Real-time column chromatography separation system based on spectrum detection

Abstract

The invention belongs to the technical field of column chromatography separation, and discloses a real-time column chromatography separation system based on spectrum detection, which comprises a light source, a data acquisition unit, a spectrum detection unit and a core control unit which are connected in sequence, the data acquisition unit comprises a chromatographic column, an iron stand, a beaker, a cross clamp, a flexible rubber hose, two light-passing flow cells, a flow regulator, a micro sprayer and a black box, and the chromatographic column, the iron stand, the beaker and the cross clamp are assembled and fixed on an operation table; the light source comprises a visible light source and an ultraviolet light source; the spectrum detection unit comprises a spectrum sensor and an ultraviolet sensor; the core control unit comprises a power supply, a microprocessor and a microcontroller. The concentration change of each component is monitored in real time, and a signal is transmitted to the spectrum sensor through the optical fiber, so that the accurate detection of the solution separation process is realized. And internal programs of the system are controllable, communication with other equipment is convenient, and the complexity of operation is reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of column chromatography separation, and relates to a column chromatography device used in a solution separation process, in particular to a real-time column chromatography separation system based on spectral detection. Background Art

[0002] Column chromatography is a separation technology widely used in chemical analysis, preparation and purification. It uses the distribution difference of different components between the stationary phase and the mobile phase to separate the sample components. Column chromatography separation technology is widely used in chemical analysis, pharmaceuticals, environmental monitoring and other fields due to its ability to efficiently separate complex mixtures. However, traditional column chromatography systems usually rely on offline detection equipment, resulting in detection delays, complex operations and lack of real-time performance. At the same time, the separation effect is not obvious, making it difficult to meet the needs of modern industry and scientific research for online monitoring.

[0003] Spectral detection technology has become an important means of online analysis because of its advantages of high sensitivity and rapid detection by utilizing the unique absorption or emission characteristics of substances under specific wavelengths of light. Combining spectral detection with column chromatography can realize real-time analysis of effluent components and dynamically adjust separation parameters, thereby improving separation efficiency and accuracy. In recent years, with the rapid development of miniaturized spectroscopic devices, high-sensitivity detectors and embedded systems, the application potential of real-time column chromatography separation systems in industrial quality control, environmental monitoring and pharmaceutical processes has continued to expand, becoming an important development direction of modern analytical technology.

[0004] However, it is not easy to combine spectral detection technology with column chromatography separation system to achieve real-time detection. On the one hand, it is necessary to solve the compatibility problem between the spectral detection probe and the chromatographic separation column to ensure that the detection signal can accurately reflect the actual situation of each component in the separation column; on the other hand, it is also necessary to consider the processing and analysis methods of spectral detection data to achieve fast and accurate qualitative and quantitative analysis.

[0005] Therefore, developing a real-time column chromatography separation system based on spectral detection has important practical significance and application value. This system can not only achieve efficient separation of mixtures, but also monitor the concentration changes of each component in real time during the separation process, providing more comprehensive and accurate analytical data for researchers and engineers, and helping to promote the further development of chromatographic separation technology.

[0006] At present, although some chromatographic separation systems based on spectral detection have been reported, these systems still have some shortcomings, such as insufficient detection sensitivity and insufficient intelligence of data processing algorithms. Therefore, developing a real-time column chromatography separation system based on spectral detection with higher sensitivity and more intelligent data processing algorithms will be an important research direction in this field in the future. Summary of the invention

[0007] In order to overcome the defects of existing traditional column chromatography separation devices, such as difficulty in real-time detection during the separation process, difficulty in achieving accurate separation effect and flow rate control, and inconvenience in data recording and analysis, the present invention provides a real-time column chromatography separation system based on spectral detection, which uses Raspberry Pi as the control core and monitors the concentration changes in the solution separation process in real time through spectral sensors and flow control modules.

[0008] The technical solution adopted by the present invention to solve its technical problem is:

[0009] A real-time column chromatography separation system based on spectral detection comprises a light source, a data acquisition unit, a spectral detection unit and a core control unit, which are connected in sequence. The data acquisition unit comprises a chromatography column, an iron frame, a beaker, a cross clamp, a soft rubber tube, a two-way light flow pool, a flow regulator micro-sprinkler and a black box. The chromatography column, the iron frame, the beaker and the cross clamp are assembled and fixed on an operating table. The light source comprises a visible light source and an ultraviolet light source. The spectral detection unit comprises a spectral sensor and an ultraviolet sensor. The core control unit comprises a power supply, a microprocessor and a microcontroller.

[0010] In the above-mentioned real-time column chromatography separation system based on spectral detection, the chromatography column and the two-way light flow cell are fixed with a beaker clamp, and the flow regulator micro-sprinkler and the beaker are connected with a soft hose, and the above-mentioned are assembled and fixed in sequence from top to bottom. The two-way light flow cell is a quartz flow cuvette flow cell with an outer diameter of 8mm, an inner diameter of 6mm, a total length of 70mm, and an optical path of 10mm. The caliber of the flow regulator micro-sprinkler is 4nm.

[0011] The above-mentioned real-time column chromatography separation system based on spectral detection uses the hose to connect one end of the two-way light flow cell to the mobile phase outlet of the chromatography column to form a passage, and the other end is connected to the flow regulator micro-sprinkler to form a passage.

[0012] In the above-mentioned real-time column chromatography separation system based on spectral detection, the flow regulator micro-sprinkler precisely drips the liquid into the beaker.

[0013] In the above-mentioned real-time column chromatography separation system based on spectral detection, the light source can be a 5mm flat-head red, green and blue full-color common cathode transparent LED lamp or an ultraviolet light source.

[0014] In the above-mentioned real-time column chromatography separation system based on spectral detection, the two-pass light flow cell is placed in a black box, which is modified by drilling holes, and optical fibers are used to allow the light source to pass through, while the other end of the black box is connected to the spectral detection unit to transmit the signal to the control system.

[0015] In the above-mentioned real-time column chromatography separation system based on spectral detection, the spectral detection unit is connected to the cuvette through an optical fiber, and the two-way light flow cell is placed in a black box, and the light-transmitting hole of the black box is used to guide the light emitted by the light source.

[0016] In the above-mentioned real-time column chromatography separation system based on spectral detection, the power supply of the core control unit is a DC power supply with a voltage of 5V and a current of 3.5A, the microprocessor of the core control unit is a Raspberry Pi, and the microcontroller of the core control unit is an Arduino UNO rev 3.

[0017] In the above-mentioned real-time column chromatography separation system based on spectral detection, the spectral sensors of the spectral detection unit are respectively AS7341 sensor and S12SD ultraviolet sensor, which are used to detect the concentration changes of each component in the solution within a specified wavelength range.

[0018] In the above-mentioned real-time column chromatography separation system based on spectral detection, the Raspberry Pi is connected to multiple external components, including a spectral sensor module, a voice control module and a button switch. The spectral sensor module, the voice control module and the button switch are all external components connected to the Raspberry Pi. The spectral sensor module detects light or color, the voice control module recognizes sound, and the button switch provides a simple interactive function. The spectral sensor module communicates with the Raspberry Pi through a specific interface or protocol. The power supply (VCC) of the spectral sensor is powered and grounded (GND). The SDA and SCL pins are connected to the I2C interface (GPIO2 and GPIO3) of the main control chip, and the GPIO pin is connected to the GPIO0 pin of the main control chip. When the voice control module detects sound, it sends a signal to the main control chip, and the main control chip responds according to the sound signal. Additional control is achieved through the button switch for human intervention or debugging.

[0019] In the above-mentioned real-time column chromatography separation system based on spectral detection, the control steps of the core control unit 11 are as follows:

[0020] S1: According to the optical and electrical line wiring, start the core control unit and wait for three minutes until the system is stable;

[0021] S2: Turn off the light source, test the dark current under the test environment, and obtain the corresponding light intensity data I dark ;

[0022] S3: Turn on the light source, use deionized water as a blank solvent, flow through the cuvette, measure the light intensity of the deionized water, start the test program, test the blank current, and obtain the corresponding light intensity data I blank ;

[0023] S4: The solution separated by the chromatography column flows through the cuvette, the test program is started, and the corresponding light intensity data I is obtained. sample , according to the formula:

[0024]

[0025] Calculate the absorbance A.

[0026] The absorption coefficient can also be further calculated according to the formula,

[0027]

[0028] l is the optical path length, that is, the width of the sample pool, and c is the solution concentration.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The spectral detection unit in the column chromatography separation system of the present invention can monitor the concentration changes of each component in real time during the column chromatography separation process, and transmit the signal to the spectral sensor through the optical fiber to realize the accurate detection of the solution separation process. The spectral detection unit is equipped with a precision flow regulator, which can dynamically adjust the flow rate according to the concentration changes detected in real time through the control of the Raspberry Pi, thereby ensuring the stability and accuracy of the separation process. The core control unit uses a microprocessor and a microcontroller. Under the premise of ensuring low power consumption and portability, it can also communicate with other devices anytime and anywhere through the network, and can perform cluster operations of various sizes according to needs. The column chromatography separation system provided by the present invention has a controllable system internal program, convenient communication with other devices, and reduces the complexity of operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 It is a schematic diagram of the system structure;

[0032] Figure 2 This is a schematic diagram of the circuit connection structure.

[0033] In the figure: 1. light source, 2. chromatography column, 3. iron stand, 4. cross clamp, 5. black box, 6. two-way light flow pool, 7. flow regulator micro-sprinkler, 8. soft rubber tube, 9. beaker, 10. data acquisition unit, 11. core control unit. DETAILED DESCRIPTION

[0034] The following will be combined with the present invention Figure 1-2 , a clear and complete description of the technical solution of the present invention is given. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work are within the scope of protection of the present invention.

[0035] In the description of the present invention, it should be noted that the terms "upper", "lower", "inner", "outer", "front end", "rear end", "two ends", "one end", "the other end" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0036] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "provided with", "connected", etc. should be understood in a broad sense. For example, "connected" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0037] Example 1

[0038] This embodiment provides a real-time column chromatography separation system based on spectral detection, including: a light source 1 , a spectral detection unit 10 , a data acquisition unit and a core control unit 11 .

[0039] The data acquisition unit is composed of a chromatography column 2, an iron stand 3, a beaker 9, a flask clamp, two corner claws, a cross clamp 4, a soft rubber tube 8, a frosted plug, a two-way light flow cell 6, a flow regulator micro-sprinkler 7, and a black box 5; the assembled chromatography column 2, the iron stand 3, the beaker 9, and the cross clamp 4 are fixed on the operating table. The two-way light flow cell 3 is a quartz flow cuvette flow cell, and the black box 5 is a black antistatic storage box.

[0040] The quartz flow cuvette flow cell is a two-pass light flow cell with an outer diameter of 8 mm, an inner diameter of 6 mm, a total length of 70 mm, and an optical path of 10 mm. The flow regulator micro-sprinkler 7 has a caliber of 4 nm and controls the flow rate of the outflow liquid.

[0041] The soft rubber tube 8 connects one end of the square flow cell of the flowing quartz cuvette to the mobile phase outlet of the chromatography column 2 device to form a passage, and the other end is connected to the micro nozzle 7 of the flow regulator to form a passage.

[0042] The separated liquid droplets flowing out of the mobile phase outlet enter the square flow cell of the mobile quartz cuvette, and the sample cell is placed in the modified black box 5, and the left and right side walls of the black box 5 can pass through the optical fiber. The separated components will flow out from the bottom of the column in turn, and the mobile phase outlet is connected to the flow regulator micro nozzle 7 through a hose to control the flow rate of the outflowing liquid.

[0043] The light source 1 is a 5mm flat-head red, green and blue full-color common cathode transparent LED lamp. In order to reduce the influence of ambient light, the light source is transmitted through optical fibers to the flowing quartz cuvette square circulation pool.

[0044] like Figure 2 As shown, the spectral detection unit 10 is an AS7341 spectral sensor, which provides 8 visible light channels (VIS, 410-680nm) and one near infrared channel (NIR, 940nm), and an additional 4 Flicker channels for light source stroboscopic detection. The volume is only 2.0mm×2.0mm×1.0mm. Its spectral range: 410-940nm, can collect the complete spectrum. The resolution is 16-bit ADC, with high sensitivity, and converts the optical signal into a digital signal;

[0045] The core control unit 11 is based on Raspberry Pi, whose CPU is a multi-core processor based on ARM, supports microSD card as the main storage device, has USB port, HDMI, Ethernet, Wi-Fi, Bluetooth, audio output, camera interface (CSI) and display interface (DSI) interface. It has 40 general input and output pins for connecting sensors, controllers and other devices.

[0046] The control steps of the core control unit 11 are as follows:

[0047] S1: Start the core control unit 11 according to the optical and electrical line wiring, and wait for three minutes until the system is stable;

[0048] S2: Turn off light source 1, test the dark current under the test environment, and obtain the corresponding light intensity data;

[0049] S3: Turn on light source 1, use deionized water as blank solvent, flow through the quartz flow cuvette, measure the light intensity of deionized water, start the test program, test the blank current, and obtain the corresponding light intensity data;

[0050] S4: The solution separated by chromatography column 2 flows through the quartz flow cuvette, the test program is started, and the corresponding light intensity data is obtained according to the formula:

[0051]

[0052] Calculate the absorbance.

[0053] The spectrum detection unit 10, the laser light source module, the data acquisition unit and the core control unit 11 are connected in sequence to collect the sample concentration signal and obtain the photophysical characteristic data such as the absorbance of the sample.

[0054] The absorption coefficient can also be further calculated according to the formula.

[0055]

[0056] l is the optical path length, that is, the width of the sample pool, and c is the solution concentration.

[0057] Example 2

[0058] This embodiment further provides another real-time column chromatography separation system based on spectral detection, including: a light source module, a data acquisition unit and a core control unit 11.

[0059] Light source 1 is ultraviolet light. To reduce the influence of ambient light, the light source is transmitted through optical fiber to the square flow cell of the flowing quartz cuvette.

[0060] The data acquisition unit 10 is a S12SD ultraviolet sensor, the detection spectrum wavelength range is 240m-370mm, the power supply is 2.7V-5V, and the ultraviolet detection module adopts a Schottky photodiode made of nitride-based material;

[0061] The core control unit 11 uses the Raspberry Pi as the core, turns on the light source 1 and the power supply, uses the rich GPIO interfaces on the Raspberry Pi to connect other devices, and controls sampling.

[0062] The spectrum detection unit 10, the laser light source module, the data acquisition unit and the core control unit 11 are connected in sequence to collect the sample concentration signal and obtain the photophysical characteristic data such as the absorbance of the sample.

[0063] Example 3

[0064] This embodiment provides a user interface development solution for a real-time column chromatography separation system based on spectral detection, integrating a practical Web interface to achieve remote control, real-time monitoring and data management. The system includes a front-end interactive interface, back-end services, data storage modules and network connection modules. Through the interaction between the Raspberry Pi and the experimental hardware, it supports parameter setting, experimental data recording and remote access, significantly improving experimental efficiency and data reliability.

[0065] System workflow

[0066] System startup: Users access the interface through a web browser and enter the system main interface after logging in.

[0067] Experimental parameter setting: Set relevant parameters according to experimental requirements and save them.

[0068] Real-time data monitoring: During the experiment, spectral data is acquired in real time and the absorbance curve is updated dynamically.

[0069] Data analysis and storage: After the experiment is completed, the system automatically analyzes the data and saves it to the database.

[0070] Report generation and export: Users can generate experimental reports with one click and support data file download.

[0071] Equipment and technical parameters

[0072] Use a 5mm flat-head red, green and blue full-color common cathode transparent LED lamp or ultraviolet light source. The light source is transmitted to the quartz flow cuvette circulation pool through an optical fiber to reduce interference from ambient light. The AS7341 spectral sensor has a detection range of 410-940nm and a resolution of 16-bit ADC. The S12SD ultraviolet sensor has a detection wavelength range of 240-370nm and supports high-sensitivity ultraviolet detection. The core device is the Raspberry Pi, which has a rich GPIO interface for connecting light sources, sensors and flow regulation devices. Provide a network communication interface (Wi-Fi / Ethernet) to support user remote operation. Front-end framework: Developed based on HTML, CSS and JavaScript. Back-end framework: Use Flask to build back-end services to communicate with the Raspberry Pi.

Claims

1. A real-time column chromatography separation system based on spectral detection, characterized in that: The invention comprises a light source (1), a data acquisition unit, a spectrum detection unit (10) and a core control unit (11), which are connected in sequence. The data acquisition unit comprises a chromatography column (2), an iron frame (3), a beaker (9), a cross clamp (4), a soft rubber tube (8), a two-way light flow pool (6), a flow regulator micro-sprinkler (7) and a black box (5). The chromatography column (2), the iron frame (3), the beaker (9) and the cross clamp (4) are assembled and fixed on an operating table. The light source (1) comprises a visible light source and an ultraviolet light source. The spectrum detection unit (10) comprises a spectrum sensor and an ultraviolet sensor. The core control unit (11) comprises a power supply, a microprocessor and a microcontroller.

2. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The chromatography column (2) and the two-way light flow cell (6) are fixed using a beaker clamp, and the flow regulator micro-sprinkler (7) and the beaker (9) are connected using a soft rubber tube (8), and are assembled and fixed in sequence from top to bottom.

3. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The two-pass light flow cell (6) is a quartz flow cuvette flow cell.

4. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The light source (1) is a 5mm flat-head red, green and blue full-color common cathode transparent LED lamp or an ultraviolet light source.

5. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The spectrum detection unit (10) is connected to the cuvette via an optical fiber, and the two-way light flow cell (6) is placed in a black box (5). The light-transmitting hole of the black box (5) is used to guide the light emitted by the light source (1).

6. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The core control unit (11) has a microprocessor of RaspberryPi and a microcontroller of Arduino UNO rev 3.

7. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The spectrum sensors of the spectrum detection unit (10) are respectively an AS7341 sensor and an S12SD ultraviolet sensor, which are used to detect the concentration changes of various components in the solution within a specified wavelength range.

8. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The core control unit (11) is based on the Raspberry Pi.

9. The real-time column chromatography separation system based on spectral detection according to claim 1, characterized in that: The control steps of the core control unit (11) are as follows: S1: Start the core control unit (11) according to the optical and electrical line wiring, and wait for three minutes until the system is stable; S2: Turn off the light source (1), test the dark current under the test environment, and obtain the corresponding light intensity data I dark ; S3: Turn on the light source (1), use deionized water as a blank solvent, flow through the cuvette, measure the light intensity of the deionized water, start the test program, test the blank current, and obtain the corresponding light intensity data I blank ; S4: The solution separated by the chromatography column (2) flows through the cuvette, the test program is started, and the corresponding light intensity data I is obtained. sample , according to the formula: Calculate the absorbance A.

10. The real-time column chromatography separation system based on spectral detection according to claim 9, characterized in that: The absorption coefficient can also be further calculated according to the formula, l is the optical path length, that is, the width of the sample pool, and c is the solution concentration.

Images