CN107976467B - Thermal power measuring device with Raman spectrum measuring function - Google Patents
Thermal power measuring device with Raman spectrum measuring function Download PDFInfo
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- CN107976467B CN107976467B CN201711261481.1A CN201711261481A CN107976467B CN 107976467 B CN107976467 B CN 107976467B CN 201711261481 A CN201711261481 A CN 201711261481A CN 107976467 B CN107976467 B CN 107976467B
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N25/00—Investigating or analyzing materials by the use of thermal means
- G01N25/20—Investigating or analyzing materials by the use of thermal means by investigating the development of heat, i.e. calorimetry, e.g. by measuring specific heat, by measuring thermal conductivity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
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Abstract
The invention relates to a thermal power measuring device with a Raman spectrum measuring function, which comprises a thermal measuring system, a Raman spectrum measuring system and a data acquisition and control system, wherein the thermal measuring system is connected with the Raman spectrum measuring system through a thermal sensor; the heat measuring system is used for measuring the titration heat released in the titration process; the Raman spectrum measuring system comprises a laser source, monochromatic light emitted by the laser source is divided into two paths through a Y-shaped branched optical fiber and respectively enters an optical fiber attenuator, optical signals emitted by the first optical fiber attenuator are focused at the center of a sample reaction tank through a first optical fiber and a first optical fiber Raman probe, and Raman signals generated by solution in the sample reaction tank return to the first optical fiber Raman probe and are sequentially emitted to a grating and a Raman spectrometer; similarly, the optical signal emitted by the second optical fiber attenuator is focused on the reference reaction tank through the second optical fiber and the second optical fiber Raman probe, and the Raman signal generated by the solution in the reference reaction tank returns to the second optical fiber Raman probe and is sequentially transmitted to the grating and the Raman spectrometer. The invention can measure thermal power and Raman spectrum simultaneously, and can also measure thermal power and Raman spectrum independently.
Description
Technical Field
The invention relates to a thermal power measuring device, in particular to a thermal power measuring device with a Raman spectrum measuring function, and relates to the technical field of research and application of weak interaction between molecules such as biomacromolecules and molecular self-assembly and other solution reaction systems.
Background
The thermal power measuring device (i.e. calorimeter) is widely applied to the scientific fields of physics, chemistry, biology and the like and the technical fields of oil extraction, chemical engineering, batteries and the like. Among them, the isothermal titration calorimeter is especially suitable for the research of biological system and weak interaction between molecules. Due to the complexity of such systems, raman spectroscopy is typically used to obtain material structural information. Isothermal titration calorimetry can perform continuous titration at multiple points, and has the characteristic of concentration scanning. At present, instruments for structure research such as Raman spectrum and the like do not have the scanning function and can only test point by point respectively. Therefore, in the face of complicated and variable calorimetric curves and finite raman spectrum results which cannot be measured continuously, researchers often cannot determine structures and phase states corresponding to different calorimetric curve change intervals. Especially for biological macromolecules and molecular self-assembly systems, the interaction, the structure and the phase state conversion have strong kinetic dependence, and even small differences of temperature, stirring mode, stirring speed, time and the like can result in different structures. This makes it often difficult to obtain a close correspondence between interaction and material structure by performing titration calorimetry and raman spectroscopy separately.
International has combined differential scanning calorimetry and fiber Raman measurement of the simultaneous change of thermal and Raman signals during sample temperature change (k.p.menard, e.l.dz and r.spragg: "DSC-Raman analytical system and method", United States Patent Application No. us2011/0170095), but no instrument is available at home and abroad for more general chemical or biological reaction processes, especially for the simultaneous measurement of on-line thermal and Raman signals in solution systems.
Disclosure of Invention
In view of the above problems, the present invention provides a thermal power measuring device with raman spectroscopy measurement function, which can ensure that both thermal measurement and raman signal measurement can work normally without affecting each other and operate synchronously.
In order to achieve the purpose, the invention adopts the following technical scheme: a thermal power measuring device with a Raman spectrum measuring function is characterized by comprising a thermal measuring system, a Raman spectrum measuring system and a data acquisition and control system; the heat measuring system is used for measuring the titration heat released in the titration process; the Raman spectrum measurement system comprises a laser light source, a Y-shaped branched optical fiber, two optical fiber Raman probes, corresponding electric translation tables, a grating and a Raman spectrometer; monochromatic light emitted by the laser source is divided into two paths through the Y-shaped branched optical fiber and enters an optical fiber attenuator respectively, an optical signal emitted by the first optical fiber attenuator is focused on one point on the axis of the sample reaction tank through the first optical fiber and the first optical fiber Raman probe, and a Raman signal generated by a solution in the sample reaction tank returns to the first optical fiber Raman probe and is transmitted to the grating and the Raman spectrometer in sequence; similarly, the optical signal emitted by the second optical fiber attenuator is focused on one point on the axis of the reference reaction tank through the second optical fiber and the second optical fiber Raman probe, and the Raman signal generated by the solution in the reference reaction tank returns to the second optical fiber Raman probe and is sequentially transmitted to the grating and the Raman spectrometer; the data acquisition and control system controls the corresponding electric translation table to move back and forth, so that the distances between the two optical fiber Raman probes and the sample reaction tank and the distance between the two optical fiber Raman probes and the reference reaction tank are adjusted, and an optical signal is guaranteed to be focused on one point on the axis of the sample reaction tank or the reference reaction tank.
Further, the heat measuring system comprises a constant temperature system, a calorimetric pool and a heat calibration system; the constant temperature system comprises a constant temperature bath and a heat sink, the heat sink is arranged in the constant temperature bath, the calorimetric pool is arranged in the heat sink, the calorimetric pool comprises a sample pool and a reference pool which are made of completely same materials and have the same size, the sample pool and the reference pool both comprise a heat conducting piece, a thermoelectric pile and a reaction pool, the sample reaction pool and the reference reaction pool are both made of quartz glass tubes and are respectively fixed at the center of the heat conducting piece, the heat conducting piece is contacted with the heat sink through the thermoelectric pile, and the output ends of the thermoelectric pile are reversely connected; the thermal calibration system comprises a program-controlled direct-current power supply and a thermal resistor fixed in the heat conducting piece, and the program-controlled direct-current power supply is connected with the thermal resistor.
Further, the data acquisition and control system comprises a computer, a nanovoltmeter, a data acquisition card or a digital multimeter with a scanning card, a calorimetric module, a Raman spectrum module and a calorimetric-Raman spectrum combined module, wherein the computer is used for controlling the movement of each electric translation stage and monitoring and recording a thermal signal and a Raman spectrum signal; the nanovoltmeter collects a heat flow signal generated by the thermopile; the data acquisition card or the digital multimeter with the scanning card acquires current and voltage signals of the constant temperature bath temperature and the program control direct current power supply; the calorimetric module is used for calibrating the thermal measurement system and measuring thermal power; the Raman spectrum module is used for receiving signals of a Raman spectrometer and measuring Raman spectra, the calorimetric-Raman spectrum combined module drives the program-controlled direct-current power supply to apply pulse refrigerating current on the thermoelectric pile during spectrum measurement so as to cool the sample reaction tank and the reference reaction tank, meanwhile, the nanovoltmeter measures the voltage of the thermoelectric pile and the current applied by the program-controlled direct-current power supply to the thermoelectric pile to work in a time-sharing mode, the refrigerating power applied by the program-controlled direct-current power supply is slightly smaller than the laser heating power, after the Raman spectrum measurement is finished, the calorimetric-Raman spectrum combined module continues to apply cooling power through a PID (proportion integration differentiation) module so as to rapidly cool the thermoelectric pile until the output thermal signals on the thermoelectric pile are reduced to be less than 3 times of the standard deviation absolute value of baseline noise, it is then allowed to cool naturally to restore thermal equilibrium.
Further, the measuring device also comprises a constant-temperature and constant-humidity chamber, wherein the thermal measuring system, the Raman spectrum measuring system and the data acquisition and control system are all placed in the constant-temperature and constant-humidity chamber with the temperature controlled at 20-25 ℃, the temperature control precision of +/-0.5 ℃ and the relative humidity of (45-70)% RH.
Further, the heat conducting piece is made of aluminum alloy or copper with high heat conduction coefficient and is oxidized into black, and a through hole is further formed in the heat conducting piece for placing the optical fiber Raman probe.
Further, the temperature of the constant-temperature bath is required to be controlled between 15 and 60 ℃, the temperature control precision is +/-0.0001 ℃, and the constant-temperature bath adopts a water bath or an oil bath.
due to the adoption of the technical scheme, the invention has the following advantages: 1. the invention arranges the optical path outside the reaction tank, and the operation is simpler, more convenient and faster. 2. The invention adopts the sample cell and the reference cell to simultaneously carry out the double-photon cell thermal measurement and the double-optical-path Raman spectrum measurement, influences the thermal signal generated by light absorption in the offset Raman spectrum measurement on the thermal measurement, and effectively improves the measurement efficiency. 3. The invention further reduces the influence of light measurement on heat measurement through active cooling of Peltier effect, accelerates heat balance and improves the overall measurement efficiency. 4. After Raman spectrum measurement is finished, the calorimetric-Raman spectrum combined module continuously applies cooling power through the PID module to rapidly cool until the output thermal signal on the thermopile is reduced to be close to a baseline, and the photoinduced thermal signal is effectively eliminated. 5. In order to achieve more accurate heat balance, the sample cell and the reference cell are simultaneously and respectively cooled so as to derive heat generated by the Raman excitation light irradiating the calorimetric cell. 6. The invention can measure thermal power and Raman spectrum simultaneously, and can also measure thermal power and Raman spectrum independently.
Drawings
FIG. 1 is a schematic view showing the structure of a thermal power measuring apparatus having a Raman spectroscopy measurement function according to the present invention;
FIG. 2 is a schematic diagram of a Raman spectroscopy measurement system according to the present invention;
FIG. 3 shows the results of thermal and Raman spectroscopy measurements during the reaction in which Sodium Dodecyl Sulfate (SDS) was added dropwise to Bovine Serum Albumin (BSA) in the measurement examples of the present invention, in which (a) shows the results of thermal measurement during the reaction, and (b) shows the results of Raman spectroscopy measurement of the sample base solution (curve No. 0 in the figure) and after each titration reaction.
Detailed Description
the present invention is described in detail below with reference to the attached drawings. It is to be understood, however, that the drawings are provided solely for the purposes of promoting an understanding of the invention and that they are not to be construed as limiting the invention. In the description of the present invention, it is to be understood that the terms "first," "second," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
As shown in fig. 1 and fig. 2, the thermal power measuring device with raman spectroscopy measurement function provided by the present invention includes a thermal measurement system 1, a raman spectroscopy measurement system 2, a data acquisition and control system 3, and a constant temperature and humidity chamber 4. The heat measuring system 1 is an isothermal titration type calorimeter (but not limited to titration type), and comprises a constant temperature bath 11, a heat sink 111, a calorimetric pool 12, a heat calibration system 13 and a calorimetric module 34 used in cooperation. A heat sink 111 is placed in the constant temperature bath 11 to further improve temperature stability, and the heat measuring bath 12 is fixed on the heat sink 111. The calorimeter cell 12 comprises a sample cell 12A and a reference cell 12B which are identical in size and material, and the sample cell 12A and the reference cell 12B each comprise a heat conducting member 121, a thermopile 122 and a reaction cell 123 made of a quartz glass tube. The sample reaction cell 123A and the reference reaction cell 123B are both made of quartz glass tubes and are fixed at the centers of the heat conducting members 121A and 121B, respectively, and the two cells contain the same components and mass of reaction base liquid to be used. The heat conducting member 121 is made of aluminum alloy or copper with high thermal conductivity and oxidized to black to eliminate stray light. The thermal power measurement is realized by the thermopile 122 using the Seebeck (Seebeck) effect, the thermopile 122 may be assembled by using a commercial semiconductor thermoelectric module, and different models may be selected according to the specific requirements of the thermal measurement. In thermal measurements, the outputs of the thermopile 122 are reversed (e.g., the positive terminals of the two stacks are connected to the positive terminal and the negative terminal is connected to the output, or the negative terminals of the two stacks are connected to the negative terminal and the positive terminal is connected to the output) to affect the ambient and photo-thermal signals. The thermal calibration system 13 includes a programmable dc power supply 131 and a thermal resistor 132 fixed in the heat conducting member 121, the programmable dc power supply 131 outputs different constant currents (or constant powers) to the thermal resistor 132, and the thermopile 122 outputs voltage signals with different amplitudes, so as to obtain the instrument constant of the thermal measurement system, i.e. the ratio between the input thermal power and the output voltage signal, and the relaxation time of the thermal signal response.
The Raman spectrum measuring system 2 is used for measuring a Raman spectrum and comprises a laser monochromatic source 21, a Y-shaped branched optical fiber 22, optical fiber attenuators 23A and 23B, optical fibers 24A and 24B, optical fiber Raman probes 25A and 25B, a grating 26 and a Raman spectrometer 27, wherein the two optical fiber Raman probes 25A and 25B are respectively fixed on an electric translation table, the Raman spectrum can be measured in the direction of 180 degrees of incident light, or can be measured at any angle between 90 and 180 by adopting two optical fiber coupling lenses, but the optical focuses of the two optical fiber Raman probes are ensured to be confocal at the same point on the axis in a reaction tank. Monochromatic light emitted by a laser source 21 is divided into two paths through a Y-shaped branched optical fiber 22 and respectively enters optical fiber attenuators 23A and 23B, optical signals emitted by the optical fiber attenuator 23A are focused at the center of a sample reaction tank 123A through an optical fiber 24A and an optical fiber Raman probe 25A, and Raman signals generated by a solution in the sample reaction tank 123A return to the optical fiber Raman probe 25A and are sequentially emitted to a grating 26 and a Raman spectrometer 27; similarly, the optical signal emitted from the optical fiber attenuator 23B is focused on the center of the reference reaction cell 123B through the optical fiber 24B and the fiber raman probe 25B, and the raman signal generated from the solution in the reference reaction cell 123B returns to the fiber raman probe 25B and is sequentially emitted to the grating 26 and the raman spectrometer 27. Wherein, the fiber Raman probe 25 is fixed in a prepared hole of the heat conducting member 121. In addition, the data acquisition and control system can control the back and forth movement of the corresponding electric translation table, so as to adjust the distances between the fiber Raman probes 25A and 25B and the sample reaction pool 123A and the reference reaction pool 123B respectively, and ensure that the optical signal is focused on one point on the axis of the sample reaction pool 123A or the reference reaction pool 123B. The raman spectroscopy of the present invention utilizes a sample cell and a reference cell to realize dual optical path measurement, and the optical fiber attenuator 23 is used to make the intensities of two optical signals consistent, i.e. adjust the relative intensities of the optical signals to make the thermal signals offset to a minimum value, so as to eliminate the influence of the optical measurement on the thermal measurement as much as possible, and the two raman spectral signals can be simultaneously measured through different channels of the raman spectrometer 27.
The data acquisition and control system comprises a computer 31, a nanovoltmeter 32, a data acquisition card or a digital multimeter 33 with a scanning card, a calorimetric module 34, a Raman spectrum module 35 and a calorimetric-Raman spectrum combination module 36. The computer 31 is used for controlling each electric translation stage to move, and monitoring and recording a thermal signal and a Raman spectrum signal at the same time; the nanovoltmeter 32 collects a heat flow signal generated by the thermopile 122; a data acquisition card or a digital multimeter 33 with a scanning card acquires the temperature of the constant temperature bath 11 and current and voltage signals of the program-controlled direct current power supply 131; the calorimetric module 34 is used for calibration and thermal power measurement of a thermal measurement device; the raman spectrum module 35 is configured to receive signals from the raman spectrometer and measure a raman spectrum. During raman spectrum measurement, an incident light signal heats the sample reaction cell 123A and the reference reaction cell 123B, at this time, the thermopile 122A (122B) generates a thermoelectromotive force on the nanovoltmeter 32A (32B), and in order to eliminate a photo-induced thermal signal, the calorimetric-raman spectrum combination module 36 drives the direct-current power supply 131A (131B) to apply a pulse cooling current on the thermopile 122A (122B) during spectrum measurement so as to cool the sample reaction cell 123A (reference reaction cell 123B). Meanwhile, the nanovoltmeter 32 measures the voltage of the thermopile 122 and the direct current power supply 131 applies current to the thermopile 122 to work in a time-sharing manner so as to avoid mutual interference, and the refrigeration power applied by the direct current power supply 131 is slightly smaller than the laser heating power so as to avoid supercooling of the sample reaction cell 123A or the reference reaction cell 123B. After the raman spectroscopy measurement is finished, the calorimetric-raman spectroscopy combined module 36 continues to apply cooling power through a PID (proportional-integral-derivative) module to rapidly cool the sample cell and the reference cell until the output thermal signal on the thermopile 122 is reduced to less than 3 times of the absolute value of the standard deviation of the baseline noise, and then is naturally cooled to restore thermal balance.
In a preferred embodiment, the thermal measurement system 1, the Raman spectrum measurement system 2 and the data acquisition and control system 3 are all placed in a constant temperature and humidity chamber 4 with the temperature controlled at 20 to 25 ℃, the temperature control precision of +/-0.5 ℃ and the relative humidity of (45-70)% RH, and electromagnetic interference is avoided.
In a preferred embodiment, the temperature of constant temperature bath 11 is controlled to be between 15 and 60 ℃, the temperature control precision is +/-0.0001 ℃, and constant temperature bath 11 can be a water bath or an oil bath.
The following describes in detail the procedure of using the thermal power measuring apparatus having raman spectroscopy measurement function of the present invention, with reference to the titration reaction procedure of Sodium Dodecyl Sulfate (SDS) and Bovine Serum Albumin (BSA):
Before the titration reaction, the raman spectrum of 0.6 ml of 50mg/ml BSA aqueous solution system (i.e., the curve numbered 0 in fig. 3 (b)) was measured, the measurement wavelength was 785nm, the integration time of each sample was 60s, 10 μ l of 0.2M SDS was added dropwise each time, the titration heat was measured, the raman spectrum was measured once after the titration heat was completed each time, and the heat measurement and the raman spectrum measurement were performed alternately. The results of 15 times of titration and 16 times of Raman spectroscopy are shown in FIGS. 3(a) and (b), respectively, and it can be seen from the figure that the thermal power measurement device of the present invention can simultaneously measure the thermal signal and Raman spectroscopy signal of the system, and certain characteristic Raman spectral lines are indeed changed during the titration process.
The above embodiments are only used for illustrating the present invention, and the structure, connection mode, manufacturing process, etc. of the components may be changed, and all equivalent changes and modifications performed on the basis of the technical solution of the present invention should not be excluded from the protection scope of the present invention.
Claims (5)
1. A thermal power measuring device with a Raman spectrum measuring function is characterized by comprising a thermal measuring system, a Raman spectrum measuring system and a data acquisition and control system; the heat measuring system is used for measuring the titration heat released in the titration process; the Raman spectrum measurement comprises a laser light source, a Y-shaped branched optical fiber, two optical fibers, two optical fiber Raman probes, a corresponding electric translation table, a grating and a Raman spectrometer; monochromatic light emitted by the laser source is divided into two paths through the Y-shaped branched optical fiber and enters an optical fiber attenuator respectively, an optical signal emitted by the first optical fiber attenuator is focused at the center of the sample reaction tank through the first optical fiber and the first optical fiber Raman probe, and a Raman signal generated by a solution in the sample reaction tank returns to the first optical fiber Raman probe and is transmitted to the grating and the Raman spectrometer in sequence; similarly, the optical signal emitted by the second optical fiber attenuator is focused at the center of the reference reaction tank through the second optical fiber and the second optical fiber Raman probe, and the Raman signal generated by the solution in the reference reaction tank returns to the second optical fiber Raman probe and is sequentially emitted to the grating and the Raman spectrometer; the data acquisition and control system controls the back and forth movement of the corresponding electric translation table, so as to adjust the distances between the optical fiber Raman probe and the sample reaction tank and the reference reaction tank respectively, and ensure that an optical signal is focused on the axis of the sample reaction tank or the reference reaction tank;
the data acquisition and control system comprises a computer, a nanovoltmeter, a data acquisition card or a digital multimeter with a scanning card, a calorimetric module, a Raman spectrum module and a calorimetric-Raman spectrum combined module, wherein the computer is used for controlling the movement of each electric translation table and monitoring and recording a thermal signal and a Raman spectrum signal at the same time; the nano-voltmeter is used for collecting a heat flow signal generated by the thermoelectric pile; the data acquisition card or the digital multimeter with the scanning card is used for acquiring current and voltage signals of the constant-temperature bath temperature and the program-controlled direct-current power supply; the calorimetric module is used for calibrating the thermal measurement system and measuring thermal power; the Raman spectrum module is used for receiving signals of a Raman spectrometer to measure Raman spectra, and the calorimetric-Raman spectrum combination module drives the program-controlled direct-current power supply to apply pulse refrigeration current to the thermoelectric pile during spectrum measurement so as to cool the sample reaction tank and the reference reaction tank; simultaneously, the nanovoltmeter measures the voltage of the thermoelectric thermopile, and the program-controlled direct-current power supply applies current to the thermoelectric thermopile, and the two processes work in a time-sharing manner; the refrigeration power applied by the program control direct current power supply is slightly smaller than the laser heating power, after Raman spectrum measurement is finished, the calorimetric-Raman spectrum combined module continues to apply cooling power through the PID module to rapidly cool until the output thermal signal on the temperature difference thermopile is reduced to be less than 3 times of the standard deviation absolute value of baseline noise, and then the temperature difference thermopile is naturally cooled to restore thermal balance.
2. The thermal power measuring device with raman spectroscopy measurement according to claim 1, wherein the thermal measurement system includes a constant temperature system, a calorimetric cell, and a thermal calibration system; the constant temperature system comprises a constant temperature bath and a heat sink, the heat sink is arranged in the constant temperature bath, the calorimetric pool is arranged in the heat sink, the calorimetric pool comprises a sample pool and a reference pool which are made of completely same materials and have the same size, the sample pool and the reference pool both comprise a heat conducting piece, a thermoelectric pile and a reaction pool, the sample reaction pool and the reference reaction pool are both made of quartz glass tubes and are respectively fixed at the center of the heat conducting piece, the heat conducting piece is contacted with the heat sink through the thermoelectric pile, and the output ends of the thermoelectric pile are reversely connected; the thermal calibration system comprises a program-controlled direct-current power supply and a thermal resistor fixed in the heat conducting piece, and the program-controlled direct-current power supply is connected with the thermal resistor.
3. The thermal power measuring device with raman spectroscopy according to claim 1, further comprising a constant temperature and humidity chamber, wherein the thermal measuring system, the raman spectroscopy measuring system and the data acquisition and control system are all placed at a temperature of 20 to 25 ℃ with a temperature control accuracy0.5 ℃ and relative humidity within the constant temperature and humidity chamber of (45-70)% RH.
4. The thermal power measuring apparatus with raman spectroscopy according to claim 2, wherein the thermal conductive member is made of aluminum alloy or copper with high thermal conductivity and oxidized to black, and a through hole is formed in the thermal conductive member for accommodating the fiber raman probe.
5. The thermal power measuring apparatus with raman spectroscopy according to claim 2, wherein the temperature control of the constant-temperature bath is required to be between 15 and 60 ℃, the temperature control accuracy is ± 0.0001 ℃, and the constant-temperature bath is a water bath or an oil bath.
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| CN113552109B (en) * | 2020-04-23 | 2023-12-29 | 中国石油化工股份有限公司 | Memory, and method, device and equipment for testing and analyzing reaction thermal effect based on Raman spectrum |
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| CN101576520B (en) * | 2009-06-03 | 2011-08-31 | 中国科学院化学研究所 | Thermal power measurement device |
| CA2780725A1 (en) * | 2009-11-25 | 2011-06-03 | Perkinelmer Health Sciences, Inc. | Analytical system and method with combined differential scanning calorimetry (dsc) and raman spectroscopy |
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