CN107037169B - A kind of full two-dimensional highly effective liquid phase chromatographic instrument and chromatography separating method with Multifunctional switching valve - Google Patents
A kind of full two-dimensional highly effective liquid phase chromatographic instrument and chromatography separating method with Multifunctional switching valve Download PDFInfo
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- CN107037169B CN107037169B CN201610938634.0A CN201610938634A CN107037169B CN 107037169 B CN107037169 B CN 107037169B CN 201610938634 A CN201610938634 A CN 201610938634A CN 107037169 B CN107037169 B CN 107037169B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
- G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
- G01N30/463—Flow patterns using more than one column with serial coupling of separation columns for multidimensional chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
- G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
- G01N30/465—Flow patterns using more than one column with serial coupling of separation columns with specially adapted interfaces between the columns
Abstract
The present invention provides a kind of full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve, the full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve are made of one-dimensional chromatogram flow phase liquid storage device M1, one-dimensional chromatography high-pressure pump P1, one-dimensional chromatography column feed materials valve V1, one-dimensional chromatography chromatographic column C1, one-dimensional chromatographic detector D1, Two way chromatograms mobile phase liquid storage device M2, Two way chromatograms high-pressure pump P2, Multifunctional switching valve V, multiple two dimensional separation column C2-1 to C2-8, Two way chromatograms detector D, fraction collector F.Protrusion effect of the invention is: 1. carry out two dimensional separation using multiple Two way chromatograms columns, and the optional type of stationary phase is more, and separative efficiency is high, and good separating effect is, it can be achieved that the full ingredient of complex sample separates and analysis.2. system is equipped with multiple detectors, it is ensured that separation process accurately controls, and improves separating effect.
Description
Technical field
The present invention relates to a kind of full two-dimensional highly effective liquid phase chromatographic instrument and for the method for comprehensive two dimensional gas chromatography separation, belong to instrument
Equipment manufacturing technology field is a kind of high performance liquid chromatography separation analytical equipment.
Background technique
High performance liquid chromatography is examined with high-pressure delivery mobile phase using high-effective fixed phase and higher sensitivity detector
Survey, have the characteristics that efficiently, quickly, be widely used, at present oneself developed into solve organic substance research in it is various separation, divide
The key technology for analysing problem, obtains extensively in fields such as pharmacy, life science, medicine, Food Science, petrochemical industry and environmental protections
General application.
Common high performance liquid chromatograph is by groups such as liquid storage device, high-pressure pump, sample injector, chromatographic column, detector, recorders
At mostly using a root chromatogram column to be separated in application process, that is, one-dimensional chromatographic isolation.In one-dimensional chromatographic separation process
In, the eluent in liquid storage device is conveyed through sample injector through high-pressure pump, and the sample in carrier band sample injector enters chromatographic column and divided
From, by separation component successively from chromatographic column outflow enter detector.For complex systems such as natural products, body fluid, usual one
Tens even several hundred kinds of components are contained in sample, and structure is similar between each component, the difference of amount is usually again very big, by one-dimensional
The limitation of Resolution of Liquid Chromatography and peak capacity etc., is unable to satisfy the needs of complex separations.
Two-dimensional liquid chromatography is the technology to grow up on the basis of high performance liquid chromatography, it by technically and
One-dimensional chromatographic column separate fraction is introduced second using an one-dimensional chromatographic column and a Two way chromatograms column by the improvement in equipment
Chromatographic column carries out two dimensional separation.Compared with one-dimensional liquid chromatographic system, the advantage of Two way chromatograms is the provision of bigger peak and holds
Amount, so as to more effectively solve the problem analysis of complex system sample.
Jorgenson et al. realized on the basis of research " heartcut " technology " complete for the first time early in 1990
Two dimension cutting ", two quantity tubes are connected in an eight ways valve by they, and two quantity tube alternating samplings, sample introductions realize two
Dimension separation.Chinese patent " a kind of preparative multidimensional liquid chromatograph " (application number 201110256302.1), it is one-dimensional using one
Two enriching columns are connected to by splitter, a two dimensional separation column, two enriching columns and two ten-way valves, two dimensional separation column
On two ten-way valves, the alternately enrichment, sample introduction of two quantity tubes realizes two dimensional separation.A kind of Chinese patent " point of natural products
From method " (application number 201210022094.3) using an one-dimensional chromatographic column, a Two way chromatograms column, an enriching column and
One two six-way valve is replaced using enriching column enrichment, sample introduction by shutting down for high-pressure pump, realizes two dimensional separation.It is Chinese special
Sharp " a kind of full bidimensional capillary liquid chromatography separation system " (application number 200410098443.5), using an one-dimensional separation
Column, two two dimensional separation columns, an enriching column, two six-way valves, two ten-way valves, realize two dimensional separation.It is Chinese special
Benefit " full two dimension super-pressure high performance liquid chromatography separation system " (application number 201010256470.6), using a conventional high-pressure
Liquid-phase chromatographic column, a ultrahigh pressure liquid phase chromatographic column, two quantitative loops, two ten-way valves, conventional high-pressure liquid-phase chromatographic column
For one-dimensional splitter, ultrahigh pressure liquid phase chromatographic column is two dimensional separation column, hands over two quantitative loops using the rotation of two ten-way valves
For sampling, sample introduction, two dimensional separation is realized.
For now, full two dimension or multi-dimensional chromatograph mode are by with the interfaces mould such as six-way valve, eight ways valve, ten-way valve
One-dimensional chromatographic column separate fraction is introduced the second chromatographic column and carries out two dimensional separation by formula.Two dimensional separation column is required during the separation process
Short, analysis speed is fast, this is restricted entire separating effect.For the preparative separation of large capacity, the prior art can not
Enough meet demands not can be used directly the separation preparation in ingredient.The prior art uses an one-dimensional chromatographic column, a two-dimentional color
Spectrum column is separated, and most two kinds of stationary phase chromatographic columns can only be used to be separated every time in separation process, it is difficult to be met complicated
The separation of sample needs.
Summary of the invention
To solve defect present in existing Two way chromatograms technology, the present invention provides a kind of with the complete of Multifunctional switching valve
Two-dimensional highly effective liquid phase chromatographic instrument is switched by the conversion of valve, so that one-dimensional separation efflux is sequentially switched to multiple Two way chromatograms
Two dimensional separation is carried out in column, the part that Two way chromatograms separate is collected by fraction collector.
To achieve the goals above, the technical solution adopted by the present invention are as follows:
A kind of full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve, the full two-dimensional highly effective liquid with Multifunctional switching valve
Chromatography is by one-dimensional chromatogram flow phase liquid storage device M1, one-dimensional chromatography high-pressure pump P1, one-dimensional chromatography column feed materials valve V1, one-dimensional chromatography color
Compose column C1, one-dimensional chromatographic detector D1, Two way chromatograms mobile phase liquid storage device M2, Two way chromatograms high-pressure pump P2, Multifunctional switching valve
V, multiple two dimensional separation column C2-1 to C2-8, Two way chromatograms detector D, fraction collector F composition.
Connection relationship between each component is: No. 0 channel of Multifunctional switching valve V and M1 → P1 → V1 → D1 pipeline connect
It connects, No. 1-No. 8 channels of Multifunctional switching valve V are connect with the input end of Two way chromatograms column C2-1-C2-8 respectively, Two way chromatograms
The outlet end of column C2-1 to C2-8 respectively connects a detector D, and the outlet of detector D is connected to fraction collector F.It is multi-functional
No. 9 of switching valve V are connected together to No. 16 channels with the outlet of a multi-pass, the import and M2 → P2 piping connection of multi-pass.
Flow path in use process are as follows: the M1 → P1 → V1 → D1 → channel Multifunctional switching valve V(0) → Multifunctional switching valve V
(the successively channel 1-8), the M2 → P2 → channel Multifunctional switching valve V(9-16) → Two way chromatograms column (C2-1- C2- respectively
8) → detector D(difference) → fraction collector F.
The Multifunctional switching valve V port number and channel number are not limited only to the channel number of above-mentioned fixation, also include benefit
Similar realize that other of situation of the present invention connect situation with designing with Multifunctional switching valve V.
Described liquid storage device M1, M2 can be the corrosion resistant glass container of organic solvent-resistant or organic material container etc..
Described high-pressure pump P1, P2 can be the constant flow pump that can provide steady pressure and flow velocity or constant pressure pump etc..
The sampling valve V1 can be sampling valve or sampling pump of fixed volume etc..
The chromatographic column C1 and multiple two dimensional separation column C2-1, C2-2, C2-3, C2-4, C2-5, C2-6, C2-7, C2-8 can
To be glass, stainless steel, PEEK etc..The filler of filling can be filled out for ion exchange material, normal-phase chromatography filler, reverse-phase chromatography
Material, gel filler etc., the amount of filler can be analytic type, semi-preparative, preparative etc..
The Multifunctional switching valve V is one one into the multidigit multi-pass selector valve having more.
Described detector D1, D can be the detection of the flow types such as UV detector, fluorescence detector, differential refraction detector
Device etc..The detection cell of detector D1 is high pressure resistant detection cell.
The invention also provides the chromatography separating methods of the full two-dimensional highly effective liquid phase chromatographic instrument: the elution in liquid storage device M1
Liquid is conveyed through sampling valve V1 through high-pressure pump P1, carries the sample in sampling valve V1 and enters the one-dimensional separation of chromatographic column C1 progress, color
The efflux of spectrum column C1 enters No. 0 through-hole of Multifunctional switching valve V, is successively switched by multifunctional rotary switching valve V
Be connected to chromatographic column C2-1, C2-2 on the 1-8 through-hole of Multifunctional switching valve V, C2-3, C2-4, C2-5, C2-6, C2-7,
C2-8.Eluent in liquid storage device M2 is conveyed through 9-16 through-hole, the 1-8 through-hole of Multifunctional switching valve V through high-pressure pump P2
Into chromatographic column C2-1, C2-2, C2-3, C2-4, C2-5, C2-6, C2-7, C2-8, two dimensional separation, two dimensional separation efflux are carried out
It is detected into detector D, the efflux of detector D enters fraction collector F and is collected.The one-dimensional separation eluent of separation process
Glide path are as follows: M1 → P1 → V1 → D1 → V(0) → V(1-8) → C2-1 to C2-8(successively), two dimensional separation eluent flowing
Route are as follows: M2 → P2 → V(9-16) → V(1-8) → C2-1 to C2-8(is parallel) → D(is parallel) → F(is parallel).
A kind of full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve of the present invention and for comprehensive two dimensional gas chromatography separation
Method, separating capacity with super strength.
In addition to this, protrusion effect of the invention is:
1. carrying out two dimensional separation using multiple Two way chromatograms columns, the optional type of stationary phase is more, and separative efficiency is high, separation
Effect is good, it can be achieved that the full ingredient of complex sample separates and analysis.
2. system is equipped with multiple detectors, it is ensured that separation process accurately controls, and improves separating effect.
3. carrying out two dimensional separation using multiple Two way chromatograms columns, there is no to one-dimensional column size and flow rate of mobile phase
Limitation, can be used for large sample amount preparative separation.
4. carrying out two dimensional separation using multiple Two way chromatograms columns, there is the sufficient time to carry out two dimensional separation, there is no to two
Dimension chromatographic column must be the limitation of short column.
5. system design is simple, good separating effect, suitable for isolating and purifying for all kinds of complex mixtures, practicability is extremely strong.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of the full two-dimensional highly effective liquid phase chromatographic instrument of the embodiment of the present invention.
Fig. 2 is embodiment one-dimensional detector output signal schematic diagram.
Fig. 3 is embodiment two-dimensional detector output signal schematic diagram.
Specific embodiment
Below with reference to embodiment and attached drawing, the present invention will be described in detail, but the scope of protection is not limited by this.
Embodiment 1 as shown in Figure 1, a kind of full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve and be used for complete two
The method for tieing up chromatographic isolation, full two-dimensional highly effective liquid phase chromatographic instrument is by one-dimensional chromatogram flow phase liquid storage device M1, one-dimensional chromatography high-pressure pump
P1, one-dimensional chromatography column feed materials valve V1, one-dimensional chromatography chromatographic column C1, one-dimensional chromatographic detector D1, Two way chromatograms mobile phase liquid storage device M2,
Two way chromatograms high-pressure pump P2, Multifunctional switching valve V, Two way chromatograms chromatographic column C2-1 to C2-8, Two way chromatograms detector D, fraction
Collector F composition.
Eluent in liquid storage device M1 is conveyed through sampling valve V1 through high-pressure pump P1, and the sample carried in sampling valve V1 enters
Chromatographic column C1 carries out one-dimensional separation, and the efflux of chromatographic column C1 enters No. 0 through-hole of Multifunctional switching valve V, by rotating more function
Energy switching valve V successively switches into chromatographic column C2-1, C2-2, the C2- being connected on the 1-8 through-hole of Multifunctional switching valve V
3,C2-4,C2-5,C2-6,C2-7,C2-8.Eluent in liquid storage device M2 is conveyed through Multifunctional switching valve V through high-pressure pump P2
9-16 through-hole, 1-8 through-hole enter chromatographic column C2-1, C2-2, C2-3, C2-4, C2-5, C2-6, C2-7, C2-8, carry out
Two dimensional separation, two dimensional separation efflux enter detector D detection, and the efflux of detector D enters fraction collector F and received
Collection.The one-dimensional separation eluent glide path of separation process are as follows: M1 → P1 → V1 → D1 → V(0) → V(1-8) → C2-1-8, two dimension
Separation eluent glide path are as follows: M2 → P2 → V(9-16) → V(1-8) → C2-1 to C2-8(is parallel) → D(is parallel) → F(is flat
Row).
By above-mentioned connection and mode of operation, the comprehensive two dimensional gas chromatography separation of Fructus Sophorae extract is carried out:
First dimension chromatographic column is C18 column, length 10cm.Second dimension chromatographic column C2-1, C2-2, C2-3, C2-4, C2-5,
C2-6, C2-7, C2-8 are followed successively by C4 column, C4 column, benzene column, benzene column, C8 column, C8 column, C18 column, C18 column, length 10cm.It is one-dimensional
Mobile phase is methanol-water (volume ratio 40/60), and flow velocity 1ml/min, two-dimensional flow is mutually that -0.1% formic acid gradient of acetonitrile is washed
De-, 0-30min, acetonitrile becomes 80% from 20%, and the flow velocity of two-dimensional flow phase is 1.0ml/min.
One-dimensional chromatogram and two-dimensional chromatogram when the embodiment of the present invention is separated are shown in attached drawing 2,3, illustrate the present invention
Separative efficiency is high, practical, is particularly suitable for the separation of full ingredient and the chipal compounds, knot of the complex systems such as Chinese medicine, body fluid
The separation of the hardly possible separate substance such as structure analogue compounds.
The above is only embodiments of the present invention, it is noted that for those skilled in the art,
Under the premise of not departing from present inventive concept, several improvements and modifications can also be made, these improvements and modifications also should be regarded as this hair
In bright protection scope.
Completion of the invention has benefited from grant of national natural science foundation (project approval number: 21675071).
Claims (3)
1. a kind of full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve, characterized in that by one-dimensional chromatogram flow phase liquid storage
Device M1, one-dimensional chromatography high-pressure pump P1, one-dimensional chromatography column feed materials valve V1, one-dimensional chromatography chromatographic column C1, one-dimensional chromatographic detector D1, two dimension
Chromatogram flow phase liquid storage device M2, Two way chromatograms high-pressure pump P2, Multifunctional switching valve V, two dimensional separation column C2-1 to C2-8, two-dimentional color
Compose detector D, fraction collector F composition；No. 0 channel of Multifunctional switching valve V and M1 → P1 → V1 → D1 piping connection,
No. 1-No. 8 channels of Multifunctional switching valve V are connect with the input end of Two way chromatograms column C2-1-C2-8 respectively, Two way chromatograms column
The outlet end of C2-1 to C2-8 respectively connects a detector D, and the outlet of detector D is connected to fraction collector F；Multifunction cutting
No. 9 to No. 16 channels for changing valve V are connected together with the outlet of a multi-pass, the import and M2 → P2 piping connection of multi-pass；Institute
Stating liquid storage device M1, M2 is the corrosion resistant glass container of organic solvent-resistant or organic material container；Described high-pressure pump P1, P2 are to mention
For the constant flow pump or constant pressure pump of steady pressure and flow velocity；The sampling valve V1 is the sampling valve or sampling pump of fixed volume；It is described
The material of chromatographic column C1 and two dimensional separation column C2-1, C2-2, C2-3, C2-4, C2-5, C2-6, C2-7, C2-8 is glass, stainless
Steel or PEEK, the filler of filling are ion exchange material, normal-phase chromatography filler, reverse-phase chromatography filler or gel filler；It is described more
Function switch valve V is one one into the multidigit multi-pass selector valve having more；Described detector D1, D are UV detector, fluorescence inspection
Survey device or differential refraction detector；The detection cell of detector D1 is high pressure resistant detection cell.
2. the full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve as described in claim 1, characterized in that the amount of filler
It is analytic type, semi-preparative or preparative.
3. the chromatography separating method of the full two-dimensional highly effective liquid phase chromatographic instrument with Multifunctional switching valve as described in claim 1,
Be characterized in: the eluent in liquid storage device M1 is conveyed through sampling valve V1 through high-pressure pump P1, and the sample carried in sampling valve V1 enters
Chromatographic column C1 carries out one-dimensional separation, and the efflux of chromatographic column C1 enters No. 0 through-hole of Multifunctional switching valve V, by rotating more function
Energy switching valve V successively switches into chromatographic column C2-1, C2-2, the C2- being connected on the 1-8 through-hole of Multifunctional switching valve V
3,C2-4,C2-5,C2-6,C2-7,C2-8；Eluent in liquid storage device M2 is conveyed through Multifunctional switching valve V through high-pressure pump P2
9-16 through-hole, 1-8 through-hole enter chromatographic column C2-1, C2-2, C2-3, C2-4, C2-5, C2-6, C2-7, C2-8, carry out
Two dimensional separation, two dimensional separation efflux enter detector D detection, and the efflux of detector D enters fraction collector F and received
Collection.
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| CN101169393A (en) * | 2006-10-25 | 2008-04-30 | 中国科学院大连化学物理研究所 | Two-dimensional array liquid phase chromatogram-mass spectrometry combination method |
| CN103063753A (en) * | 2012-08-31 | 2013-04-24 | 西安奥岚科技开发有限责任公司 | Multidimensional liquid phase chromatography separation system and separation method for protein separation |
| CN103884811A (en) * | 2014-03-04 | 2014-06-25 | 中国人民解放军第二军医大学 | Biological chromatography comparing and screening system and application thereof |
| CN206146889U (en) * | 2016-11-01 | 2017-05-03 | 聊城大学 | Take full multidimensional high performance liquid chromatography appearance of multi -functional diverter valve |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US6802967B2 (en) * | 2002-03-06 | 2004-10-12 | Shimadzu Corporation | Multi-dimension liquid chromatography separation system |
| US20050090013A1 (en) * | 2003-10-28 | 2005-04-28 | Joel Myerson | Separation channel with transverse pump extraction |
| US8621915B2 (en) * | 2007-05-29 | 2014-01-07 | Waters Technologies Corp. | Apparatus and methods for multidimensional analysis |
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2016
- 2016-11-01 CN CN201610938634.0A patent/CN107037169B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101169393A (en) * | 2006-10-25 | 2008-04-30 | 中国科学院大连化学物理研究所 | Two-dimensional array liquid phase chromatogram-mass spectrometry combination method |
| CN103063753A (en) * | 2012-08-31 | 2013-04-24 | 西安奥岚科技开发有限责任公司 | Multidimensional liquid phase chromatography separation system and separation method for protein separation |
| CN103884811A (en) * | 2014-03-04 | 2014-06-25 | 中国人民解放军第二军医大学 | Biological chromatography comparing and screening system and application thereof |
| CN206146889U (en) * | 2016-11-01 | 2017-05-03 | 聊城大学 | Take full multidimensional high performance liquid chromatography appearance of multi -functional diverter valve |
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