CN105999765B - A kind of natural medicinal ingredients system separator and its method of work based on separation and concentration pattern - Google Patents
A kind of natural medicinal ingredients system separator and its method of work based on separation and concentration pattern Download PDFInfo
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- CN105999765B CN105999765B CN201610494265.0A CN201610494265A CN105999765B CN 105999765 B CN105999765 B CN 105999765B CN 201610494265 A CN201610494265 A CN 201610494265A CN 105999765 B CN105999765 B CN 105999765B
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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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- 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
- B01D15/42—Selective adsorption, e.g. chromatography characterised by the development mode, e.g. by displacement or by elution
- B01D15/424—Elution mode
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Abstract
A kind of natural medicinal ingredients system separator and its method of work based on separation and concentration pattern, it belongs to the continuous separation equipment technical field of drug ingedient, applied to Chinese medicine, plant, the research and development of marine drug, production field.Device is by 3 high-pressure pumps, main splitter, 10 20 1 grade of enriching columns, 32 grades of splitters and 10 20 2 grades of enriching columns compositions.The chromatogram intercolumniation of two-stage separation is connected with 2 4-way valves, realizes that separation, enrichment, desorption and the stream of composition switch；Connecting detection device and blender after main splitter and 2 grades of splitters, with solid phase extraction concentration natural drug component and pure compound.System is controlled with PLC, single-chip microcomputer and host computer software, realizes the automation of chromatographic separation process, greatly improves efficiency prepared by the separation of the complex systems such as Chinese medicine.
Description
Technical field
The present invention relates to a kind of based on the natural medicinal ingredients system separator of separation-Enrichment Mode and its work side
Method, it belongs to continuous separation equipment technical field, applied to the research and development such as Chinese medicine, autonomic drug, marine drug and food, production neck
Domain.
Background technology
Natural products from plant, marine organisms, Chinese medicine and bioengineering etc. has become original new drug, function food
Product, the source of new fine chemicals innovation.Natural Medicine Chemistry composition separation including polypeptide, Plant Secondary Metabolites
Technology of preparing is the key of original new drug research and development, and formulates the emphasis and difficult point of modern Chinese herbal medicine, scale, repeatable, system
The active component that the separation of property is prepared in natural products is the key based on natural drug Development of New Drugs.
Separation preparation of the chromatographic technique to the approximate target compound of the physicochemical properties such as molecular weight, structure has prominent
Advantage, it has also become separate and prepare autonomic drug, the important method and means of marine drug isoreactivity composition, and modern Chinese herbal medicine and grind
The key technology studied carefully.The separation preparation and large-scale production for the active ingredient in natural products and Chinese medicine of being particularly suitable for use in.
Natural drug composition is complicated, and single chromatographic isolation effect is limited, it is necessary to by separating for several times, makes in separation process
It can be only achieved with different chromatographic isolation patterns and be satisfied with separating effect.During conventional Natural products research, although having accumulated
Rich experience, but still have the suspicion for lacking systematicness for natural medicinal ingredients such as systematic Study Chinese medicines.In recent years, it is two-dimentional
The development of chromatographic technique, road is undoubtedly paved for solving this problem using the complementary separation of several modes filler.And
Automation is realized, after position separation is realized, carrying out directly carrying out pure compound after on-line preconcentration preparing can simplify point
From preparation process, operating efficiency is improved to greatest extent.Shao Ping（CN101791491A）Devise a set of online basic, normal, high pressure
With reference to two-dimentional preparing chromatography system, two preparing chromatography systems are connected by six-way valve and software, first dimension prepare
Sample overload loading operation in chromatographic isolation, interception target fraction prepare chromatogram into the second dimension and further separated, be adapted to
In separating principal component and overloading the contradiction between sample introduction and separating capacity for solution, chromatogram separating capacity is improved.Zhang Weibing is set
A kind of Quasi dynamic two dimension preparing chromatography system and material separation method are counted（CN102914610A, CN202870047U）, pass through
Triple valve prepares chromatographic column connection by two, and the component that the first dimension is prepared to chromatogram post separation by Vavle switching is transferred to the second dimension
Prepare chromatographic column to be separated, be equally used for the contradiction for solving between overload sample introduction and separating capacity.Li Heran, Xiao Hongbin are disclosed
（CN102961892A、CN104713973A）Day is separated using the on-line preconcentration chromatograph of 2 10 port valves and multiple 2 six-way valves
The method of right drug ingedient.Solve problem to a certain extent, but in the repeatable system for obtaining active component, active component
Property separation and separation process Automated condtrol on still suffer from deficiency.
The content of the invention
To solve problems of the prior art, the present invention provides a kind of natural drug based on separation-Enrichment Mode
Component system separator and its method of work, the device are that the complex material system such as autonomic drug, marine drug and food is entered
Automated system prepared by row systematicness, scale and continuity separation, not only improves and shortens the time used in separation, improve work
Efficiency, meanwhile, the active component obtained, active ingredient monomer are repeated, is developed for such as new Chinese medicine, pharmacology activity research
It is significant.
The technical solution adopted by the present invention is：A kind of natural medicinal ingredients system separation dress based on separation-Enrichment Mode
Put, it includes liquid-way system, splitter, enriching column, detector and automation control system.The automation control system includes
PLC system, HMI interfaces and PC terminals and running control software；It is defeated that the PLC system includes Boolean quantity input module, Boolean quantity
Go out module and Analog input mModule, the device includes one-level separation and enrichment system and the second-order separation and enrichment system, described
One-level separates is connected to first through the first solvent pump, the second solvent tank using the first solvent tank with enrichment system through the second solvent pump
Blender composition gradient separates fluid feed system；First blender is connected to through the first blender efferent duct, loading column valve
Sample post；Loading post is through loading post efferent duct to one-level splitter, and one-level splitter is through one-level splitter selector valve, the one one fraction
One-level enriching column group is connected to from post branch pipe, detector, the second blender valve, the branch pipe of blender first, in one-level enriching column group
10-20 one-level enriching column is in series with, each one-level enriching column is provided with 1 two-position four-way valve；Flow container is diluted through diluting pump
It is connected to the second blender, there is provided enrichment of the dilution by one-level separation component on one-level enriching column；Stream after example enrichment
Go out the second-order separation post selector valve of the waste liquid through the second-order separation post, the second-order separation valve, two level enriching column input pipe, two level enrichment column valve
Waste liquid port into fraction collector is discharged.The second-order separation includes the first solvent tank through the first solvent with enrichment liquid-way system
Pump, the second solvent tank are connected to the first blender composition gradient through the second solvent pump and separate fluid feed system through the first blender
Efferent duct, loading column valve, the branch pipe of blender first, through any one Vavle switching in two-position four-way valve, by one-level enriching column
Sample be used as loading post successively, the selection through one-level enriching column selector valve, any one of connection one-level enriching column is switched to
Separated on suitable 1 the second-order separation post, the liquid after separation is through the second-order separation valve, the second one-level splitter branch pipe, one
Level splitter selector valve, the first one-level splitter branch pipe, detector, the second blender valve, the branch pipe of blender second, two level enrichment
Post input pipe is connected to two level enriching column group, is in series with 10-20 enriching column in two level enriching column group, on each two level enriching column
Be enriched with column valve provided with 1 two level, simultaneously, dilution flow container be connected to the second blender through diluting pump, there is provided diluent liquid is with reality
Existing enrichment of the second-order separation component on two level enriching column；Waste liquid after two level enrichment enters cut through two level enriching column Vavle switching
The waste liquid port discharge of collector；In one-level enriching column the collection stream of component by the first solvent tank through the first solvent pump, second molten
Agent tank is connected to the first blender composition gradient through the second solvent pump and collects fluid feed system offer, and it is through first to collect stream
The liquid of blender enters the switching of one-level enriching column selector valve through loading column valve, and the sample in 20 enriching columns is washed successively
It is de-, through one-level enriching column efferent duct, the second-order separation post selector valve, the second-order separation valve, two level enriching column input pipe, two level enriching column
Valve switches into fraction collector；The collection of two level enriching column sample is connected to the second mixing by dilution flow container through diluting pump
The selection of device, the second blender valve, the branch pipe of blender second, two level enriching column input pipe and two level enrichment column valve is discharged sample and entered
Enter fraction collector.The PLC system, HMI interfaces and PC terminals and the first solvent pump, the second solvent pump, the 3rd solvent pump, on
Sample column valve, one-level splitter selector valve, the second-order separation valve, the second-order separation post selector valve, the second blender valve, the choosing of one-level enriching column
Select valve, two level enrichment column valve is electrically connected, PLC system, HMI interfaces and PC terminals and the first solvent pump of software control, second
Solvent pump, the 3rd solvent pump, loading column valve, one-level splitter selector valve, the second-order separation valve, the second-order separation post selector valve, second
Break-make conversion, lock out operation and the stopping of blender valve, one-level enriching column selector valve, two level enrichment column valve.
A kind of method of work of the natural medicinal ingredients system separator based on separation-Enrichment Mode, including following step
Suddenly：
（1）Natural medicinal ingredients one-level separation process
The gradient liquid chromatography solvent system being made up of the first solvent pump of PLC system startup, the second solvent pump, the first blender
System, and open loading column valve and bring the blend sample in loading post into and chromatographic isolation is carried out in one-level splitter, detect
The second blender is flowed into after device detection chromatographic isolation；Meanwhile start diluting pump injection dilution and change in the second blender
The characteristic of efflux, by way of one-level enriching column selector valve is according to time-switching, by mixture according to polarity or compound
Category Properties distinguish solid phase extraction concentration in one-level enriching column, obtain active ingredient position；Outflow after example enrichment it is useless
The second-order separation post selector valve of the liquid through the second-order separation post, the second-order separation valve, two level enriching column input pipe, two level enrichment column valve enter
The waste liquid port discharge of fraction collector；；
The target of one-level separation process is to separate the natural medicinal ingredients of complexity according to compound polarity or category Properties
Into 20 parts of repeatable active ingredient positions obtained, for pharmaceutical activity system research；
（2）Natural medicinal ingredients the second-order separation process
PLC system is by controlling one-level enriching column selector valve using an one-level enriching column as sample introduction post, the first solvent tank warp
First solvent pump, second the second solvent pump of solvent tank are connected to the gradient liquid chromatography solvent system of the first blender composition, warp
First blender efferent duct, loading column valve, the branch pipe of blender first, the switching of one-level enriching column selector valve is controlled with an one-level
Enriching column is sample introduction post；Separated through the second-order separation post selector valve on the second-order separation post, the liquid after separation is through two fractions
Mixed from valve, the second-order separation post branch pipe, one-level splitter selector valve, one-level splitter branch pipe, detector, the second blender, second
Clutch valve, the branch pipe of blender second, two level enriching column input pipe are connected to two level enriching column group；Meanwhile start dilution infusion
Enter the characteristic that dilution changes efflux in the second blender, by the way that two level enrichment column valve is collected according to chromatographic peak, solid phase extracts
The mode taken, mixture is enriched in two level enriching column respectively according to polarity or compounds category characteristic, obtains natural drug
Pure active ingredient, waste liquid are enriched with column valve by two level and discharged；
The target of the second-order separation is that the active ingredient position of one-level separation is separated into monomeric compound respectively；
（3）The desorption of active component and pure compound
After the completion of the one-level separation of each natural drug or the second-order separation at position, position is carried out automatically with strong solution analysis solvent
Or the desorption of pure compound.
The one-level splitter, the second-order separation post use dynamic axial compression chromatographic column or static axial compression column；It is described
Loading post, one-level enriching column, two level enriching column use static axial compression columns or common performance liquid chromatographic column；Described one
Level splitter, one-level enriching column, the second-order separation post, the filler of two level enrichment chromatographic column are selected from silica gel, aluminum oxide, Bonded Phase silicon
Glue, macroporous absorbent resin, ion exchange resin or polyamide.
The detector is selected from preparative UV-detector, EISD or differential refraction detector.
The loading column valve, one-level splitter selector valve, the second-order separation valve, the second blender valve, the selection of one-level enriching column
Valve, two level enrichment column valve use two-position four-way valve.
The separator is by 3 efficient liquid-phase chromatographic pumps, 1 loading post or injector, 1 main splitter, 10-20 individual one
Level enriching column, 3 alternative the second-order separation posts, 10-20 2 grades of enriching columns, UV-detector and automation control systems
Composition.
The device realizes separation process automatic running in a manner of Automated condtrol runtime.2 height of separator
Effect liquid phase chromatogram pump provides chromatographic isolation mobile phase, 1 enrichment diluting pump, UV-detector detection chromatographic isolation, Vavle switching
Realize that chromatographic isolation, enrichment and stream switching etc. carry out Automated condtrol by with PLC, HMI and upper PCs software
Realize.
The device realizes that systemic, the repeatable method of work obtained of natural medicinal ingredients mainly includes the following steps that：
（1）On analytic type liquid chromatograph, using with main splitter identical chromatograph packing material establish and optimize crude drug
The flow visualizing and method program of thing component gradient chromatography separating method.
（2）By in the separable programming embedded device of foundation, under the control of the computer, 1,2 grade is carried out to natural medicinal ingredients
Automate chromatographic isolation.
（3）Natural drug mixture is separated into 10-20 active component of repeatable acquisition by 1 grade of chromatogram, can after parsing
Active ingredient system for Chinese medicine is studied.
（5）Change solvent orange 2 A and B ratio, 20 the repeatable active components obtained or monomeric compound mixture are led to
The switching of 2 4-way valves is crossed, automatic elution is placed in fraction collector, obtains active component or monomeric compound.
（6）Loading is repeated, after carrying out 1 grade of separation, then 2 grades is carried out and separates the monomeric compound that can obtain each position.
The main splitter of the device, 2 grades of splitters can use dynamic axial compression chromatographic column, static axial compression column；On
Sample post, 1 grade of enriching column, 2 grades of enriching columns use static axial compression columns or common performance liquid chromatographic column；Main splitter, 1
Level enriching column, 2 grades of splitters, the fillers of 2 grades of enrichment chromatographic columns may be selected from silica gel, aluminum oxide, bonded-phase silica, C18, C8, big
Macroporous adsorbent resin, ion exchange resin or polyamide.
The separation example enrichment method of device is online solid phase extraction method, one-level splitter, one-level enriching column, two level
Splitter, two level enriching column can use the filler of identical or different type to coordinate, and realize the two-dimentional color of natural drug active ingredient
It is prepared by spectra system separation.
The separator realizes systemic 1 grade separated of natural drug active ingredient, 2 grades of separation, cut and position and list
In the collecting processes of body compound, its separate and sample collection pattern switching have with computer control loading column valve,
The switching of one-level splitter selector valve, the second-order separation valve and second blender valve these 2 4-way valves realizes automatically, meanwhile, loading
The switching of column valve, one-level splitter selector valve, the second-order separation valve and the second blender valve also realizes that two-stage pillar layer separation shares 1
The target of platform UV-detector detection.
The control system of the equipment carries out Automated condtrol by PLC, single-chip microcomputer and upper PCs software, shows, sets
Put and be made up of PLC+HMI+PC.Modularization PLC system, in satisfaction, small-scale control requires；The module of various performances can expire
Foot and adaptation Automated condtrol task；Simple and practical distributed frame and general network capabilities so that application is very flexible；
Boolean quantity input module（I）：Acquisition system dispersed placement, external signal is detected and collects processing；Boolean quantity exports mould
Block (O)：For control system external drive vigour part, perform corresponding control and require, realize the unattended of system；Analog quantity is defeated
Enter module (AN)：Chromatogram work system state is monitored, is responsible for effective collecting work of process variable.Can be according to technique
It is required that the different working times is set, and working method, the flow of liquid phase pump, maximum working pressure (MWP)；Effectively can intuitively it show
Chromatogram, and can effectively be operated according to chromatogram.
The beneficial effects of the invention are as follows：The device uses the system that two-stage separation-Enrichment Mode realizes natural medicinal ingredients
Property separation, sample separates post separation by solid loading post loading, by one-level, by sample according to polarity size separation into 20 groups
The repeatable cut obtained, is enriched in one-level enriching column, completes one-level separation, and the cut of enrichment can carry out biology as needed
Activity research or further separating monomer compound；Loading post of the one-level enriching column as the second-order separation during the second-order separation,
According to compound property difference, select suitable filler to carry out stepwise gradient elution separation on the second-order separation post, detector with
Track is detected, and isolated monomer or simple component are enriched on two level enriching column, is received with strong elution property liquid wash to cut
In storage, the systematicness separation of compound is realized.
Brief description of the drawings
A kind of process charts of the natural medicinal ingredients system separator based on separation-Enrichment Mode of Fig. 1.
In figure：1st, the first solvent tank, the 2, second solvent tank, 3, dilution flow container, the 4, first solvent pump, the 5, second solvent pump,
6th, diluting pump, the 7, first blender, 7a, the first blender efferent duct, 8, loading post, 8a, loading column valve, 9, one-level separation
Post, 9a, one-level splitter selector valve, 9b, the first one-level splitter branch pipe, 9c, the second one-level splitter branch pipe, 10, loading post
Efferent duct, 11, detector, the 12, second blender, 12a, the branch pipe of blender first, 12b, the branch pipe of blender second, 13, one-level
Enriching column, 13a, one-level enriching column selector valve, 13b, one-level enriching column efferent duct, the 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 level enriching column, 16a, two level enriching column input pipe,
16b, two level enrichment column valve, 17, fraction collector.
Specific embodiment
With reference to specific embodiment, the present invention is further elaborated：
A kind of natural medicinal ingredients system separator based on separation-Enrichment Mode, it includes liquid-way system, separation
Post, enriching column, detector and automation control system.Automation control system include PLC system, HMI interfaces and PC terminals and
Running control software；PLC system includes Boolean quantity input module, Boolean quantity output module and Analog input mModule, the device
Including one-level separation and enrichment system and the second-order separation and enrichment system, one-level separation uses the first solvent tank 1 with enrichment system
Through the first solvent pump 4, the second solvent tank 2 composition gradient of the first blender 7 separation fluid supply system is connected to through the second solvent pump 5
System；First blender 7 is connected to loading post 8 through the first blender efferent duct 7a, loading column valve 8a；Loading post 8 is defeated through loading post
Outlet pipe 10 is to one-level splitter 9, and one-level splitter 9 is through one-level splitter selector valve 9a, the first one-level splitter branch pipe 9b, detection
Device 11, the second blender valve 14, the first branch pipe of blender 12a are connected to one-level enriching column group, are in series with one-level enriching column group
10-20 one-level enriching column 13, each one-level enriching column 13 are provided with 1 one-level enriching column selector valve 13a；Dilution flow container 3 passes through
Diluting pump 6 is connected to the second blender 12, there is provided enrichment of the dilution by one-level separation component on one-level enriching column；Sample
The second-order separation post selector valve of the outflow waste liquid through the second-order separation post, the second-order separation valve, two level enriching column input pipe after enrichment, two
The waste liquid port that level enrichment column valve enters fraction collector 17 is discharged.The second-order separation passes through with enrichment liquid-way system including the first solvent tank
First solvent pump, the second solvent tank are connected to the composition gradient of the first blender 7 separation fluid feed system warp through the second solvent pump
First blender efferent duct 7a, loading column valve 8a, the first branch pipe of blender 12a, through any one valve in two-position four-way valve 13a
Switching, using the sample in one-level enriching column successively as loading post, the selection through one-level enriching column selector valve 13a, connect one-level
Any one of enriching column 13 is switched on suitable 1 the second-order separation post 15 and separated, and the liquid after separation is through two fractions
From valve 15a, the second one-level splitter branch pipe 9c, one-level splitter selector valve 9a, the first one-level splitter branch pipe 9b, detector
11st, the second blender valve 14, the second branch pipe of blender 12b, two level enriching column input pipe 16a are connected to two level enriching column group, and two
10-20 enriching column 16 is in series with level enriching column group, each two level enriching column 16 is provided with 1 two level and is enriched with column valve 16b, together
When, dilution flow container 3 be connected to the second blender 12 through diluting pump 6, there is provided diluent liquid is to realize the second-order separation component two
Enrichment on level enriching column；Waste liquid after two level enrichment switches into the waste liquid of fraction collector 17 through two level enrichment column valve 16b
Mouth discharge；The collection stream of component is molten through second through the first solvent pump, the second solvent tank by the first solvent tank in one-level enriching column
Agent pump is connected to the composition gradient of the first blender 7 and collects fluid feed system offer, and it is the liquid through the first blender 7 to collect stream
Body enters one-level enriching column selector valve 13a switching through loading column valve 8a, the sample in 20 enriching columns is eluted successively, through one
Level enriching column efferent duct 13b, the second-order separation post selector valve 15b, the second-order separation valve 15a, two level enriching column input pipe 16a, two level
Enrichment column valve 16b's switches into fraction collector 17；The collection of two level enriching column sample is by dilution flow container 3 through diluting pump 6
It is connected to the second blender 12, the second blender valve 14, the second branch pipe of blender 12b, two level enriching column input pipe 16a and two level
Enrichment column valve 16b selection discharge sample enters fraction collector 17.PLC system, HMI interfaces and PC terminals and the first solvent pump
4th, the second solvent pump 5, the 3rd solvent pump 6, loading column valve 8a, one-level splitter selector valve 9a, the second-order separation valve 15a, two fractions
It is electrically connected from post selector valve 15b, the second blender valve 14, one-level enriching column selector valve 13a, two level enrichment column valve 16b,
PLC system, HMI interfaces and PC terminals and software control the first solvent pump 4, the second solvent pump 5, the 3rd solvent pump 6, loading column valve
8a, one-level splitter selector valve 9a, the second-order separation valve 15a, the second-order separation post selector valve 15b, the second blender valve 14, one-level are rich
Clustered column selector valve 13a, two level enrichment column valve 16b break-make conversion, lock out operation and stopping.
The separator is made up of by structure liquid-way system, detecting system and automatic control system.Wherein liquid-way system is by 3
Platform efficient liquid-phase chromatographic pump, a solid-state loading post, an one-level splitter, three the second-order separation posts, 20 one-level SPE enrichments
Post, 10 two level SPE enriching columns compositions.Control system PLC uses the S7 series of products of Siemens Company.Chromatogram monitoring system is adopted
With 1 on-line monitoring of all-wave length UV230II the UV-detectors, -600nm of wave-length coverage 190.
The gradient liquid chromatography solvent system being made up of the first solvent pump of PLC system startup, the second solvent pump, the first blender
System, and open loading column valve and bring the blend sample in loading post into and chromatographic isolation is carried out in one-level splitter, detect
The second blender is flowed into after device detection chromatographic isolation；Meanwhile start diluting pump injection dilution and change in the second blender
The characteristic of efflux, by way of one-level enriching column selector valve is according to time-switching, by mixture according to polarity or compound
Category Properties distinguish solid phase extraction concentration in one-level enriching column, obtain active ingredient position, waste liquid is selected by one-level enriching column
Select valve discharge；The target of one-level separation process is to separate the natural medicinal ingredients of complexity according to compound polarity or category Properties
Into 20 parts of repeatable active ingredient positions obtained, for pharmaceutical activity system research.
PLC system starts the first solvent by controlling one-level enriching column selector valve using an one-level enriching column as sample introduction post
The gradient liquid chromatography solvent system of pump, the second solvent pump and the first blender composition, by the mixing in one-level enriching column
Thing sample is brought into and chromatographic isolation is carried out in the second-order separation post, and the second blender is flowed into after detector detection chromatographic isolation；Meanwhile open
Dynamic diluting pump injection dilution changes the characteristic of efflux in the second blender, and column valve is enriched with according to chromatographic peak by two level
Collect, the mode of SPE, mixture is enriched in two level enriching column respectively according to polarity or compounds category characteristic, obtained
The active ingredient pure to natural drug, waste liquid are enriched with column valve by two level and discharged；The target of the second-order separation is to separate one-level
Active ingredient position is separated into monomeric compound respectively.
After the completion of the one-level separation of each natural drug or the second-order separation at position, position is carried out automatically with strong solution analysis solvent
Or the desorption of pure compound.The device realizes separation process automatic running in a manner of Automated condtrol runtime.
2 efficient liquid-phase chromatographic pumps of separator provide chromatographic isolation mobile phase, 1 enrichment diluting pump, ultraviolet inspection
Survey device detection chromatographic isolation, Vavle switching realize chromatographic isolation, enrichment and stream switching etc. by with PLC, single-chip microcomputer and host computer
Computer software carries out Automated condtrol to realize, technologist or operator station（DCS）Mainly by computer and corresponding WCC host computers
Control software and S7 V5.5 Automated condtrols software composition, realize that the signal acquisition to system, processing and control, parameter are set
Fixed and modification, the detection of the system failure, the running status of observing system.Whole process can be realized within 24-48 hours.
Application Example 1
Medicine-food two-purpose fungus-Poria cocos is chosen as separation object, takes purification on normal-phase silica gel filler silica gel to carry out gradient separations.
10kg Poria cocos medicinal materials are weighed, dry powder 0.184kg is obtained after methanol eddy extraction；Extract, obtain through ethyl acetate after dry powder is water-soluble
Ethyl acetate phase 34g.Direct pump loading after ethyl acetate phased soln, with 10:0-9:1……1:9-0:10 chlorofonn-ethylacetates
As eluent system, gradient elution is carried out through one-level splitter, n-hexane obtains as enrichment dilution after the enrichment of one-level enriching column
To 20 groups of different fractions.Loading post of each one-level enriching column as the second-order separation simultaneously, according to different component compound pole
Property size, choose chlorofonn-ethylacetate as eluent system, fine gradient is carried out on the second-order separation post using different proportion
Elution separation.Simultaneously use UV detector tracing detections, isolated component is enriched on two level enriching column, with methanol elute into
Enter in fraction collector.Concentrated after TLC and HPLC is detected simultaneously, 47 components, the wherein singulation of content more than 90% are obtained
Compound 27, the simple component of two compounds 19, the multicomponent of 3 compounds 21, respectively more than ten milligrams Dao several grams
.
Application Example 2
Moutan bark is chosen as separation object, take the positive phase filling silica gel of one-level and two level reverse phase filler C18 is combined
Method carries out stepwise gradient separation.2kg moutan bark medicinal materials are weighed, dry powder 0.456kg is obtained after methanol eddy extraction.Silica gel mixed sample
It is upper afterwards to arrive solid loading post, with 10:0-9:1……1:9-0:10 chloroform-methanols are as eluent system, through one-level silica gel splitter
Gradient elution is carried out, n-hexane obtains the component of 15 opposed polarities as enrichment dilution after the enrichment of one-level enriching column.According to
The difference of compound property, 1-8 group components continue two level silica gel post separation, and 1-4 groups eluent system is n-hexane-acetone,
5-8 groups eluent system is methylene chloride-methanol；9-15 groups component carries out the second-order separation, eluent system first with reverse phase filler C18
Alcohol-water, the methanol of elution ratio 0%-10%...... 100%.After TLC and HPLC is detected simultaneously, 65 components are obtained, wherein
Monomeric compound 16 (content more than 90%), simple component 21, multicomponent 28, respectively tens milligrams to more than ten grams not
Deng.
Application Example 3
Red rooted salvia 10kg is taken to be extracted with 70% ethanol water, ethanol extract uses chlorine successively after an alcohol precipitation is handled
Imitative, ethyl acetate extraction, respectively obtains chloroform extract layer（Red sage root fat-soluble compound）With ethyl acetate extract layer.To acetic acid second
Ester extract layer carries out system and prepares chromatographic isolation.One-level splitter and the second-order separation post use C18 reverse phase fillers, with not year-on-year
Example methanol-water carries out gradient elution, and 10 kinds of pressure differential selfs are obtained after the second-order separation, and content is more than 95%, and quality exists
Between 10-500mg；Identified through mass spectrum, nuclear-magnetism etc., respectively salviandic acid A, tanshin polyphenolic acid B, salvianolic acid D, danshensu, former catechu
Aldehyde, alkannic acid, 7- methyl alkannic acid, alkannic acid C, Rosmarinic acid, 4-（3,4- dihydroxyphenyls）- 2- butenoic acids etc..
Claims (5)
1. a kind of natural medicinal ingredients system separator based on separation-Enrichment Mode, it include liquid-way system, splitter,
Enriching column, detector and automation control system, the automation control system include PLC system, HMI interfaces and PC terminals and
Running control software；The PLC system includes Boolean quantity input module, Boolean quantity output module and Analog input mModule, its
It is characterised by：The device includes one-level separation and enrichment system and the second-order separation and enrichment system, the one-level separation and enrichment
System uses the first solvent tank（1）Through the first solvent pump（4）, the second solvent tank（2）Through the second solvent pump（5）It is mixed to be connected to first
Clutch（7）Composition gradient separates fluid feed system；First blender（7）Through the first blender efferent duct（7a）, loading column valve
（8a）It is connected to loading post（8）；Loading post（8）Through loading post efferent duct（10）To one-level splitter（9）, one-level splitter（9）
Through one-level splitter selector valve（9a）, the first one-level splitter branch pipe（9b）, detector（11）, the second blender（12）, second
Blender valve（14）, the branch pipe of blender first（12a）One-level enriching column group is connected to, 10-20 is in series with one-level enriching column group
Individual one-level enriching column（13）, each one-level enriching column（13）It is provided with 1 one-level enriching column selector valve（13a）；Dilute flow container（3）
Through diluting pump（6）It is connected to the second blender（12）；The second-order separation uses 1 in one-level enriching column group with enrichment system
Individual one-level enriching column（13）As loading post, through one-level enriching column efferent duct（13b）It is connected to the second-order separation post（15）, two fractions
From post（15）Through the second-order separation valve（15a）, the second one-level splitter branch pipe（9c）, one-level splitter selector valve（9a）, detector
（11）, the second blender（12）, the branch pipe of blender second（12b）, two level enriching column input pipe（16a）It is connected to two level enriching column
Group, fraction collector（17）, 10-20 two level enriching column is in series with two level enriching column group（16）, two level enriching column（16）On
Column valve is enriched with provided with two level（16b）；The PLC system and the first solvent pump（4）, the second solvent pump（5）, diluting pump（6）, on
Sample column valve（8a）, one-level splitter selector valve（9a）, the second-order separation valve（15a）, the second-order separation post selector valve（15b）, it is second mixed
Clutch valve（14）, one-level enriching column selector valve（13a）, two level enrichment column valve（16b）It is electrically connected, PLC system control loading
Column valve（8a）, one-level splitter selector valve（9a）, the second-order separation valve（15a）, the second blender valve（14）, one-level enriching column selection
Valve（13a）, two level enrichment column valve（16b）Break-make conversion, PLC system control the first solvent pump（4）, the second solvent pump（5）, it is dilute
Release liquid pump（6）Operating and stopping.
A kind of 2. work of natural medicinal ingredients system separator based on separation-Enrichment Mode according to claim 1
Make method, it is characterised in that comprise the following steps：
（1）Natural medicinal ingredients one-level separation process
First solvent pump is started by PLC system（4）, the second solvent pump（5）, the first blender（7）The gradient liquid chromatography of composition
Solvent system, and open loading column valve（8a）Loading post will be in（8）In blend sample bring one-level splitter into（9）In enter
Row chromatographic isolation, detector（11）The second blender is flowed into after detection chromatographic isolation（12）；Meanwhile start diluting pump（6）Note
Enter dilution in the second blender（12）The middle characteristic for changing efflux, passes through one-level enriching column selector valve（13a）According to the time
The mode of switching, mixture is distinguished into solid phase extraction concentration in one-level enriching column according to polarity or compounds category characteristic（13）
In, obtain active ingredient position；The second-order separation post selector valve of the waste liquid of outflow after example enrichment through the second-order separation post
（15b）, the second-order separation valve（15a）, two level enriching column input pipe（16a）, two level enrichment column valve（16b）Into fraction collector
（17）Waste liquid port discharge；
The target of one-level separation process is that the natural medicinal ingredients of complexity are separated into 10- according to compound polarity or category Properties
20 parts of repeatable active ingredient positions obtained, for pharmaceutical activity system research；
（2）Natural medicinal ingredients the second-order separation process
PLC system is by controlling one-level enriching column selector valve（13a）With an one-level enriching column（13）For sample introduction post, the first solvent
Tank（1）Through the first solvent pump（4）, the second solvent tank（2）Second solvent pump（5）It is connected to the first blender（7）The gradient of composition
Liquid chromatography solvents system, through the first blender efferent duct（7a）, loading column valve（8a）, the branch pipe of blender first（12a）, control
One-level enriching column selector valve（13a）Switching with an one-level enriching column（13）For sample introduction post；Through the second-order separation post selector valve
（15b）In the second-order separation post（15）On separated, the liquid after separation is through the second-order separation valve（15a）, the second one-level splitter
Branch pipe（9c）, one-level splitter selector valve（9a）, the first one-level splitter branch pipe（9b）, detector（11）, the second blender
（12）, the second blender valve（14）, the branch pipe of blender second（12b）, two level enriching column input pipe（16a）It is connected to two level enrichment
Post group；Meanwhile start diluting pump（6）Dilution is injected in the second blender（12）The middle characteristic for changing efflux, passes through two
Level enrichment column valve（16b）Collected, in the way of SPE by chromatographic peak, by mixture according to polarity or compounds category characteristic
Two level enriching column is enriched in respectively（16）In, the pure active ingredient of natural drug is obtained, waste liquid is enriched with column valve by two level（16b）
Discharge；
The target of the second-order separation is that the active ingredient position of one-level separation is separated into monomeric compound respectively；
（3）The desorption of active component and pure compound
After the completion of the one-level separation of each natural drug or the second-order separation at position, position or pure is carried out automatically with strong solution analysis solvent
The desorption of compound.
3. a kind of natural medicinal ingredients system separator based on separation-Enrichment Mode according to claim 1, it is special
Sign is：The one-level splitter（9）, the second-order separation post（15）It is compressed axially using dynamic axial compression chromatographic column or static state
Post；The loading post（8）, one-level enriching column（13）, two level enriching column（16）Using static axial compression columns or common high
Effect liquid phase chromatogram post；The one-level splitter（9）, one-level enriching column（13）, the second-order separation post（15）, two level enrichment chromatographic column
（16）Filler be selected from silica gel, aluminum oxide, bonded-phase silica, macroporous absorbent resin, ion exchange resin or polyamide.
4. a kind of natural medicinal ingredients system separator based on separation-Enrichment Mode according to claim 1, it is special
Sign is：The detector（11）Selected from preparative UV-detector, EISD.
5. a kind of natural medicinal ingredients system separator based on separation-Enrichment Mode according to claim 1, it is special
Sign is：The loading column valve（8a）, one-level splitter selector valve（9a）, the second-order separation valve（15a）, the second blender valve
（14）, one-level enriching column selector valve（13a）, two level enrichment column valve（16b）Using two-position four-way valve.
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| EP0245940A1 (en) * | 1986-04-16 | 1987-11-19 | Imperial Chemical Industries Plc | Liquid flow analysis |
| CN101791491A (en) * | 2010-03-29 | 2010-08-04 | 苏州汇通色谱分离纯化有限公司 | Online low-pressure, middle-pressure and high-pressure combined two-dimension preparation liquid chromatographic system |
| CN102961892A (en) * | 2011-09-01 | 2013-03-13 | 李贺然 | Preparative multidimensional liquid chromatographic instrument |
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