and enrichment system and the second-order separation and enrichment system, and flash trapping stage uses with enrichment system the first solvent tank 1 to be connected to the first blender 7 composition gradient through first solvent pump the 4, second solvent tank 2 through the second solvent pump 5 and separates fluid feed system；First blender 7 is connected to loading post 8 through the first blender outlet tube 7a, loading column valve 8a；Loading post 8 is through loading post outlet tube 10 to flash trapping stage post 9, flash trapping stage post 9 is connected to one-level enriching column group through flash trapping stage post selector valve 9a, the first flash trapping stage post arm 9b, detector the 11, second blender valve 14, blender the first arm 12a, being in series with 10-20 one-level enriching column 13 in one-level enriching column group, each one-level enriching column 13 is provided with 1 one-level enriching column selector valve 13a；Dilution flow container 3 is connected to the second blender 12 through diluting pump 6, it is provided that diluent is by the enrichment on one-level enriching column of the flash trapping stage component；The waste liquid port that waste liquid enters fraction collectors 17 through the second-order separation post selector valve of the second-order separation post, the second-order separation valve, two grades of enriching column input pipes, two grades of enrichment column valves that flow out after example enrichment are discharged.nullWith enrichment liquid-way system, the second-order separation includes that the first solvent tank is through the first solvent pump、Second solvent tank is connected to the first blender 7 composition gradient separation fluid feed system through the first blender outlet tube 7a through the second solvent pump、Loading post selector valve 8a、Blender the first arm 12a、Any one Vavle switching in two-position four-way valve 13a，Using the sample in one-level enriching column successively as loading post，Selection through one-level enriching column selector valve 13a，Any one of connection one-level enriching column 13 is switched on suitable 1 the second-order separation post 15 separate，Liquid after separation is through the second-order separation valve 15a、Second flash trapping stage post arm 9c、Flash trapping stage post selector valve 9a、First flash trapping stage post arm 9b、Detector 11、Second blender valve 14、Blender the second arm 12b、Two grades of enriching column input pipe 16a are connected to two grades of enriching column groups，Two grades of enriching column groups are in series with 10-20 enriching column 16，Each two grades of enriching columns 16 are provided with 1 two grades enrichment column valve 16b，Simultaneously、Dilution flow container 3 is connected to the second blender 12 through diluting pump 6，There is provided diluent liquid to realize the enrichment on two grades of enriching columns of the second-order separation component；The waste liquid port that waste liquid after two grades of enrichments switches into fraction collector 17 through two grades of enrichment column valve 16b is discharged；In one-level enriching column, the collection stream of component is connected to first blender 7 composition gradient through the second solvent pump by the first solvent tank collects fluid feed system offer through the first solvent pump, the second solvent tank, collect stream be through the first blender 7 liquid through loading column valve 8a enter one-level enriching column selector valve 13a switching, by the sample eluting successively in 20 enriching columns, through one-level enriching column outlet tube 13b, the second-order separation post selector valve 15b, the second-order separation valve 15a, two grades of enriching column input pipe 16a, two grades of enrichment column valve 16b switch into fraction collector 17；Sample entrance fraction collector 17 is discharged in the selection being connected to second blender the 12, second blender valve 14, blender the second arm 12b, two grades of enriching column input pipe 16a and two grades of enrichment column valve 16b through diluting pump 6 by dilution flow container 3 of collecting of two grades of enriching column samples.PLC system, HMI interface and PC terminal and the first solvent pump 4, second solvent pump 5, 3rd solvent pump 6, loading column valve 8a, flash trapping stage column valve 9a, the second-order separation valve 15a, the second-order separation post selector valve 15b, second blender valve 14, one-level enrichment column valve 13a, two grades of enrichment column valve 16b are electrically connected, PLC system, HMI interface and PC terminal and software control the first solvent pump 4, second solvent pump 5, 3rd solvent pump 6, loading column valve 8a, flash trapping stage column valve 9a, the second-order separation column valve 15a, the second-order separation post selector valve 15b, second blender valve 14, one-level enrichment column valve 13a, the break-make conversion of two grades of enrichment column valve 16b, lock out operation and stopping.
This segregation apparatus is made up of liquid-way system, detecting system and automatic control system by structure.Wherein liquid-way system is made up of 3 efficient liquid-phase chromatographic pumps, solid-state loading post, flash trapping stage post, three the second-order separation posts, 20 one-level SPE enriching columns, 10 two grades of SPE enriching columns.Control system PLC uses the S7 series of products of Siemens Company.Chromatogram monitoring system uses all-wave 1 on-line monitoring of long UV230II UV-detector, wave-length coverage 190 600nm.
Started the first solvent pump by PLC system, gradient liquid chromatography solvent system that the second solvent pump, the first blender form, and open loading column valve and the blend sample being in loading post is brought into flash trapping stage post carries out chromatographic isolation, flow into the second blender after detector detection chromatographic isolation；Simultaneously, start diluting pump injection diluent in the second blender, change the characteristic of effluent, by the way of one-level enrichment column valve is according to time-switching, mixture is distinguished solid phase extraction concentration in one-level enriching column according to polarity or compounds category characteristic, obtaining effective ingredient position, waste liquid is discharged by one-level enrichment column valve；The target of flash trapping stage process is complicated natural medicinal ingredients to be separated into the effective ingredient position of 20 parts of repeatable acquisitions according to compound polarity or category Properties, for pharmaceutically active systematic study.
PLC system is by controlling one-level enrichment column valve with an one-level enriching column for sample introduction post, start the first solvent pump, the second solvent pump and the gradient liquid chromatography solvent system of the first blender composition, the blend sample being in one-level enriching column is brought in the second-order separation post and carry out chromatographic isolation, after detector detection chromatographic isolation, flow into the second blender；Simultaneously, start diluting pump injection diluent in the second blender, change the characteristic of effluent, by the way of two grades of enrichment column valves are according to chromatographic peak collection, Solid-Phase Extraction, mixture is enriched in two grades of enriching columns respectively according to polarity or compounds category characteristic, obtaining the effective ingredient that natural drug is pure, waste liquid is discharged by two grades of enrichment column valves；The target of the second-order separation is that the effective ingredient position of flash trapping stage is individually separated into monomeric compound.
After the flash trapping stage of each natural drug or the second-order separation at position complete, automatically carry out the desorption of position or pure compound with strong solution analysis solvent.This device realizes separation process in the way of Automated condtrol runs system and automatically runs.
2 efficient liquid-phase chromatographic pumps of segregation apparatus provide chromatographic isolation flowing phase, 1 enrichment diluting pump, UV-detector detection chromatographic isolation, Vavle switching realizes chromatographic isolation, enrichment and stream switching etc. are by with PLC, single-chip microcomputer and upper PCs software carry out Automated condtrol and realize, technologist or operator station (DCS) are mainly made up of computer and corresponding WCC PC control software and S7 V5.5 Automated condtrol software, realize the signals collecting to system, process and control, the setting of parameter and amendment, the detection of the system failure, the running status of viewing system.Whole process can realize in 24-48 hour.
Application Example
1
Choose medicine-food two-purpose fungus Poria as separate object, take purification on normal-phase silica gel filler silica gel to carry out gradient separations.Weighing 10kg Poria medical material, methanol eddy obtains dry powder 0.184kg after extracting；Extract through ethyl acetate after dry powder is water-soluble, obtain ethyl acetate phase 34g.Direct pump loading after ethyl acetate phased soln, with 10:0-9:1 ... 1:9-0:10 chlorofonn-ethylacetate, as eluent system, carries out gradient elution through flash trapping stage post, normal hexane, as enrichment diluent, obtains 20 groups of different fractions after the enrichment of one-level enriching column.Each one-level enriching column is as the loading post of the second-order separation simultaneously, according to different component compound polarity size, chooses chlorofonn-ethylacetate as eluent system, uses different proportion to carry out on the second-order separation post fine gradient elution separation.Simultaneously with UV detector tracing detection, the component of isolated is enriched on two grades of enriching columns, enters in fraction collector by methanol-eluted fractions.Concentrate simultaneously after TLC and HPLC detects, there are 47 components, wherein content more than 90% monomeric compound 27, the simple component of two compounds 19, the multicomponent of 3 compounds 21, respectively tens milligrams Dao several grams.
Application Example
2
Choosing Cortex Moutan as separating object, the method taking one-level positive phase filling silica gel and two grades of reverse phase filler C18 to combine carries out stepwise gradient separation.Weighing 2kg Cortex Moutan medical material, methanol eddy obtains dry powder 0.456kg after extracting.Above to solid loading post after silica gel mixed sample, with 10:0-9:1 ... 1:9-0:10 chloroform-methanol, as eluent system, carries out gradient elution through one-level silica gel detached dowel, normal hexane, as enrichment diluent, obtains the component of 15 opposed polarities after the enrichment of one-level enriching column.According to the difference of compound property, 1-8 group component proceeds two grades of silicagel columns and separates, and 1-4 group eluent system is normal hexane-acetone, and 5-8 group eluent system is methylene chloride-methanol；9-15 group component reverse phase filler C18 carries out the second-order separation, eluent system methanol-water, eluting ratio 0%-10%...... 100% methanol.After TLC and HPLC detects simultaneously, there are 65 components, wherein monomeric compound 16 (content more than 90%), simple component 21, multicomponent 28, respectively tens milligrams to tens grams.
Application Example
3
Taking red rooted salvia 10kg to extract with 70% ethanol water, ethanol extraction, after a precipitate with ethanol processes, successively with chloroform, ethyl acetate extraction, respectively obtains chloroform extract layer (Radix Salviae Miltiorrhizae liposolubility compound) and ethyl acetate extract layer.Ethyl acetate extract layer is carried out system preparative hplc separation.Flash trapping stage post and the second-order separation post all use C18 reverse phase filler, and methanol-water carries out gradient elution in varing proportions, there are 10 kinds of pressure differential self after the second-order separation, and content is more than 95%, and quality is between 10-500mg；Identify through mass spectrum, nuclear-magnetism etc., respectively salvianolic acid A, salvianolic acid B, salvianolic acid D, danshensu, protocatechualdehyde, alkannic acid, 7-crystal violet oxalic acid, alkannic acid C, rosmarinic acid, 4-(3,4-dihydroxyphenyl) acid of-2-butylene etc..
Claims (5)
1. a natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode, it includes that liquid-way system, detached dowel, enriching column, detector and automation control system, described automation control system include PLC system, HMI interface and PC terminal and running control software；Described PLC system includes Boolean quantity input module, Boolean quantity output module and Analog input mModule, it is characterized in that: this device includes flash trapping stage and enrichment system and the second-order separation and enrichment system, and described flash trapping stage uses the first solvent tank (1) to be connected to the first blender (7) composition gradient through the second solvent pump (5) separate fluid feed system through the first solvent pump (4), the second solvent tank (2) with enrichment system；First blender (7) is connected to loading post (8) through the first blender outlet tube (7a), loading column valve (8a)；Loading post (8) is through loading post outlet tube (10) to flash trapping stage post (9), flash trapping stage post (9) is connected to one-level enriching column group through flash trapping stage post selector valve (9a), the first flash trapping stage post arm (9b), detector (11), the second blender valve (12), the second blender valve (14), blender the first arm (12a), being in series with 10-20 one-level enriching column (13) in one-level enriching column group, each one-level enriching column (13) is provided with 1 one-level enriching column selector valve (13a)；Dilution flow container (3) is connected to the second blender (12) through diluting pump (6)；Described the second-order separation and enrichment system use 1 one-level enriching column (13) in one-level enriching column group as loading post, it is connected to the second-order separation post (15) through one-level enriching column outlet tube (13b), the second-order separation post (15) is through the second-order separation valve (15a), second flash trapping stage post arm (9c), flash trapping stage post selector valve (9a), detector (11), second blender (12), blender the second arm (12b), two grades of enriching column input pipes (16a) are connected to two grades of enriching column groups, fraction collector (17), two grades of enriching column groups are in series with 10-20 two grades of enriching columns (16), two grades of enriching columns (16) are provided with two grades of enrichments column valve (16b)；Described PLC system and the first solvent pump (4), second solvent pump (5), diluting pump (6), loading column valve (8a), flash trapping stage post selector valve (9a), the second-order separation valve (15a), the second-order separation post selector valve (15b), second blender valve (14), one-level enrichment column valve (13a), two grades of enrichments column valve (16b) are electrically connected, PLC system controls loading column valve (8a), flash trapping stage post selector valve (9a), the second-order separation valve (15a), second blender valve (14), one-level enrichment column valve (13a), the break-make conversion of two grades of enrichments column valve (16b), PLC system controls the first solvent pump (4), second solvent pump (5), the operating of diluting pump (6) and stopping.
The method of work of a kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode the most according to claim 1, it is characterised in that comprise the following steps:
(1) natural medicinal ingredients flash trapping stage process
Started the first solvent pump (4) by PLC system, gradient liquid chromatography solvent system that the second solvent pump (5), the first blender (7) form, and open loading column valve (8a) and the blend sample being in loading post (8) is brought into flash trapping stage post (9) carries out chromatographic isolation, flow into the second blender (12) after detector (11) detection chromatographic isolation；Simultaneously, start diluting pump (6) and inject diluent characteristic of change effluent in the second blender (12), by the way of one-level enrichment column valve (13a) is according to time-switching, mixture in one-level enriching column (13), is obtained effective ingredient position according to polarity or compounds category characteristic solid phase extraction concentration respectively；The waste liquid port that the waste liquid of the outflow after example enrichment enters fraction collector (17) through the second-order separation post selector valve (15b) of the second-order separation post, the second-order separation valve (15a), two grades of enriching column input pipes (16a), two grades of enrichments column valve (16b) are discharged；；
The target of flash trapping stage process is complicated natural medicinal ingredients to be separated into the effective ingredient position of the repeatable acquisition of 10-20 part according to compound polarity or category Properties, for pharmaceutically active systematic study；
(2) natural medicinal ingredients the second-order separation process
PLC system is sample introduction post by controlling one-level enrichment column valve (13a) with an one-level enriching column (13), first solvent tank (1) is connected to, through the first solvent pump (4), the second solvent tank (2) second solvent pump (5), the gradient liquid chromatography solvent system that the first blender (7) forms, through the first blender outlet tube (7a), loading post selector valve (8a), blender the first arm (12a), the switching controlling one-level enrichment column valve (13a) is sample introduction post with an one-level enriching column (13)；Separating on the second-order separation post (15) through the second-order separation post selector valve (15b), the liquid after separation is connected to two grades of enriching column groups through the second-order separation valve (15a), the second flash trapping stage post arm (9c), flash trapping stage column valve (9a), the first flash trapping stage post arm (9b), detector (11), the second blender (12), the second blender valve (14), blender the second arm (12b), two grades of enriching column input pipes (16a)；Simultaneously, start diluting pump (6) and inject diluent characteristic of change effluent in the second blender (12), by the way of two grades of enrichments column valve (16b) are according to chromatographic peak collection, Solid-Phase Extraction, mixture is enriched in two grades of enriching columns (16) respectively according to polarity or compounds category characteristic, obtaining the effective ingredient that natural drug is pure, waste liquid is discharged by two grades of enrichments column valve (16b)；
The target of the second-order separation is that the effective ingredient position of flash trapping stage is individually separated into monomeric compound；
(3) effective site and the desorption of pure compound
After the flash trapping stage of each natural drug or the second-order separation at position complete, automatically carry out the desorption of position or pure compound with strong solution analysis solvent.
A kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode the most according to claim 1 and 2, it is characterised in that: described flash trapping stage post (9), the second-order separation post (15) use dynamic axial compression chromatographic column or static axial compression column；Described loading post (8), one-level enriching column (13), two grades of enriching columns (16) use static axial compression columns or common performance liquid chromatographic column；Described flash trapping stage post (9), one-level enriching column (13), the second-order separation post (15), the filler of two grades of enrichments chromatographic column (16) are selected from silica gel, aluminium oxide, bonded-phase silica, macroporous adsorbent resin, ion exchange resin or polyamide.
A kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode the most according to claim 1 and 2, it is characterised in that: described detector (11) is selected from preparative UV-detector, evaporative light scattering detector.
A kind of natural medicinal ingredients system segregation apparatus based on separation-Enrichment Mode the most according to claim 1 and 2, it is characterised in that: described loading column valve (8a), flash trapping stage post selector valve (9a), the second-order separation valve (15a), the second blender valve (14), one-level enrichment column valve (13a), two grades of enrichments column valve (16b) use two-position four-way valve.
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| CN110152353A (en) * | 2019-06-18 | 2019-08-23 | 大连博迈科技发展有限公司 | A kind of continuous chromatography device and arasaponin production method |
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