CN204405613U - A kind of Two-dimensional Liquid Chromatography - Google Patents

Abstract

The utility model provides a two-dimensional liquid chromatograph. The two-dimensional liquid chromatograph comprises a first flow path (L3), a second flow path (L11), an analysis flow Channel (L14), waste liquid channel (L4), intermediate chromatographic column (C2), multi-channel switching valve (V1) and storage valve (V2), etc., the two-dimensional liquid chromatography through the multi-channel switching valve ( The valve switch between V1) and the storage valve (V2) can change the position of the intermediate chromatographic column in the flow channel to realize the storage function of the intermediate chromatographic column. The two-dimensional liquid chromatograph uses only one first chromatographic column to realize the parallel operation of the first-dimensional liquid chromatography and the second-dimensional liquid chromatography for processing different samples.

Description

A two-dimensional liquid chromatograph

technical field

The utility model belongs to the field of liquid chromatography, in particular to a two-dimensional liquid chromatography with the function of parallel operation.

Background technique

Two-dimensional liquid chromatography is a chromatographic instrument developed on the basis of ordinary liquid chromatography. It improves the separation ability by increasing the number of chromatographic separation stages and chromatographic columns. Usually, the first-dimensional liquid chromatography is responsible for the concentration and primary separation of samples. The second-dimensional liquid chromatography is responsible for the further separation and detection of the sample, so that the sample can be processed online to separate the target components from the complex matrix. Compared with ordinary liquid chromatography, ordinary liquid chromatography often requires cumbersome and complicated liquid-liquid extraction or solid-phase extraction processes when analyzing target components in blood samples and urine. material extracted. Two-dimensional liquid chromatography can complete the concentration, transfer and separation of samples with a high degree of automation, which greatly improves the accuracy and automation of chromatographic separation.

In conventional two-dimensional chromatography, the first-dimensional liquid chromatography system of the sample performs preliminary separation and concentration processing on the sample, and then passes it to the second-dimensional liquid chromatography for subsequent processing, so the analysis time of the sample is the first-dimensional liquid phase The sum of the chromatographic processing time and the second-dimension liquid chromatography processing time. In order to increase the speed of sample analysis, there are currently two-dimensional chromatographs that operate in parallel. The first-dimensional liquid chromatography of this instrument has two identical sample concentration and primary separation columns, which are used to alternately process and transfer the target components in the sample. , that is, when a chromatographic column of the first-dimensional liquid phase is connected to a chromatographic column of the second-dimensional liquid phase and is in the working state of separation and detection of samples, the other first-dimensional chromatographic column is at the concentration or primary level of the next sample Separating the working state, this operation mode makes the first-dimensional liquid chromatography and the second-dimensional liquid chromatography work at the same time, so the analysis speed is faster and more efficient than the conventional two-dimensional liquid chromatography.

However, there is such a problem in the two-dimensional chromatography of the above technologies: the two chromatographic columns of the first-dimensional chromatographic column must have the same specifications, properties, and advantages and disadvantages in order to obtain consistent analysis results. After analyzing a large number of samples, the chromatographic columns will both There are different degrees of deterioration, so the same two first-dimensional chromatographic columns have inconsistent changes in retention capacity and chromatographic peak shape after a period of use, resulting in differences in analysis results, and in severe cases, continuous analysis cannot be performed Another problem is that when analyzing samples in batches, it is necessary to specify which one of the two first-dimensional chromatographic columns corresponds to the sample, which is very troublesome when compiling the sample processing sequence, and increases the complexity of compiling the sample injection sequence list.

Utility model content

Aiming at the deficiencies of the prior art, the utility model of the two-dimensional liquid chromatograph aims to use only one first-dimensional chromatographic column to realize the parallel operation of the first-dimensional liquid chromatography and the second-dimensional liquid chromatography to process samples . The two-dimensional liquid chromatograph changes the position of the intermediate chromatographic column in the flow channel by switching the valve between the multi-channel switching valve and the storage valve, so that the intermediate chromatographic column can be in the downstream path of the first flow channel or in the second flow channel. In the downstream path of the flow channel or neither in the downstream path of the first flow channel nor in the downstream path of the second flow channel, the storage function of the intermediate chromatographic column is realized, so as to achieve the first-dimensional liquid chromatography and the second-dimensional liquid chromatography. The purpose of running jobs in parallel for sample jobs.

In order to achieve the above object, the technical solution of the utility model is:

A two-dimensional liquid chromatograph, comprising: a first flow channel connected with a first chromatographic column, used to transport the first mobile phase, and perform preliminary separation on the sample; a second flow channel, used to transport the second mobile phase; The analysis flow channel is used to separate and detect the captured substances; the waste liquid flow channel is used to discharge waste liquid; it also includes a multi-channel switching valve with multiple interfaces and a storage valve with multiple ports; all An intermediate chromatographic column is connected between any two ports of the storage valve; any three ports of the multi-channel switching valve are respectively connected to a section of the first connecting pipeline, the second connecting pipeline and the third connecting pipeline, so The other ends of the first connecting pipeline and the second connecting pipeline are respectively communicated with the idle port of the register valve; the other end of the third connecting pipeline is communicated with the second connecting pipeline through a tee I; The first flow channel, the second flow channel, the analysis flow channel and the waste liquid flow channel are respectively connected to any other ports on the multi-channel switching valve.

A preferred solution: the two-dimensional liquid chromatograph further includes a modulating channel for transporting a modulating solution; the modulating channel is connected to the second connecting pipeline through a tee II. The prepared solution can be an acidic, neutral, alkaline solution, or a high proportion of organic solvent. By adjusting the pH of the first-dimensional mobile phase, the proportion of solvent components, and the ionic strength of the first-dimensional mobile phase entering the intermediate column, the target components in the sample can be captured by the intermediate chromatographic column to enhance the ability to prevent the loss of the target component. In addition, the first-dimensional chromatographic column The eluted components reduce the diffusion as much as possible on the intermediate column, so when the first-dimensional chromatography column deteriorates and deteriorates after a period of use, the diffusion and change of the target substance in the sample on the intermediate column is not obvious , after being transferred to the second-dimensional chromatographic column, the analysis result and chromatographic peak shape can be kept stable.

A further preferred solution: the multi-channel switching valve includes interface a, interface b, interface c, interface d, interface e, interface f, interface g, and interface j; the interface a is connected to the first flow channel, and the interface b It is connected to the waste liquid channel, the interface c is connected to one end of the first connecting pipeline, the interface d is connected to the analysis flow channel, the interface e is connected to the second flow channel, and the interface f is connected to the second One end of the pipeline is connected, and the interface j is connected with one end of the third connecting pipeline.

The modulation channel can also be connected to the interface g of the multi-channel switching valve.

The multi-channel switching valve may further include an interface h and an interface i. If the interface h and the interface i exist, the interface g and the interface i are in a blocked state.

Further preferred solution: the deposit valve includes port a, port b, port c, port d, port e, and port f; an intermediate chromatographic column is arranged between the port a and the port d, and the port e is connected to the second The other end of the pipeline is connected, and the port f is connected with the other end of the first connecting pipeline.

Preferred solution: the intermediate chromatographic column is a chromatographic column that has a stronger effect on the target component flowing out of the first chromatographic column.

A preferred solution: the first chromatographic column and the intermediate chromatographic column are chromatographic columns or quantitative loops capable of storing target components in the sample.

Unless otherwise specified, the multi-channel switching valve and storage valve described in the present invention are generally connected to only one pipeline with one port or interface. The multi-channel switching valve and the storage valve described in the utility model are two-position switching valves.

Except for the ports or interfaces that are in the closed state, the ports or interfaces that are not connected to pipelines are called idle ports or idle interfaces.

Compared with the prior art, the utility model has the advantages of:

1. The two-dimensional liquid chromatograph has only one first-dimensional chromatographic column, which reduces the complexity of instrument operation, and can reduce the impact of the deterioration of the first-dimensional chromatographic column on the separation and detection of the second-dimensional chromatographic column.

2. Since the two-dimensional liquid chromatograph has only one first-dimensional chromatographic column, it is not necessary to specify the corresponding relationship between the sample and the first-dimensional chromatographic column when programming the sample injection sequence, which reduces the complexity of processing sequence programming.

3. Strengthen the holding capacity of the target components in the sample eluted from the first dimension chromatographic column on the intermediate chromatographic column through the added modulation function, and make the transfer of the target components complete and stable.

Description of drawings

Fig. 1 is the liquid chromatograph structural representation of embodiment 1;

Fig. 2 is a kind of working status figure of the two-dimensional liquid chromatography of embodiment 1;

Fig. 3 is a kind of working state diagram of the two-dimensional liquid chromatography of embodiment 1;

Fig. 4 is a kind of working state figure of the two-dimensional liquid chromatography of embodiment 1;

Fig. 5 is a kind of working status diagram of the two-dimensional liquid chromatography of embodiment 1;

Fig. 6 is a kind of working status figure of the two-dimensional liquid chromatography of embodiment 1;

Fig. 7 is a kind of working status diagram of the two-dimensional liquid chromatography of embodiment 1;

Fig. 8 is a kind of working status diagram of the two-dimensional liquid chromatography of embodiment 2;

Among them, 1 is port a, 2 is port b, 3 is port c, 4 is port d, 5 is port e, 6 is port f, 7 is port g, 8 is port h, 9 is port i, and 10 is port j, 11 is port a, 12 is port b, 13 is port c, 14 is port d, 15 is port e, 16 is port f; AS is injector, W1 is waste liquid port, DE is detector; S1 is the first mobile phase, S2 is the second mobile phase, S3 is the preparation solution; P1 is the transfer pump I, P2 is the transfer pump II, P3 is the transfer pump III; C1 is the first chromatographic column, C2 is the middle chromatographic column, C3 is the second chromatographic column; L3 is the first flow channel, L11 is the second flow channel, L14 is the analysis flow channel, L4 is the waste liquid flow channel, L9 is the first connecting line, L12 is the second connecting line, L5 is the The third connecting pipeline, L17 is the modulation channel; T1 is the three-way I, T2 is the three-way II; V1 is the multi-channel switching valve, and V2 is the storage valve. The solid dots in the figure represent the blocking state, and the thick solid line represents the flow route of the mobile phase.

Detailed ways

Below in conjunction with accompanying drawing and embodiment the utility model is further explained and illustrated

Example 1

As shown in Figure 1, a two-dimensional liquid chromatograph includes:

The first flow channel L3 connected with the sample injector AS and the first chromatographic column C1 in sequence is used to transport the first mobile phase S1 and perform preliminary separation on the sample;

The second channel L11 is used to transport the second mobile phase S2;

Analytical channel L14, used for separation and detection of captured substances;

The waste liquid channel L4 is used to discharge the waste liquid;

It also includes a multi-channel switching valve V1 with multiple interfaces and a storage valve V2 with multiple ports; an intermediate chromatographic column C2 is connected between any two ports of the storage valve V2;

Any three ports of the multi-channel switching valve V1 are respectively connected to the first connecting pipeline L9, the second connecting pipeline L12 and the third connecting pipeline L5, and the first connecting pipeline L9 and the second connecting pipeline The pipelines L12 are respectively communicated with idle ports of the register valve V2; the third connecting pipeline L5 is communicated with the second connecting pipeline L12 through the tee IT1;

The first flow channel L3, the second flow channel L11, the analysis flow channel L14 and the waste liquid flow channel L4 are respectively connected to any other ports on the multi-channel switching valve V1.

The multi-channel switching valve V1 includes a port a1, a port b2, a port c3, a port d4, a port e5, a port f6, a port g7, a port h8, a port i9 and a port j10; the port a1 is connected to the first flow channel L3, The interface b2 is connected to the waste liquid channel L4, the interface c3 is connected to the first connection pipeline L9, the interface d4 is connected to the analysis channel L14, the interface e5 is connected to the second channel L11, and the The interface f6 is connected to the second connection pipeline L12, and the interface j10 is connected to the third connection pipeline L5. When working, the interface g7 and the interface i9 are in a blocked state.

The storage valve V2 includes a port a11, a port b12, a port c13, a port d14, a port e15, and a port f16; an intermediate chromatographic column C2 is provided between the port a11 and the port d14, and the port e15 is connected to the second connecting pipeline L12 is connected, and the port f16 is connected to the first connecting pipeline L9.

Function description:

As shown in Figure 2, the separation function of the first chromatographic column: turn on the delivery pump IP1, so that the first mobile phase moves downstream under the push of IP1, and the sample is introduced into the first mobile phase through the sample injector AS, and the second mobile phase containing the sample A mobile phase enters the first chromatographic column C1, and each component in the sample begins to separate under the chromatographic separation mechanism of the first mobile phase and the first chromatographic column, and the port a1 and port b2 of the multi-channel switching valve V1 are connected, so that the first The mobile phase flows to the waste liquid channel L4 and is discharged.

As shown in Figure 3, the capture function of the middle chromatographic column: turn on the delivery pump IP1, transport the first mobile phase to the downstream, pass through the sample injector AS and the first chromatographic column C1 in sequence, and the sample is introduced into the first mobile phase through AS, Then the target component in the sample is separated by the chromatographic force of the first mobile phase and the first chromatographic column. When the target component in the sample is about to flow out of the first chromatographic column, the multi-channel switching valve V1 port a1 and port j10, so that the first mobile phase containing the target component flows into the storage valve V2 through the third connecting line L5 and the second connecting line L12, and then flows through the second chromatographic column C2, which has a strong effect on the second chromatographic column C2 The target components can be captured by the second chromatographic column C2, and the uncaptured components pass through the first connecting pipeline L9 with the first mobile phase, and move through the connected multi-channel switching valve V1 port c3 and port b2, Flow to the waste liquid channel L4 and discharge.

As shown in Figure 4, the registration function: switch the registration valve V2, so that the intermediate column is connected to the ports b12 and c13. Since there is no fluid entering or flowing out from the ports b12 and c13, the intermediate chromatographic column C2 is neither in the downstream path of the first flow channel The medium is also not in the downstream path of the second flow channel; at this time, the components retained on the intermediate column are in a static state, waiting to be washed down when the next fluid passes, so as to realize the storage function of the target components in the intermediate column C2.

As shown in Figure 5, the function of parallel operation: in the same period of time, the transfer pump IP1 is started, and the first mobile phase is sent to the downstream pipeline, passing through the sampler AS and the first chromatographic column C1 in turn, and the sample is introduced into the second chromatographic column through AS. After a mobile phase, the first chromatographic column C1 starts to separate the components in the sample; at the same time, the solution in the second flow channel is driven by the infusion pump IIP2, and then passes through the intermediate chromatographic column C2 on the multi-channel switching valve V1 and the storage valve V2 , push the target components stored in the middle column C2 into the second column C3, and the target components stored in C3 start to separate, so that the first column C1 and the second column C3 are in the sample processing work at the same time running in parallel.

As shown in Figure 6, when the target component in the solution to be analyzed on the first chromatographic column C1 is about to be pushed out by the solution in the first flow channel L3, the positions of the register valve and the multi-channel switching valve are selected so that the middle chromatographic column C2 and the first chromatographic column Column C1 is connected, so that the target component is pushed into the middle column C2, and the component with strong retention force with C2 will be captured by the C2 column; at this time, the second column C3 is also working to separate the previous sample component state; thereby realizing the function of parallel operation of the capture of the middle chromatographic column C2 and the separation of the second chromatographic column C3.

As shown in Figure 7, after the target component flowing out of the first chromatographic column C1 enters the intermediate chromatographic column C2, select the register valve position so that the intermediate chromatographic column C2 is neither connected to the first flow channel L3 nor to the second flow channel L11 connected, so that the target component is stored in the middle chromatographic column C2; at this time, the second chromatographic column C3 is in the working state of separating the previous sample component at the same time; the storage of the middle chromatographic column C2 is separated from the second chromatographic column C3 functions that run in parallel.

Repeat the working process shown in Fig. 5-Fig. 7 to realize the continuous parallel operation function of the continuous two-dimensional liquid chromatograph.

Example 2

A two-dimensional liquid chromatograph, on the basis of Embodiment 1, also includes a modulation flow path L17 for delivering the modulation solution S3; the modulation flow path L17 is connected to the second connection line L12 through a tee IIT2 Or connect directly to port g7 on the multi-port valve. And the intermediate chromatographic column C2 selects a chromatographic stationary phase capable of retaining the target components in the sample.

Functional description: As shown in Figure 8, while the intermediate chromatographic column C2 capture function is performed in Example 1, the transfer pump P3 is turned on, and the modulation solution S3 is transferred to the second connecting pipeline L12 to merge with the first mobile phase, and through the modulation Solution S3 changes the pH value, ionic strength or organic phase ratio of the first mobile phase S1, so as to enhance the retention capacity of the target component on the intermediate column C2 and prevent the target component from flowing out in the C2 capture process.

Claims (6)

1. A two-dimensional liquid chromatograph, comprising: The first flow channel (L3) connected with the first chromatographic column (C1) is used to deliver the first mobile phase (S1) and perform preliminary separation of the sample; The second channel (L11) is used to transport the second mobile phase (S2); Analytical channel (L14) for separation and detection of captured substances; Waste liquid flow channel (L4), used to discharge waste liquid; It is characterized in that it also includes a multi-channel switching valve (V1) with multiple interfaces and a storage valve (V2) with multiple ports; any two ports of the storage valve (V2) are connected with an intermediate chromatography Column(C2); Any three ports of the multi-channel switching valve (V1) are respectively connected to one end of the first connecting pipeline (L9), the second connecting pipeline (L12) and the third connecting pipeline (L5). The other ends of the first connecting pipeline (L9) and the second connecting pipeline (L12) are respectively communicated with the free port of the storage valve (V2); the other end of the third connecting pipeline (L5) is connected through the tee I ( T1) communicates with the second connecting pipeline (L12); The first flow channel (L3), the second flow channel (L11), the analysis flow channel (L14) and the waste liquid flow channel (L4) are respectively connected to any other interface on the multi-channel switching valve (V1). 2. The two-dimensional liquid chromatograph according to claim 1, characterized in that it also includes a modulating channel (L17) for delivering the modulating solution (S3); the modulating channel (L17) passes through the tee II ( T2) is connected to the second connecting pipeline (L12) or the modulation channel (L17) is connected to the idle port of the multi-channel switching valve (V1). 3. The two-dimensional liquid chromatograph according to claim 1 or 2, characterized in that the multi-channel switching valve (V1) includes interface a (1), interface b (2), interface c (3), Interface d(4), interface e(5), interface f(6), interface g(7) and interface j(10); the interface a(1) is connected to the first flow channel (L3), and the interface b (2) Connected to the waste liquid channel (L4), the interface c(3) is connected to one end of the first connecting pipeline (L9), the interface d(4) is connected to the analysis channel (L14), the The interface e(5) is connected to the second flow channel (L11), the interface f(6) is connected to one end of the second connecting pipeline (L12), and the interface j(10) is connected to the third connecting pipeline ( L5) is connected at one end. 4. The two-dimensional liquid chromatograph according to claim 2, characterized in that, the modulation channel (L17) is connected to the interface g (7) of the multi-channel switching valve (V1). 5. The two-dimensional liquid chromatograph according to claim 1 or 2, characterized in that, the register valve (V2) includes port a (11), port b (12), port c (13), port d ( 14), port e (15), port f (16); an intermediate chromatographic column (C2) is provided between the port a (11) and port d (14), and the port e (15) is connected to the second The other end of the pipeline (L12) is connected, and the port f(16) is connected with the other end of the first connecting pipeline (L9). 6. The two-dimensional liquid chromatograph according to claim 1 or 2, characterized in that, the first chromatographic column (C1) and the intermediate chromatographic column (C2) are capable of storing the target components in the sample column or loop.