JP5397352B2 - Fluorescence spectrophotometer - Google Patents

Description

  The present invention relates to a fluorescence spectrophotometer, and more particularly to a fluorescence spectrophotometer capable of easily inspecting the wavelength accuracy of an excitation side spectroscope and a fluorescence side spectroscope of the fluorescence spectrophotometer.

  In a fluorescence spectrophotometer, light emitted from a light source having a continuous spectrum such as a xenon lamp is dispersed by an excitation-side spectroscope, and excitation light having a target wavelength is irradiated onto a sample in a sample cell. The light generated from the sample enters the fluorescence side spectroscope, and only the fluorescence generated from the excitation sample is dispersed by the excitation side spectroscope and detected by the detector (Patent Document 1).

  The wavelengths of the excitation side spectroscope and the fluorescence side spectroscope are set by adjusting the rotation angle of the diffraction grating in each spectroscope. Conventionally, in order to confirm the accuracy of the set wavelength of a spectrometer, light having an emission line at a specific known wavelength (about 254 nm), such as a mercury lamp, is incident on each spectrometer, and the wavelength of each spectrometer. Are used to detect the emitted light and compare the peak position of the detected bright line with the set wavelength.

  In such a conventional inspection method, it is necessary to use a light source having an emission line at a specific wavelength in order to confirm the wavelength accuracy of the spectrometer. However, as described above, since a light source having a continuous spectrum is used in normal fluorescence spectrometry, it is necessary to replace the light source from a xenon lamp to a mercury lamp when performing wavelength inspection of the spectrometer.

  Since the accuracy of the set wavelength of the spectrometer is greatly related to the accuracy of the fluorescence spectroscopic analysis, it is necessary to perform wavelength inspection of the spectrometer with a certain frequency. However, the replacement work of the lamp is troublesome for the user, and there has been a risk that the accuracy of the analysis deteriorates because the axis of the lamp is displaced at the time of replacement.

JP 2001-83093 A

  This invention is made | formed in view of said problem, and makes it a subject to provide the fluorescence spectrophotometer which can test | inspect wavelength accuracy of a spectrometer easily.

The first invention of the present application made to solve the above problems is
A light source having a continuous spectrum;
An excitation-side spectroscope that splits the light emitted from the light source as excitation light and irradiates the sample cell;
A fluorescence-side spectroscope comprising a fluorescence-side diffraction grating that diverges and emits light emitted from the sample cell when irradiated with excitation light, and a fluorescence-side scanning means that rotates the fluorescence-side diffraction grating and performs wavelength scanning;
A detector for detecting the intensity of light emitted from the fluorescence side spectroscope;
In a fluorescence spectrophotometer comprising:
The rotation angle of the fluorescence-side diffraction grating that emits the n-order diffracted light of the inspection light from the fluorescence-side diffraction grating when the inspection light of a specific wavelength is incident on the fluorescence-side diffraction grating, and the n + 1-order diffracted light of the inspection light Fluorescence side inspection light rotation angle difference storage means for storing a difference in rotation angle of the fluorescence side diffraction grating to be emitted; and
Based on the detection result of the detector when the set wavelength of the excitation side spectrometer is set to the specific wavelength, the light from the light source is incident on the excitation side spectrometer, and the fluorescence side diffraction grating is rotated. A fluorescence side rotation angle difference detecting means for detecting a difference between a rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light having the set wavelength is obtained and a rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light is obtained;
By comparing the rotation angle difference stored in the fluorescence side inspection light rotation angle difference storage unit with the rotation angle difference detected by the fluorescence side rotation angle difference detection unit, the accuracy of the set wavelength of the excitation side spectroscope is compared. It is a fluorescence spectrophotometer characterized by comprising excitation side inspection means for inspecting.

  In this case, an inspection light source that emits inspection light of a specific wavelength is detachably provided, and the fluorescence-side inspection light rotation angle difference storage means causes the light from the inspection light source to enter the fluorescence-side diffraction grating. Based on the detection result of the detector, the difference between the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the inspection light is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light of the inspection light is obtained is stored. You may make it do.

In addition, the second invention
A light source having a continuous spectrum;
An excitation side spectroscope comprising an excitation side diffraction grating that splits the light emitted from the light source as excitation light and irradiates the sample cell, and excitation side scanning means that rotates the excitation side diffraction grating to perform wavelength scanning;
A fluorescence-side spectroscope that splits and emits light emitted from the sample cell;
A detector for detecting the intensity of light emitted from the fluorescence side spectroscope;
In a fluorescence spectrophotometer comprising:
The rotation angle of the excitation side diffraction grating that emits the nth order diffracted light of the inspection light from the excitation side diffraction grating when the inspection light of a specific wavelength enters the excitation side diffraction grating, and the n + 1st order diffracted light of the inspection light. Excitation-side inspection light rotation angle difference storage means for storing a difference in rotation angle of the excitation-side diffraction grating to be emitted;
Based on the detection result of the detector when the set wavelength of the fluorescence side spectrometer is set to the specific wavelength and the light from the light source is incident on the fluorescence side spectrometer and the excitation side diffraction grating is rotated, Excitation-side rotation angle difference detection means for detecting a difference between a rotation angle of the excitation-side diffraction grating from which the n-th order diffracted light having the set wavelength is obtained and a rotation angle of the excitation-side diffraction grating from which the n + 1-order diffraction light is obtained;
By comparing the rotation angle difference stored in the excitation side inspection light rotation angle difference storage means with the rotation angle difference detected by the excitation side rotation angle difference detection means, the accuracy of the set wavelength of the fluorescence side spectroscope is compared. Fluorescent side inspection means for inspecting
It is a fluorescence spectrophotometer characterized by comprising.

  In this case, the excitation-side inspection light rotation angle difference storage means is detachably provided with an inspection optical path for directing incident light from the excitation-side spectrometer to the detector and an inspection light source for emitting inspection light of a specific wavelength. The nth-order diffracted light of the inspection light is obtained based on the detection result of the light that has passed through the inspection optical path and entered the detector when light from the inspection light source is incident on the excitation-side diffraction grating. You may make it memorize | store the difference of the rotation angle of the said excitation side diffraction grating, and the rotation angle of the said excitation side diffraction grating from which the n + 1st order diffracted light of the said inspection light is obtained.

On the other hand, the third invention of the present application is directed to a light source having a continuous spectrum, an excitation-side spectroscope that divides the light emitted from the light source as excitation light and irradiates the sample cell, and the sample cell when irradiated with excitation light. Fluorescence side spectrometer comprising a fluorescence side diffraction grating that divides and emits emitted light and a fluorescence side scanning means that rotates the fluorescence side diffraction grating to perform wavelength scanning, and detects the intensity of the emitted light from the fluorescence side spectrometer A wavelength setting method for an excitation-side spectroscope in a fluorescence spectrophotometer comprising a detector to perform,
The rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the inspection light is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1st order diffracted light is obtained when inspection light of a specific wavelength is incident on the fluorescence side diffraction grating. Memorize the difference in advance,
Rotating the fluorescence side diffraction grating while setting the set wavelength of the excitation side spectrometer to the specific wavelength and making the light from the light source enter the excitation side spectrometer,
Based on the detection result of the detector at that time, the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the set wavelength is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light of the set wavelength is obtained are obtained. Detect the difference,
This rotation angle difference is compared with the difference between the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the inspection light stored in advance is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light is obtained, The accuracy of the set wavelength of the excitation-side spectrometer is inspected.

According to a fourth aspect of the present invention, a light source having a continuous spectrum, an excitation-side diffraction grating for irradiating a sample cell with light emitted from the light source as excitation light, and rotating the excitation-side diffraction grating for wavelength scanning Fluorescence provided with an excitation-side spectroscope provided with excitation-side scanning means, a fluorescence-side spectroscope that divides and emits light emitted from the sample cell, and a detector that detects the intensity of the emitted light from the fluorescence-side spectroscope. A setting wavelength inspection method for a fluorescence side spectroscope in a spectrophotometer,
The rotation angle of the excitation side diffraction grating from which the nth order diffracted light of the inspection light is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1st order diffracted light is obtained when inspection light of a specific wavelength is incident on the excitation side diffraction grating. Memorize the difference in advance,
Rotating the excitation side diffraction grating while setting the set wavelength of the fluorescence side spectrometer to the specific wavelength and making the light from the light source enter the excitation side spectrometer,
Based on the detection result of the detector at that time, the rotation angle of the excitation side diffraction grating from which the nth order diffracted light of the set wavelength is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1st order diffracted light of the set wavelength is obtained are obtained. Detect the difference,
This rotation angle difference is compared with the difference between the rotation angle of the excitation side diffraction grating from which the nth order diffracted light of the inspection light stored in advance is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1 order diffraction light is obtained, The accuracy of the set wavelength of the fluorescence side spectroscope is inspected.

The present invention is a fluorescence spectrophotometer and a set wavelength inspection method capable of easily inspecting the accuracy of the set wavelength of the fluorescence side spectrometer or the excitation side spectrometer.
In the fluorescence spectrophotometer of the present invention, when inspection light having a specific wavelength is incident on the diffraction grating included in the fluorescence-side spectrometer or the diffraction grating included in the excitation-side spectrometer in advance, the fluorescence-side diffraction grating Alternatively, the difference between the rotation angle of each diffraction grating that emits the nth order diffracted light of the inspection light from the excitation side diffraction grating and the rotation angle of each diffraction grating that emits the n + 1st order diffracted light of the inspection light is the fluorescence side inspection light rotation angle. It is stored in the difference storage means or the excitation side inspection light rotation angle difference storage means.
The processing for storing the rotation angle difference in the fluorescence side inspection light rotation angle difference storage means and the excitation side inspection light rotation angle difference storage means is usually performed in the factory before or at the time of shipment of the product.

On the other hand, the inspection of the accuracy of the set wavelength of the fluorescence side spectrometer or the excitation side spectrometer is performed at the time of product delivery or after delivery.
For example, in the fluorescence spectrophotometer according to the first invention, light from a light source having a continuous spectrum is sequentially incident on the excitation-side spectroscope and the sample cell. Then, the light emitted from the sample cell is incident on the fluorescence side spectroscope, and the light emitted from the fluorescence side spectroscope is detected by the detector. At this time, a part of the excitation light having a set wavelength incident on the sample cell from the excitation side spectroscope is incident on the fluorescence side spectroscope by Rayleigh scattering inside the sample cell. Therefore, when inspecting the accuracy of the setting wavelength of the excitation side spectrometer, the setting wavelength of the excitation side spectrometer is set to be the same as the inspection light wavelength, and the fluorescence side rotation angle difference detecting means is based on the detection result of the detector. By setting so as to detect the difference between the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the set wavelength is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1st order diffracted light is obtained, it is caused by Rayleigh scattering from the sample cell. The difference in the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light and n + 1 order diffracted light of the excitation light incident on the fluorescence side spectroscope is detected is detected by the fluorescence side rotation angle difference detecting means.

  The rotation angle difference thus obtained has a unique value due to the characteristics of the diffraction grating. That is, the diffraction grating rotation angle difference Δθ1 between the nth order diffracted light and the n + 1st order diffracted light when the light of wavelength λ1 is incident, and the diffraction grating rotation angle of the nth order diffracted light and the n + 1st order diffracted light when light of other wavelength λ2 is incident. The difference Δθ2 always takes a different value. Therefore, if the rotation angle difference detected at the time of wavelength inspection is the same as the rotation angle difference at the time of inspection light incidence stored in the fluorescence side inspection light rotation angle difference storage means in advance, the wavelength of the excitation side spectrometer is accurate. Can be confirmed.

  The fluorescence spectrophotometer according to the second aspect of the invention is for easily inspecting the wavelength accuracy of the fluorescence side spectroscope, and the accuracy of the set wavelength of the excitation side spectroscope in the fluorescence spectrophotometer according to the first aspect of the invention. The basic principle is the same as the method of testing. However, unlike the first invention, in the second invention, the setting wavelength of the fluorescence side spectroscope is set to be the same as the inspection light wavelength, and the excitation is performed in a state where the fluorescence side spectroscope splits and emits only the set wavelength light. The wavelength of the side spectroscope is scanned, and the difference between the rotation angle at which the nth order diffracted light is obtained and the rotation angle at which the n + 1st order diffracted light is obtained is detected. That is, in the fluorescence spectrophotometer according to the second aspect of the invention, when inspecting the set wavelength of the fluorescence side spectroscope, light from a light source having a continuous spectrum is sequentially incident on the excitation side spectroscope and the sample cell. The difference between the rotation angle of the excitation-side diffraction grating from which the n-order diffracted light is obtained and the rotation angle of the excitation-side diffraction grating from which the n + 1-order diffracted light is obtained is determined on the excitation side. It is detected by the rotation angle difference detecting means.

  As described above, in the fluorescence spectrophotometer according to the present invention, prior to the wavelength inspection of each spectrometer, the rotation angle difference storage means stores the diffraction grating rotation angle difference from which the diffracted light of the inspection light having the specific wavelength is obtained. . Therefore, the operation of replacing the light source with the inspection light source is only necessary when storing the rotation angle difference. In addition, if the rotation angle difference data of the inspection light is stored in the rotation angle difference storage means before or at the time of shipment of the fluorescence spectrophotometer, it is necessary for each user to replace the light source after the shipment of the fluorescence spectrophotometer. Absent. Therefore, it is possible to greatly reduce the labor of wavelength inspection, and it is possible to easily inspect the accuracy of the set wavelength of each spectrometer. Further, it is possible to avoid the problem that the accuracy of the analysis deteriorates due to the deviation of the axis of the light source lamp when the light source is replaced.

  Also in the set wavelength inspection methods according to the third invention and the fourth invention of the present application, the same effects as those of the first invention and the second invention described above can be obtained.

The schematic block diagram of the fluorescence spectrophotometer which concerns on one Embodiment of this invention. The schematic block diagram of the fluorescence spectrophotometer used in the preparatory stage of Example 1 of this invention. The flowchart figure which shows the process in the preparatory stage of Example 1 of this invention. The flowchart which shows the process in the wavelength test | inspection stage of Example 1 of this invention. The figure which shows the example of the spectrum acquired in the preparation stage of Example 1 of this invention. The figure which shows the example of the spectrum obtained in the wavelength test | inspection stage of Example 1 of this invention. The schematic block diagram of the fluorescence spectrophotometer used in the preparatory stage of Example 2 of this invention. The schematic block diagram of the fluorescence spectrophotometer used in the wavelength test | inspection stage of Example 2 of this invention.

  Hereinafter, an embodiment of the present invention will be described with reference to FIGS.

FIG. 1 is a schematic configuration diagram of a fluorescence spectrophotometer according to the present embodiment. The fluorescence spectrophotometer according to the present embodiment includes a xenon lamp 1, an excitation-side spectrometer 2, a sample cell 3, a fluorescence-side spectrometer 4, a detector 5, a calculation unit 6, and a storage unit 7 as light sources having a continuous spectrum. Have. The light from the xenon lamp 1 enters the sample cell 3 via the excitation side spectroscope 2, and thereby the light emitted from the sample cell 3 enters the fluorescence side spectroscope 4.
The excitation-side spectrometer 2 includes an excitation-side diffraction grating 21 and an excitation-side drive mechanism 22, and the excitation-side diffraction grating 21 is driven to rotate by the excitation-side drive mechanism 22, thereby scanning the wavelength of incident light. Do.
The fluorescence side spectroscope 4 includes a fluorescence side diffraction grating 41 and a fluorescence side drive mechanism 42. By rotating the fluorescence side diffraction grating 41 by the fluorescence side drive mechanism 42, wavelength scanning is performed on incident light. Do.
The light emitted from the fluorescence side spectroscope 4 is detected by the detector 5. The detector 5 measures the intensity of the emitted light, the measurement result is calculated by the calculation unit 6, and necessary data is stored in the storage unit 7.
Hereinafter, the inspection method of the set wavelength of the excitation side spectroscope 2 in the spectrophotometer of this embodiment will be described as Example 1, and the inspection method of the set wavelength of the fluorescence side spectroscope 4 will be described as Example 2.

  Here, the inspection method of the set wavelength of the excitation side spectroscope 2 will be described separately in the preparation stage and the wavelength inspection stage.

(Preparation stage)
In the preparation stage, light from an inspection light source having an emission line at a specific wavelength is directly incident on the fluorescence side spectroscope 4 and wavelength scanning of the fluorescence side diffraction grating 41 is performed to obtain diffraction light of a specific wavelength. In this step, the rotation angle difference of 41 is stored in the storage unit 7. The preparation stage is performed in a fluorescence spectrophotometer having the configuration shown in FIG. That is, light from a mercury lamp 8 as an inspection light source enters the fluorescence side spectroscope 4. In the fluorescence side spectroscope 4, the fluorescence side diffraction grating 41 is rotationally driven by the fluorescence side drive mechanism 42, whereby wavelength scanning is performed on the incident light. The light emitted from the fluorescence side spectroscope 4 enters the detector 5 and the intensity thereof is measured. The measurement result of the detector 5 is input to the calculation unit 6, and the calculation result is stored in the storage unit 7.

  The spectrophotometer used in the preparation stage may be a spectrophotometer different from the spectrophotometer of the present embodiment shown in FIG. 1, but the spectrophotometer of the present embodiment can also be used. When the spectrophotometer shown in FIG. 1 is used, the spectrophotometer includes a mercury lamp 8 as an inspection light source in place of the xenon lamp 1 so that light can enter the excitation-side spectroscope 2 from the light source. And an optical path incident on the fluorescence spectrometer 4 are switchable. Alternatively, by setting the setting wavelength of the excitation side spectroscope 2 to the emission line wavelength of the inspection light source or the 0th order light (light including all wavelengths), the emission line from the inspection light source passes through the excitation side spectroscope 2 and the sample cell. 3, and then a part of the light irradiated to the sample cell 3 is scattered (Rayleigh scattering) by the buffer in the sample cell 3 and enters the fluorescence side spectroscope 4.

The preparation stage is performed according to the flowchart shown in FIG.
First, the fluorescence side spectroscope 4 is directly irradiated with the light from the mercury lamp 8 (step S11). Then, the diffraction grating 41 in the fluorescence side spectroscope 4 is rotated by the fluorescence side drive mechanism 42 to perform wavelength scanning, and the spectrum of the radiated light from the diffraction grating 41 is based on the detection signal obtained by the detector 5 at that time. It is created (step S12). In the next step 13, the rotation angle of the diffraction grating 41 from which the peaks of the first-order diffracted light and the second-order diffracted light are obtained is detected from the created radiation spectrum, and the difference between the rotation angles is calculated. The processing of step 12 and step 13 is performed by the calculation unit 6.

FIG. 5 shows an example of a radiation spectrum created by the calculation unit 6. According to this spectrum, the peak of the first-order diffracted light appears when the rotation angle of the fluorescent-side diffraction grating 41 is θi, and the peak of the second-order diffracted light appears when θj. Accordingly, the calculation unit 6 detects the rotation angles (θi, θj) from which the peaks of the first-order diffracted light and the second-order diffracted light are obtained from the spectrum shown in FIG. 5, and calculates the difference between the rotation angles Δθ _Em254 = | _θj −θi | calculate. The calculated rotation angle difference Δθ _Em254 is stored in the storage unit 7 (step S14).

(Wavelength inspection stage)
In the wavelength inspection stage, the accuracy of the set wavelength of the excitation-side spectroscope 2 is inspected using the rotation angle difference stored in the storage unit 7 in the preparation stage. The wavelength inspection step is performed without replacing the light source in the spectrophotometer of the present embodiment shown in FIG.

The wavelength inspection stage is performed according to the flowchart shown in FIG.
First, the spectral wavelength of the excitation side spectroscope 2 is set to the same wavelength (254 nm) as the emission line wavelength of the mercury lamp 8 (step S21), and the excitation side spectroscope 2 is irradiated with the light of the xenon lamp 1 (step S22). . Unlike the mercury lamp 8, the light of the xenon lamp 1 has a continuous spectrum. However, since the setting wavelength of the excitation side spectroscope 2 is the same as the emission line wavelength of the mercury lamp, the wavelength setting of the excitation side spectroscope 2 should be accurate. For example, the light split by the excitation-side spectroscope 2 should have the same wavelength (254 nm) as when the mercury lamp was irradiated. The light split by the excitation side spectroscope 2 is irradiated to the sample cell 3, and the light emitted from the sample cell 3 enters the fluorescence side spectroscope 4. At this time, a part of the excitation light incident on the sample cell 3 is scattered (Rayleigh scattering) by the buffer in the sample cell 3 and enters the fluorescence side spectroscope 4.

In this state, the diffraction grating 41 in the fluorescence side spectroscope 4 is rotated to perform wavelength scanning. At that time, based on the detection signal obtained by the detector 5, the calculation unit 6 creates a spectrum of the radiated light from the diffraction grating 41 (step S23). FIG. 6 shows an example of the radiation spectrum obtained in step S23. According to this spectrum, it can be seen that the peak of the first-order diffracted light appears when the rotation angle of the fluorescence side diffraction grating 41 is θk, and the peak of the second-order diffracted light appears when θl. Based on the result thus obtained, the calculation unit 6 calculates the rotation angle difference Δθ _Em between the first-order diffracted light and the second-order diffracted light having the set wavelength of the excitation-side spectrometer 2 (step S24). That is, the rotation angle difference Δθ _Em between the first-order diffracted light and the second-order diffracted light of the excitation light that has entered the fluorescence side spectroscope 4 due to Rayleigh scattering inside the sample cell 3 is calculated.

Next, the calculation unit 6 _{compares the} rotation angle difference Δθ _Em254 stored in the storage unit 7 with the rotation angle difference Δθ _Em calculated in step S24 (step S25), and sets the wavelength of the set wavelength of the excitation-side spectrometer 2. The accuracy is determined (step S26). For example, when the rotation angle difference Δθ _Em254 and the rotation angle difference Δθ _Em are the same, or when the difference between the two is equal to or less than a preset threshold, it is determined that the wavelength setting of the excitation-side spectrometer 2 is accurate. .

  Here, the method of inspecting the set wavelength of the fluorescence side spectroscope 4 will be described separately in the preparation stage and the wavelength inspection stage, as in the first embodiment.

(Preparation stage)
The preparation stage of Example 2 is an excitation in which light from an inspection light source having an emission line at a specific wavelength is incident on the excitation-side spectrometer 2 and wavelength scanning of the excitation-side diffraction grating 21 is performed to obtain diffracted light of a specific wavelength. In this stage, the rotational angle difference of the side diffraction grating 21 is stored in the storage unit 7. Here, using the spectrophotometer shown in FIG. 7, the light from the mercury lamp 8 is incident on the excitation side spectroscope 2, and the excitation side diffraction grating 21 is rotationally driven to perform wavelength scanning, while the excitation side spectroscope 2. The light emitted from is detected by the detector 5. The calculation unit 6 creates a radiation spectrum based on the detection result of the detector 5, and calculates a rotation angle difference Δθ _EX254 between the first-order diffracted light and the second-order diffracted light having a specific wavelength from the spectrum. The calculated rotation angle difference Δθ _EX254 is stored in the storage unit 7.

  Note that the spectrophotometer used in the preparation stage may be a spectrophotometer different from the spectrophotometer according to the present embodiment shown in FIG. 1, but the spectrophotometer shown in FIG. 1 can also be used. When the spectrophotometer of FIG. 1 is used, the spectrophotometer is provided so that the light path emitted from the excitation side spectroscope 2 can be switched between the light path incident on the sample cell and the light path incident on the detector 5. Alternatively, by setting the setting wavelength of the fluorescence side spectroscope 4 to the emission line wavelength of the inspection light source or the 0th order light (light including all wavelengths), a part of the inspection light emitted from the excitation spectroscope 2 is a sample. The light enters the fluorescence side spectroscope through scattering (Rayleigh scattering) by the buffer in the cell 3, and then the inspection light enters the detector 5 through the fluorescent spectroscope.

(Wavelength inspection stage)
In the wavelength inspection stage of the second embodiment, the accuracy of the set wavelength of the fluorescence side spectroscope 4 is inspected using the rotation angle difference stored in the storage unit 7 in the preparation stage. In Example 2, as shown in FIG. 8, the rotation angle of the diffraction grating 41 in the fluorescence side spectroscope 4 is fixed and the spectral wavelength is set to 254 nm.

Then, the light from the xenon lamp 1 is irradiated onto the excitation side spectroscope 2, and the diffraction grating 21 in the excitation side spectroscope 2 is rotated by the drive mechanism 22 to perform wavelength scanning. A part of the excitation light that is split by the excitation side spectroscope 2 and applied to the sample cell 3 is scattered by the buffer in the sample cell 3 and enters the fluorescence side spectroscope 4. Since the fluorescence side spectroscope 4 is set to 254 nm, if the wavelength setting of the fluorescence side spectroscope 4 is accurate, only the light having a wavelength of 254 nm among the scattered light should be detected by the detector 5. The calculation unit 6 detects the rotation angle at which the first-order diffracted light and the second-order diffracted light of the scattered light (Rayleigh scattered light) are obtained, and calculates the rotation angle difference Δθ _EX ).

Finally, the rotation angle difference Δθ _EX254 stored in the storage unit 7 in step S114 is compared with the rotation angle difference Δθ _EX calculated in step S124. If these are the same, the wavelength setting of the excitation-side spectrometer 2 is set. Can be determined to be accurate.

  In addition, this invention is not limited to the said Example, A change is accept | permitted within the range of the meaning of invention. For example, in the present embodiment, a mercury lamp is used as a light source for inspection at the preparation stage, but any kind of light source may be used as long as it has a bright line with a specific wavelength. In the first and second embodiments, the wavelength inspection is performed using the difference in rotation angle between the first-order diffracted light and the second-order diffracted light. However, the detected diffracted light may be any order, for example, third-order diffracted light. It is also possible to use the difference in rotation angle from which the fourth-order diffracted light is obtained.

DESCRIPTION OF SYMBOLS 1 ... Xenon lamp 2 ... Excitation side spectroscope 3 ... Sample cell 4 ... Fluorescence side spectroscope 5 ... Detector 6 ... Calculation part 7 ... Memory | storage part 8 ... Mercury lamp 21 ... Excitation side diffraction grating 22 ... Excitation side diffraction grating drive mechanism 41 ... Fluorescent side diffraction grating 42 ... Fluorescent side diffraction grating drive mechanism

Claims (6)

A light source having a continuous spectrum;
An excitation-side spectroscope that splits the light emitted from the light source as excitation light and irradiates the sample cell;
A fluorescence-side spectroscope comprising a fluorescence-side diffraction grating that diverges and emits light emitted from the sample cell when irradiated with excitation light, and a fluorescence-side scanning means that rotates the fluorescence-side diffraction grating and performs wavelength scanning;
A detector for detecting the intensity of light emitted from the fluorescence side spectroscope;
In a fluorescence spectrophotometer comprising:
The rotation angle of the fluorescence-side diffraction grating that emits the n-order diffracted light of the inspection light from the fluorescence-side diffraction grating when the inspection light of a specific wavelength is incident on the fluorescence-side diffraction grating, and the n + 1-order diffracted light of the inspection light Fluorescence side inspection light rotation angle difference storage means for storing a difference in rotation angle of the fluorescence side diffraction grating to be emitted; and
Based on the detection result of the detector when the set wavelength of the excitation side spectrometer is set to the specific wavelength, the light from the light source is incident on the excitation side spectrometer, and the fluorescence side diffraction grating is rotated. A fluorescence side rotation angle difference detecting means for detecting a difference between a rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light having the set wavelength is obtained and a rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light is obtained;
By comparing the rotation angle difference stored in the fluorescence side inspection light rotation angle difference storage unit with the rotation angle difference detected by the fluorescence side rotation angle difference detection unit, the accuracy of the set wavelength of the excitation side spectroscope is compared. Excitation-side inspection means for inspecting
A fluorescence spectrophotometer comprising: Removably equipped with an inspection light source that emits inspection light of a specific wavelength,
The fluorescence side inspection light rotation angle difference storage means obtains the nth order diffracted light of the inspection light based on the detection result of the detector when the light from the inspection light source is incident on the fluorescence side diffraction grating. The fluorescence spectrophotometer according to claim 1, wherein a difference between a rotation angle of the fluorescence side diffraction grating and a rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light of the inspection light is obtained is stored. A light source having a continuous spectrum;
An excitation side spectroscope comprising an excitation side diffraction grating that splits the light emitted from the light source as excitation light and irradiates the sample cell, and excitation side scanning means that rotates the excitation side diffraction grating to perform wavelength scanning;
A fluorescence-side spectroscope that splits and emits light emitted from the sample cell;
A detector for detecting the intensity of light emitted from the fluorescence side spectroscope;
In a fluorescence spectrophotometer comprising:
The rotation angle of the excitation side diffraction grating that emits the nth order diffracted light of the inspection light from the excitation side diffraction grating when the inspection light of a specific wavelength enters the excitation side diffraction grating, and the n + 1st order diffracted light of the inspection light. Excitation-side inspection light rotation angle difference storage means for storing a difference in rotation angle of the excitation-side diffraction grating to be emitted;
Based on the detection result of the detector when the set wavelength of the fluorescence side spectrometer is set to the specific wavelength and the light from the light source is incident on the fluorescence side spectrometer and the excitation side diffraction grating is rotated, Excitation-side rotation angle difference detection means for detecting a difference between a rotation angle of the excitation-side diffraction grating from which the n-th order diffracted light having the set wavelength is obtained and a rotation angle of the excitation-side diffraction grating from which the n + 1-order diffraction light is obtained;
By comparing the rotation angle difference stored in the excitation side inspection light rotation angle difference storage means with the rotation angle difference detected by the excitation side rotation angle difference detection means, the accuracy of the set wavelength of the fluorescence side spectroscope is compared. Fluorescent side determination inspection means for inspecting
A fluorescence spectrophotometer comprising: An inspection optical path for directing incident light from the excitation side spectroscope to the detector;
Removably equipped with an inspection light source that emits inspection light of a specific wavelength,
The excitation side inspection light rotation angle difference storage means is based on the detection result of the light that has passed through the inspection optical path and entered the detector when the light from the inspection light source is incident on the excitation side diffraction grating, The difference between the rotation angle of the excitation side diffraction grating from which the n-th order diffracted light of the inspection light is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1 order diffracted light of the inspection light is obtained is stored. 3. The fluorescence spectrophotometer according to 3. A light source having a continuous spectrum;
An excitation-side spectroscope that splits the light emitted from the light source as excitation light and irradiates the sample cell;
A fluorescence-side spectroscope comprising a fluorescence-side diffraction grating that diverges and emits light emitted from the sample cell when irradiated with excitation light, and a fluorescence-side scanning means that rotates the fluorescence-side diffraction grating and performs wavelength scanning;
A detector for detecting the intensity of light emitted from the fluorescence side spectroscope;
A wavelength setting method for an excitation-side spectroscope in a fluorescence spectrophotometer comprising:
The rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the inspection light is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1st order diffracted light is obtained when inspection light of a specific wavelength is incident on the fluorescence side diffraction grating. Memorize the difference in advance,
Rotating the fluorescence side diffraction grating while setting the set wavelength of the excitation side spectrometer to the specific wavelength and making the light from the light source enter the excitation side spectrometer,
Based on the detection result of the detector at that time, the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the set wavelength is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light of the set wavelength is obtained are obtained. Detect the difference,
This rotation angle difference is compared with the difference between the rotation angle of the fluorescence side diffraction grating from which the nth order diffracted light of the inspection light stored in advance is obtained and the rotation angle of the fluorescence side diffraction grating from which the n + 1 order diffraction light is obtained, An excitation-side spectrometer setting wavelength inspection method for inspecting the accuracy of the setting wavelength of the excitation-side spectrometer. A light source having a continuous spectrum;
An excitation side spectroscope comprising an excitation side diffraction grating that splits the light emitted from the light source as excitation light and irradiates the sample cell, and excitation side scanning means that rotates the excitation side diffraction grating to perform wavelength scanning;
A fluorescence-side spectroscope that splits and emits light emitted from the sample cell;
A detector for detecting the intensity of light emitted from the fluorescence side spectroscope;
A method for inspecting a setting wavelength of a fluorescence spectrometer in a fluorescence spectrophotometer comprising:
The rotation angle of the excitation side diffraction grating from which the nth order diffracted light of the inspection light is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1st order diffracted light is obtained when inspection light of a specific wavelength is incident on the excitation side diffraction grating. Memorize the difference in advance,
Rotating the excitation side diffraction grating while setting the set wavelength of the fluorescence side spectrometer to the specific wavelength and making the light from the light source enter the excitation side spectrometer,
Based on the detection result of the detector at that time, the rotation angle of the excitation side diffraction grating from which the nth order diffracted light of the set wavelength is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1st order diffracted light of the set wavelength is obtained are obtained. Detect the difference,
This rotation angle difference is compared with the difference between the rotation angle of the excitation side diffraction grating from which the nth order diffracted light of the inspection light stored in advance is obtained and the rotation angle of the excitation side diffraction grating from which the n + 1 order diffraction light is obtained, Fluorescence side spectrometer setting wavelength inspection method for inspecting the accuracy of the setting wavelength of the fluorescence side spectrometer.

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