Estimation method of isoflavone amount in beans and estimation device for isoflavone amount in beans

Abstract

To provide an estimation method of an isoflavone amount in beans and an estimation device for an isoflavone amount in beans capable of more correctly estimating an isoflavone amount included in beans.SOLUTION: An estimation method of an isoflavone amount in beans comprises: an isoflavone amount measurement step for measuring an isoflavone amount of isoflavone included in beans by quantitative analysis; a reference EEM acquisition step for acquiring an excitation fluorescence matrix (EEM) of beans used in the isoflavone amount measurement step; a standard curve acquisition step for acquiring a standard curve of the isoflavone amount by the isoflavone amount included in beans measured by quantitative analysis and multi variable analysis of EEM of the beans; a sample bean EEM acquisition step for acquiring EEM of sample beans whose isoflavone amount is estimated; and an isoflavone amount estimation step for estimating the isoflavone amount included in the sample beans, by applying the EEM of the sample beans to the standard curve.SELECTED DRAWING: Figure 3

Description

[Technical field]

[0001] The present invention relates to a method for estimating the amount of isoflavones contained in beans and an apparatus for estimating the amount of isoflavones contained in beans. [Background technology]

[0002] Non-Patent Document 1 discloses a method for estimating the amount of isoflavones contained in soybeans by measuring diffuse reflectance in the near-infrared band with wavelengths of 1000 to 2500 nm. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Berhow, MA, Singh, M., Bowman, MJ, Price, NP, Vaughn, SF, & Liu, SX, 2020, Quantitative NIR determination of isoflavone and saponin content of ground soybeans, Food Chemistry, 317, 126373 Summary of the Invention [Problem to be solved by the invention]

[0004] According to the above-mentioned method, in the wavelength region of 1000 to 1700 nm, the absorption wavelengths of OH (oxygen-hydrogen bond), CH (carbon-hydrogen bond), and NH (nitrogen-hydrogen bond) derived from isoflavones contained in soybeans overlap with the absorption wavelengths of OH derived from water and CH and NH derived from proteins contained in soybeans. Therefore, the absorption wavelengths of OH, CH, and NH derived from water and proteins become noise, making it difficult to accurately estimate the amount of isoflavones contained in soybeans.

[0005] The present invention has been made in consideration of the above circumstances. That is, an object of the present invention is to provide a method for estimating the amount of isoflavones contained in beans and an apparatus for estimating the amount of isoflavones contained in beans, which can more accurately estimate the amount of isoflavones contained in beans. [Means for solving the problem]

[0006] The present invention is as follows. [1] The amount of isoflavones contained in beans can be estimated using the following method: an isoflavone amount measuring step for measuring the amount of isoflavone contained in the beans by quantitative analysis; A reference EEM acquisition step for obtaining an excitation emission matrix (EEM) of the beans used in the isoflavone amount measurement step; A calibration curve acquisition step of obtaining a calibration curve of the amount of isoflavones by multivariate analysis of the amount of isoflavones contained in the beans measured by the quantitative analysis and the EEM of the beans; A sample bean EEM acquisition step for obtaining an EEM of a sample bean for estimating the amount of isoflavone contained therein; and an isoflavone amount estimating step of estimating the amount of isoflavone contained in the sample beans by fitting the EEM of the sample beans to the calibration curve.

[0007] [2] The beans and the sample beans may be beans belonging to the legume family.

[0008] [3] The beans and the sample beans may be powdered beans.

[0009] [4] The quantitative analysis method may be liquid chromatography.

[0010] [5] The EEM of the beans may be EEM measured by a spectrofluorometer.

[0011] [6] The EEM of the sample beans may be EEM measured by a spectrofluorometer.

[0012] [7] The bean isoflavone content estimation device is A calibration curve acquisition means for obtaining a calibration curve of the amount of isoflavones by multivariate analysis of the amount of isoflavones contained in the beans measured by quantitative analysis and the excitation emission matrix (EEM) of the beans; A sample bean EEM acquisition means for obtaining an EEM of a sample bean for estimating the amount of isoflavone contained therein; and an isoflavone amount estimating means for estimating the amount of isoflavone contained in the sample beans by fitting the EEM of the sample beans to the calibration curve. Effect of the Invention

[0013] According to the present invention, it is possible to provide a method for estimating the amount of isoflavones contained in beans and an apparatus for estimating the amount of isoflavones contained in beans, which can more accurately estimate the amount of isoflavones contained in beans. [Brief description of the drawings]

[0014] [Figure 1] 1 is a block diagram showing the configuration of a bean-containing isoflavone amount estimation device according to an embodiment. [Diagram 2] 1 is a flowchart of a process for estimating the amount of isoflavones contained in beans. [Diagram 3] FIG. 1 shows data on the isoflavone amounts of 34 varieties of soybeans measured by an HPLC system, and calibration curves obtained by multivariate analysis of the EEM data of these soybeans. [Figure 4] This figure shows a calibration curve obtained by substituting data on the reflectance (R) of the near-infrared reflectance spectra of 34 soybean varieties, subjecting the data to second-order differentiation of log(1 / R), and then subjecting the data to multivariate analysis. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Hereinafter, the form for carrying out the present invention (hereinafter referred to as the embodiment) will be described in detail. The following embodiment is an example for explaining the present invention, and is not intended to limit the present invention to the following content. The present invention can be carried out with appropriate modifications within the scope of the gist. Note that "isoflavones" used in the present invention is a collective term for daidzin, daidzein, genistin, and genistein.

[0016] [Method for estimating the amount of isoflavones contained in beans] A method for estimating the amount of isoflavones contained in beans according to this embodiment will be described. The method for estimating the amount of isoflavones contained in beans includes an isoflavone amount measuring step, a reference EEM obtaining step, a calibration curve obtaining step, a sample bean EEM obtaining step, and an isoflavone amount estimating step.

[0017] (Isoflavone amount measurement process) The amount of isoflavone contained in the beans is measured by quantitative analysis. Quantitative analysis methods include, for example, electrophoresis and liquid chromatography. As the quantitative analysis method, liquid chromatography is preferred, and high performance liquid chromatography (HPLC) is more preferred, from the viewpoint of measuring the amount of isoflavone contained in the beans in a short time.

[0018] The beans used in the isoflavone amount measuring step are not limited as long as they belong to the legume family. Examples of beans that belong to the legume family include beans of the genus Vigna such as adzuki beans, beans of the genus Phaseolus such as kidney beans, beans of the genus Vicia such as broad beans, beans of the genus Pisum such as peas, beans of the genus Chickpea such as chickpeas, beans of the genus Lentil such as peas, beans of the genus Glycine soja such as soybeans, and beans of the genus Arachis such as peanuts.

[0019] In addition, the beans used in the isoflavone amount measurement step are preferably beans of a single genus and a single type, such as soybeans of the soybean genus, from the viewpoint of creating a highly accurate calibration curve of isoflavone amounts. Furthermore, in order to further improve the accuracy of the calibration curve of isoflavone amounts, it is preferable to use multiple beans with different isoflavone contents, such as beans with high isoflavone contents and beans with low isoflavone contents.

[0020] The beans are preferably in a powder form. By using powdered beans, isoflavones can be efficiently extracted from the beans, for example, in a liquid chromatography method. Powdered beans are obtained by crushing beans using a crushing device such as a mill or a mixer, and sieving the beans. The powdered beans are preferably beans that have passed through a sieve with an opening of 500 μm or less, preferably a sieve with an opening of 400 μm or less, and more preferably a sieve with an opening of 300 μm or less.

[0021] (Reference EEM acquisition process) An excitation emission matrix (EEM) of beans used in the isoflavone amount measurement step is obtained. The EEM is obtained by measuring with a spectrofluorophotometer. That is, the spectrofluorophotometer irradiates the beans with excitation light while gradually changing the wavelength of the excitation light, i.e., the excitation wavelength (Ex, Excitation wavelength), and measures the fluorescence intensity according to the wavelength of the fluorescence emitted from the isoflavones contained in the beans, i.e., the fluorescence wavelength (Em, Emission wavelength). This measurement provides the EEM of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity. The region in which the excitation wavelength is scanned is 200 to 750 nm, preferably 200 to 500 nm. The region in which the fluorescence wavelength of the fluorescence is scanned is 220 to 750 nm, preferably 220 to 600 nm. The bandwidth of the excitation wavelength is 10 nm or less, more preferably 5 nm or less, from the viewpoint of increasing the accuracy of the calibration curve. The bandwidth of the fluorescence wavelength is 10 nm or less, more preferably 5 nm or less, from the viewpoint of increasing the accuracy of the calibration curve.

[0022] The beans used in the reference EEM acquisition step are preferably in a powder form, which makes it easy to mold and process the beans into a sample for measurement with a spectrofluorometer, for example.

[0023] When excitation light is irradiated onto beans, elastically scattered light such as primary and secondary light of the excitation light and stray light are generated in addition to the fluorescence emitted from the isoflavones contained in the beans. Noise caused by these lights appears in the EEM. This noise is a factor that reduces the accuracy of the calibration curve. For this reason, it is preferable to remove data related to the elastically scattered light and stray light that cause this noise from the data that constitutes the EEM.

[0024] In addition, the beans used in the reference EEM acquisition step are the same as those used in the isoflavone amount measurement step, from the viewpoint of creating a highly accurate calibration curve. By using the same beans, a correlation can be obtained between the amount of isoflavones contained in the beans measured in the isoflavone amount measurement step and the EEM of the beans obtained in the reference EEM acquisition step, making it possible to create a highly accurate calibration curve.

[0025] (Calibration curve acquisition process) A calibration curve of the isoflavone amount is obtained by multivariate analysis of the amount of isoflavone contained in the beans obtained in the isoflavone amount measurement step and the EEM of the beans obtained in the reference EEM acquisition step. Examples of multivariate analysis methods include partial least squares regression, discriminant analysis, and multiple regression. The calibration curve is created using partial least squares regression, for example, so that when the data of the EEM of the beans obtained in the reference EEM acquisition step is input, the output value approaches the amount of isoflavone contained in the beans obtained in the isoflavone amount measurement step (i.e., the actual measured value of isoflavones contained in the beans). In this way, since the calibration curve is created so that the input EEM data approaches the actual measured value data, which is the output value, the obtained calibration curve has higher accuracy than the calibration curve created only from the EEM of the beans obtained in the reference EEM acquisition step. Examples of the calibration curve include a linear function calibration curve. The vertical axis (y-axis) of this calibration curve represents the estimated amount of isoflavone, and the horizontal axis (x-axis) represents the actual measured value of isoflavone.

[0026] Cross-validation may be used when creating a calibration curve using multivariate analysis. This can improve the accuracy of the calibration curve. In addition, from the viewpoint of further improving the accuracy of the calibration curve, it is preferable to determine a latent variable that minimizes the validation error. Here, cross-validation is a method of determining a target variable when creating a calibration curve model, dividing the obtained data into training data and validation data, and validating the calibration curve. For example, in 5-fold cross-validation, the obtained data is divided into training data and validation data in a ratio of 4:1 to validate the calibration curve. In this validation process, the number of latent variables that minimize the validation error is determined.

[0027] (Sample bean EEM acquisition process) The EEM of the sample beans for estimating the amount of isoflavones contained therein is obtained. The sample beans are beans of the same genus and species as the beans used in the isoflavone amount measurement step and the reference EEM acquisition step, and are beans for which the amount of isoflavones has not yet been measured. The EEM of the sample beans is obtained by measurement with a spectrofluorophotometer, in the same manner as the EEM obtained by measurement with a spectrofluorophotometer in the reference EEM acquisition step.

[0028] (Isoflavone amount estimation process) The amount of isoflavones contained in the sample beans is estimated by fitting the EEM of the sample beans to the calibration curve. For example, the amount of isoflavones contained in the sample beans is output by inputting the EEM data of the sample beans obtained in the sample bean EEM acquisition step into the calibration curve obtained in the calibration curve acquisition step.

[0029] As described above, the amount of isoflavones contained in beans measured by quantitative analysis and the calibration curve of the amount of isoflavones obtained by multivariate analysis of the EEM of the beans are correlated with the measured amount of isoflavones and EEM. In addition, by using beans of the same genus and species as the beans used to prepare the calibration curve as sample beans, the amount of isoflavones contained in beans can be estimated with high accuracy.

[0030] The order of each step can be determined arbitrarily. The location where each step is performed can also be determined arbitrarily. For example, in the order of steps, in this embodiment, the isoflavone amount measurement step was performed first, but the reference EEM acquisition step may be performed first. Also, the isoflavone amount measurement step and the reference EEM acquisition step may be performed simultaneously. Also, in this embodiment, the sample bean EEM acquisition step was performed after the isoflavone amount measurement step and the reference EEM acquisition step, but the sample bean EEM acquisition step may be performed first. Also, the sample bean EEM acquisition step may be performed between the isoflavone amount measurement step and the reference EEM acquisition step. Also, the sample bean EEM acquisition step may be performed simultaneously with the isoflavone amount measurement step and the reference EEM acquisition step.

[0031] [Estimation device for isoflavone content in beans] Next, the bean-containing isoflavone amount estimating device according to this embodiment will be described. The bean-containing isoflavone amount estimating device comprises a calibration curve obtaining means, a sample bean EEM obtaining means, and an isoflavone amount estimating means.

[0032] The bean-containing isoflavone amount estimation device is configured, for example, by an information processing device 1 as shown in Fig. 1. The information processing device 1 includes a storage device 101 for storing various programs and various data, a communication device 102 for communicating with the HPLC device 2 and the spectrofluorophotometer 3, an input device 103 for receiving user operation input, a display device 104 for displaying various data, a memory 105 for expanding various programs, and a processor 106 for executing various programs. The storage device 101, the communication device 102, the input device 103, the display device 104, the memory 105, and the processor 106 are mutually connected via a data bus 107.

[0033] The storage device 101 stores various programs executed by the processor 106, and display data such as images and characters to be displayed on the display device 104. The storage device 101 can be configured using a storage device such as an HDD (Hard Disk Drive) or an SSD (Solid State Drive).

[0034] The communication device 102 is a device capable of transmitting and receiving various data from the HPLC device 2 and the fluorescence spectrophotometer 3 via the Internet 100.

[0035] The input device 103 is an input unit through which the user inputs various data. The display device 104 is configured using an LCD (Liquid Crystal Display) monitor or the like, and displays various screens under the control of the processor 106 .

[0036] The memory 105 is a device for expanding various programs stored in the storage device 101. The memory 105 can be configured using a storage element and a storage medium, for example, a volatile or non-volatile semiconductor memory such as a RAM (Random Access Memory) or a flash memory.

[0037] The processor 106 reads out various programs stored in the storage device 101, develops them in the memory 105, and executes them. The processor 106 can be configured using a processing device such as a CPU (Central Processing Unit) or an MPU (Micro-processing Unit).

[0038] Next, an operation of estimating the amount of isoflavone contained in beans executed by the information processing device 1 will be described.

[0039] FIG. 2 is a flow chart of a process for estimating the amount of isoflavones contained in beans. The processor 106 acquires data on the amount of isoflavones contained in the beans measured by the HPLC device 2 via the communication device 102, and stores the data in the memory 105 (step S11). The processor 106 acquires data on the EEM of the beans measured by the spectrofluorophotometer 3 via the communication device 102, and stores the data in the memory 105 (step S12). Here, the beans measured by the spectrofluorophotometer 3 are the same beans as those measured by the HPLC device 2.

[0040] The processor 106 loads the multivariate analysis program stored in the storage device 101 into the memory 105, executes calculations based on the data obtained in steps S11 and S12, and obtains a calibration curve (step S13). For example, the multivariate analysis program is a program using partial least squares regression that is calculated by the processor 106 when the processor 106 receives data on the EEM of the beans measured by the spectrofluorophotometer 3, so that the output value approaches the amount of isoflavone contained in the beans measured by the HPLC device 2, i.e., the actual measured value of isoflavone contained in the beans. The above corresponds to the calibration curve obtaining means.

[0041] The processor 106 acquires the EEM data of the sample beans measured by the spectrofluorophotometer 3 via the communication device 102, and stores it in the memory 105 (step S14). The sample beans are beans of the same genus and species as the beans used in steps S11 and S12, and are beans for which the amount of isoflavones has not yet been measured. The above corresponds to the sample bean EEM acquisition means.

[0042] The processor 106 estimates the amount of isoflavone contained in the sample beans by fitting the EEM data of the sample beans acquired in step S14 to the calibration curve obtained in step S13 via the input device 103 (step S15). This corresponds to the isoflavone amount estimation means.

[0043] The information processing device 1 may be composed of one device or may be composed of multiple devices. When the information processing device 1 is composed of multiple devices, the multiple devices may be installed in separate locations. Also, the multiple devices may operate at different times.

[0044] The amount of isoflavones contained in beans measured by quantitative analysis and the calibration curve of the amount of isoflavones obtained by multivariate analysis of the EEM of the beans are correlated with the amount of isoflavones actually measured and EEM. In addition, the bean-containing isoflavone amount estimation device of this embodiment can estimate the amount of isoflavones contained in beans with high accuracy by using the same genus and type of beans as the sample beans used to create the calibration curve. EXAMPLES

[0045] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples in any way.

[0046] (Example) The beans used in the examples were soybeans. Thirty-four varieties of soybeans were used, including soybeans from Japan, the United States, Canada, Russia, and China. The weight of 100 kernels of one variety of soybean was 24.2±8.8g, and the moisture content was 10.3±1.5%.

[0047] (soybean grinding) For each soybean variety, 60 g of soybeans were weighed and crushed using a vibrating mill (CMT, TI-100). The crushed soybeans were stored frozen at -40°C until various measurements were performed.

[0048] (Quantitative analysis of soy isoflavone content) In the embodiment, high performance liquid chromatography (HPLC) was used as the quantitative analysis. First, a measurement sample was prepared for measurement with an HPLC device. The crushed soybeans were sieved through a 300 μm mesh sieve to make the size uniform, and impurities such as skin were removed to obtain powdered soybeans (hereinafter, powdered soybeans). Next, 1.0 g of precisely weighed powdered soybeans was placed in a beaker, 5 mL of 80% methanol was added thereto, and the mixture was stirred for 2 minutes so that the methanol and powdered soybeans were uniformly mixed. The stirred mixture was transferred to a container dedicated to the stirrer and centrifuged at 6000 rpm for 5 minutes to obtain a supernatant. Only the supernatant was transferred to another container, and 5 mL of 80% methanol was newly added to the container containing the remaining solids, and the mixture was stirred for 2 minutes so that the methanol and powdered soybeans were uniformly mixed. The stirred mixture was centrifuged again at 6000 rpm for 5 minutes to obtain a supernatant. The resulting supernatants were pooled together and the measurement sample was subjected to HPLC to measure the amount of isoflavones contained in the powdered soybean.

[0049] The HPLC device used was an LC-20AD manufactured by Shimadzu Corporation. The measurement conditions were as follows, and HPLC measurements were performed on each of the 34 varieties of soybeans. Measured components: daidzin, daidzein, genistin, genistein. Phase A: 100 mmol sodium phosphate buffer, Phase B: 90% methanol. Gradient conditions: 0 min (phase A 66.7%, phase B 33.3%), 30 min (phase A 44.4%, phase B 55.6%), 35 min (phase A 66.7%, phase B 33.3%). Flow rate: 1.0 mL / min, measurement time: 50 min, detector: UV detection (measurement wavelength: 254 nm), column: Shim-pack VP-ODS 150L-4.6 (Shimadzu Corporation), oven: 40°C.

[0050] (Measurement of soybeans using a spectrofluorophotometer) 2.0 g of the powdered soybeans prepared in the quantitative analysis above was precisely weighed and a sample cell for the spectrofluorophotometer was prepared. This sample cell was set in the spectrofluorophotometer, and measurements were performed to obtain EEM.

[0051] The spectrofluorophotometer used was a JASCO FP-8300. The measurement conditions were as follows, and each of the 34 varieties of soybeans used in the quantitative analysis above was measured using the spectrofluorophotometer. In addition, for each of the 34 varieties of soybeans obtained, EEMs in which noise was generated due to elastic scattered light or stray light were removed from the EEM. Excitation wavelength (Ex) 200-500 nm, Fluorescence wavelength (Em) 220-600nm, Bandwidth of excitation wavelength (Ex) and emission wavelength (Em) 5.0 nm, Scanning speed: 5000 nm / min. Response 50ms, Measurement method: Surface fluorescence measurement (Front-Face). The intensity was normalized by the integrated Raman scattering intensity of water. The excitation wavelength (Ex) was 350 nm, and the emission wavelength (Em) was 371 to 428 nm.

[0052] (Creating a calibration curve) The data on the isoflavone content of 34 varieties of soybeans obtained by the measurement with an HPLC device and the EEM data of 34 varieties of soybeans obtained by the measurement with a spectrofluorophotometer were input into a computer, and a calibration curve was obtained by multivariate analysis. In this embodiment, partial least squares regression was used as the multivariate analysis. In addition, when creating a calibration curve using partial least squares regression, the objective variable, i.e., the variable to be predicted (estimated), was set to the amount of isoflavone contained in soybeans, and a calibration curve was obtained after performing 5-fold cross-validation. The number of latent variables that minimized the validation error in this validation was 5. Figure 3 shows the calibration curve obtained by multivariate analysis of the data on the isoflavone content of 34 varieties of soybeans measured by an HPLC device and the EEM data of 34 varieties of soybeans. The y-axis represents the estimated amount of isoflavone. The x-axis represents the actual value of the amount of isoflavone contained in soybeans with water removed. The coefficient of determination (R 2) was 0.87 and the RPD was 2.73. The coefficient of determination is an index that indicates the accuracy of a prediction model; the closer it is to 1, the better the accuracy. RPD (Residual Prediction Deviation) is the ratio of the standard deviation to the standard error of prediction, and is an index for evaluating the practicality of a calibration curve. The RPD criteria are as follows: RPD < 2.0 Cannot be used for screening. RPD 2.0 or more and less than 2.5 Can be used for simple screening. RPD 2.5 or greater but less than 3.0 Can be used for screening. RPD 3.0 or higher Can be used for more detailed screening. From the above, it was found that the calibration curve obtained in the examples is a calibration curve that can be used for screening.

[0053] Comparative Example 1 The beans used in Comparative Example 1 were the same 34 varieties of soybeans as those used in the Examples. The powdered soybeans used were the same soybean powder as those used in the Examples.

[0054] (Measurement of isoflavone content in soybeans using near-infrared spectroscopy) Near-infrared spectroscopy (NIR) was used in Comparative Example 1. First, 2.0 g of powdered soybeans was precisely weighed, and a sample cell for a near-infrared spectrophotometer was prepared. This sample cell was set in the near-infrared spectrophotometer, and measurements were performed to obtain a near-infrared reflectance spectrum.

[0055] The near-infrared spectrophotometer used was a V-670 manufactured by JASCO Corp. The measurement conditions were as follows, and measurements were performed with the near-infrared spectrophotometer for each of the 34 varieties of soybeans. Measurement wavelength 800~1800nm, Bandwidth in the UV-Vis region: 5nm Bandwidth in the NIR region is 8.0 nm, data interval is 0.5 nm. An integrating sphere unit (ISN-923) was used, and a reference was measured with a Spectralon (standard reflector) for each sample.

[0056] (Creating a calibration curve) The near-infrared reflectance spectrum data of 34 varieties of soybeans obtained by measurement with a near-infrared spectrophotometer was input into a computer. Next, the reflectance (R) data of the near-infrared reflectance spectrum was substituted into the formula log(1 / R) to convert it into data that represents the absorption of the near-infrared reflectance spectrum. The obtained data (log(1 / R)) was then subjected to second-order differential processing (d 2 log(1 / R)). Then, partial least squares regression was used to obtain a calibration curve of the data subjected to the second derivative processing. When creating the calibration curve using partial least squares regression, the objective variable was the amount of isoflavone contained in soybeans as in the embodiment, and a calibration curve was obtained after performing 5-fold cross validation. The number of latent variables that minimized the validation error in this validation was 4. Figure 4 shows the calibration curve obtained by subjecting the log(1 / R) into which the reflectance (R) data of the near-infrared reflectance spectrum data of 34 soybean varieties was substituted and subjecting the data to multivariate analysis. The y-axis represents the estimated amount of isoflavone, and the x-axis represents the measured value of the amount of isoflavone. The coefficient of determination (R 2 ) was 0.62, and the RPD was 1.62. The calibration curve obtained in Comparative Example 1 had improved estimation accuracy by the second derivative process, and the RPD was 1.62. However, according to the above-mentioned RPD standard, the RPD was 2 or less, and it was found that it is difficult to use it for screening. 2 log(1 / R) 2 By performing this second derivative process, it is possible to remove baseline noise present in the obtained near-infrared reflectance spectrum, thereby improving the estimation accuracy.

[0057] Comparative Example 2 A calibration curve was obtained by using partial least squares regression for the data obtained in Comparative Example 1 that represents the absorption of the near-infrared reflectance spectra of 34 varieties of soybeans. As in Comparative Example 1, the objective variable was the amount of isoflavone contained in soybeans, and a calibration curve was obtained by performing 5-fold cross-validation. In this validation, the number of latent variables that minimized the validation error was 3. The coefficient of determination (R 2 ) was 0.21, and the RPD was 1.12. The RPD of the calibration curve obtained in Comparative Example 2 was lower than the RPD of 1.62 in Comparative Example 1 because no second derivative treatment was performed, and it was found that use for screening was difficult.

[0058] As described above, according to the method for estimating the amount of isoflavones contained in beans and the device for estimating the amount of isoflavones contained in beans of the present embodiment, the amount of isoflavones contained in beans can be estimated more accurately.

[0059] Although the present invention has been described above using the embodiments, various embodiments and modifications of the present invention are possible without departing from the broad spirit and scope of the present invention. The above-described embodiments are for the purpose of explaining the present invention and do not limit the scope of the present invention. In other words, the scope of the present invention is indicated by the claims, not the embodiments. Various modifications made within the scope of the claims and within the scope of the meaning of the invention equivalent thereto are considered to be within the scope of the present invention. [Explanation of symbols]

[0060] 1 Information processing device, 2 HPLC device, 3 Spectrofluorometer, 100 Internet, 101 Storage device, 102 Communication device, 103 Input device, 104 Display device, 105 Memory, 106 Processor, 107 Data bus.

Claims

1. an isoflavone amount measuring step for measuring the amount of isoflavone contained in the beans by quantitative analysis; a reference EEM acquisition step for obtaining an excitation emission matrix (EEM) of the beans to be used in the isoflavone amount measurement step; a calibration curve acquisition step of obtaining a calibration curve of the amount of isoflavones by multivariate analysis of the amount of isoflavones contained in the beans measured by the quantitative analysis and the EEM of the beans; A sample bean EEM obtaining step of obtaining an EEM of a sample bean for estimating the amount of isoflavone contained therein; an isoflavone amount estimation step of estimating the amount of isoflavone contained in the sample beans by fitting the EEM of the sample beans to the calibration curve; A method for estimating the amount of isoflavones contained in beans, comprising:

2. The method for estimating an amount of isoflavones contained in beans according to claim 1 , wherein the beans and the sample beans are beans belonging to the legume family.

3. The method for estimating an amount of isoflavones contained in beans according to claim 1 , wherein the beans and the sample beans are in powder form.

4. The method for estimating the amount of isoflavones contained in beans according to claim 1 , wherein the quantitative analysis is performed by liquid chromatography.

5. The method for estimating the amount of isoflavones contained in beans according to claim 1 , wherein the EEM of the beans is EEM measured by a spectrofluorophotometer.

6. The method for estimating the amount of isoflavones contained in beans according to claim 1 , wherein the EEM of the sample beans is EEM measured by a spectrofluorophotometer.

7. A calibration curve acquisition means for obtaining a calibration curve of the amount of isoflavones by multivariate analysis of the amount of isoflavones contained in the beans measured by quantitative analysis and the excitation emission matrix (EEM) of the beans; A sample bean EEM acquisition means for acquiring an EEM of a sample bean for estimating the amount of isoflavone contained therein; an isoflavone amount estimating means for estimating the amount of isoflavone contained in the sample beans by fitting the EEM of the sample beans to the calibration curve; A bean-containing isoflavone amount estimation device comprising:

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