JP3448135B2 - Optical axis moving type fluorescence photometer and measurement method - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a fluorescence spectrophotometer for measuring a fluorescent substance in a solution in which a plurality of substances coexist.

[0002]

2. Description of the Related Art FIG. 7 is a block diagram of a conventional fluorescence spectrophotometer.
Shown in. The fluorescence method is a method in which a quartz sample cell into which a sample solution containing a fluorescent substance is injected is irradiated with excitation light, and quantitative and qualitative analysis is performed from the intensity and spectrum of fluorescence generated thereby.

In FIG. 7, the light emitted from the light source 1 is split into monochromatic light by the excitation-side spectroscope 2 and applied to the sample solution in the sample cell 3. The electrons excited by the energy of this light generate fluorescence when transitioning to the ground state again. This light is taken into the fluorescence side spectroscope 4 installed in the direction perpendicular to the optical axis of the excitation light, and only the target wavelength is photomultiplier tube 5.
To introduce. Light energy is converted into an electric signal by the photomultiplier tube 5, and an output corresponding to the fluorescence intensity is displayed on the recorder 6.

Since substances that emit fluorescence are limited to uranium U and some lanthanoids, substances that do not emit fluorescence need to be converted into fluorescent substances by a chemical reaction for analysis. This fluorometric method is an analysis method that is very sensitive to the properties of the solution (presence or absence of coexisting substances, etc.),
In the measurement, it is necessary to pay sufficient attention to the pretreatment of the sample.

[0005]

The fluorescence photometric method has a higher sensitivity than the absorptiometry method and the substances that emit fluorescence are limited, so that the target component can be selectively measured. However, this fluorescence spectrum is often affected by the quenching effect of coexisting substances and the properties of the sample solution.

[0006] For example, when a sample solution is irradiated with excitation light of a specific wavelength, if a substance that absorbs the excitation light coexists, the energy of light consumed for excitation of the fluorescent substance varies depending on the amount of the substance present. However, the fluorescence intensity also changes accordingly.

Similarly, when a substance that absorbs similarly generated fluorescence coexists, the fluorescence intensity varies depending on the amount of the substance present. Therefore, in order to obtain an accurate analysis value using the fluorometric method, there is a problem that a pretreatment for removing the substance causing such a quenching action is necessary.

An object of the present invention is to provide an apparatus and method capable of solving the above problems, that is, a fluorometric apparatus and method capable of performing fluorescence analysis without pretreatment for removing interfering substances. And

[0009]

Optical axis mobile fluorometer measuring apparatus according to the present invention SUMMARY OF THE INVENTION is an apparatus for analyzing the dissolved liquid sample fluorometrically, the sample cell to put the solution sample, from a light source an excitation side spectroscope for dispersing the light to a particular wavelength, the excitation light side optical fiber for transmitting light from the excitation side spectroscope to specimen cell
When a fluorescent-side optical fiber for transmitting the fluorescence generated in the specimen cell to the fluorescence side spectroscope was dispersed by fluorescence side spectroscope fluorescence
There is input to include a photomultiplier tube to be output to the recorder
It Then, it is necessary to change the optical path length in the sample cell.
Ku, at least one of the optical axes of the fluorescent optical fiber of the excitation light side optical fiber, it moves to the sample cell.
Or, to change the optical path length in the sample cell,
Sample cell, to at least one of the optical axes of the fluorescent optical fiber of the excitation light side optical fiber, it moves. Fluorescence spectrophotometric method according to the present <br/> invention is a method of analyzing a soluble liquid sample fluorometrically, the excitation side spectroscope for dispersing the light from the light source to a specific wavelength, excitation side the excitation light side optical fiber for transmitting light from the spectroscope to the sample cell, fluorescent-side optical fiber for transmitting the fluorescence generated in the specimen cell to the fluorescence side spectroscope
When, to enter the fluorescence dispersed by fluorescence side spectroscope, using a device comprising a photomultiplier tube to be output to the recorder, to move the optical axis of the excitation Okoshiko side optical fiber to the sample cell by, or by the optical axis of the fluorescent optical fiber to move with respect to the sample cell, the relationship between the optical path length and the fluorescence intensity in the sample cell determined Me, the optical path length in the sample cell in the horizontal axis, vertical axis to create a relationship view taken fluorescence intensity, this relationship diagram, by obtaining a fluorescence intensity when the optical path length in the sample cell is 0, the fluorescence absorption or excitation light by the coexisting materials in the sample cell Obtain the fluorescence intensity under the condition without absorption. Also, the position of the optical fiber on the excitation light side is moved.
In this case, keep the position of the optical fiber on the fluorescent side constant and
When moving the position of the fiber,
Keep the position of the bar constant. In addition, as a coexisting substance in the sample cell
To obtain the fluorescence intensity under the condition that there is no absorption of fluorescence by
The optical axis of the excitation light passing through the solution sample is
Go to a direction away from the side surface of the sample cell Le there is bus, trial
Change the optical path length of fluorescence from the side of the material cell to the fluorescence measurement position
The optical axis movement type fluorescence photometric device is used. Or sample
Fluorescence under the condition that excitation light is not absorbed by coexisting substances in the cell
Measurement position of fluorescence emitted from solution sample to obtain light intensity
The fluorescence side optical fiber in the direction in which the
Excitation from edge of sample cell to fluorescence measurement position
An optical axis movement type fluorescence spectrophotometer in which the optical path length of light changes.
It

That is, according to the present invention, in an apparatus and method for analyzing a sample solution by a fluorescence method, the optical axis of the excitation light passing through the sample solution and the measurement position of the fluorescence generated from the sample can be freely changed. The system and the pretreatment operation to remove the coexisting substance that interferes with the fluorescence measurement by applying the optical axis movement type fluorescence spectrophotometer equipped with the method (program) that can correct the quenching effect by the coexisting substance from the measurement data It was unnecessary.

Therefore, it operates as follows. The present invention is
In a fluorescence spectrophotometer, when analyzing a sample solution that absorbs excitation light and fluorescence due to coexisting substances, the optical axis is moved, multiple data are acquired, and by analyzing them, quenching by coexisting substances is performed. Correct the effect.

Conventionally, when an unknown sample is measured with a fluorescence spectrophotometer, it is necessary to confirm the presence or absence of a substance that causes a quenching action in the sample, and pretreatment for removing the substance when these substances coexist. Although an operation was required, the use of the apparatus and method of the present invention made it possible to eliminate the above operation and obtain an analytical value quickly and with higher reliability.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION An embodiment of the present invention is shown in FIGS.
Shown in. FIG. 1 shows an overall configuration diagram of an optical axis movement type fluorescence photometric device of the present invention. The basic configuration of the present invention includes a light source 1,
Excitation side spectroscope 2, sample cell 3, fluorescence side spectroscope 4,
It is composed of a photomultiplier tube 5, a recorder 6 and optical fibers 7 and 8 for arbitrarily selecting an optical axis.

The excitation light side optical fiber 7 is for transmitting light from the excitation side spectroscope to the sample cell, and a lens for collimating the light is installed at the tip of the exit side. A laser was used as a light source to increase the intensity of excitation light.

FIG. 2 is a detailed view of the sample cell section. The excitation light side optical fiber 7 and the fluorescence side optical fiber 8 are installed vertically centering on the sample cell. These optical fibers can be moved so that their irradiation position and photometric position can be arbitrarily selected.

In FIG. 3, the position of the excitation light side optical fiber 7 is made constant (that is, y in FIG. 2 is made constant), and the position of the fluorescence side optical fiber 8 is moved (that is, x in FIG. 2 is changed). A conceptual diagram for measuring fluorescence is shown, and the measurement results are shown in FIG.

When the position of the optical fiber 8 on the fluorescence side is farther from the exit of the excitation light (for example, when it moves from position a to position b in FIG. 3), the fluorescence intensity tends to decrease. It is considered that this is because the substance coexisting in the sample absorbs the excitation light, and the intensity of the excitation light reaching the measurement position of the fluorescence decreases.

Therefore, the vertical axis of the graph is the fluorescence intensity, and the horizontal axis is the optical path length of the excitation light from the end of the sample cell to the fluorescence measurement position (that is, the optical path length corresponding to x in FIG. 2), and the decreasing straight line Then, by obtaining the point where the vertical axis intersects, the fluorescence intensity can be obtained under the condition that the coexisting substance does not absorb the excitation light.

In FIG. 4, the position of the optical fiber 8 on the fluorescence side is kept constant (that is, x in FIG. 2 is kept constant), and the position of the optical fiber 7 on the excitation light side is moved (that is, y in FIG. 2 is changed). A conceptual diagram for measuring fluorescence is shown, and the measurement results are shown in FIG.

The fluorescence intensity tends to decrease as the optical axis of the excitation light becomes farther from the side surface of the sample cell where the fluorescence-side optical fiber is located (for example, in FIG. 4, it moves from position c to position d).

This is also considered to be because the substance coexisting in the sample absorbs the fluorescence, and in the same manner as above, the vertical axis represents the fluorescence intensity,
The horizontal axis indicates the optical path length of fluorescence from the side surface of the sample cell (that is, FIG.
, The optical path length corresponding to y), and the decreasing straight line,
If the intersection with the vertical axis is obtained, the fluorescence intensity can be obtained under the condition that there is no fluorescence absorption by the coexisting substance.

Therefore, by performing the above operation, even if a substance that absorbs excitation light or fluorescence coexists in the sample solution, its influence can be corrected. In the above-described embodiment, the method of moving the optical axis by moving the position of the excitation light side optical fiber 7 or the position of the fluorescence side optical fiber 8 is selected, but the property of the sample, the device There is also a method of changing the optical path length by moving the sample cell depending on the configuration conditions.

[0023]

Since the present invention is constructed as described above, it has the following effects. (1) The pretreatment operation for removing coexisting interfering substances becomes unnecessary. Therefore, the analysis work can be speeded up and the analysis waste liquid can be reduced. (2) The quenching effect of coexisting substances can be corrected. Therefore, a more reliable analysis value can be obtained. (3) The optical path length can be freely set. Therefore, the measurement range can be easily changed.

[Brief description of drawings]

FIG. 1 is an overall configuration diagram of an optical axis movement type fluorophotometer according to a first embodiment of the present invention.

FIG. 2 is a detailed view of a sample cell unit.

FIG. 3 is an explanatory diagram of a sample cell unit in which a fluorescent side optical fiber 8 is moved.

FIG. 4 is an explanatory view of a sample cell unit in which an optical fiber 7 on the excitation light side is moved.

FIG. 5 is a diagram showing the relationship between fluorescence intensity and the optical path length of excitation light.

FIG. 6 is a diagram showing a relationship between fluorescence intensity and an optical path length of fluorescence.

FIG. 7 is an overall configuration diagram of a conventional fluorescence photometric device.

[Explanation of symbols]

1 ... light source, 2 ... Excitation-side spectrometer, 3 ... Sample cell, 4 ... Fluorescence side spectroscope, 5 ... Photomultiplier tube, 6 ... recorder, 7 ... Excitation light side optical fiber, 8 ... Fluorescent side optical fiber.

─────────────────────────────────────────────────── ─── Continuation of front page (58) Fields surveyed (Int.Cl. ⁷ , DB name) G01N 21/00-21/61 G01N 21/62-21/74 PATOLIS

Claims (7)

(57) [Claims] The method according to claim 1] solution sample in the device for analyzing fluorometrically, the sample cell to put the solution sample, the excitation side spectroscope for dispersing the light of the light source or al to a particular wavelength, before Symbol excitation side spectroscope the excitation-side fiber-optic transmitting pressurized et al light to the sample cell Le, the fluorescent side optical Fiber for transmitting fluorescence generated in the previous SL in the sample cell Le to the fluorescent side spectroscope before Symbol fluorescence side spectroscope in spectrally fluorescence is input, recorder
Comprises a photomultiplier tube to be output to said order to change the optical path length in the sample cell, at least between the optical axis of the <br/> excitation light side optical Fiber the fluorescent side optical Fiber optical axis on the other hand but the optical axis mobile fluorometer measuring device according to claim <br/> Rukoto to move against to the sample cell. 2. A method solution sample in the device for analyzing fluorometrically, the sample cell to put the solution sample, the excitation side spectroscope for dispersing the light of the light source or al to a particular wavelength, before Symbol excitation side spectroscope the excitation-side fiber-optic transmitting pressurized et al light to the sample cell Le, the fluorescent side optical Fiber for transmitting fluorescence generated in the previous SL in the sample cell Le to the fluorescent side spectroscope before Symbol fluorescence side spectroscope in spectrally fluorescence is input, recorder
Comprises a photomultiplier tube to be output to, so as to change the optical path length within the sample cell, the sample cell Le is, the optical axis of the fluorescence side optical Fiber optical axis of the excitation light side light Fiber optical axis mobile fluorometer measuring device according to claim move to <br/> Rukoto to at least one of. 3. A method of analyzing a solution sample fluorometrically, the excitation side spectroscope for dispersing the light of the light source or al to a particular wavelength, the light of the excitation side spectroscope or al the sample cell Le the excitation light side fiber-optic transmitting and fluorescent side optical Fiber for transmitting fluorescence generated in the sample cell Le to the fluorescent side spectrometer, type the fluorescence dispersed by the fluorescent side spectroscope, the recorder out <br/> using a device having a photomultiplier tube to force, by the optical axis of the front Symbol excitation light side light Fiber moved against the sample cell Le, or light of a fluorescent side optical Fiber by the axial movement against the sample cell Le obtains the relationship between the optical path length and the fluorescence intensity in the sample cell, the optical path length in the sample cell in the horizontal axis, the relationship view taken fluorescence intensity on the vertical axis create, by pre-Symbol relationship diagram, the fluorescence intensity when the optical path length in the sample cell is zero determined The Rukoto, fluorescence spectrophotometric method characterized by obtaining a fluorescence intensity at the absorption without conditions fluorescence absorption or excitation light by the coexisting materials in the sample cell. 4. The position of the optical fiber on the pumping light side is moved.
If so, keep the position of the optical fiber on the fluorescent side constant.
The optical axis shift type fluorescence intensity according to claim 1, wherein
measuring device. 5. The position of the optical fiber on the fluorescent side is moved.
If so, keep the position of the optical fiber on the pumping light side constant
The optical axis shift type fluorescence intensity according to claim 1, wherein
measuring device. 6. An apparatus for analyzing a solution sample by a fluorometric method
In the above, a sample cell for containing the solution sample, an excitation side spectroscope for separating light from a light source into a specific wavelength, and an excitation side for transmitting light from the excitation side spectroscope to the sample cell.
The optical fiber on the originating side and the fluorescence generated in the sample cell are transmitted to the spectroscope on the fluorescence side.
The fluorescence side optical fiber and the fluorescence dispersed by the fluorescence side spectroscope are input, and the recorder
A photomultiplier for output to the sample cell and no fluorescence absorption by coexisting substances in the sample cell
Excitation to pass through the solution sample to obtain the fluorescence intensity at
The optical axis of the light emission is the sample cell where the fluorescent side optical fiber is located.
Move away from the side of the sample cell
To change the optical path length of the fluorescence from the
A characteristic optical axis movement type fluorescence photometric device. 7. An apparatus for analyzing a solution sample by a fluorometric method
In the above, a sample cell for containing the solution sample, an excitation side spectroscope for separating light from a light source into a specific wavelength, and an excitation side for transmitting light from the excitation side spectroscope to the sample cell.
The optical fiber on the originating side and the fluorescence generated in the sample cell are transmitted to the spectroscope on the fluorescence side.
The fluorescence side optical fiber and the fluorescence dispersed by the fluorescence side spectroscope are input, and the recorder
A photomultiplier tube for outputting the light to the sample cell, and the excitation light is not absorbed by the coexisting substance in the sample cell.
Generated from the solution sample in order to obtain the fluorescence intensity under the conditions
In front of the fluorescence measurement position away from the excitation light emission port
The optical fiber on the fluorescence side moves, and the fluorescent fiber is moved from the end of the sample cell.
Characterized by changing the optical path length of the excitation light up to the light measurement position
An optical axis movement type fluorescence spectrophotometer.