CN208060444U - A kind of moveable mobile phase control system with double gradient regulatory functions - Google Patents

Abstract

The utility model provides a movable mobile phase control system with dual-gradient adjustment functions. The system includes a control assembly, a syringe pump group, a gradient mixer and a sampling valve connected in sequence. The mobile phase control system described in the utility model is suitable as a mobile phase system of an analysis and characterization instrument in biological material research to simulate a complex solid-liquid interface environment, and is applied in the field of interface analysis, characterization and detection.

Description

A Mobile Phase Control System with Dual Gradient Adjustment Function

technical field

The utility model belongs to the field of liquid phase pumps, and relates to a mobile phase control system, in particular to a movable mobile phase control system with dual gradient adjustment functions, which is suitable for analysis and characterization instruments in biological material research The mobile phase system is used to simulate the complex solid-liquid interface environment and is used in the field of interface analysis, characterization and detection.

Background technique

In recent years, the interface analysis and detection technology has been developing continuously, and its main development direction is to increase the scanning rate, improve the detection accuracy and provide more types of detection chips. At present, the widely used interface analysis instruments in biomaterial research include atomic force microscope (AFM), dual polarization interference interface analysis system (DPI) and quartz crystal microbalance analyzer (QCM). AFM is a typical interface characterization and mechanical measurement tool, and is widely used in surface imaging, mechanical characterization of interface molecular layers, single-molecule force spectroscopy and interface electromagnetic characterization. In a liquid environment, AFM can maintain a good working condition, so it can be used to study biological macromolecules at the solid-liquid interface, even living biological cells and the forces between them. DPI is an interface characterization technology developed in recent years, which relies on a special detection chip to observe the changing interface layer in real time. The principle of DPI detection is: two polarized lights pass through the optical channel in the middle of the detection chip in parallel, forming interference fringes on the detector, and the change of the molecular layer on the chip surface will affect the interface wave field (evanescent wave field) of the optical path, thereby changing the interference fringes , through comparison and fitting, the adsorption amount of interface molecules and the thickness of the adsorption layer can be deduced. The detection sensitivity of DPI is less than 1pg/mm ² , and the detection interface layer thickness reaches 100nm; in addition to the adsorption-desorption behavior, the quality and thickness data of the adsorption layer can also be obtained. QCM has a mobile phase control system similar to DPI, relies on a special quartz crystal plate, can observe the changing interface layer in real time, and can obtain data on the quality, thickness and viscoelasticity of the interface molecular layer. The core component of QCM is a quartz crystal sensor, which consists of a quartz crystal sandwiched between two electrodes. Adding AC voltage at both ends of the electrodes can cause weak vibration of the sensor; when the AC voltage is turned off, the vibration decays exponentially, and the interface molecular layer will change the attenuation process. Through comparison and fitting, the structure information and physical properties of the interface layer can be obtained. Compared with optical-based interface analysis methods, QCM has a relatively low sensitivity of about 20pg/mm ² , but it is more suitable for the measurement of large-scale interface layers and can characterize physical properties such as interface viscoelasticity. The above-mentioned interface analysis instrument has been applied to study the interaction of biological macromolecules on the solid-liquid interface, providing useful scientific research data for the research work of the preparation, purification and formulation of biological materials.

However, in the face of the rapid development of the field of biomaterials research, especially the urgent need for the analysis and characterization of complex interface environments, the existing instruments still have many shortcomings. Mainly in terms of mobile phase control, the development is relatively slow. DPI uses a unit, a dual-channel syringe pump and a six-way valve injector, which cannot perform gradient changes in the mobile phase; QCM uses a one-way peristaltic pump and relies on pipeline switching to achieve sample injection. ; AFM has only a liquid sample cell, without a pump system, relying on a manual syringe to advance the mobile phase. Therefore, the mobile phase control systems of existing instruments and equipment lack precise control of the liquid environment, and cannot realize gradient changes in solution composition, such as ionic strength and pH, let alone adjust various solution parameters. In the field of biomaterials research, many research contents involve the dynamic change of the interface environment. For example, in the process of protein separation and purification and the process of administering protein pharmaceutical preparations, the interface environment of protein molecules is dynamically changing, and the existing interface Characterization equipment cannot effectively simulate the solid-liquid interface environment in the actual system.

Some existing liquid phase control systems, such as liquid phase pumps, not only have a large dead volume, but cannot meet the needs of existing interface analysis equipment for rapid and accurate mobile phase gradient changes; moreover, large shear forces can easily cause biomacromolecules to be inactivated, so it is not possible to A mobile phase control system suitable for biomacromolecule characterization equipment.

Utility model content

Aiming at the problems existing in the prior art, the utility model provides a movable mobile phase control system with dual-gradient adjustment functions to realize the dual-gradient adjustment of the mobile phase. The mobile phase control system is suitable as a mobile phase system of an analysis and characterization instrument in biological material research to simulate a complex solid-liquid interface environment, and is applied in the field of interface analysis, characterization and detection.

For this purpose, the utility model adopts the following technical solutions:

The utility model provides a mobile phase control system with dual-gradient adjustment functions. The system includes a control assembly, a syringe pump group, a gradient mixer and a sampling valve connected in sequence.

The system of the utility model controls the flow of the mobile phase in the injection pump group through the control component, so that the mobile phase in the injection pump group is pushed into the gradient mixer at a certain ratio for mixing, and then enters the injection valve to realize the flow of the mobile phase Accuracy and stability of gradient adjustment, thus forming gradient changes of mobile phase concentration and other parameters.

The control components described in the utility model can adopt existing components in the prior art, typical but non-limitative ones can be controlled by a computer, and the programs used can adopt existing programs in the prior art (such as CN 103698428 A and CN 105771037 A etc. are all introduced). Command control, data collection and collation of the entire system are carried out through the control components.

In the utility model, the outlet of the sampling valve is connected to the inlet of the mobile phase of the interface characterization analysis instrument, thereby forming an interface characterization analysis experiment with a gradient change of the mobile phase.

The mobile phase control system described in the utility model can be used in connection with existing interface analysis instruments, and the interface analysis instruments include atomic force microscope (AFM), dual polarization interference interface analysis system (DPI) and quartz crystal microbalance analyzer ( QCM).

The following is the preferred technical solution of the utility model, but not as a limitation of the technical solution provided by the utility model. Through the following technical solutions, the technical purpose and beneficial effects of the utility model can be better achieved and realized.

As a preferred technical solution of the present invention, the control assembly includes a control device and a main controller connected in sequence, and the control device and the main controller are connected by electronic circuits, such as network cables.

As a preferred technical solution of the present invention, the control device includes a computer, but is not limited to a computer. Those skilled in the art can also select other devices as required, as long as they can realize the control function.

As a preferred technical solution of the utility model, the main controller is composed of a core control board and an IO board, and the core control board and the IO board are integrated together, wherein the core control board is an ARM scientific research board.

In the utility model, the core control board is used for data processing, and the IO board is used for controlling the switch of the sampling valve.

As a preferred technical solution of the present invention, the control assembly is connected to the syringe pump group through an electronic circuit, and the syringe pump group, the gradient mixer and the sampling valve are sequentially connected through a liquid connection pipe.

As a preferred technical solution of the present invention, the syringe pump group includes at least two groups of syringe pump groups, and each group of syringe pump groups is arranged in parallel; each group of syringe pump groups is equipped with a gradient mixer, and each gradient mixer is connected with a Injection valve connection.

In the utility model, each group of injection pumps can be started at the same time to realize multi-group dual-gradient regulation, and a certain group can also be started separately to perform dual-gradient regulation.

As a preferred technical solution of the present invention, each group of syringe pumps includes two syringe pumps, the two syringe pumps are respectively connected to the feed ports of the gradient mixer, and the two syringe pumps are arranged in parallel.

In the utility model, the gradient mixer includes two different feeding ports, and the two injection pumps are respectively connected to the two different feeding ports, and are mixed in the gradient mixer. The uniform and stable mixing of the mobile phase is achieved by the gradient mixer to ensure the accuracy and stability of the gradient adjustment.

In the utility model, the dual-gradient adjustment of mobile phase parameters is realized through the set injection pump group and the two injection pumps and syringes set in each group.

As a preferred technical solution of the present invention, the syringe pump is a single-channel syringe pump, and a syringe is placed in the syringe pump, and the volume of the syringe is 0.5mL to 200mL, such as 0.5mL, 1mL, 10mL, 30mL, 50mL, 70mL, 100mL, 130mL, 150mL, 170mL or 200mL, etc., but not limited to the listed values, other unlisted values within this range are also applicable.

As a preferred technical solution of the present invention, the single-channel syringe pump includes a split syringe pump and/or a series push-pull syringe pump.

As a preferred technical solution of the utility model, the sampling valve is a six-way valve.

In the utility model, the precise sampling of the sample can be satisfied by switching the sampling valve.

The processing method of mobile phase control system described in the utility model comprises the following steps:

The flow of the mobile phase in the syringe pump group is controlled by the control component, so that the mobile phase enters the gradient mixer equipped with each syringe pump group from the syringe pump group at a set ratio, and after the mobile phase is uniformly mixed in the gradient mixer Enter the injection valve, and inject the sample through the switching of the injection valve.

In the utility model, parameters such as the gradient of the injection pump group, the flow rate, the injection time, and the diameter of the syringe can be set on the control assembly, so that the mobile phase flows according to the set parameters.

Before the flow of the mobile phase in the syringe pump group is controlled by the control component, the mobile phase is respectively loaded into the syringes in each group of the syringe pump group and placed on the syringe pump.

Compared with the prior art, the utility model has the following beneficial effects:

(1) The system described in this utility model can realize the dual-gradient adjustment of the mobile phase through the injection pump group, realize the uniform and stable mixing of the mobile phase through the gradient mixer, and then satisfy the precise sampling of the sample through the switching of the sampling valve, The mobile phase control precision is high and the stability is good. The gradient error Tci=1% measured with reference to the detection method of GB/T 26792-2011 meets the standard requirements;

(2) The control components of the system described in the utility model can control multiple groups of injection pumps at the same time, can realize parallel connection with multiple interface characterization instruments or set up reference channels, and simultaneously detect and obtain data; or control one group of injection pumps alone , the number of injection pump groups can be added or reduced according to actual needs;

(3) The system described in the utility model is conveniently connected with the instrument, and can meet the connection interface requirements of various interface characterization instruments;

(4) The system described in the utility model can be integrated on a mobile sample platform, which can be connected with the instrument anytime and anywhere;

(5) The components of the system described in the utility model are simple in structure, easy to disassemble and easy to maintain.

Description of drawings

Fig. 1 is the structural representation of the mobile phase control system with double gradient adjustment function in the utility model embodiment 1;

Fig. 2 is the structural representation of the main controller in the mobile phase control system with double gradient adjustment function in the utility model embodiment 1;

Fig. 3 is the gradient error test result of the mobile phase control system with dual gradient adjustment function in Example 1 of the present invention;

Fig. 4 is that mobile phase control system described in embodiment 4 is connected with quartz crystal microbalance and observes protein BSA elution frequency change diagram through mobile phase salt gradient change;

Fig. 5 is that mobile phase control system described in embodiment 4 is connected with quartz crystal microbalance through mobile phase salt gradient change and observes protein BSA elution mass change figure;

Among them, 11-control device, 21-main controller, 31-syringe pump group, 41-gradient mixer, 51-injection valve, 211-core control board, 212-IO board, 311-single-channel syringe pump, 312 -Single-channel syringe pump, 313-single-channel syringe pump, 314-single-channel syringe pump, 511-mobile phase outlet, 512-sample inlet.

Detailed ways

In order to better illustrate the utility model and facilitate understanding of the technical solution of the utility model, the utility model is further described in detail below. However, the following embodiments are only simple examples of the utility model, and do not represent or limit the protection scope of the utility model, and the protection scope of the utility model shall be determined by the claims.

Part of the specific embodiment of the utility model provides a mobile phase control system with dual gradient adjustment functions and its processing method, the system includes a control assembly, a syringe pump group 31, a gradient mixer 41 and a sampling valve 51 connected in sequence .

The processing method comprises the following steps:

The flow of the mobile phase in the syringe pump group 31 is controlled by the control assembly, so that the mobile phase enters the gradient mixer 41 equipped with each syringe pump group 31 from the syringe pump group 31 at a set ratio, and the mobile phase is in the gradient mixer. After mixing evenly in 41, it enters the injection valve 51, and the sample is injected by switching the injection valve 51.

The following are typical but non-limiting embodiments of the utility model:

Example 1:

This embodiment provides a mobile phase control system with dual gradient adjustment functions and a processing method thereof. The system, as shown in FIG. Valve 51.

Wherein, the control assembly includes a control device 11 and a main controller 21 connected in sequence, the control device 11 and the main controller 21 are connected by a network cable, the control assembly and the injection pump group 31 are connected by an electronic circuit, and the injection pump group 31. The gradient mixer 41 and the sampling valve 51 are sequentially connected through a liquid connecting pipe.

Described control device 11 selects computer, main controller 21 is made up of core control board 211 and IO board 212, as shown in Figure 2, core control board 211 and IO board 212 are integrated together, and wherein described core control board 211 is ARM research board.

The syringe pump group 31 includes two groups of syringe pump groups, and the two groups of syringe pump groups are connected in parallel. Each group of syringe pump groups includes two single-channel syringe pumps. The single-channel syringe pump 311 and the single-channel syringe pump 312 form a set The syringe pump group, the single-channel syringe pump 313 and the single-channel syringe pump 314 form a group of syringe pumps, each group of syringe pumps is equipped with a gradient mixer 41 , and each gradient mixer 41 is connected to a sampling valve 51 . The single-channel syringe pump is a series push-pull syringe pump, and a syringe is placed in the single-channel syringe pump, and the volume of the syringe is 100 mL.

The parameters of each single-channel syringe pump pump are: the injection volume per microstep is 0.026uL (10mL syringe), the injection distance per microstep is 0.156um, the flow rate range is 0.1uL/h～681.73mL/min, and the maximum linear velocity is 17.33mm/s, the minimum linear speed is 0.00001583mm/s, the maximum step speed is 6933 steps/s, the minimum step speed is 0.02533 steps/s, the linear thrust is >30Kgf, and the control accuracy is when >30% of the full stroke, the control error ≤0.1.

The tube diameter of the gradient mixer 41 is 1/16in, the hole diameter is 0.75mm, and the material is CTFE.

The sampling valve 51 is a six-way valve, which includes a mobile phase outlet 511 and a sampling port 512, and is made of PEEK.

The liquid connection tube is a PEEK tube with an inner diameter of 0.25mm and an outer diameter of 1/16in.

The processing method of the system includes the following steps:

The system controls the main controller 21 and the syringe pump group 31 through the control device 11, and the mobile phase is respectively loaded into the syringe and placed in the single-channel syringe pump 311, the single-channel syringe pump 312, the single-channel syringe pump 313 and the single-channel syringe pump 314, set the gradient, flow rate, injection time, and syringe diameter of the syringe pump group 31 on the control device 11, and click Run to make the mobile phase pass through the liquid connection tube from the syringe at a set ratio and enter into each pump group. In the gradient mixer 41, after the mobile phase is mixed evenly, it enters the injection valve 51 through the connecting pipe. Through the switching of the injection valve 51, the precise injection of the sample to be tested can be satisfied. The mobile phase of the injection valve The outlet 511 is connected to the mobile phase inlet of the interface characterization instrument, thereby forming an interface characterization analysis experiment with a gradient change of the mobile phase.

The gradient error Tci of the system is carried out with reference to the detection method of GB/T 26792-2011, and the test results are shown in FIG. 3 .

In this embodiment, the sample injection control error of the sample to be tested is ≤0.1.

Example 2:

This embodiment provides a mobile phase control system with dual-gradient adjustment functions and its processing method. The structure and processing method of the system refer to Embodiment 1, the difference is that the single-channel syringe pump is a split type syringe pump, The volume of the syringe is 0.5 mL.

Example 3:

This embodiment provides a mobile phase control system with dual gradient adjustment functions and its processing method. The structure and processing method of the system refer to Embodiment 1, except that the volume of the syringe is 200 mL.

Example 4:

This embodiment provides the use of a mobile phase control system with dual gradient adjustment functions. The mobile phase control system described in Example 1 is connected to a quartz crystal microbalance analyzer, and the protein BSA is observed through the change of the mobile phase salt gradient. Elution condition. During the detection process of the quartz crystal microbalance analyzer, the change trend of the frequency F can be used to calculate the change in the quality of the protein, the frequency increases, the protein quantity decreases, and the frequency decreases, the protein quantity increases. The results are shown in Figure 4 and As shown in Fig. 5, the abscissa in Fig. 4 is the elution time Time (s), and the ordinate is the frequency F (Hz); the abscissa in Fig. 5 is the gradient (%), and the ordinate is the mass (ng/cm ² ).

Among them, the setting conditions of the mobile phase control system are: select one group of syringe pump groups 31, one syringe is equipped with 20mM pH8.0 Tris-Hcl buffer solution, and the other syringe is equipped with 1M NaCl 20mM pH8.0 Tris-Hcl buffer solution solution, set 25%, 50% and 100% gradient changes on the control module 11, the flow rate is 50 μ L/min, and each gradient holding time is 10 min, and the injection valve 51 is controlled by the control device 11 to first make the protein BSA adsorb on the quartz On the crystal microbalance chip, switch the gradient mixed mobile phase into the quartz crystal microbalance, so as to observe the elution behavior of BSA with the change of mobile phase salt concentration in real time. BSA is adsorbed on the QCM chip, the frequency decreases and the mass increases. After the adsorption balance, the concentration of salt ions is gradually changed through the mobile phase control system, so that the protein bound to the weak anion exchange surface begins to elute. With the gradual increase of the concentration of salt ions As the temperature increases, the quality of BSA on the chip gradually decreases, and finally almost completely falls off from the chip.

The applicant declares that the utility model illustrates the detailed method of the utility model through the above-mentioned embodiments, but the utility model is not limited to the above-mentioned detailed method, that is, it does not mean that the utility model must rely on the above-mentioned detailed method to be implemented. Those skilled in the art should understand that any improvement of the utility model, the equivalent replacement of each raw material of the utility model product, the addition of auxiliary components, the selection of specific methods, etc., all fall within the scope of protection and disclosure of the utility model within.

Claims (10)

1. A mobile phase control system with dual gradient adjustment function, characterized in that, the system comprises a control assembly, a syringe pump group (31), a gradient mixer (41) and a sampling valve (51) connected in sequence. 2. mobile phase control system according to claim 1, is characterized in that, described control assembly comprises control device (11) and main controller (21) that are connected in sequence, control device (11) and main controller (21) ) are connected by electronic circuits. 3. The mobile phase control system according to claim 2, characterized in that the control device (11) comprises a computer. 4. mobile phase control system according to claim 2, is characterized in that, described master controller (21) is made up of core control board (211) and IO board (212), and core control board (211) and IO board (212) integrated together, wherein the core control board (211) is an ARM scientific research board. 5. mobile phase control system according to claim 1, is characterized in that, is connected by electronic circuit between described control assembly and syringe pump group (31), syringe pump group (31), gradient mixer (41) and The sampling valves (51) are sequentially connected through liquid connecting pipes. 6. The mobile phase control system according to claim 1, characterized in that, the syringe pump group (31) comprises at least two groups of syringe pump groups, each group of syringe pump groups are arranged in parallel; each group of syringe pump groups is equipped with A gradient mixer (41), each gradient mixer (41) is connected with a sampling valve (51). 7. mobile phase control system according to claim 6, is characterized in that, every group of syringe pump groups comprises two syringe pumps, and two syringe pumps are respectively connected with the feeding port of gradient mixer (41), two injection pumps The pumps are set up in parallel. 8. The mobile phase control system according to claim 7, wherein the syringe pump is a single-channel syringe pump, and a syringe is placed in the syringe pump, and the volume of the syringe is 0.5mL-200mL. 9. The mobile phase control system according to claim 8, wherein the single-channel syringe pump comprises a split type syringe pump and/or a series push-pull type syringe pump. 10. The mobile phase control system according to claim 1, characterized in that, the sampling valve (51) is a six-way valve.