Real-time temperature control high-flux ultraviolet-visible spectrophotometer and use method thereof

Abstract

The invention discloses a real-time temperature control high-flux ultraviolet-visible spectrophotometer and a use method thereof, and belongs to the technical field of spectral analysis. A sample chamber of the photometer body is internally provided with a multi-connected pool bracket, a plurality of sample pools are arranged in the multi-connected pool bracket, and the bottoms of the sample pools are provided with liquid runners; after the cuvette is arranged in the sample pool, the liquid flow channel is in contact with the bottom of the cuvette; a liquid flow channel inlet and a liquid flow channel outlet are respectively formed in the bottom ends of the head number sample pool and the tail number sample pool and are respectively communicated with a liquid outlet and a liquid inlet of the circulating water bath temperature control system; thermistor ends of two temperature measuring cables of the multi-channel temperature measuring instrument extend into cuvettes in the head number sample pool and the tail number sample pool and are arranged below the liquid level of liquid in the cuvettes. The spectrophotometer can accurately measure ultraviolet-visible light spectral absorption spectrums of 16 samples in a program temperature change process in real time at most, and has a good application prospect in related research fields such as temperature-sensitive biomacromolecule conformation or structural transformation, chemical reaction and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of spectral analysis, and in particular to a high-throughput ultraviolet-visible spectrophotometer capable of performing real-time temperature control on a sample and a use method thereof. Background Art

[0002] UV-visible spectrophotometer is an instrument based on the principle of UV-visible spectrophotometry, which uses the absorption of radiation in the UV-visible spectrum by molecules to analyze. In the fields of chemistry and biomedicine, UV-visible spectrophotometer can be used to measure the absorbance of molecules to study their structure and function. Variable temperature absorption spectroscopy is widely used in studying the conformation or structural transformation of biomacromolecules and chemical reactions, especially in analyzing the thermal stability of nucleic acids or proteins.

[0003] Traditional ultraviolet spectrophotometers are easily affected by ambient temperature and are difficult to obtain the absorption spectrum of temperature-variable samples. In order to solve this problem, technicians in this field have designed temperature-controllable analysis equipment, such as the ultraviolet-visible spectrophotometer disclosed in Chinese patent CN209198310 U, which fixes a sample pool inside the sample chamber, installs a constant temperature sensor inside the sample chamber at one end of the sample pool, and fixes a heating block at the end of the sample pool away from the constant temperature sensor and one side of the sample pool, and connects a heat conductive sheet between the heating block and the sample pool to facilitate monitoring and heating of the sample solution and maintain the temperature of the sample solution. Although this device solves the problem of sample temperature being uncontrollable to a certain extent, it still has several obvious shortcomings:

[0004] 1) Low throughput: Usually, only absorbance data of no more than 8 samples can be collected in one experiment. For the time-consuming temperature-varying experiment (single data collection time is about 2-20 hours), the experimental efficiency needs to be improved;

[0005] 2) The temperature measurement position is not accurate enough: the temperature sensing device is located outside the sample pool, and mainly collects the temperature of the sample pool bracket rather than the actual temperature of the sample;

[0006] 3) The number of detection wavelengths is limited: During the temperature change process, only the absorbance at a few individual wavelengths can be measured instead of the absorption spectrum within a wide wavelength range. The full spectrum is crucial for characterizing the production of new reaction substances or changes in the secondary structure of biomacromolecules during the reaction. Summary of the invention

[0007] The purpose of the present invention is to solve the problems existing in the prior art, provide a real-time temperature-controlled high-throughput ultraviolet-visible spectrophotometer and disclose its specific use method. The spectrophotometer can be used to synchronously perform real-time temperature-controlled spectral scanning on 16 samples, significantly improving the experimental efficiency of variable temperature spectral scanning and the accuracy of temperature measurement.

[0008] In order to achieve the above technical objectives, the present invention is implemented through the following technical solutions: a high-throughput ultraviolet-visible spectrophotometer with real-time temperature control, the photometer body includes a detection module and a sample chamber, the sample chamber is arranged at one end inside the photometer body, and is characterized in that a multi-cell bracket is provided in the sample chamber, a plurality of sample cells numbered in sequence are provided in the multi-cell bracket, a liquid flow channel is provided at the bottom of the sample cell, and the liquid flow channel extends along the sample cells arranged in sequence; after the cuvette is placed in the sample cell, the liquid flow channel contacts the bottom of the cuvette; the liquid flow channel inlet is provided at the bottom of the first sample cell, and the outlet is provided at the bottom of the last sample cell, and the liquid flow channel inlet and outlet are respectively connected to the liquid outlet and liquid inlet of a circulating water bath temperature control system; the thermistor ends of the two temperature measuring cables of the multi-channel thermometer extend into the cuvettes in the first and last sample cells and are placed below the liquid surface of the liquid in the cuvette.

[0009] Furthermore, a total of 18 sample pools are arranged in sequence on the multi-pool holder, sample pool No. 1 is the first sample pool, sample pool No. 18 is the last sample pool, and sample pools No. 2 to No. 17 are called intermediate sample pools.

[0010] Furthermore, the sample pools are distributed in two rows, the sample pools in the left and right rows are staggered, and the contact area between the cuvette placed in the middle sample pool and the liquid flow channel at its bottom is the same.

[0011] Furthermore, a sealing cover is provided on the cuvette, and a temperature measuring through hole is provided on the sealing cover used to cover the cuvettes in the first and last number sample pools, and the thermistor end of the temperature measuring cable extends from the temperature measuring through hole into the cuvettes in the first and last number sample pools.

[0012] Furthermore, a first pipe connection port and a second pipe connection port are provided at one end of the multi-cell bracket, the liquid flow channel inlet is connected to the first pipe connection port, and the liquid flow channel outlet is connected to the second pipe connection port through a pipeline; a third pipe connection port and a fourth pipe connection port are provided on the outer shell of the photometer body, the first and third pipe connections and the second and fourth pipe connections are respectively connected by a pipeline, the third pipe connection port is connected to the liquid outlet of the circulating water bath temperature control system through a pipeline, and the fourth pipe connection port is connected to the liquid inlet of the circulating water bath temperature control system through a pipeline, so as to realize the circulation of the condensing medium.

[0013] Furthermore, light-transmitting grooves are provided on both sides of the multi-cell bracket, the multi-cell bracket is movably connected to the bottom plate, the bottom plate is fixed in the sample chamber, a driving member is provided on the bottom plate, and the multi-cell bracket moves under the action of the driving member to adjust the position of the sample cell.

[0014] Furthermore, the sample chamber is connected to an air compression pump, the end of the ventilation pipeline of the air compression pump is connected to a three-way diverter, and the two air outlet ends of the three-way diverter are respectively connected to a diverter pipe. The two diverter pipes pass through the side wall of the sample chamber and extend into the interior thereof, and are correspondingly fixed on the inner walls on both sides of the opposite sides of the sample chamber.

[0015] Furthermore, the shunt tube is fixed in the sample chamber to avoid the optical path of the spectrophotometer.

[0016] The method of using the above-mentioned real-time temperature-controlled high-throughput UV-visible spectrophotometer is as follows:

[0017] 1) After adding ultrapure water to two cuvettes, cover them with sealing covers with temperature measuring through holes, and place them in the head and tail sample pools respectively. Extend the thermistor ends of the two temperature measuring cables of the multi-channel thermometer into the corresponding cuvettes through the temperature measuring through holes on a sealing cover, and place them below the liquid surface;

[0018] 2) Add the samples to be tested into different cuvettes, cover them with sealing covers without temperature measurement holes, clean the surface of the cuvettes and place them into the middle sample pool;

[0019] 3) Start the circulating water bath temperature control system, set the temperature change range and change rate, and transport the condensing medium to the liquid flow channel through the circulating pump;

[0020] 4) Set thermistor parameters and reading time interval, and monitor the temperature data of the multi-channel thermometer through the computer;

[0021] 5) The temperature data of the first sample pool and the last sample pool at each time point read by the multi-channel thermometer are processed arithmetic difference and assigned to the middle sample pool respectively, that is, the real-time temperature of the samples in the middle sample pool at different time points is obtained;

[0022] 6) The sample temperature is regulated and recorded in real time while setting scanning parameters, and the spectrophotometer is started to perform a full spectrum scan of the sample in the middle sample pool to obtain a full spectrum scan diagram of the sample melting process;

[0023] 7) Matching the measured continuous spectrum data and temperature data of the sample to obtain the variable temperature spectrum data of the sample to be measured.

[0024] The beneficial effects of the present invention are:

[0025] 1. The UV-visible spectrophotometer disclosed in this application can detect the temperature-varying spectra of 16 samples at the same time, which not only greatly improves the detection efficiency, but also ensures the reliability and stability of the measurement;

[0026] 2. This application uses a multi-channel thermometer to directly measure the real-time temperature of the samples in the head and tail sample pools, and performs arithmetic processing on the acquired data and assigns it to the middle sample pool to obtain the real-time temperature of the samples to be tested in different sample pools at different time points. This application does not obtain the temperature information of the samples by measuring the temperature of the rack. The accuracy of temperature detection is significantly improved, which is more conducive to accurate research on sample configuration changes and stability;

[0027] 3. By using the spectrophotometer disclosed in the present application to analyze samples and matching the measured continuous spectrum data with the temperature data measured by the multi-channel thermometer, the variable temperature spectrum data of the sample to be tested can be obtained, and the variable temperature full spectrum scanning of the sample between 0 and 100°C and the wavelength range of 190-1100nm can be realized. Compared with the traditional spectrophotometer that can only obtain the variable temperature absorbance data of a single or a few wavelengths of the sample, it can more comprehensively reflect the changes in sample properties caused by temperature changes;

[0028] 4. The spectrophotometer disclosed in the present application has a simple detection principle, is easy to operate, and has a wide range of applications. It is suitable for use in fields such as chemistry, materials, biology, and pharmacy that require analysis of temperature-controlled absorption spectra. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a schematic diagram of the structure of a high-throughput UV-visible spectrophotometer capable of real-time temperature control;

[0030] Figure 2 This is the axonometric drawing of the multi-tank bracket;

[0031] Figure 3 The figure is a schematic diagram of the distribution of the liquid flow channels arranged in the multi-cell support, and the arrows in the figure indicate the flow direction of the circulating medium in the liquid flow channels;

[0032] Figure 4 It is a schematic diagram of the connection between the circulating water bath temperature control system and the sample chamber;

[0033] Figure 5 It is a schematic diagram of the connection between the multi-cell bracket and the multi-channel temperature measuring instrument;

[0034] Figure 6 The experimental results obtained after temperature matching and full spectrum scanning using the UV-visible spectrophotometer disclosed in Example 1, wherein, panel a is a full spectrum scan of the melting process of the sample in sample pool No. 2; panel b is a melting curve drawn after normalizing the absorbance data of the samples in sample pools No. 2 to No. 17 at 295 nm; panel c is the T obtained after parallel measurement of the same sample three times using sample pools No. 2 to No. 17 m values ​​and their mean and standard deviation data;

[0035] Figure 7This is a schematic diagram of the connection between the sample chamber and the air compression pump;

[0036] Among them, 1-photometer body, 2-circulating water bath temperature control system, 3-multi-channel temperature measuring instrument, 4-multi-cell bracket, 5-bottom plate, 6-driving part, 7-computer, 8-air compression pump;

[0037] 11- the third pipe connection port, 12- the fourth pipe connection port;

[0038] 21-liquid outlet, 22-liquid inlet;

[0039] 31-thermistor;

[0040] 41 - sample pool, 42 - cuvette, 43 - light-transmitting slot, 44 - first connecting port, 45 - second connecting port, 46 - liquid flow channel, 47 - sealing cover;

[0041] Sample pool No. 411-1, sample pool No. 412-18;

[0042] 81-Three-way splitter. DETAILED DESCRIPTION

[0043] The following examples further illustrate the content of the present invention, but should not be construed as limiting the present invention. Without departing from the essence of the present invention, modifications and substitutions made to the methods, steps or conditions of the present invention all fall within the scope of the present invention.

[0044] In order to solve the problems of inaccurate temperature control and low throughput of existing ultraviolet spectrophotometers, this embodiment discloses a high-throughput ultraviolet-visible spectrophotometer capable of real-time temperature control.

[0045] The photometer body 1 includes a circulating water bath temperature control system 2 and a multi-channel temperature measuring instrument 3. The photometer body 1 is composed of a detection module and a sample chamber. The detection module is connected to a computer 7 for data analysis. The detection module part is a prior art and will not be described in detail here. The sample chamber is arranged at one end of the interior of the photometer body 1. A multi-cell bracket 4 is arranged inside the sample chamber. 18 sample cells 41 are arranged inside the multi-cell bracket 4 for placing cuvettes 42. The sample cells 41 are numbered in the order of 1-18, and are sequentially recorded as sample cell No. 1 411, sample cell No. 2 ... sample cell No. 18 412. In order not to affect the light path, the sample cells 41 on the left and right sides need to be staggered (see Figure 2 ).

[0046] The multi-cell support 4 is provided with light-transmitting grooves 43 on both sides for easy detection. The multi-cell support 4 is slidably connected to the bottom plate 5, which is fixed in the sample chamber. A driving member 6 is provided at one end of the bottom plate 5. The multi-cell support 4 moves along the length direction of the bottom plate 5 under the action of the driving member 6 to facilitate the movement of the position of the sample pool 41, thereby facilitating the analysis of the information of the samples in the cuvettes 42 placed in different sample pools 41. The driving member 6 can be a common driving element such as a cylinder, an oil cylinder or a motor screw, which will not be described in detail here.

[0047] A first pipe connection port 44 and a second pipe connection port 45 are provided at one end of the multi-cell support 4, and a zigzag liquid flow channel 46 extending in the order in which the sample cells 41 are arranged is provided at the bottom of the sample cells 41. After the cuvette 42 is placed in the sample cell 41, the liquid flow channel 46 contacts the bottom of the cuvette 42, and the contact area between the cuvettes 42 in sample cells 41 No. 2 to 17 and the liquid flow channel 46 at their bottom is the same, so that the condensing medium in the liquid flow channel 46 can be used to exchange heat with the sample in the cuvette 42 to achieve temperature control of the sample; the inlet of the liquid flow channel 46 is provided at the bottom of sample cell No. 1 411 and is directly connected to the first pipe connection port 44, and the outlet of the liquid flow channel 46 is provided at the bottom of sample cell No. 18 412 and is connected to the second pipe connection port 45 through a pipeline. A third pipe connection port 11 and a fourth pipe connection port 12 are provided on the outer shell of the photometer body 1. The first and third pipe connections 44 / 11 and the second and fourth pipe connections 45 / 12 are respectively connected by a pipe. The third pipe connection port 11 is connected to the liquid outlet 21 of the circulating water bath temperature control system 2 through a pipe, and the fourth pipe connection port 12 is connected to the liquid inlet 22 of the circulating water bath temperature control system 2 through a pipe. With this design, the condensed medium flowing out of the circulating water bath temperature control system 2 will flow into the liquid flow channel 46 after passing through the third and first pipe connection ports 11 / 44 in sequence, and then pass through the bottom of the sample cells No. 1-2-3...17-18 in sequence ( Figure 3 The direction marked by the arrow in the middle is the flow direction of the condensing medium) to adjust the temperature of the samples in each sample pool 41, and then flow out from the flow channel outlet at the bottom of sample pool No. 18 412, and then flow out of the photometer body 1 after passing through the second and fourth connecting ports 45 / 12 in sequence, and finally return to the circulating water bath temperature control system 2 from the liquid inlet 22 to realize the circulation flow of the condensing medium. The circulating water bath temperature control system 2 can adjust the temperature of the condensing medium, and finally achieve the effect of controlling the temperature of the sample circulating water bath.

[0048] In this embodiment, a suitable sealing cover 47 is configured for each cuvette 42, and the sealing cover 47 used to cover the cuvette 42 in sample pool No. 1 411 and sample pool No. 18 412 is provided with a temperature measuring through hole. After the sample to be tested or the temperature measuring liquid is added to all the cuvettes 42, the top of the cuvette 42 is covered with the sealing cover 47, and the surface of the cuvette 42 is cleaned with lens paper before the cuvette 42 is placed in the sample pool 41.

[0049] The multi-channel thermometer 3 can measure the temperature information of different samples at the same time. The wiring terminals of the two temperature measuring cables are respectively connected to the multi-channel thermometer 3, and the other ends with the thermistor 31 respectively pass through the temperature measuring through holes adapted on the sealing covers 47 on the cuvettes 42 in the No. 1 sample pool 411 and the No. 18 sample pool 412 and then extend into the cuvettes 42. The thermistor 31 is placed below the liquid surface of the liquid in the cuvettes 42. In this embodiment, the multi-channel thermometer 3 is used to detect the temperature of the liquid in the cuvettes 42 placed in the No. 1 and No. 18 sample pools 411 / 412 (the temperature measuring liquid usually filled in these two cuvettes is ultrapure water, which is only used for temperature measurement), so the thermistors 31 at the ends of the two temperature measuring cables are respectively inserted into the interiors of the cuvettes 42 placed in the No. 1 and No. 18 sample pools 411 / 412 and extend below the liquid surface to directly contact the liquid. The parameters of each thermistor 31 are set, the reading time interval is 0.12s, and the temperature data of the multi-channel temperature measuring instrument 3 is monitored by the computer 7.

[0050] The process of temperature matching and full spectrum scanning using the above spectrophotometer is as follows:

[0051] The circulating water bath temperature control system 2 is started, the temperature change range and change rate (such as 95-5°C, 0.2°C / min) are set, the circulating water bath temperature control device is started, and the condensation medium is transported to the liquid flow channel 46 through the circulating pump.

[0052] Real-time temperature detection: set the temperature measurement time, save the detection file, monitor the data of the multi-channel thermometer 3 through the computer 7, and ensure that the temperature of the sample pool 41 is uniform and stable within the preset range. Since the water bath circulation uses the condensing medium to flow through the 18 sample pools 41 at a uniform speed from the No. 1 sample pool 411 in sequence, the temperature of the sample in the cuvette in the corresponding sample pool 41 will show a phenomenon of gradually increasing or decreasing from the No. 1 to the No. 18 sample pool, and since the contact area of ​​the cuvette 42 in the No. 2 to No. 17 sample pools 41 and the liquid flow channel 46 at the bottom thereof is the same, the temperature of the samples in these 16 sample pools shows a regular increase or decrease phenomenon, so the data of the No. 1 and No. 18 sample pools 411 / 412 at each time point read by the multi-channel thermometer 3 can be processed by arithmetic difference and assigned to the No. 2 to No. 17 sample pools respectively, which is the real-time temperature of the samples in these 16 sample pools 41 at different time points. This type of temperature matching method is accurate to the temperature change of each sample, and is more accurate in studying the configuration change and stability of the sample.

[0053] Sample preparation and full spectrum scanning:

[0054] The sequence of the test DNA sample is: CCCTAACCCTAACCCTAACCC. This DNA sample can form a semi-protonated C·C +The i-motif structure based on bases is easily affected by environmental factors. The full spectrum scanning of the real-time temperature-controlled UV-spectrophotometer can better reflect the folding state of the structure in a complex environment. Take 1 μM DNA sample and add it to BR buffer with a pH of 6.15 and a potassium ion strength of 0.1 M. Add the dissolved samples to the cuvettes respectively, cover them with sealing covers and place them in sample pools 41 No. 2 to 17 respectively. Set the parameters on the spectrophotometer: full spectrum scanning range 220-335nm, scanning speed 1200nm / min, scanning interval 3nm, and total scanning time 825min. Start the spectrophotometer for full spectrum scanning, and at the same time, the computer 7 starts to record the real-time temperature.

[0055] Data Analysis:

[0056] The collected data are analyzed and processed by computer 7 to obtain data on the change of absorbance of the samples in the 16 sample pools No. 2 to No. 17 with their own temperature. Figure 6 , taking sample pool No. 2 as an example, Figure 6 The small figure a in the middle is a full spectrum scan of the melting process of the sample in the No. 2 sample pool. The melting curve is drawn after the absorbance data of the sample at 295nm is normalized. The data processing method for other samples is the same. Figure 6 Panel b also shows the melting curves drawn after the absorbance data of the samples in sample pools 3 to 17 were normalized, and then the turning point temperature of the melting curve, i.e., the melting temperature (T m ). Figure 6 The middle panel c shows the T values ​​obtained after 16 sample pools 2 to 17 measured the same sample three times in parallel. m The T values ​​and their mean and standard deviation data of the same group of 16 sample pools are shown in Figure 2. m The standard deviations of the values ​​(0.09 / 0.09 / 0.07°C) are all less than 0.1°C, which proves that the present invention has a high measurement accuracy; the average T m The fluctuation range between the values ​​(36.10 / 35.98 / 36.08℃) does not exceed 0.1℃, which proves that the spectrophotometer has high measurement reliability and stability.

[0057] Example 2

[0058] The difference from Example 1 is that in this embodiment, the sample chamber is connected to the air compression pump 8, mainly to prevent the condensation of water vapor in the cuvette 42 at a low temperature when the temperature of the sample is controlled, thereby affecting the detection.

[0059] Specifically, the end of the ventilation pipeline of the air compression pump 8 is connected to the three-way diverter 81, and the two air outlet ends of the three-way diverter 81 are connected to the diverter tubes. The two diverter tubes respectively pass through a circular hole on the side wall of the sample chamber (the diameter of the circular hole is adapted to the outer diameter of the diverter tube) and then extend into the interior of the sample chamber, and are respectively fixed on the left inner wall and the right inner wall of the sample chamber (be careful to avoid the light-transmitting groove 43, that is, to avoid the optical path of the spectrophotometer). The pressure of the air compression pump 8 is adjusted to 60MPa to ensure that the airflow is stable and strong enough, and can continuously output dry and stable air, forming a positive pressure in the cabin, removing humid air, and avoiding the condensation of water vapor in the cuvette 42 in the sample pool in a low temperature environment, which can improve the spectral data quality of the low temperature experiment.

[0060] The above shows and describes the basic principles, main features and advantages of the present invention. However, the above is only a specific embodiment of the present invention, and the technical features of the present invention are not limited thereto. Any other implementation methods derived by any technician in the field without departing from the technical solution of the present invention should be included in the patent scope of the present invention.

Claims

1. A high-throughput UV-visible spectrophotometer with real-time temperature control, wherein the photometer body comprises a detection module and a sample chamber, wherein the sample chamber is arranged at one end of the interior of the photometer body, It is characterized in that A multi-cell support is provided in the sample chamber, and a plurality of sample cells numbered in sequence are provided in the multi-cell support. A liquid flow channel is provided at the bottom of the sample cells, and the liquid flow channel extends along the sample cells arranged in sequence. After the cuvette is placed in the sample cell, the liquid flow channel contacts the bottom of the cuvette. The inlet of the liquid flow channel is arranged at the bottom of the first sample pool, and the outlet is arranged at the bottom of the last sample pool. The inlet and outlet of the liquid flow channel are respectively connected with the liquid outlet and liquid inlet of the circulating water bath temperature control system; The thermistor ends of the two temperature measuring cables of the multi-channel thermometer extend into the cuvettes in the first and last number sample pools and are placed below the liquid surface of the liquid in the cuvettes.

2. A high-throughput UV-visible spectrophotometer with real-time temperature control as claimed in claim 1, It is characterized in that There are 18 sample pools on the multi-pool holder and they are arranged in sequence. Sample pool No. 1 is the first sample pool, sample pool No. 18 is the last sample pool, and sample pools No. 2 to No. 17 are called intermediate sample pools.

3. A high-throughput UV-visible spectrophotometer with real-time temperature control as claimed in claim 2, It is characterized in that The sample pools are distributed in two rows, the sample pools in the left and right rows are staggered, and the contact area of ​​the cuvette placed in the middle sample pool and the liquid flow channel at its bottom is the same.

4. The high-throughput UV-visible spectrophotometer with real-time temperature control according to claim 1, It is characterized in that The cuvette is provided with a sealing cover, and the sealing cover used for covering the cuvettes in the first and last number sample pools is provided with a temperature measuring through hole, and the thermistor end of the temperature measuring cable extends from the temperature measuring through hole into the cuvettes in the first and last number sample pools.

5. The high-throughput UV-visible spectrophotometer with real-time temperature control according to claim 1, It is characterized in that A first pipe connection port and a second pipe connection port are provided at one end of the multi-cell bracket, the liquid flow channel inlet is connected to the first pipe connection port, and the liquid flow channel outlet is connected to the second pipe connection port through a pipeline; a third pipe connection port and a fourth pipe connection port are provided on the outer shell of the photometer body, the first and third pipe connections and the second and fourth pipe connections are respectively connected through a pipeline, the third pipe connection port is connected to the liquid outlet of the circulating water bath temperature control system through a pipeline, and the fourth pipe connection port is connected to the liquid inlet of the circulating water bath temperature control system through a pipeline, so as to realize the circulation of the condensing medium.

6. The high-throughput UV-visible spectrophotometer with real-time temperature control according to claim 1, It is characterized in that Light-transmitting grooves are provided on both sides of the multi-cell bracket, which is movably connected to the bottom plate, which is fixed in the sample chamber, and a driving member is provided on the bottom plate. The multi-cell bracket moves under the action of the driving member to adjust the position of the sample cell.

7. The high-throughput UV-visible spectrophotometer with real-time temperature control according to claim 1, It is characterized in that The sample chamber is connected to an air compression pump, the end of the ventilation pipeline of the air compression pump is connected to a three-way splitter, and the two air outlet ends of the three-way splitter are respectively connected to a splitter pipe. The two splitter pipes extend through the side wall of the sample chamber to the interior thereof and are correspondingly fixed on the inner walls on the opposite sides of the sample chamber.

8. A high-throughput UV-visible spectrophotometer with real-time temperature control according to claim 7, It is characterized in that The splitter tube should be fixed in the sample chamber in a position away from the optical path of the spectrophotometer.

9. A method for using the high-throughput UV-visible spectrophotometer with real-time temperature control according to any one of claims 1 to 8, It is characterized in that The steps include: 1) After adding ultrapure water to two cuvettes, cover them with sealing covers with temperature measuring through holes, and place them in the head and tail sample pools respectively. Extend the thermistor ends of the two temperature measuring cables of the multi-channel thermometer into the corresponding cuvettes through the temperature measuring through holes on a sealing cover, and place them below the liquid surface; 2) Add the samples to be tested into different cuvettes, cover them with sealing covers without temperature measurement holes, clean the surface of the cuvettes and place them into the middle sample pool; 3) Start the circulating water bath temperature control system, set the temperature change range and change rate, and transport the condensing medium to the liquid flow channel through the circulating pump; 4) Set thermistor parameters and reading time interval, and monitor the temperature data of the multi-channel thermometer through the computer; 5) The temperature data of the first sample pool and the last sample pool at each time point read by the multi-channel thermometer are processed arithmetic difference and assigned to the middle sample pool respectively, that is, the real-time temperature of the samples in the middle sample pool at different time points is obtained; 6) The sample temperature is regulated and recorded in real time while setting scanning parameters, and the spectrophotometer is started to perform a full spectrum scan of the sample in the middle sample pool to obtain a full spectrum scan diagram of the sample melting process; 7) Matching the measured continuous spectrum data and temperature data of the sample to obtain the variable temperature spectrum data of the sample to be measured.

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