JP5093380B2 - Chromatographic quantitative measurement method - Google Patents

Description

  The present invention relates to a chromatographic quantitative measurement method, and more particularly to a chromatographic quantitative measurement method that performs optical signal detection to measure the concentration of an analyte.

  Hereinafter, a conventional chromatographic quantitative measurement apparatus will be described.

  FIG. 11 is a schematic configuration diagram of a conventional chromatographic quantitative measurement apparatus. FIG. 11 (a) is a schematic block diagram of the measuring apparatus using a spectrophotometer, and FIG.11 (b) is a block diagram of a chromatography test piece.

  In FIG. 11A, the light beam 33 emitted from the lamp 1 is incident on the diffraction grating 3 through the reflector 2. A light wavelength of the light beam 33 incident on the diffraction grating 3 is selected, and the light beam 33 is further narrowed by the opening 4 and then incident on the glass plate 5. The light beam 33 reflected from the glass plate 5 is received as the reference light 6 by the first photomultiplier tube 7. On the other hand, the light beam 33 transmitted through the glass plate 5 irradiates a part of the chromatographic test piece 8 and the scattered light 9 from the chromatographic test piece 8 is received by the second photomultiplier tube 10. The output signals of the first photomultiplier tube 7 and the second photomultiplier tube 10 are each subjected to Log conversion, and the Log conversion value of the first photomultiplier tube 7 is used to calculate the Log of the second photomultiplier tube 10. The converted value is subtracted and output as an absorbance signal.

  As shown in FIG. 11 (b), the chromatographic test piece 8 includes a test solution adding section 11 for adding a test solution, a development layer 14 for developing the test solution, and a part of the development layer 14. The reagent immobilization unit 13 formed by immobilizing the reagent to the measurement target in the solution to be inspected and the labeling reagent that can be eluted by the development of the solution to be inspected are held in the other part of the development layer 14. The labeling reagent holding part 12 formed in this way, and the dough part 15 which is a part obtained by removing the reagent fixing part 13 and the labeling reagent holding part 12 from the development layer 14 are configured.

  The operation of the chromatographic quantitative measurement apparatus configured as described above will be described with reference to FIG.

  First, when a test solution is added to the test solution addition unit 11, the test solution is developed on the development layer 14. At this time, when the test solution reaches the labeling reagent holding unit 12, it specifically binds to the analyte contained in the test solution while eluting the labeling reagent. Subsequently, the bound substance is immobilized in the reagent immobilization unit 13, and the remaining labeling reagent that is not immobilized flows out downstream of the development layer 14 without being immobilized.

  Therefore, in order to measure the concentration of the analyte contained in the test solution, the reagent immobilization unit 13 and the dough part 15 of the chromatographic test piece 8 are immobilized by the spectrophotometer from the absorbance of the dough part 15. The concentration of the analyte can be obtained by subtracting the absorbance of the unit 13 to detect the absorbance signal and converting it from a known calibration curve.

Japanese Patent Laid-Open No. 8-240591

  However, in the conventional chromatographic quantitative measurement apparatus, the reagent immobilization unit 13 of the chromatographic test piece 8 slowly develops the development layer 14 with the solution to be inspected, so that the value of the detection signal gradually increases with time. Will change. In other words, in order to obtain a more stable measurement result, it is important to manage the measurement time, so there is no function to manage the time in measurement using a conventional spectrophotometer. In addition, it is necessary for the measurer to perform time management manually, and there is a problem that the measurement work becomes troublesome.

  Further, depending on the state of the solution to be inspected and the chromatographic test piece 8, there may be a test piece that hinders normal measurement. In measurement using a conventional spectrophotometer, Since there is no function for detecting the state of the photographic test piece 8, the measurer has to bother to manually determine whether it is good or bad, and the measurement work is troublesome.

  Further, since the labeling reagent remains in the labeling reagent holding part 12 of the chromatographic test piece 8 even after elution, it is necessary to reduce the influence of the residual labeling reagent in order to improve the accuracy of quantitative measurement. However, in the measurement using a conventional spectrophotometer, since there is no function to identify the residual labeling reagent, it is necessary for the measurer to manually identify it, and there is a problem that the measurement work becomes troublesome. It was.

  The present invention has been made in order to solve such problems. Time management until measurement, the quality of the test piece, the correction of the quantitative measurement is automatically performed, and the operability of the apparatus is facilitated. An object of the present invention is to provide a chromatographic quantitative measurement method.

  In order to solve this problem, the chromatographic quantitative measurement method of the present invention includes a development layer for developing a test solution, and a reagent for the measurement object in the test solution is immobilized on a part of the development layer. A reagent immobilization part formed by the above, a labeled reagent holding part formed by holding a labeling reagent that can be eluted by the development of the solution to be inspected in another part of the development layer, and the development layer. The chromatographic test piece provided with the reagent immobilization part and the dough part excluding the labeling reagent holding part is irradiated with a light beam emitted from a light source, and transmitted light from the chromatography test piece. Alternatively, the present invention relates to a chromatographic quantitative measurement method for performing optical signal detection using reflected light and quantitatively measuring the concentration of an analyte contained in the test solution from the signal. From the detection signal obtained by irradiating a light beam to the downstream end of the dough on the chromatography test piece to which the test solution is added, the addition amount of the test solution is insufficient, and the chromatography test piece The method includes a step of determining a development failure.

  Further, the chromatographic quantitative measurement method of the present invention is characterized in that, in the chromatographic quantitative measurement method, the light beam is scanned from the upstream end to the downstream end of the dough part on the chromatographic test piece. Is.

  The chromatographic quantitative measurement method according to the present invention includes a development layer for developing a solution to be tested, and a reagent immobilization formed by immobilizing a reagent for a measurement object in the solution to be tested on a part of the development layer. A labeling reagent holding part formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, the reagent immobilization part from the development layer, and A chromatography test piece provided with a dough portion that is a portion excluding the labeling reagent holding portion is irradiated with a light beam emitted from a light source, and transmitted light or reflected light from the chromatography test piece is used. In the chromatographic quantitative measurement method for performing optical signal detection and quantitatively measuring the concentration of an analyte contained in the test solution from the signal, the chromatograph A step of adding a test solution to the test piece, and a step of measuring the concentration of the analyte contained in the test solution after a predetermined time after detecting the development of the test solution at an arbitrary place; As a result, it is no longer necessary for the measurer to manually manage the time.

  Further, in the chromatographic quantitative measurement method according to the present invention, in the chromatographic quantitative measurement method, the development of the solution to be tested is detected by detecting a change in a detection signal generated by elution of the labeling reagent. Therefore, it is not necessary for the measurer to manually manage the time. In addition, since the elution of the labeling reagent is detected and the measurement is performed, it is possible to distinguish from the used test piece in which the labeling reagent has already been eluted.

  Further, the chromatographic quantitative measurement method according to the present invention is the chromatographic quantitative measurement method, wherein the test solution is added to the chromatography test piece, and the development of the test solution is performed from the labeling reagent holding unit to the reagent. Since the detection is performed in the area up to the immobilization unit, it is not necessary for the measurer to manually manage the time. In addition, since the elution of the labeling reagent is detected and the measurement is performed, it is possible to distinguish from the used test piece in which the labeling reagent has already been eluted.

  Further, the chromatographic quantitative measurement method according to the present invention is the above-mentioned chromatographic quantitative measurement method, wherein at least one of temperature and humidity is monitored, and a predetermined fixed time during which the concentration measurement of the analyte is performed is performed. Since the correction is made, the influence of the ambient temperature and humidity on the change in the development speed of the solution to be inspected on the chromatographic test piece can be reduced.

  Further, in the chromatographic quantitative measurement method according to the present invention, in the chromatographic quantitative measurement method, while the development of the solution to be tested is detected, the light source is turned on and off alternately. It is possible to prevent the performance deterioration of the chromatographic test piece accompanying the temperature rise of the laser beam irradiation part.

  Further, in the chromatographic quantitative measurement method according to the present invention, in the chromatographic quantitative measurement method, the light source is turned off until immediately before detection of development of the solution to be inspected. It is possible to prevent the performance deterioration of the chromatographic test piece accompanying the temperature rise of the laser beam irradiation part.

  In addition, the chromatographic quantitative measurement method according to the present invention is such that, in the chromatographic quantitative measurement method, the output of the light source is made smaller than when measuring the concentration of the analyte while the development of the solution to be tested is detected. In addition, it is possible to prevent the deterioration of the performance of the chromatographic test piece accompanying the increase in the temperature of the laser light irradiation portion on the chromatographic test piece.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method, optical signal detection is performed, and the concentration of the analyte contained in the test solution is quantitatively measured from the signal. Since it includes a step of adding a test solution to the luffy test piece, and a step of detecting the development speed after the addition of the test solution and judging the quality of the chromatographic test piece from the development speed. In addition, it is possible to determine a defect such as an abnormal clogging of a chromatographic test piece.

  Further, the chromatographic quantitative measurement method according to the present invention is the above-described chromatographic quantitative measurement method, wherein the development speed is generated by an outflow of the labeling reagent accompanying the development of the solution to be inspected on the chromatographic test piece. Since the time change of the value of γ is detected and calculated, it is possible to determine a defect such as an abnormal clogging of the chromatographic test piece.

  Further, the chromatographic quantitative measurement method according to the present invention is the detection signal generated in the chromatographic quantitative measurement method, wherein the development speed is generated by elution of a labeling reagent accompanying the development of the solution to be tested on the chromatographic test piece. Since the light beam is scanned so as to keep the rising value constant, and the calculation is made from the scanning speed of the light beam, it is possible to determine a defect such as an abnormal clogging of the chromatographic test piece.

  Further, the chromatographic quantitative measurement method according to the present invention is the chromatographic quantitative measurement method, wherein at least one of ambient temperature and humidity at the time of development of the solution to be tested on the chromatographic test piece is measured. From the result, since the discriminating value of the development speed for determining the quality of the chromatography test piece is corrected, the erroneous determination of the quality due to the influence of temperature and humidity can be prevented.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method for performing optical signal detection and quantitatively measuring the concentration of the analyte contained in the test solution from the signal, the test solution Since the step of discriminating the type of the solution to be inspected is included from the detection signal in the dough on the chromatography test piece to which is added, the type of the solution to be inspected added to the chromatographic test piece is discriminated. Can do.

  Further, in the chromatographic quantitative measurement method according to the present invention, in the chromatographic quantitative measurement method, since the dough part for measuring the detection signal is located downstream in the development direction from the reagent immobilization part, the downstream of the reagent immobilization part. Compared to the side fabric portion, it is possible to suppress erroneous determination of the type of the solution to be inspected due to the influence of the labeling reagent that tends to remain in the upstream fabric portion.

  Further, the chromatographic quantitative measurement method according to the present invention is such that, in the chromatographic quantitative measurement method, a calibration curve suitable for the test solution can be selected in advance, so when measuring a plurality of types of test solutions, There is no need for the user to manually input the type of solution to be inspected into the apparatus, and automatic measurement is possible.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method for performing optical signal detection and quantitatively measuring the concentration of the analyte contained in the test solution from the signal, the test solution From the detection signal obtained by irradiating the downstream end portion of the dough on the chromatography test piece to which light is added with a light beam, the lack of the addition amount of the solution to be tested and the poor development of the chromatography test piece are determined. Since the process is included, it is possible to detect a development failure of the chromatographic test piece that occurs due to an insufficient amount of the solution to be inspected added to the chromatographic test piece or clogging.

  Further, in the chromatographic quantitative measurement method according to the present invention, in the chromatographic quantitative measurement method, the light beam is scanned from the upstream end to the downstream end of the dough portion on the chromatography test piece. In order to detect insufficient addition of the solution to be inspected and poor development of the chromatographic test piece, it does not require a new light source, and has the effect of suppressing the increase in size and price of the device due to the addition of functions.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method for performing optical signal detection and quantitatively measuring the concentration of the analyte contained in the test solution from the signal, the reagent fixation The detection signal at the reagent immobilization unit when the detection signal at the position downstream of the unit in the development direction and at the position where there is no influence of the reagent immobilization unit is used as the reference signal. Since the process is included, the influence of the absorbance measurement error due to the labeling reagent that tends to remain in the upstream fabric portion can be suppressed as compared with the downstream fabric portion of the reagent immobilization portion.

  Further, the chromatographic quantitative measurement method according to the present invention is the chromatographic quantitative measurement method, wherein the concentration measurement detection signal is an average value of values before and after the extreme value of the reagent immobilization unit, and becomes the reference value. Since the detection signal is the average value of the values before and after the position where the reagent immobilization unit is not influenced by the downstream side of the reagent immobilization unit in the development direction of the test solution, Even when electrical noise is added, it is possible to reduce the influence on the calculation result for obtaining the concentration of the analysis object.

  In the chromatographic quantitative measurement method according to the present invention, the concentration measurement detection signal is an intermediate value between values before and after the extreme value of the reagent immobilization unit, and becomes the reference value. Since the detection signal is set to an intermediate value before and after the position where there is no influence of the reagent immobilization unit on the downstream side in the development direction of the solution to be inspected from the reagent immobilization unit, Even when static noise is added, the influence on the calculation result for obtaining the concentration of the analysis object can be made smaller than when the average value is used.

  Further, the chromatographic quantitative measurement method according to the present invention is the chromatographic quantitative measurement method, wherein the value before and after the extreme value of the detection signal in the reagent immobilization part is compared, and when the difference exceeds the discriminant value, Since it is determined that the test piece has poor performance, erroneous measurement due to immobilization unevenness of the labeling reagent in the reagent immobilization section, scratches on the surface of the chromatographic test piece, and the like can be avoided.

  Further, the chromatographic quantitative measurement method according to the present invention is the chromatographic quantitative measurement method, wherein the values before and after the position where there is no influence of the reagent immobilization part are compared downstream of the reagent immobilization part in the development direction. When the difference exceeds the discriminant value, it is determined that the performance of the chromatographic test piece is poor, so the development of the solution to be inspected due to clogging in the dough part, scratches on the surface of the chromatographic test piece The wrong measurement can be avoided.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method for performing optical signal detection and quantitatively measuring the concentration of the analyte contained in the test solution from the signal, Since the step of performing the measurement by removing the labeled reagent holding part on the chromatography test piece is included, the absorbance measurement value in the labeled reagent holding part is not included, so that the misrecognition of the absorbance peak position does not occur and is normal. This makes it possible to detect the concentration of a simple analysis object.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method, optical signal detection is performed, and the concentration of the analyte contained in the test solution is quantitatively measured from the signal. The region where the value of the detection signal on the luffy test piece is flat is regarded as the region of the labeling reagent holding part, so that misrecognition of the absorbance peak position does not occur and normal analyte concentration detection is possible Become.

  Further, the chromatographic quantitative measurement method according to the present invention is the chromatographic quantitative measurement method, wherein the width of the region where the value of the detection signal on the chromatographic test piece is flat is calculated, and the width is defined as the labeling reagent holding unit. Therefore, the amount of the labeling reagent retained can be confirmed, and the poor performance of the chromatographic test piece can be discriminated.

  Further, the chromatographic quantitative measurement method according to the present invention is such that, in the chromatographic quantitative measurement method, a value at which the detection signal on the chromatographic test piece becomes flat is detected, and the amount of the residual labeling reagent is confirmed from the value. Therefore, it can be confirmed whether the labeling reagent has flowed normally.

  Further, the chromatographic quantitative measurement method according to the present invention is formed by developing a developing layer for developing the solution to be tested, and immobilizing a reagent for the measurement object in the solution to be tested on a part of the developing layer. A reagent immobilization unit, a labeled reagent holding unit formed by holding a labeling reagent that can be eluted by the development of the solution to be tested in another part of the development layer, and the reagent immobilization unit from the development layer And irradiating a chromatography test piece having a dough part which is a part excluding the labeling reagent holding part with a light beam emitted from a light source, and using transmitted light or reflected light from the chromatography test piece In the chromatographic quantitative measurement method for performing optical signal detection and quantitatively measuring the concentration of the analyte contained in the test solution from the signal, It recognizes the rising and falling, since to include the step of determining an extreme value of the detection signal, without generating false recognition of the absorbance peak position, it is possible to density detection normal analyte.

  Further, the chromatographic quantitative measurement method according to the present invention, in the chromatographic quantitative measurement method, recognizes the rise and fall of the detection signal, calculates an interval between the rise and fall, Since the size is compared with the width of the prescribed reagent immobilization part, the width of the reagent immobilization part can be confirmed. Accordingly, it is possible to determine the poor performance of the chromatographic test piece.

1 is a schematic configuration diagram of a chromatographic quantitative measurement apparatus according to Embodiment 1 of the present invention. The figure of the change of the light absorbency accompanying development of the solution to be examined in Embodiment 1 of the present invention Diagram of absorbance change in standby state in Embodiment 2 of the present invention The figure of the light absorbency measurement result of the chromatography test piece by the difference in the to-be-tested solution in Embodiment 3 of this invention The figure which shows expansion | deployment of the to-be-tested solution of the chromatography test piece in Embodiment 4 of this invention. The figure which shows the light-absorbency difference measurement of the chromatography test piece in Embodiment 5 of this invention. The figure which shows the electrical noise of the light absorbency signal of the chromatography test piece in Embodiment 6 of this invention The figure which shows the optical noise of the light absorbency signal of the chromatography test piece in Embodiment 7 of this invention. The figure which shows the light absorbency including the labeling reagent holding | maintenance part of the chromatography test piece in Embodiment 8 of this invention. The figure which shows the light absorbency peak value detection method of the chromatography test piece in Embodiment 9 of this invention. Schematic configuration diagram of conventional chromatography test equipment

  Embodiments of the present invention will be described below.

(Embodiment 1 (reference example))
Hereinafter, Embodiment 1 of the present invention will be described with reference to FIGS. 1 and 2.

  FIG. 1 is a schematic configuration diagram of a chromatographic quantitative measurement apparatus according to Embodiment 1 of the present invention. FIG. 1A is a schematic configuration diagram of a measuring apparatus using a spectrophotometer, and FIG. 1B is a configuration diagram of a chromatographic test piece.

  In FIG. 1A, light emitted from the semiconductor laser 17 is converted into a parallel beam by passing through a collimating lens 18. The parallel beam passes through the opening 4 and enters the beam splitter 19. Here, a part of the light beam reflected by the beam splitter 19 is received by the first photodiode 20 as the reference light 6. On the other hand, the remaining light beam that has passed through the beam splitter 19 is condensed only in the long side direction of the chromatographic test piece 8 by the cylindrical lens 21 and irradiated onto the chromatographic test piece 8 as an elliptical beam 16. Further, scattered light 9 is generated from the surface of the chromatographic test piece 8 and is received by the second photodiode 22.

  Next, the output of the first photodiode 20 that has received the reference light 6 and the output of the second photodiode 22 that has received the scattered light 9 are subjected to Log conversion, and the log conversion value of the first photodiode 20 is The Log conversion value of the second photodiode 22 is subtracted and output as an absorbance signal.

  As shown in FIG. 1B, the chromatographic test piece 8 includes a test solution adding part 11 for adding a test solution, a developing layer 14 for developing the test solution, and a part of the developing layer 14. The reagent immobilization unit 13 formed by immobilizing the reagent to the measurement target in the solution to be inspected and the labeling reagent that can be eluted by the development of the solution to be inspected are held in the other part of the development layer 14. The labeling reagent holding part 12 formed in this way, and the dough part 15 which is a part obtained by removing the reagent fixing part 13 and the labeling reagent holding part 12 from the development layer 14 are configured.

  The operation of the thus-configured chromatographic quantitative measurement apparatus will be described with reference to FIG.

  First, when a test solution is added to the test solution addition unit 11, the test solution is developed on the development layer 14. At this time, when the test solution reaches the labeling reagent holding unit 12, it specifically binds to the analyte contained in the test solution while eluting the labeling reagent. Subsequently, the bound substance is immobilized in the reagent immobilization unit 13, and the remaining labeling reagent that is not immobilized flows out downstream of the development layer 14 without being immobilized.

  The concentration of the analyte contained in the solution to be inspected can be obtained by detecting the difference between the absorbance signals of the reagent immobilization part 13 and the dough part 15 of the chromatographic test piece 8 and converting it from a known calibration curve. it can.

  Here, by scanning the chromatographic test piece 8 in the long side direction, it is possible to measure the difference between the absorbance signals of the fabric portion 15 and the reagent immobilization portion 13 by a single light beam. Moreover, the influence of the nonuniformity by the position of the short side direction of the chromatographic test piece 8 is relieved by making a light beam into an ellipse.

  Next, the absorbance measurement will be described.

  FIG. 2 is a diagram showing absorbance measurement of the chromatographic test piece according to Embodiment 1 of the present invention. FIG. 2A shows the state of development of the solution to be inspected on the chromatographic test piece and the irradiation position of the light beam. FIG. 2B shows the change in the absorbance signal with respect to the measurement time.

  The chromatographic test piece 8 is attached to the measuring device, and the test solution 23 is added to the test solution adding section 11. Next, along with the development of the test solution 23, the analysis target contained in the test solution 23 is pushed away while being bound to the eluted labeling reagent, and the bound substance is fixed in the reagent immobilization unit 13. When the absorbance measurement is performed in a state where the light beam 16 is continuously irradiated onto the reagent immobilization unit 13, the absorbance signal 24 rapidly rises and falls due to the passage of the labeling reagent and then gradually rises, and the solution to be tested As it dries, it gradually decreases again.

  Therefore, in order to suppress an error in the absorbance measurement, the light beam 16 is kept in a state of being irradiated between the labeling reagent holding unit 12 and the reagent immobilization unit 13 to develop the solution to be inspected or to elute the labeling reagent. The change in absorbance due to was detected, and measurement was automatically started after a lapse of a certain time from this detection.

  Moreover, the fixed time mentioned above has a possibility of affecting the expansion | deployment speed | velocity | rate of a to-be-tested solution with the temperature and humidity around a measuring apparatus. Therefore, the temperature and humidity were monitored and corrected for a certain period of time from the elution of the labeling reagent accompanying the development of the solution to be inspected until the concentration of the analyte was measured. The operation of the light beam is to alternately turn on and off the light beam while detecting the development of the solution to be inspected. Alternatively, the time for detecting the development of the solution to be inspected is predicted, and the light beam is turned off until just before the expected arrival time. Alternatively, while detecting the development of the solution to be inspected, the output of the light beam was set to be lower than that at the time of measurement.

  As described above, according to the chromatographic quantitative measurement apparatus according to the first embodiment, the test solution is added to the chromatographic test piece, and the development of the test solution is detected. Since the concentration of the contained analyte is measured, there is no need for the measurer to manually manage the time. In addition, since the elution of the labeling reagent is detected and the measurement is performed, it is possible to distinguish from the used test piece in which the labeling reagent has already been eluted.

  Further, according to the chromatographic quantitative measurement apparatus according to the first embodiment, the ambient temperature and humidity are monitored, and the time from the detection of the elution of the labeling reagent until the measurement is performed is corrected. And the influence of humidity on the change in the development rate of the solution to be tested on the chromatographic test piece can be reduced.

  Further, according to the chromatographic quantitative measurement apparatus according to the first embodiment, while detecting the development of the solution to be inspected, the light beam is repeatedly turned on and off alternately, or the time for detecting the development of the solution to be inspected is set. Estimate and turn off the laser until just before the estimated arrival time, or set the laser output lower than the measurement while detecting the development of the solution to be inspected on the chromatographic test piece, or those methods Therefore, it is possible to prevent the deterioration of the performance of the chromatographic test piece accompanying the temperature increase of the laser light irradiation part on the chromatographic test piece.

  In the first embodiment, the detection of the elution of the labeling reagent has been described. However, the same effect can be obtained by detecting the development of the solution to be inspected.

(Embodiment 2 (reference example))
Next, Embodiment 2 of the present invention will be described with reference to FIG.

  FIG. 3 is a diagram showing absorbance measurement of a chromatographic test piece according to Embodiment 2 of the present invention. FIG. 3A shows the development state of the solution to be inspected on the chromatographic test piece and the irradiation position of the light beam. Moreover, FIG.3 (b) is an enlarged view of the part which shows a rapid raise among the light absorbency changes in the standby state in Embodiment 2 of this invention.

  The light beam 16 is made to stand by while being irradiated between the labeling reagent holding unit 12 and the reagent immobilization unit 13. The absorbance signal at this time increases monotonously with the elution of the labeling reagent.

  Therefore, the development rate of the solution 23 to be tested is calculated by obtaining the slope θ of the absorbance signal with time, and the quality of the chromatographic test piece is determined from the development rate. Alternatively, the light beam is scanned so that the increase in absorbance due to elution of the labeling reagent is kept constant, the development speed of the solution to be tested 23 is calculated from the scanning speed, and the performance of the chromatographic test piece is calculated from the development speed. Judge the quality.

  Moreover, the discrimination | determination value of the expansion | deployment speed | velocity | rate was corrected from the result of having measured at least one of the ambient temperature at the time of expansion | deployment of the to-be-tested solution to a chromatography test piece, and humidity.

  Thus, according to the chromatographic quantitative measurement apparatus according to the second embodiment, the development speed after addition of the test solution is detected, and the quality of the chromatographic test piece is judged from the development speed. It is possible to determine a defect such as a clogging abnormality.

  In addition, since the development rate of the test solution is calculated after detecting the time change of the detection signal value that occurs due to the elution of the labeling reagent accompanying the development of the test solution, Defect determination such as clogging abnormality can be performed.

  In addition, the light beam is scanned so that the rising value of the detection signal generated by elution of the labeling reagent accompanying the development of the test solution is kept constant, and the development speed of the test solution is calculated from the scanning speed of the light beam. Therefore, it is possible to determine a defect such as an abnormal clogging of a chromatographic test piece.

  In addition, since the determination value of the development speed is corrected based on the result of measuring at least one of the ambient temperature and humidity at the time of development of the solution to be inspected, it is possible to prevent erroneous determination of quality due to the influence of temperature and humidity.

(Embodiment 3 (reference example))
Hereinafter, Embodiment 3 of the present invention will be described with reference to FIG.

  FIG. 4 is a diagram showing the difference in the absorbance measurement result by the chromatographic test piece according to Embodiment 3 of the present invention depending on the solution to be inspected. FIG. 4 (a) shows the scanning state of the light beam on the chromatographic test piece, and FIG. 4 (b) shows the change in the absorbance signal with respect to the position of the light beam.

  The light beam 16 scans from the upstream fabric portion 15a through the reagent immobilization portion 13 to the downstream fabric portion 15b on the chromatographic test piece 8. The absorbance signal at this time varies depending on the type of the solution to be inspected. For example, in the case of a plasma sample and a whole blood sample, the whole blood sample has a higher absorbance overall. Further, the magnitude of the absorbance in the dough portion 15 is constant regardless of the concentration of the analysis target contained in the solution to be inspected.

  Therefore, the signal detection position for discriminating the type of solution to be inspected is located downstream of the reagent immobilization unit 13, and the absorbance in the dough portion 15 is detected. Compared. Further, the type of the solution to be inspected was discriminated from the magnitude of the absorbance in the dough part 15, a calibration curve corresponding to the type was selected, and the concentration of the analyte contained in the solution to be inspected was converted.

  As described above, according to the chromatographic quantitative measurement apparatus according to the third embodiment, since the type of the solution to be inspected is determined from the detection signal in the fabric portion on the chromatography test piece to which the solution to be inspected is added, the chromatography test The type of the solution to be inspected added to the piece can be discriminated.

  In addition, the fabric portion for measuring the detection signal is located downstream in the development direction from the reagent immobilization portion. Therefore, a label that tends to remain in the upstream fabric portion compared to the downstream fabric portion of the reagent immobilization portion. Misidentification of the type of solution to be inspected due to the influence of the reagent can be suppressed.

  In addition, the type of solution to be inspected is determined from the detection signal of the dough part, and a calibration curve suitable for the solution to be inspected can be selected in advance. Thus, it is not necessary to input the type of solution to be inspected into the apparatus, and automatic measurement is possible.

(Embodiment 4)
Hereinafter, a fourth embodiment of the present invention will be described with reference to FIG.

  FIG. 5 is a diagram showing the flow of the solution to be inspected in the chromatographic test strip according to Embodiment 4 of the present invention.

  FIG. 5A shows a case where a sufficient test solution 23 is added to the test solution adding section 11. The added solution 23 to be inspected expands the labeling reagent holding part 12, the upstream dough part 15a, the reagent immobilization part 13, and the downstream dough part 15b of the chromatography test piece 8, and further the downstream dough part 15b. It reaches the downstream end.

  FIG. 5B shows a case where the test solution 23 added to the test solution adding part 11 is insufficient, and the added test solution 23 is located at the downstream end of the downstream dough part 15b. It has not reached the department.

  Therefore, the downstream end of the downstream fabric portion 15b was irradiated with a light beam, and the value of the detection signal obtained in that case was determined. Further, in order to measure the concentration of the analysis object, the same light beam 16 as that scanned before and after the reagent immobilization unit 13 was further scanned to the downstream end of the downstream fabric portion 15b.

  Thus, according to the chromatographic quantitative measurement apparatus according to the fourth embodiment, from the detection signal obtained by irradiating the downstream end of the dough part on the chromatography test piece to which the solution to be inspected is added with the light beam. Chromatographic test piece generated due to insufficient addition of the test solution added to the chromatographic test piece, clogging, etc. Can be detected.

  In addition, since the light beam was scanned to the downstream end of the dough on the chromatographic test piece, a new light source was required to detect insufficient addition of the solution to be inspected and poor development of the chromatographic test piece. In addition, it has the effect of suppressing the increase in size and price of the device due to the function addition.

(Embodiment 5 (reference example))
The fifth embodiment of the present invention will be described below with reference to FIG.

  FIG. 6 is a diagram showing absorbance measurement of a chromatographic test piece according to Embodiment 5 of the present invention. FIG. 6A shows the state of scanning of the light beam on the chromatographic test piece, and FIG. 6B shows the change in the absorbance signal with respect to the position of the light beam.

  The light beam 16 scans from the upstream fabric portion 15a through the reagent immobilization portion 13 to the downstream fabric portion 15b on the chromatographic test piece 8.

  The magnitude of the absorbance corresponding to the concentration of the analyte is a distance D from a position that is not affected by the absorbance of the labeled reagent immobilized on the reagent immobilization unit 13, that is, a position T where the absorbance signal 24 peaks. Only the absorbance value at the downstream position U can be used as a reference, and a difference value E from the peak position T at that time can be obtained. In other words, the absorbance value at the peak position T includes the absorbance component of the solution to be inspected itself, but this becomes an error in the absorbance measurement of the labeled reagent immobilized on the reagent immobilization unit 13, and thus the absorbance at the position U. By using the value (corresponding to the light absorption component of the solution under test itself) as a reference, the influence of this error can be eliminated. Further, since the reference position is not the upstream side of the reagent immobilization unit 13 but the downstream side U, the absorbance measurement error due to the labeling reagent that tends to remain in the upstream fabric portion 15a (F in FIG. 6B). Part) can be removed.

  As described above, according to the chromatographic quantitative measurement apparatus according to the fifth embodiment, at a position downstream of the reagent immobilization part on the chromatographic test piece in the development direction of the solution to be inspected, there is no influence of the reagent immobilization part. The detection signal at the reagent immobilization unit when the detection signal is used as a reference value is the detection signal of the concentration to be measured, so compared with the downstream dough part of the reagent immobilization unit, the upstream dough part The influence of the absorbance measurement error due to the labeling reagent that tends to remain can be suppressed.

(Embodiment 6 (reference example))
Hereinafter, Embodiment 6 of the present invention will be described with reference to FIG.

  FIG. 7 is a diagram showing absorbance measurement of a chromatographic test piece according to Embodiment 6 of the present invention. FIG. 7A shows a state of scanning of a light beam on a chromatographic test piece, and FIG. 7B shows a state where a steep electrical noise 25 is added to the change in the absorbance signal with respect to the position of the light beam. Is shown.

  The light beam 16 scans from the upstream fabric portion 15a through the reagent immobilization portion 13 to the downstream fabric portion 15b on the chromatographic test piece 8. At this time, the data of the absorbance signal 24 is stored at an interval G at which a smooth change can be sufficiently detected.

  The electrical noise 25 added to the absorbance signal is generated from a power supply (for example, a switching power supply) added to the electric circuit or generated from a circuit that performs digital processing, and is compared with the scanning speed of the light beam 16. Thus, it shows a very steep change.

  Therefore, when the value of the position U on the downstream side by the distance D from the peak positions T and T of the absorbance signal 24 is obtained, the average value of the number data before and after is associated with each position. Further, when the value of the position U downstream from the peak positions T and T of the absorbance signal 24 by the distance D is obtained, the intermediate values of the number data before and after (rearranged in order of the size of each data) for each position. Data value located in the middle of the case).

  The number of data for obtaining the average value and the intermediate value described above is within a range that does not hinder the reading of the smooth change in the absorbance signal 24.

  Thus, according to the chromatographic quantitative measurement apparatus according to the sixth embodiment, the detection signal in the reagent immobilization unit is the average value of the values before and after the extreme value, and the detection signal serving as the reference value is the reagent immobilization. Since the average value before and after the position where there is no influence of the reagent immobilization part is downstream of the development direction of the test solution from the part, even if electrical noise is accidentally added to the detection signal It is possible to reduce the influence on the calculation result for obtaining the concentration of an object.

  The detection signal at the reagent immobilization unit is an intermediate value between the values before and after the extreme value, and the reference detection signal is downstream of the reagent immobilization unit in the direction of development of the solution to be tested. Since it is an intermediate value before and after the position where there is no influence, even when electrical noise is accidentally added to the absorbance signal, the influence on the calculation result to obtain the concentration of the analyte is more than that when using the average value. It can be further reduced.

(Embodiment 7 (reference example))
Hereinafter, a seventh embodiment of the present invention will be described with reference to FIG.

  FIG. 8 is a diagram showing absorbance measurement of a chromatographic test piece according to Embodiment 7 of the present invention. FIG. 8A shows a state of scanning of a light beam on a chromatographic test piece, and FIG. 8B shows a state in which optical noises 31 and 32 are added to the change in absorbance signal with respect to the position of the light beam. Is shown. FIG. 8C shows a change in the normal absorbance signal with respect to the position of the light beam.

  The light beam 16 scans from the upstream fabric portion 15a through the reagent immobilization portion 13 to the downstream fabric portion 15b on the chromatographic test piece 8. At this time, the data of the absorbance signal 24 is stored at an interval G at which a smooth change can be sufficiently detected. Further, the amount of change K before and after the peak position T (interval I) and the amount of change L before and after the downstream position U (interval J) of the absorbance signal 24 obtained by measuring the absorbance with a normal chromatographic test piece 8 in advance are obtained. , K + α, and L + α (α is the tolerance of the noise component) are stored as discriminant values at the respective positions.

  The optical noises 31 and 32 added to the absorbance signal are caused by the labeling reagent immobilization unevenness 26 (commonly referred to as color unevenness) in the reagent immobilization unit 13 or the labeling reagent clogging in the downstream fabric portion 15b. This is due to development unevenness 27 and scratches on the surface of the chromatographic test piece 8. The optical noises 31 and 32 prevent a gradual change in the absorbance signal 24, and depending on the magnitude, normal absorbance measurement is impossible.

  Therefore, the values before and after the peak position T (interval I) of the absorbance signal 24 were compared, and when the difference between the maximum value and the minimum value exceeded the discriminant value, it was determined that the performance of the chromatographic test piece 8 was poor. Further, when the values before and after the position U downstream (distance J) by the distance D from the peak position T of the absorbance signal 24 are compared, and the difference between the maximum value and the minimum value exceeds the discriminant value, the chromatographic test piece 8 It was determined that the performance was poor.

  Thus, according to the chromatographic quantitative measurement apparatus according to the seventh embodiment, in the reagent immobilization part on the chromatographic test piece, the values before and after the extreme value of the detection signal are compared, and when the difference exceeds the discrimination value, Since it was determined that the performance of the chromatographic test piece was poor, erroneous measurement due to immobilization unevenness of the labeling reagent in the reagent immobilization section, scratches on the surface of the chromatographic test piece, and the like can be avoided.

  Also, compare the values before and after the position where there is no influence of the reagent immobilization part on the downstream side of the reagent immobilization part on the chromatographic test piece, and if the difference exceeds the discriminant value, the performance of the chromatographic test piece is poor. Therefore, erroneous measurement due to uneven development of the solution to be inspected due to clogging in the dough portion, scratches on the surface of the chromatographic test piece, and the like can be avoided.

(Embodiment 8 (reference example))
Hereinafter, an eighth embodiment of the present invention will be described with reference to FIG.

  FIG. 9 shows the scanning state of the light beam on the chromatographic test piece according to the eighth embodiment of the present invention and the change of the absorbance signal with respect to the position of the light beam.

  In the labeling reagent holding unit 12, the labeling reagent remains even after passing through the test solution. That is, in the measurement of an analyte having a low concentration, when the light beam 16 is scanned from the upstream side of the labeling reagent holding unit 12, the absorbance of the labeling reagent remaining in the labeling reagent holding unit 12 is the peak position. There is. Further, since the residual labeled reagent in the labeled reagent holding unit 12 is uniformly distributed, the absorbance 28 in this region is a flat signal.

  Therefore, in order to avoid erroneous recognition of the peak position, the light beam 16 was scanned from the position of the upstream fabric portion 15a excluding the labeling reagent holding portion 12, and measurement was started. Alternatively, a flat absorbance signal 28 was detected and distinguished from the absorbance peak position T corresponding to the reagent immobilization unit 13.

  Further, the width H of the labeling reagent holding part 12 was obtained from the width of the flat absorbance signal 28 and compared with a prescribed width. Further, the value of the flat absorbance signal 28 was detected, and the amount of the remaining labeling reagent was determined.

  As described above, according to the chromatographic quantitative measurement apparatus according to the eighth embodiment, since the measurement was performed excluding the labeling reagent holding part on the chromatography test piece during the concentration measurement, the absorbance measurement value in the labeling reagent holding part was Since it is not included, there is no misrecognition of the absorbance peak position, and normal concentration detection of the analyte is possible.

  In addition, the region where the value of the detection signal on the chromatographic test piece is flat is regarded as the region of the labeling reagent holding part, so that no misrecognition of the absorbance peak position occurs and normal analyte concentration detection is possible. It becomes possible.

  In addition, the width of the region where the value of the detection signal on the chromatographic test piece is flat is calculated, and the width is compared with the width of the specified labeling reagent holding part, so the amount of the labeling reagent held can be confirmed, The poor performance of the chromatographic test piece can be identified.

  Further, since the value at which the detection signal on the chromatographic test piece becomes flat is detected and the amount of the residual labeled reagent is confirmed from the value, it can be confirmed whether or not the labeled reagent has flowed normally.

(Embodiment 9 (reference example))
The following describes Embodiment 9 of the present invention with reference to FIG.

  FIG. 10 shows the state of scanning of the light beam on the chromatographic test piece according to the ninth embodiment of the present invention and the change in the absorbance signal with respect to the position of the light beam.

  In the labeling reagent holding unit 12, the labeling reagent remains even after passing through the test solution. That is, in the measurement of an analyte having a low concentration, when the light beam 16 is scanned from the position of the labeling reagent holding unit 12, the absorbance of the labeling reagent remaining in the labeling reagent holding unit 12 may be a peak position. is there.

  Therefore, the rising portion 29 and the falling portion 30 are detected from the change in the slope of the absorbance signal 24, and the maximum position in the region sandwiched between the rising portion 29 and the falling portion 30 is recognized as the peak position T.

  Further, the interval between the rising portion 29 and the falling portion 30 was obtained and compared with the width of the prescribed reagent immobilization portion 13.

  As described above, according to the chromatographic quantitative measurement apparatus according to the ninth embodiment, since the extreme value of the detection signal is obtained by recognizing the rise and fall of the detection signal, erroneous recognition of the absorbance peak position can occur. Therefore, it is possible to detect the concentration of a normal analyte.

  Also, the rise and fall of the detection signal is recognized, the interval between the rise and fall is calculated, and the size of the interval is compared with the width of the specified reagent immobilization unit. The width can be confirmed. Accordingly, it is possible to determine the poor performance of the chromatographic test piece.

  Since the chromatographic quantitative measurement method according to the present invention can detect insufficient development of the chromatographic test piece, which occurs due to insufficient addition of the test solution added to the chromatographic test piece or clogging, It is useful for chromatographic quantitative measurement devices with good operability.

DESCRIPTION OF SYMBOLS 1 Lamp 2 Reflecting plate 3 Diffraction grating 4 Opening part 5 Glass plate 6 Reference light 7,10 Photomultiplier tube 8 Chromatographic test piece 9 Scattered light 11 Test solution addition part 12 Labeling reagent holding part 13 Reagent immobilization part 14 Development Layer 15 Fabric part 15a Upstream fabric part 15b Downstream fabric part 16, 33 Light beam 17 Semiconductor laser 18 Collimating lens 19 Beam splitter 20, 22 Photo diode 21 Cylindrical lens 23 Test solution 24 Absorbance signal 25 Electrical noise 26 Color Unevenness 27 Unevenness of development 28 Flat part 29 Rising part 30 Falling part 31, 32 Optical noise

Claims (2)

A spreading layer for spreading the solution to be tested, a reagent immobilization part formed by immobilizing a reagent for the measurement object in the solution to be tested on a part of the spreading layer, and another one of the spreading layers. A labeling reagent holding part formed by holding a labeling reagent that can be eluted by the development of the solution to be inspected, and a part obtained by removing the reagent immobilization part and the labeling reagent holding part from the development layer. A chromatographic test piece provided with a fabric portion is irradiated with a light beam emitted from a light source, optical signal detection is performed using transmitted light or reflected light from the chromatographic test piece, and from the signal In a chromatographic quantitative measurement method for quantitatively measuring the concentration of an analyte contained in the test solution,
From the detection signal obtained by irradiating a light beam to the downstream end of the dough on the chromatography test piece to which the test solution is added, the addition amount of the test solution is insufficient, and the development of the chromatography test piece Including a step of determining a defect,
A chromatographic quantitative measurement method characterized by the above. The chromatographic quantitative measurement method according to claim 1,
The light beam scans from the upstream end to the downstream end of the dough on the chromatography specimen,
A chromatographic quantitative measurement method characterized by the above.

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