KR100254812B1 - Array-type optode device for the quantitative analysis of electrolytes and their related species in multiple samples - Google Patents

Abstract

PURPOSE: A sample analyzer with an optical senor is provided to simultaneously analyze a lot of samples simultaneously, and analyze the optical sensor membrane solvent composite in such analyzers precisely. CONSTITUTION: Optical sensor membrane solvent composite having a quick response time to analyze a sample is spread evenly on an absorbance measuring substrate(70). An optical sensor measures changes of color in adding the sample. A multiarray type UV detector measures the color change from reacting the sample and the color material, and measures the sample with the optical sensor absorbance measuring substrate composed of ion-selective polymer color material. The ion-selective polymer color material such as sensing material(20) has similar components and form to the sensing material in an ion-selective electrode with optimizing for installing in the analyzer. The absorbance measuring substrate measures absorbance and transmittance on a lot of samples simultaneously. The optical sensor membrane solvent includes chromoionophore, KTpCIPB, ionophore, plasticizer, polymer and solvent.

Description

Optical sensor measuring device and method for quantitative analysis of a large number of samples at the same time

The present invention is to apply a large number of light sensor membrane solution composition to the simultaneous absorbance measurement substrate which is a device that can measure the absorbance (absorbance or optical density (OD) or transmittance in the UV / visible region) at the same time A new device capable of simultaneously measuring analytical samples, an optical sensor film solution composition suitable for application to such a new device, and an analytical method using such device and composition.

According to the present invention, an optical sensor that can be analyzed by two-point calibration can be designed for an uncorrected optical sensor or an individual optical sensor that does not need correction, and a sample (environmental sample, blood, serum, whole blood) to be analyzed using such a device , Biological samples including urine, initial and intermediate chemical processes, or resultant substances, food chemicals, etc. can be quantitatively analyzed.

Recently, the necessity of quantitatively measuring a large number of analytical samples in each field such as medicine, environment, and industry has been raised. Biosamples including blood and urine such as environmental samples, serum or whole blood, initial, intermediate or resultant chemical processes For food chemicals, it is necessary to take tens to hundreds of samples and measure them quantitatively in a short time. Conventionally, ion-selective electrode (ISE) is mainly used for quantitative analysis of these samples, but recently, various types of sensors are used according to the material to be quantitatively analyzed.

Ion-selective electrodes, which were first developed in the mid-1960s, have been rapidly developed and applied in the last 20 years. Until now, research on ion-selective electrodes for hundreds of dozens of ionic species has been reported. These ion-selective electrodes have been used in food chemistry and industrial chemistry such as biological fluid analysis and fermentation, And it has been widely applied to environmental analysis. The ion-selective electrode, which is being actively researched recently, is an ion-selective membrane electrode that mounts a membrane having a selectivity for a specific ion to the electrode. These ion-selective membrane electrodes have relatively good selectivity and are not affected by the color or turbidity of the sample, eliminating the need for a pretreatment step when analyzing the sample, and being relatively easy to manufacture and inexpensive compared to other analytical methods. Have Accordingly, ion-selective membrane electrodes have been used in various fields, and are generally used for analyzing various ionic species in the blood.

In the mid-1980s, Simon et al.'S team tried to change the analytical signal to color change in the membrane instead of measuring the electrical signal like the ion-selective electrode. It was found that the addition of an optical sensor membrane solvent enables the measurement of the amount of ions in the sample solution by the change in absorbance.

Component Comparison Table of Typical Ion-Selective Membrane Electrodes and Optical Sensors (wt%) Substance K ⁺ Na ⁺ Ion Selective Membrane Electrodes (ISE) Light sensor Ion Selective Membrane Electrodes (ISE) Light sensor PVC 33 33 33 33 DOS ¹⁾ 66-65 66-65 66-65 66-65 Valinomycin One One KTpClPB ²⁾ 0-1 0.5 0-1 0.5 ETH 2120 ³⁾ One 0.5 ETH 5294 ⁴⁾ 0.5 0.5 ¹⁾ plasticizer, ²⁾ anionic site fat soluble additive, ³⁾ sodium ionophore, ⁴⁾ fat soluble pH indicator

The sensitization principle of the optical sensor is based on a mechanism in which cations and anions are exchanged or coextracted between the sample solution and the sensitization membrane as shown in FIGS. 1A and 1B. That is, when the ion-selective material (L) is combined with the cation (M ⁺ ) in FIG. 1A, H ⁺ of the fat-soluble pH indicator (CH ⁺ ), which is in charge balance between the fat ^- soluble additive (R ⁻ ) and the membrane, is released. Through this process, the hydrogen ion selective indicator in the sensitizing film causes a color change, thereby changing the absorbance (see FIG. 4), and the change is measured by an ultraviolet / visible recording spectrophotometer. do.

Unlike other electrochemical sensors (eg, ion-selective electrodes, gas partial pressure sensors, ion-selective field effect transisters), these optical sensors are hardly affected by electrical noise and are necessary criteria for measuring potential difference. There is no need for a cumbersome device such as an electrode, and the advantage of using optical fibers is remote detection and field measurement.

Both ion-selective electrodes and optical sensors have similar manufacturing methods and compositional ratios, so each method itself can be applied to make an excellent sensor independently and provide complementary information in understanding the membrane response mechanism. The main materials that can be detected by such ion-selective electrodes and optical sensors are ions and materials capable of generating ions (various protein samples including urea). However, this general analyzer can only analyze one sample at a time, which causes problems in terms of time, labor, economics and reliability. In addition, existing optical sensors are virtually impossible to be used for quantitative analysis of analytical samples due to various problems, and are relatively difficult to be applied for research purposes, but are used in experiments to provide a theoretical background of ion selective electrodes. It was used for a very limited purpose.

Accordingly, the present inventors continue to study methods and apparatuses capable of quantitatively analyzing a plurality of samples at the same time for an individual optical sensor, and attach an appropriate optical sensor membrane solution composition developed by the present invention to a simultaneous absorbance measurement substrate. Thus, the present invention has been completed by developing a method and apparatus capable of quantitatively analyzing a large number of samples simultaneously, reliably, economically, and quickly.

An object of the present invention is to provide a reliable, accurate and economical analysis device capable of quantitatively analyzing a large number of samples at the same time, an optical sensor membrane solution composition suitable for mounting in such a device, and an analysis method using the device and the composition. It is.

Figure 1a shows the response mechanism of the cation selective optical sensor,

Figure 1b shows the response mechanism of the anion selective optical sensor,

Figure 2 shows an example of the optical sensor simultaneous absorbance measurement board structure,

3 is a cross-sectional view showing part A of FIG. 2 in detail;

FIG. 4 is a graph of average absorbance values when different concentrations of standard solutions are injected into different wells of FIG.

FIG. 5 is a cross-sectional view illustrating part A of FIG. 2 in detail when the optical sensor co-absorbance measurement substrate of FIG. 2 is applied to a dissolved gas volume analysis sensor.

FIG. 6 shows a case where no special correction is made for the individual optical sensor of the simultaneous absorbance measurement substrate.

7A and 7B show an example of a calibration curve of sodium that is changed by an appropriate result processing method,

8 is an uncorrected calibration curve for potassium concentration,

9 is a correlation graph of the results on the simultaneous absorbance measurement substrate and the results of the ion-selective electrode for the potassium concentration of each serum.

Explanation of symbols on the main parts of the drawings

10: Well of the simultaneous absorbance measuring substrate

20: optical sensor membrane

30: sample solution 40: gas permeable membrane

50: Hydrogel layer

60: pH selective optical sensor 70: Simultaneous absorbance substrate

80: optical fiber

90: Multiarray type UV detector

100: UV light

In order to achieve the above object, in the present invention, a device capable of simultaneously analyzing a large number of samples by applying an optical sensor membrane composition having a suitable composition to a simultaneous absorbance substrate capable of measuring absorbance or transmittance, an optical sensor membrane solution composition And a method for analyzing a sample using such an apparatus and composition.

More specifically, in the present invention, the optical sensor membrane solvent is uniformly coated on a conventional simultaneous absorbance substrate using a manual micro dispenser or an automatic dispenser, thereby quantitatively analyzing A novel analytical device having high selectivity similar to an ion selective electrode for an ion or a substance generating such an ion, an optical sensor membrane solution composition, and an analytical method using the device and the composition are provided.

Hereinafter, the present invention will be described in detail.

The present invention combines the advantages of the two sensing devices that were used completely differently, that is, the advantages of the conventional optical sensor and the advantage of the device that can simultaneously read the absorbance on the existing simultaneous absorbance measurement substrate, to quickly and quickly It provides a device that can be analyzed accurately and reliably.

The device of the present invention has the advantages of the conventional optical sensor, there is no electrical noise, high selectivity such as ion selective electrode, easy to measure, and the device can read the absorbance simultaneously on the existing simultaneous absorbance measurement substrate The fast measurement time of and the result processing program using a computer are combined, and the present invention is achieved by accurately and quickly measuring the concentration of important cations (K ⁺ , Na ⁺ and Ca ²⁺ ) in the real blood using this new sensor system. It is a quantitative analysis device suitable for use in analyzing environmental samples, industrial samples or biological samples.

More specifically, the analytical device of the present invention uniformly coats the optical sensor membrane solution composition having a fast response time available for sample analysis on a co-absorbance substrate with a thin thickness, and then adds a sample. It is a device that measures and quantifies change with an optical sensor. That is, in the present invention, by using the ion-selective polymer chromophoric material to form a light sensor simultaneous absorbance measuring substrate as shown in Figure 2 and then change the color appearing by reacting the sample and the chromophoric material with a multiarray type UV detector (Multiarray type UV detector) In order to quantify a sample by measurement, an ion-selective polymer chromophore, which is an example of a sensing material used in the present invention, has a component and a shape similar to that of the sensitive material originally mounted on an ion-selective electrode, and is mounted in the apparatus of the present invention. It is optimized for.

Simultaneous absorbance substrate used in the present invention can be any device that can measure the absorbance or transmittance of a large number of samples at the same time (see Figure 2), for example, widely used microtiter plate (microtiter plate) Can be used.

The photosensor membrane solution of the present invention comprises a fat soluble pH indicator (chromoionophore), an anionic site fat soluble additive (KTpClPB), an ion selector (ionophore), a plasticizer, a polymeric material and a suitable solvent. Among the materials constituting the optical sensor membrane solution composition of the present invention, the fat-soluble pH indicator may be used as a substance that generates H ⁺ ions by reacting with a sample. The fat-soluble pH indicator is ETH 5294 [9- (Diethylamino) -5-octadecanoylimino- 5H-benzo [a] phenoxazine], ETH 2439 [9-Dimethylamino-5- [4- (16-butyl-2,14-dioxo-3,15-dioxaeicosyl) phenylimino] benzo [a] phenoxazine], ETH 7075 ( 4 ', 5'-Dibromofluorescein octadecyl ester), ETH 2412 [3-hydroxy-4- (4-nitrophenylazo) phenyl octadecanonate], Nile Blue or Azo Violet. In addition, the anionic site lipophilic additive is present as a fat soluble anion in the membrane to act as a soluble pH indicator and ionic balance, and the available anionic site lipophilic additives include KTpClPB or NaTm (CF ₃ ) PB. As the polymer material used in the optical sensor film solution composition of the present invention, polyvinyl chloride (PVC), polyurethane (PU), and silicone rubber may be used alone or in combination. bis (2-ethylhexyl) sebecate], BBPA [bis (1-butylpentyl) adipate] or o-nitrophenyl octyl ether (Fluka AG, Buchs, Switzerland). In addition, ion selectors (ionophores) include ETH 1001, Valinomycin or sodium ionophore X; 4-tert-butylcalix [4] arene-tetraacetic acid tetraethyl ester, etc. An appropriate organic solvent may be used, but THF or cyclohexane is preferable.

One example of the optical sensor membrane solution composition suitable for use in the co-analytical substrate developed by the researchers has the composition shown in Table 2 below.

Composition of an optical sensor (% by weight) suitable for mounting on a simultaneous absorbance substrate. Furtherance Ion to measure K ⁺ Na ⁺ Ca ²⁺ Poly vinyl chloride (PVC) 13.94 13.94 20.84 Polyurethane (PU) 0 0 6.95 Fat-soluble pH indicator (Chromoionophore) 0.62 0.62 0.51 Anion site fat soluble additive (KTpClPB) 0.49 0.49 0.81 Ionphore 1.69 1.69 1.41 Plasticizer 83.26 83.26 69.47 Solvent 700 μl 700 μl 800 μl Buffer condition: HEPES (0.05M, pH 7.4)

In addition, the optical sensor membrane solution composition should be uniformly coated on the wells of the simultaneous absorbance measurement substrates with a predetermined thickness, and a manual micro dispenser may be used to drop the optical sensor membrane solution composition by a predetermined amount on the simultaneous absorbance measurement substrate.

After dropping a certain amount of the optical sensor membrane solution composition on the simultaneous absorbance measuring substrate and drying it, the color change was measured by adding a sample material to measure the concentration of potassium, sodium and potassium, which are clinically important ions. The result can be obtained (see FIG. 4). The main point in conducting this analysis is that the interface effect is caused by the sample solution injected on the simultaneous absorbance measurement substrate. It can effectively reduce the deviation of the analysis results. In addition, when the sample solution is colored, an error may be caused in the measurement. In this case, if the sample solution on the co-absorbance measurement substrate is emptied and then measured, this does not affect the analysis result. If the conditions described above are satisfied, the result can be obtained very quickly and accurately compared to the method using the ion selective electrode which is widely used in the existing biological sample analysis. In addition, these methods and devices can be analyzed for trace samples (10 μL) and can be used immediately in the field without requiring special stabilization time.

In addition, these methods and devices can be used to measure actual biological samples to obtain rapid measurement results. The ion-selective electrode, which is commercially available for the analysis of existing biomaterials, takes about 3 minutes to analyze a single biomaterial, and therefore, it is applied to 100 different biomaterials that require 5 hours to analyze with an ion-selective electrode. By using the simultaneous absorbance measurement board, the analysis can be performed in 30 minutes. Therefore, in terms of time and labor, it can be seen that using a light sensor co-absorbance substrate is a much more advantageous method. On the other hand, it also provides convenience in the result processing method, by increasing the reliability by injecting the same sample in several wells of the optical sensor simultaneous absorbance measurement substrate and averaging the results.

The above method is the most chemical species (LI ⁺ , Na ⁺ , K ⁺ , NH ^{4 +} , Rb ⁺ , Cs ⁺ , Mg ²⁺⁾ that are closely related to our lives in biological materials, industrial samples and environmental samples. , heavy metals, ClO _4, including cationic, Ag, Pd, Cu, including _{^{Ca 2+ -, IO 4 -,}} SCN -, NO 3 -, I -, Br -, Cl -, H 2 PO 4 -, HPO 4 2 ^-, HCO ₃ ^-, can be applied to negative ions and urea (urea), and so on), such as CO ₃ ^2-, it can be applied also to a gas-permeable optical sensor.

The result processing method of the sample analysis method using the simultaneous absorbance measurement substrate coated with the optical sensor film solution composition is as follows.

I. When two-point calibration is performed for individual light sensor on the simultaneous absorbance measurement board

Most existing analytical instruments employ two point calibration to obtain sufficient result reliability. Therefore, to find out how accurate the new analysis system developed by the present invention can achieve by two-point correction (see Table 3), and to use the well-known ion selective electrode to verify the reliability of this result. The results compared with are shown in FIG. The results obtained through this analytical method show that they can be immediately used for biological sample analysis.

II. When special calibration is not performed for individual light sensors on the simultaneous absorbance measurement board

The trend of modern analytical devices, along with the pursuit of miniaturization, light weight, and economics, has made the development of an analytical device easy for users to understand and use no less than the accuracy and speed of the analytical device. In this sense, minimizing calibration is very important in that the user can quickly obtain the desired result in a minimum amount of time.

All existing analyzers are usually the two point calibration analyzer described above. However, although such calibration is an essential element in an analytical device, it is inconvenient to use for an inexperienced device user, is cumbersome, and has a disadvantage in that it takes a long time to analyze a sample even for an experienced device user. Therefore, research to reduce or simplify important corrections is just as important as the importance of results in analytical instruments. However, in the case of using the simultaneous absorbance measuring substrate, by using the fact that the light sensors on each well show the same signal (absorbance) under the same conditions, the light sensors on one substrate are divided into the corrected light sensor portion and the measured light sensor portion. The analysis device does not require any special calibration by the user (see FIG. 6). In this regard, in the present invention, as shown in FIG. 6, 110 and 120 points of the wells on the simultaneous absorbance measurement substrate are set to correction points 1 and 2, respectively, and then a calibration solution is injected into each of the wells. The absorbances of the wells in the zone were averaged to be the calibration point 1 and the calibration point 2, and the remaining 80 wells were injected with sample solution to enable analysis. This method actually allowed the surface of individual optical sensors on one good sensor, the absorbance co-absorbance substrate, to quantify the ionic species in the sample even though the sample solution was only recognized once. This method uses a calibration solution, but the sensor can be used to analyze the sample without the need for special calibration. In other words, a user can purchase a simultaneous absorbance measurement board equipped with an optical sensor, inject a sample solution and a calibration solution into about 100 wells on the substrate, and analyze the concentration of a large number of sample solutions by analyzing it once. Can be. However, despite the simplicity of this method, the problem with the uncorrected analysis method is that the standard deviation of signals represented by the wells on the absorbance simultaneous measurement substrate is large. The cause of this problem is that, firstly, the polypropylene co-absorbance substrate used in the present experiment has a certain standard deviation. In order to correct this dispersion, the change in absorbance may be measured. FIG. 8 shows that the change in absorbance is similar in the range of 10 ⁻³ to 10 ⁻⁵ M even with only the change in absorbance. The second reason for the deviation is that the interface of each well after sample injection is not constant. To compensate for this, neutral surfactants can be used to greatly alleviate the variation caused by the interface. In addition, when the thickness of the optical sensor film solution composition coated on the co-absorbance measurement substrate is not constant, it may be a cause of increasing the standard deviation. This dispersion of absorbance is to first coat the thickness of the photosensor film on the simultaneous absorbance substrate by an automated process, and then to correct this difference in absorbance by a suitable formula. Indeed, an example in which the ratio of absorbance relaxed the dispersion is shown in Figs. 7A and 7B. In fact, Figure 7b is the result of dividing the absorbance of the sample shown in Figure 7a by the absorbance before sample injection (which can be commercially attached to the simultaneous absorbance measurement substrate in the form of a bar code at the factory), and then adjusted to the same scale. That is, FIG. 7B shows that the dispersion or standard deviation of the initial result value is greatly alleviated.

In addition, this system is used to obtain the appropriate calibration curve and concentration of analytical sample with two-point or no-compensation for individual optical sensors, and proposes a result processing method suitable for such a system. It is shown that the devices and methods of the invention are well suited for the analysis of biological, industrial and environmental samples.

The above-described method for quantitative analysis of a sample using the apparatus of the present invention comprises a predetermined amount of an optical sensor membrane solution suitable for a simultaneous absorbance measuring substrate capable of simultaneously measuring the absorbance of a large number of samples in the ultraviolet / visible region. And then dried to form an optical sensor film solution composition, ii) dropping a sample on the dried optical sensor film solution composition, iii) simultaneously measuring the absorbance of a plurality of samples, and iii) two-point or no-compensation The results are processed in such a way that multiple samples are quantitatively analyzed at the same time.

By the following examples will be described in detail the present invention. The examples are merely illustrative of the present invention, and the present invention is not limited by the examples.

<Example 1> Two-point correction for individual optical sensors on the simultaneous absorbance measuring substrate

After the optical sensor membrane composition of the present invention was added dropwise to a simultaneous absorbance measurement substrate using a manual micro dispenser, and left in air for 12 hours to blow off the solvent, human serum and horse serum After diluting with an appropriate magnification, the activity of K ⁺ , Na ⁺ , Ca ²⁺ was measured by two-point calibration for each well. The results are shown in Table 3 below. In the following Table 3, normal serum and abnormal serum mean human serum and normal human serum.

Determination of the Activity of Critical Cations in Serum Using Optical Sensor Co-Analysis Substrate Ionic species Measuring serum Measure average Standard Deviation ISE average value K ⁺ Normal serum 4.40mM 0.07 4.20mM Abnormal serum 5.92mM 0.46 6.12mM Horse serum 2.25mM 0.46 2.15mM Na ⁺ Normal serum 123.9mM 3.43 125mM Abnormal serum 129.1mM 3.46 133mM Horse serum 58.9mM 4.93 59mM Ca ²⁺ Normal serum 1.36mM 0.41 1.30mM Abnormal serum 2.43mM 0.67 2.45mM Horse serum 2.29mM 0.64 2.20mM

<Example 2> Special calibration is not performed for the individual light sensor on the simultaneous absorbance measurement board

After dropping the optical sensor membrane composition of the present invention uniformly dropwise to the simultaneous absorbance measurement substrate using a manual micro-dispenser and left in the air for 12 hours to blow out the solvent, human serum and horse serum containing a surfactant (horse serum activity was measured. More specifically, before injecting the unknown sample, the optical sensor remembers the absorbance of the co-absorbance measurement substrate, and then, as shown in FIG. 6, injects a solution of known concentration at 110 and 120 and rests the wells. After injecting the unknown sample, the molar concentrations of the remaining unknown samples were determined by averaging the absorbance values of the 110 and 120 points as the correction point 1 and the correction point 2, respectively. This method is characteristic in that each optical sensor film formed on the co-absorbance substrate can determine the concentration of the unknown sample after only one solution contact. The results are shown in Table 4 below.

Determination of the Activity of Critical Cations in Serum Using Optical Sensor Co-Analysis Substrate Ionic species Measuring serum Measure average Standard Deviation ISE average value K ⁺ Normal serum 4.07 mM 0.74 4.20 mM Abnormal serum 6.44 mM 1.14 6.12 mM Horse serum 2.03 mM 0.74 2.15 mM Na ⁺ Normal serum 125 mM 10.2 125 mM Abnormal serum 140 mM 11.2 133 mM Horse serum 47.1 mM 9.7 59 mM Ca ²⁺ Normal serum 1.23 mM 0.91 1.30 mM Abnormal serum 2.31 mM 0.84 2.45 mM Horse serum 2.10 mM 0.95 2.20 mM

The device of the present invention combines the advantages of two sensing devices that were used differently. That is, the device of the present invention is an advantage of the conventional optical sensor, there is no electrical noise, easy to measure with high selectivity, such as ion-selective electrode, and because the absorbance can be read simultaneously on the simultaneous absorbance measurement substrate, the measurement time is fast. In addition, the present invention is a combination of the result processing program using a computer to such a device, according to the result processing method suitable for use in the analysis of environmental samples, industrial samples or biological samples, etc. It can also be used as a non-calibration sensor.

Therefore, the apparatus and method of the present invention enables the simultaneous, quantitative analysis of a large number of samples to be performed reliably, accurately, economically, and faster than any conventional analytical device. It is a new device and method that can replace existing analytical devices including ion-selective electrodes in food chemistry.

Claims (10)

An optical sensor measuring device, characterized in that the optical sensor membrane solution composition is integrally coupled to the simultaneous absorbance substrate for quantitative analysis of a large number of samples at the same time. The optical sensor measuring apparatus according to claim 1, wherein the simultaneous absorbance measuring substrate is capable of simultaneously measuring absorbance of a large number of analyte in the ultraviolet and visible region. The optical sensor measuring apparatus according to claim 2, wherein the simultaneous absorbance measuring substrate comprises a multi-array microtiter plate. The optical sensor measuring apparatus according to claim 1, wherein the optical sensor membrane solution composition comprises an ion selective material, a fat soluble pH indicator, a plasticizer, an anionic site fat soluble additive, a polymer material, and an organic solvent. The method of claim 4, wherein the ion-selective material comprises ETH 1001, Valinomycin or sodium ionophore X; 4-tert-butylcalix [4] arene-tetraacetic acid tetraethylester Optical sensor measuring device. The method of claim 4, wherein the fat-soluble pH indicator is ETH 5294 [9- (Diethylamino) -5-octadecanoylimino-5H-benzo [a] phenoxazine], ETH 2439 [9-Dimethylamino-5- [4- (16-butyl-2) , 14-dioxo-3,15-dioxaeicosyl) phenylimino] benzo [a] phenoxazine], ETH 7075 (4 ', 5'-Dibromofluorescein octadecyl ester), ETH 2412 [3-hydroxy-4- (4-nitrophenylazo) phenyl octadecanonate ], An optical sensor measuring apparatus comprising Nile Blue or Azo Violet. 5. The apparatus of claim 4, wherein the anionic site lipophilic additive comprises KTpClPB or NaTm (CF ₃ ) PB. The optical sensor measuring apparatus according to claim 4, wherein the polymer material is polyvinyl chloride (PVC), polyurethane (PU), and silicone rubber. Large numbers of samples in the ultraviolet and visible range Iii) a predetermined amount of optical sensor film solution of appropriate composition is added to a simultaneous absorbance measurement substrate capable of simultaneously measuring absorbance and dried to form an optical sensor film solution composition. Ii) after the sample was dropped on the dried optical sensor film solution composition, Iii) simultaneously measuring the absorbance of multiple samples, Iii) a quantitative analysis method comprising the step of quantitatively analyzing a plurality of samples at the same time by processing the results in a two-point correction or no correction method. The quantitative analysis method according to claim 9, wherein a neutral surfactant is added to the analytical sample to analyze the sample when no special correction is made to the individual optical sensor of the simultaneous absorbance measurement substrate.

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