JP5890034B2 - Preprocessing determination device, preprocessing method determination method, control program, and recording medium - Google Patents

Description

  The present invention relates to determination of a sample processing method based on a database, and more particularly to determination of a pretreatment method for removing contaminants from a sample solution.

  Drug concentration measurement (bioanalysis) in biological samples for biological samples such as plasma, urine and tissue fragments obtained from non-clinical (animals) and clinical trials (humans) in the development of pharmaceuticals (Absorption, distribution, metabolism, excretion) is extremely important to know. It is commercially available after compound screening to development stage and drug approval, such as pharmacokinetic test, toxicokinetic (toxicity assessment) test and bioequivalence test. Required at every stage until further investigation.

  As an analysis method in bioanalysis, a chromatography technique such as liquid chromatograph-triple quadrupole mass spectrometer (LC / MS / MS) is used. When performing drug concentration measurement, pretreatment such as protein removal, liquid-liquid extraction, and solid phase extraction is required to remove proteins and the like in biological samples. / Used for measurement by analyzers such as MS. Therefore, in addition to the measurement method set by the parameters of the analyzer, the pretreatment method of the biological sample is important. Especially for the demands of measurement techniques at a higher level than usual, such as highly sensitive measurement, Study (analysis method development) for setting these pretreatment conditions and measurement conditions plays an even more important role.

  An example of requiring a high sensitivity measurement technique is a micro dose test. The microdose test is a test for screening an optimal compound from a new drug candidate compound, and is performed by single administration of a minute amount of a compound such as 1 / 100th dose or 100 μg dose of an effective dose. By conducting human administration tests using only simple toxicity test data compared to conventional drug development, pharmacokinetic data obtained by humans can be acquired at an early stage, resulting in an increase in the success probability of drug development. To do. In the microdose test, as described above, since the amount of the compound to be administered is very small, measurement of the blood concentration of the compound requires a very high sensitivity measurement technique. Setting of pretreatment conditions and measurement conditions is essential.

  Thus, in analytical method development, in order to determine the optimal pretreatment method and measurement method, know-how related to the physical properties of compounds and analytical techniques is required, and the knowledge and experience of analysts are also large. It is necessary to repeatedly examine the conditions before reaching the optimum pretreatment conditions and measurement conditions. In addition, as a prior art for making analysis method development easy, the following nonpatent literature 3 is mentioned, for example.

Japanese Patent Publication “JP 2005-130755 A” (published May 26, 2005) Japanese Patent Publication “Japanese Patent Laid-Open No. 2010-185719 (published on August 26, 2010)” Japanese Patent Publication “Japanese Patent Laid-Open No. 2009-222421 (published on Oct. 1, 2009)”

Mohammed Jemal, Zheng Ouyang, Yuan-Qing Xia, "Systematic LC-MS / MS bioanalytical method development that incorporates plasma phospholipids risk avoidance, usage of incurred sample and well thought-out chromatography", Biomedical Chromatography, January 2010, Volume 24, Issue 1, pages 2-19 Sebastian Dresen, Jurgen Kempf, Wolfgang Weinmann, "Electrospray-ionization MS / MS library of drugs as database for method development and drug identification", Forensic Science International, 12 September 2006, Volume 161, Issue 2, Pages 86-91 Marie-Claire Hennion, "Solid-phase extraction: method development, sorbents, and coupling with liquid chromatography", 1999, Journal of Chromatography A, 856 3-54 Hiroaki Yoshida, Katsuhiro Okamoto, Tomonori Hino, Muneyuki Hiramatsu, Hiroki Miyamoto, Masakatsu Honma, Noriyuki Watanabe, Yasuaki Enomoto, National Police Agency, National Police Research Institute, Police Department, National Police Research Institute Institute, "Development of a simple method for predicting the vapor pressure and flash point of gasoline by gas chromatography and its application to the analysis of evaporated gasoline", Analytical Chemistry, 2012, Vol. 61 No. 7 p. 569-576 Ailan Cheng, David J. Diller, Steven L. Dixon, William J. Egan, George Lauri, Kenneth M. Merz Jr., "Computation of the physio-chemical properties and data mining of large molecular collections", Journal of Computational Chemistry, 15 January 2002 Volume 23 Issue 1 pages 172-183

  With the above conventional methods, it takes a lot of time to develop an analysis method, and it is difficult for an analyst with a wealth of knowledge and experience to determine an appropriate pretreatment method and measurement method. There is.

  Therefore, in order to solve such problems, a method of study for selecting a pretreatment method based on physical property values, etc., by recording the physical property values, etc., of the compounds for which pretreatment methods have already been established as a database. Can be considered.

  However, estimation based on a database has examples in other fields (see Patent Documents 1 and 2, etc.), and it has been proposed to use a statistical analysis technique in the field of chemical analysis (Patent Document 3). Reference), based on the database, there has never been an idea of determining a pretreatment method in analytical method development. For this reason, in the field of analytical method development, a preprocessing determination method based on a database has not been established.

  The present invention has been made in view of the above-mentioned problems, and its purpose is a preprocessing determination device that makes it possible to quickly determine an appropriate preprocessing method regardless of knowledge or experience. Is to provide.

  In order to solve the above-mentioned problem, the pretreatment determination device of the present invention is a pretreatment determination device that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. Data acquisition means for acquiring retention time when subject compound is subjected to chromatography under predetermined conditions, and when each compound is subjected to chromatography under the above conditions for each of a plurality of compounds for which pretreatment methods have been established Retention time within a predetermined range from the retention time acquired by the data acquisition means based on the database data recorded in association with the physical property value indicating the physical property of the compound and the established pretreatment method Before determining the pretreatment method associated with the time compound as the pretreatment method for the target compound and outputting it together with the physical property values associated with the pretreatment method It is characterized by comprising a management decision means.

  Further, in order to solve the above-described problem, the pretreatment method determination method of the present invention is a pretreatment by a pretreatment determination device that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. A method for determining a method, the data acquisition step for acquiring a retention time when the target compound is subjected to chromatography under a predetermined condition, and each of a plurality of compounds for which a pretreatment method has been established, Based on the database data recorded in association with the retention time when subjected to chromatography under conditions, the physical property value indicating the physical property of the compound, and the established pretreatment method, the data was acquired in the data acquisition step. A pretreatment method associated with a compound having a retention time within a predetermined range from the retention time is determined as a pretreatment method for the target compound, and the pretreatment method is supported. It is characterized in that it comprises a pretreatment determination step of outputting together with the physical properties being eclipsed.

  Another pretreatment determining apparatus of the present invention is a preprocessing determining apparatus for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed in order to solve the above-described problems. The data acquisition means for acquiring the retention time when the target compound is subjected to chromatography under predetermined conditions and the plurality of compounds for which pretreatment methods have been established, are subjected to chromatography under the above conditions. When plotting on the coordinate plane the retention time when provided and the physical property values indicating the physical properties of the compound, the line segment dividing the plurality of compounds for each established pretreatment method is plotted on the coordinate plane. Preprocessing determination that determines a pretreatment method to be applied to the target compound for each range of the physical property value from the retention time acquired by the data acquisition unit and the mathematical expression. It is characterized in that it comprises a stage.

  In order to solve the above-described problem, another pretreatment method determination method of the present invention is based on a pretreatment determination device that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. A pretreatment method determination method, wherein a data acquisition step for obtaining a retention time when the target compound is subjected to chromatography under a predetermined condition, and each of a plurality of compounds for which a pretreatment method has been established, When the retention time when subjected to chromatography under the above conditions and the physical property values indicating the physical properties of the compound are plotted on a coordinate plane, the pretreatment in which the plurality of compounds are established on the coordinate plane A mathematical expression indicating a line segment to be divided for each method is acquired, and the pretreatment method to be applied to the target compound is determined from the retention time acquired in the data acquisition step and the mathematical expression. It is characterized in that it comprises a pretreatment determination step of determining for each range of values.

  Another pretreatment determining apparatus of the present invention is a preprocessing determining apparatus for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed in order to solve the above-described problems. The target compound is subjected to liquid chromatography under a second condition in which the retention time when the target compound is subjected to liquid chromatography under the first condition and the pH of the mobile phase is different from the first condition. A data acquisition means for acquiring at least the holding time when the data is acquired, a predetermined parameter calculated using at least each holding time obtained under the first and second conditions acquired by the data acquisition means, and preprocessing The above parameters and pretreatment methods obtained based on the retention times when each of a plurality of compounds for which a method has been established is subjected to liquid chromatography under the first and second conditions. Based on the correlation information indicating a correlation between, it is characterized in that it comprises a pre-processing determining means for determining a treatment prior to be applied to the target compound.

  In order to solve the above-described problem, another pretreatment determination method of the present invention uses a pretreatment determination apparatus that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. A method for determining a treatment method, wherein the target compound is subjected to liquid chromatography under the first condition and the target compound is subjected to a second condition in which the pH of the mobile phase is different from the first condition. The data acquisition step for obtaining at least the retention time when the sample was subjected to liquid chromatography, and each retention time obtained under the first and second conditions obtained in the data acquisition step was calculated using at least Based on predetermined parameters and retention times when each of a plurality of compounds for which pretreatment methods have been established are subjected to liquid chromatography under the first and second conditions. Was, based on the correlation information indicating a correlation between the parameters and the pre-processing method is characterized in that it comprises a pretreatment determination step of determining a treatment prior to be applied to the target compound.

  According to the pretreatment determination apparatus or the pretreatment method determination method of the present invention, an effective pretreatment method can be determined from the retention time when the target compound is subjected to chromatography under a predetermined condition. Regardless of whether or not there is experience, it is possible to quickly determine an appropriate pretreatment method.

It is a block diagram which shows an example of the principal part structure of the pre-processing determination apparatus concerning one Embodiment of this invention. It is a figure explaining the flow of high sensitivity in the drug quantitative analysis in comparison between the present invention and the prior art. The data regarding the compound quantified by the negative mode measurement of LC-MS / MS among the data contained in pre-processing DB with which the said pre-processing determination apparatus is provided is shown. The data regarding the compound quantified by the positive mode measurement of LC-MS / MS among the data contained in the said pretreatment DB is shown. The data regarding the compound which belongs to the statin type drug among the data contained in the said pretreatment DB is shown. It is the figure which plotted the data contained in the said preprocessing DB on the coordinate plane. It is a flowchart which shows an example of the pre-processing determination process in which the said pre-processing determination apparatus determines pre-processing from the retention time of object compound. It is a block diagram which shows an example of the principal part structure of the pre-processing determination apparatus concerning other embodiment of this invention. It is a flowchart which shows an example of the pre-process determination process in which the said pre-process determination apparatus determines the pre process for every range of a physical-property value from the retention time of a target compound. It is a block diagram which shows an example of a principal part structure of the pre-processing determination apparatus concerning further another embodiment of this invention. It is a figure which shows an example of the measurement conditions of Retention index. It is a figure (the first half) which shows the result of having measured Retention index on acidic, neutral, and basic conditions about 27 types of compounds. It is a figure (latter half) which shows the result of having measured Retention index under acidic, neutral, and basic conditions about 27 types of compounds. It is a figure which shows the response | compatibility with the pre-processing about each compound which measured Retention index, and a score. It is the figure which plotted the LLE score and the total value of Retention index of acidic, neutral, and basic conditions on a coordinate plane about a neutral and basic compound. For neutral / basic compounds, the value of (Retention index of basic condition) / (Retention index of neutral condition) and the total value of acidic, neutral and basic Retention index are plotted on the coordinate plane. FIG. It is the figure which plotted the LLE score and the value of acid / base ratio on the coordinate plane about the acidic compound. It is the figure which plotted the value of acid / base ratio, and the total value of acidic, neutral, and basic Retention index on a coordinate plane about an acidic compound. It is a flowchart which shows an example of the pre-processing determination process which determines the pre-processing of this target compound from the retention time of the target compound used as the object which determines pre-processing. It is a figure which shows the parameter calculated | required about each compound made into the object which demonstrates the effect of one Embodiment of this invention. It is the figure which showed the suitable pre-processing method and parameter about each compound classified as an acidic compound in one Embodiment of this invention. It is the figure which showed the suitable pre-processing method and parameter about each compound classified into the neutral and basic compound in one Embodiment of this invention.

[Embodiment 1]
Hereinafter, an embodiment of the present invention will be described in detail with reference to FIGS.

〔Device configuration〕
First, the configuration of the preprocessing determining apparatus according to the present invention will be described with reference to FIG. FIG. 1 is a block diagram illustrating an example of a main configuration of the preprocessing determination device 1.

  The pretreatment determination device 1 is a device that determines a pretreatment method for removing impurities from a sample solution containing a target compound. Here, an example in which human plasma containing a trace amount of the target compound is used as a sample will be described, but the target sample solution is not particularly limited as long as it is a liquid containing the target compound. For example, it may be other biological samples (including processed biological samples and pseudo biological samples) such as serum, blood, urine, tissue fluid, culture fluid, and enzyme-treated fluid containing the target compound, or biological samples (Including processed biological samples and pseudo-biological samples).

  As illustrated, the preprocessing determining device 1 includes an input unit 2, a control unit 3, an output unit 4, and a storage unit 5. Further, the control unit 3 includes a data acquisition unit 10, a preprocessing determination unit 11, and a DB update unit 12, and the storage unit 5 stores a preprocessing DB (database) 20.

  The input unit 2 receives data input to the preprocessing determination device 1. Specifically, LC (Liquid Chromatography) retention time (Retention Time) of the target compound and data to be registered in the pretreatment DB 20 (LC retention time, physical property value, etc. for a compound for which a pretreatment method has been established) Accepts input.

  The control unit 3 controls the operation of the preprocessing determination device 1 in an integrated manner. The control unit 3 includes a data acquisition unit (data acquisition unit) 10, a preprocessing determination unit (preprocessing determination unit) 11, And the DB update part 12 is included.

  The data acquisition unit 10 acquires the data received by the input unit 2 and outputs the data to the preprocessing determination unit 11 or the DB update unit 12. Specifically, the data acquisition unit 10 outputs this data to the preprocessing determination unit 11 when the retention time of the target compound, which is data for determining the preprocessing method, is input. On the other hand, when data for updating the pretreatment DB 20 (retention time, physical property value, pretreatment method, compound name of a compound registered in the pretreatment DB 20) is input, this data is output to the DB update unit 12. .

  Based on the data stored in the preprocessing DB 20, the preprocessing determination unit 11 determines a preprocessing method for the target compound from the retention time of the target compound input from the data acquisition unit 10. A method for determining the pretreatment method will be described later.

  The DB update unit 12 adds the data input from the data acquisition unit 10 to the preprocessing DB 20 and updates the preprocessing DB 20. By increasing the number of compounds registered in the pretreatment DB 20, it is possible to increase the accuracy (predicted accuracy of an appropriate pretreatment method) with which the pretreatment determination unit 11 can determine an appropriate pretreatment method.

  The output unit 4 receives data output from the control unit 3 and outputs the data to the outside of the preprocessing determination device 1. For example, the output unit 4 outputs data indicating the preprocessing method determined by the preprocessing determination unit 11. Note that the data output mode of the output unit 4 is not particularly limited. For example, data indicating the preprocessing method may be output as it is to another device or the like as it is, or may be output as a display or sound. Alternatively, it may be printed out.

  The storage unit 5 is a storage device that stores data used by the preprocessing determination device 1, and the storage unit 5 stores a preprocessing DB 20.

  The pretreatment DB 20 is a database for determining a pretreatment method, and the retention time, physical property value, pretreatment method, and compound name are registered for compounds for which an appropriate pretreatment method is known.

  The preprocessing DB 20 only needs to be stored so that the preprocessing determination device 1 can refer to it, and the storage location is not limited to the storage unit 5. For example, the preprocessing DB 20 may be stored in a server on the network, and the preprocessing determining device 1 may be referred to via the network. In this case, it becomes easy to update the preprocessing DB 20 from a plurality of devices and enrich the data.

[About high sensitivity]
Next, high sensitivity in the quantitative drug analysis will be described with reference to FIG. FIG. 2 is a diagram for explaining the flow of high sensitivity in quantitative drug analysis in comparison between the present invention and the prior art.

As shown in the figure, conventionally, a pretreatment method that enables high-sensitivity quantitative analysis is determined while repeating experiments, relying on parameters and structural formulas of physical property information. Examples of the physical property information include an acid dissociation constant (pK _a ), a water-octanol partition coefficient (Log P), a distribution coefficient obtained by adding pK _a to Log P (Log D), solubility, Dose Number, and the like. It is done.

  In such a method, the time and effort required to reach an appropriate preprocessing method are affected by the experience and knowledge of the analyst, and it becomes difficult for an unskilled analyst to reach an appropriate preprocessing method. There was also. Moreover, even if it was an expert, there existed a problem that much examination time was required.

  On the other hand, in the method of the present invention, first, a sample is subjected to LC under predetermined measurement conditions to obtain a holding time. Then, by using the above-described pretreatment DB 20, the pretreatment conditions (pretreatment method) are determined in consideration of the physical property information (physical property value) of the target compound for the obtained retention time. Based on this, it is possible to enter into the development of analytical methods including measurement conditions.

  As described above, according to the method of the present invention, an analyst can quickly determine a pretreatment method simply by subjecting a sample to LC under predetermined measurement conditions and obtaining a retention time, and has special experience and knowledge. There is an advantage that it is not necessary.

[About pre-processing DB]
Next, details of the preprocessing DB 20 will be described with reference to FIGS. FIG. 3 shows data relating to compounds quantified by LC-MS / MS negative mode measurement among data included in the pretreatment DB 20, and FIG. 4 relates to compounds quantified by LC-MS / MS positive mode measurement. Data are shown. Moreover, FIG. 5 has shown the data regarding the compound which belongs to the statin type drug among the data contained in pre-processing DB20. The internal standard substance for positive measurement is busetin, and the internal standard substance for negative measurement is tolbutamide.

  The compounds shown in FIGS. 3 to 5 are all low molecular compounds having a molecular weight of 1500 or less. Of the compounds shown in FIG. 3, warfarin, chlorzoxazone, and probenecid are CYP substrates, and digoxin is a transporter substrate. Also, among the compounds shown in FIG. 4, methotrexate is a transporter substrate, and the others are CYP substrates. Of course, the target compound is not limited to a transporter substrate, a CYP substrate, or a statin drug.

  In the column of “Pretreatment”, liquid-liquid extraction of human plasma compounds with a mixture of ethyl acetate / hexane (under acidic and alkaline conditions) and liquid-liquid extraction with ether (acidic and alkaline) And a pretreatment with a solid phase extraction cartridge Oasis HLB, Oasis MCX, Oasis MAX (all manufactured by Waters) and Empore C18 (manufactured by 3M). “Ether” was the most suitable for liquid extraction (either acidic or alkaline conditions), “HLB” was the most suitable for Oasis HLB cartridge for reversed phase solid phase extraction, “MCX” is the most suitable cartridge for solid-phase extraction of ion exchange system Oasis MCX Respectively are described. Of the compounds shown in FIGS. 3 to 5, there was no compound in which the ethyl acetate / hexane mixture was most suitable.

  In addition, the measurement conditions of “retention time” in these drawings are as follows: Column used: Ascelis (R) Express C18 (5 cm × 2.1 mm, 2.7 μm) manufactured by SUPELCO, mobile phase: methanol (0 min to 5 min) A linear gradient in which the methanol concentration is changed from 10% to 90%), and the flow rate is 0.2 ml / min.

  And in the example of these drawings, the value of LogP is used as a physical property value. Among the values of LogP, (A) shows the value of LogP calculated by the physical property value prediction function of ChemDraw (registered trademark), and (B) shows the value of LogP acquired from the Drug Bank. ChemDraw is software of PerkinElmer having a chemical structural formula drawing function, and Drug Bank is a website (URL: http://www.drugbank.ca/) that discloses physical properties of various chemical substances. ).

  As described above, in chromatographic analysis such as LC, pretreatment and measurement are often performed using the principle of reverse phase partitioning. In this case, the retention time information obtained in the reverse phase column can be a hint for studying the pretreatment method using the reverse phase solid phase extraction column. That is, an examination method for selecting a suitable pretreatment method by using measurement data obtained by LC measurement or the like can be considered. This indicates that theoretical and efficient analytical method development is possible by extracting physical property information from data obtained from LC. The present invention focuses on deriving the relationship between physical property information useful for analytical method development by creating a database of retention times obtained by LC measurement and suitable pretreatment methods. .

  For reference, these drawings show the lower limit of quantification when a human plasma sample is pretreated by the illustrated pretreatment method and the lower limit of quantification when a standard solution of the compound is used. However, since these data are not used for determining the preprocessing method, they may be omitted. The lower limit concentration of quantification is the lower limit concentration at which a calibration curve could be drawn. The lower the concentration value, the higher the sensitivity of quantification possible.

  According to FIG. 3, it can be seen that the four compounds shown in the figure can be quantified from a human plasma sample by solid phase extraction with MCX. For digoxin, since LogP could not be calculated by ChemDraw, no value is described in the (A) column of LogP.

  Moreover, according to FIG. 4, it turns out that 11 types except methotrexate among 12 types shown in the figure can be quantified from a human plasma sample by liquid-liquid extraction. Methotrexate could not be quantified by any of the liquid-liquid extraction and the solid phase extraction described above. In addition, since rifampicin was unable to calculate LogP by ChemDraw, no value is described in the (A) column of LogP.

  According to FIG. 5, it can be seen that atorvastatin, pitavastatin, and cerivastatin can be quantified from a human plasma sample by liquid-liquid extraction. It can also be seen that lovastatin and pravastatin can be quantified in human plasma samples by solid phase extraction of HLB, and rospastatin and fluvastatin by solid phase extraction of MCX.

  Simvastatin could not be quantified by either liquid-liquid extraction or solid phase extraction described above. Moreover, since LogP of pitavastatin is not disclosed in Drug Bank, no value is described in the column (B) of LogP for pitavastatin.

  As described above, in the pretreatment DB 20, for each of a plurality of compounds for which a pretreatment method has been established, the retention time when the compound is subjected to LC under predetermined conditions, the physical property values of the compound, and the establishment Are recorded in association with the preprocessing method. Note that the data stored in the pretreatment DB 20 may be classified according to the measurement method, the type of compound, and the like as shown in FIGS. 3 to 5, but may not be classified.

[Plot of pre-processing DB data]
When the data of FIGS. 3 to 5 are plotted on a coordinate plane with the vertical axis representing LogP and the horizontal axis representing the retention time, the result is as shown in FIG. FIG. 6 is a diagram in which the data of FIGS. 3 to 5 are plotted on a coordinate plane. In the figure, the value of (B) is adopted for LogP except for pitavastatin which is not disclosed in the Drug Bank.

  FIG. 6 also illustrates LC gradient conditions (methanol concentration change with time). In the same figure, according to the lower limit of quantification, while plotting in the shape of a circle, triangle, or square, the compound for which negative measurement was performed was plotted with a white figure, and the compound for which positive measurement was performed Separated. Moreover, in the same figure, the compound name is described by the abbreviation of three alphabetic characters.

  As shown in FIG. 6, on the coordinate plane, the region where the compound suitable for liquid-liquid extraction is plotted is separated from the region where the compound suitable for solid-phase extraction is plotted. .

  From this, it is considered that a compound having a close plotted position is likely to have the same suitable pretreatment method. For example, if the retention time of a target compound for which an appropriate pretreatment method is unknown is 5.5 minutes, the target compound value is close to that of GDX (digoxin), RSV (rosuvastatin), or PVS (pravastatin). When expected to have, it can be assumed that the preferred pretreatment method is solid phase extraction. In this case, when the target compound is expected to have physical properties close to those of LNS (lansoprazole) or PTV (pitavastatin), it can be presumed that the preferred pretreatment method is liquid-liquid extraction. .

  The pretreatment determining apparatus 1 of the present embodiment uses this to narrow down the pretreatment method by obtaining the LC retention time for the target compound for which the appropriate pretreatment method is unknown.

  For example, when the retention time of the target compound is 4.5 minutes, the compound having a retention time close to this in the example of FIG. 6 is only BSP (Buspirone), so that liquid-liquid extraction suitable for BSP is effective. It can be estimated that there is.

  In addition, when the retention times of a plurality of compounds having different effective pretreatments are similar, it is possible to extract a pretreatment method suitable for each compound as a candidate and allow the user to select a pretreatment method from the candidates. Good. In this case, since the compound name and the LogP value of the compound can be a clue to select an appropriate pretreatment from a plurality of candidates, it is preferable to output them together with these pieces of information.

  For example, when the retention time of the target compound is 6.0 minutes, RPG (repaglinide), ATV (atorvastatin), and WAF (warfarin) are extracted from the compounds in the example of FIG.

  In this case, LogP of RPG is 5.05, liquid-liquid extraction is effective, LogP of ATV is 4.24, liquid-liquid extraction is effective, and LogF of WAF is 2.55. Information indicating that solid-phase extraction (MCX) is effective may be output.

  Thereby, the user can determine whether the target compound has a physical property close to RPG or ATV or a physical property close to WAF, and can determine the pretreatment method. Alternatively, the pretreatment method can be determined by inferring whether the LogP value of the target compound is close to RPG, ATV, or WAF.

  Further, as shown in FIG. 6, on the coordinate plane, a region where a compound suitable for liquid-liquid extraction is plotted and a region where a compound suitable for solid-phase extraction is plotted are linearly drawn. Can be separated by

  Thus, by specifying a straight line (or a curve) that divides a region corresponding to a suitable pretreatment method, the pretreatment method can be determined for each range of physical property values from the retention time of the target compound.

In other words, when y = f (x) is used as the line segment dividing the region, the region above this line segment in the coordinate plane corresponds to liquid-liquid extraction, and the region below corresponds to solid phase extraction. If so, the range of y ≧ f (x _i ) can be determined as liquid-liquid extraction. Similarly, the range of y <f (x _i ) can be determined as solid phase extraction. Incidentally, x is the retention time, y is LogP, _{x i} is the retention time of the target compound.

  The preprocessing determining apparatus according to the second embodiment to be described later determines a preprocessing method using such a mathematical expression.

[Preprocessing decision processing]
Next, the preprocessing determining process (preprocessing method determining method) executed by the preprocessing determining apparatus 1 will be described with reference to FIG. FIG. 7 is a flowchart illustrating an example of a preprocessing determination process for determining a preprocess from the retention time of the target compound.

  First, when the input unit 2 receives an input of the retention time of the target compound (S1), the data acquisition unit 10 outputs this data to the preprocessing determination unit 11. As the retention time, a value obtained by performing LC under the same conditions as those for obtaining the retention time in the data of FIGS. 3 to 5 described above for the target compound is used.

  Next, the preprocessing determining unit 11 refers to the preprocessing DB 20 and extracts a compound having a retention time close to the retention time of the target compound output from the data acquisition unit 10 (S2). The extraction method is not particularly limited, and for example, compounds within a predetermined time before and after the retention time of the target compound (for example, within 0.2 minutes before and after) may be extracted. Compounds may be extracted.

  Then, the pretreatment determining unit 11 refers to the pretreatment DB 20 to identify the pretreatment method for the extracted compound, and determines this as a candidate for the pretreatment method for the target compound (S3).

  Finally, the preprocessing determination unit 11 notifies the output unit 4 of the preprocessing method candidate determined in S3 together with the physical property value associated with the compound in the preprocessing DB 20, and outputs the notification (S4). The process determination process ends.

  For example, when the data shown in FIGS. 3 to 5 are stored in the preprocessing DB 20, the retention time of 6.0 minutes is input in S1, and warfarin, probenecid, repaglinide, rifampicin, atorvastatin, and pitavastatin are extracted in S2. Suppose.

  In this case, in S3, the pretreatment methods of these compounds are identified as MCX (warfarin, probenecid) and ether (repaglinide, rifampicin, atorvastatin, pitavastatin), and these pretreatment methods are candidates for the pretreatment method of the target compound. As determined.

  And in S4, said candidate is output with the value of LogP of these compounds. That is, 2.55 and 1.52 are output together with “MCX”, and 5.05, 2.36, 4.24, and 3.59 are output together with “ether”.

  Thereby, the user of the pretreatment determining apparatus 1 can narrow down the pretreatment method of the target compound to “MCX” and “ether”. Further, when the LogP value of the target compound can be predicted by, for example, ChemDraw, a preprocessing method associated with a value close to the predicted value can be set as the first candidate.

  The output data is not limited to the above example as long as at least one preprocessing method is included. For example, a configuration may be adopted in which the value of LogP is not output. However, since the value of LogP is a reference for determining a preprocessing method to be applied by the user, it is preferably output. Further, since the compound name is considered to be a reference for determining the pretreatment method, the compound name may also be output.

[Embodiment 2]
Hereinafter, another embodiment of the present invention will be described in detail with reference to FIGS. 8 and 9. In addition, the same reference number is attached | subjected to the structure similar to the said embodiment, and the description is abbreviate | omitted.

〔Device configuration〕
First, the configuration of the preprocessing determination apparatus according to the present embodiment will be described with reference to FIG. FIG. 8 is a block diagram illustrating an example of a main part configuration of the preprocessing determination device 50.

  As illustrated, in the preprocessing determination device 50, the preprocessing determination unit 11 replaces the preprocessing determination unit (preprocessing determination unit) 51 in place of the preprocessing determination unit 1 of the above embodiment, and the boundary determination unit 52 includes It is different in that it has been added.

  The preprocessing determination unit 51 determines a preprocessing method for each range of physical property values using the retention time of the target compound input from the data acquisition unit 10 and the mathematical formula determined by the boundary determination unit 52.

  The boundary determination unit 52 calculates a mathematical formula indicating a line segment that divides the coordinate plane having the retention time and the physical property value as coordinate axes for each suitable preprocessing method. The calculation of the mathematical formula will be described later.

[Preprocessing decision processing]
Next, the preprocessing determination process executed by the preprocessing determination device 50 will be described with reference to FIG. FIG. 9 is a flowchart illustrating an example of a pre-processing determination process that determines a pre-process for each physical property value range from the retention time of the target compound.

  First, when the input unit 2 receives an input of the retention time of the target compound (S10), the data acquisition unit 10 outputs this data to the preprocessing determination unit 51, and the preprocessing determination unit 51 that has received the data determines the boundary. The unit 52 is instructed to determine a mathematical expression indicating the boundary. As the retention time, a value obtained by performing LC under the same conditions as those for obtaining the retention time in the data of FIGS. 3 to 5 described above for the target compound is used.

  Next, the boundary determination unit 52 that has received the above instruction, as shown in FIG. 6, coordinates corresponding to each of the compounds included in the preprocessing DB 20 with the coordinate values of the retention time and the physical property value, respectively. Plot on the plane (S11).

  Subsequently, the boundary determination unit 52 calculates a mathematical expression indicating a line segment that divides the plotted point for each established preprocessing method on the coordinate plane (S12), and which side of the line segment indicates which side is The preprocessing determination unit 51 is notified together with the data indicating whether the preprocessing method is selected. It should be noted that the mathematical formula of the line segment that divides the points on the coordinate plane according to the attribute can be obtained by using a known algorithm such as a support vector machine.

Then, the preprocessing determination unit 51 determines a preprocessing method for each range of physical property values from the holding time input in S10 and the mathematical formula calculated in S12 (S13). Specifically, the preprocessing determination unit 51 calculates the physical property value (P _TH ) corresponding to the holding time by substituting the holding time input in S10 into the calculated mathematical formula. And, for the range of P _TH or more physical properties, the pretreatment methods liquid - Determine liquid extraction and, for a range of physical properties of less than P _TH is determined with solid phase extraction.

  Finally, the preprocessing determination unit 51 notifies the output unit 4 of the preprocessing method determined in S13 and the range of the physical property values to output (S14), and ends the preprocessing determination process.

  For example, in the example of FIG. 6, when the vertical axis is y and the horizontal axis is x, the equation of the line segment that separates the liquid-liquid extraction point and the solid-phase extraction point in S12 is y = 0.6x−1. Is calculated. In this case, the boundary determination unit 52 determines that the upper side (y ≧ 0.6x−1) of the line segment is liquid-liquid extraction and the lower side (y <0.6x−1) is solid phase extraction. The boundary determining unit 52 notifies the preprocessing determining unit 51 that the region of y ≧ 0.6x−1 is liquid-liquid extraction and the region of y <0.6x−1 is solid phase extraction. . Note that it is arbitrary which pre-processing method is used on the line segment.

Here, when the holding time input in S10 is x _i , the preprocessing determination unit 51 sets the range y ≧ 0.6x _i −1) to liquid-liquid extraction, and y <0.6x _i −1. The range is determined as solid phase extraction, and the result of this determination is output to the output unit 4. Thereby, the user of the pre-processing determination apparatus 50 can determine the pre-processing method according to the physical property value of the target compound.

  In the above example, only the retention time is input. However, by inputting the LogP value of the target compound (which may be a predicted value or the like), one preprocessing method is determined and output. Can do.

  In the above example, the mathematical formula is calculated after receiving the input of the holding time. However, the mathematical formula may be calculated in advance, and the calculation timing is not particularly limited. For example, it may be calculated at the timing when the preprocessing DB 20 is updated. In this case, in the preprocessing determination process, the preprocessing method is determined using an already created mathematical expression.

  Furthermore, for example, a mathematical expression created by a user of the preprocessing device 50 or the like may be input and used to determine the preprocessing method. In this case, the boundary determining unit 52 can be omitted.

[Modification]
In each of the above embodiments, the example in which the pretreatment method is determined using the LC retention time obtained under a single condition and the physical property value (LogP) is shown, but the present invention is not limited to this example.

  For example, the LC retention times obtained under a plurality of different conditions may be measured, and the measured values may be stored in the preprocessing DB 20. In this case, the retention time of the LC can also be measured for the target compound under a plurality of different conditions described above, and the pretreatment can be determined from the correlation between the retention times. Thereby, compared with the case where LC retention time under a single condition is used, the probability that an appropriate preprocessing method can be determined can be increased.

  Further, instead of using the physical property value itself, a numerical value having a correlation with the physical property value may be used. For example, physical property values relating to a plurality of different physical properties may be used in combination. For example, LogP and LogD may be used in combination, and a value obtained by multiplying these physical property values may be used. For example, a value obtained by multiplying a physical property value by some coefficient may be used. In this way, by using numerical values that correlate with physical property values, the boundary for each optimal preprocessing method on the coordinate plane becomes clearer and the probability that an appropriate preprocessing method can be determined is increased. Is also possible.

  Furthermore, although the example which uses LC was shown above, it is not restricted to this, It is also possible to utilize other chromatography, such as a gas chromatography and a supercritical chromatography.

[Embodiment 3]
Hereinafter, another embodiment of the present invention will be described in detail with reference to FIGS. 10 to 19. In addition, the same reference number is attached | subjected to the structure similar to the said embodiment, and the description is abbreviate | omitted. Also in this embodiment, as in each of the above-described embodiments, the retention time of LC (liquid chromatography) measured in advance for a compound for which an appropriate pretreatment method is known, and a target compound for which the appropriate pretreatment method is unknown The pretreatment method of the target compound is determined based on the measured LC retention time. A major difference from the above embodiments is that a plurality of types of retention times measured in mobile phases having different pHs are combined as parameters. That is, in this embodiment, the retention time measured with the mobile phase from which pH differs is utilized as a physical-property value in the said embodiment.

  More specifically, the target compound is subjected to LC under three types of mobile phase conditions of acidic, basic, and neutral to obtain a retention time (hereinafter also referred to as a retention index). Then, based on the Retention index, the target compound is classified into an acidic compound and a neutral / basic compound, a parameter corresponding to this classification is calculated from the Retention index, and the pretreatment is determined using this.

〔Device configuration〕
First, the configuration of the preprocessing determination apparatus according to the present embodiment will be described with reference to FIG. FIG. 10 is a block diagram illustrating an example of a main configuration of the preprocessing determining apparatus 100. As illustrated, the control unit 3 of the preprocessing determination device 100 includes a data acquisition unit 10, a DB update unit 12, a preprocessing determination unit (preprocessing determination unit) 101, a boundary determination unit 102, and a classification unit (determination unit). ) 103 is included. The storage unit 5 stores a preprocessing DB 120.

  The preprocessing determination unit 101 calculates a parameter for preprocessing determination based on the Retention index of the target compound, and based on the correlation information indicating the correlation between the parameter and the preprocessing method, the preprocessing to be applied to the target compound Determine the treatment method.

  The boundary determination unit 102 stores the pre-processing method and parameters calculated for the compound stored in the pre-processing DB 120 based on data indicating the pre-processing method established and the parameters calculated for the compound. The boundary value of the parameter for determining the processing method is specified.

  The classification unit 103 classifies the target compound into an acidic compound or a neutral / basic compound based on the Retention index of the target compound. Specifically, the classification unit 102 determines whether the ratio of the Retention index in the acidic condition of the target compound to the Retention index in the basic condition is greater than or equal to a predetermined threshold for determining a parameter used for determination of pretreatment, It is determined whether it is less than the threshold value. And the classification | category part 103 will classify | categorize as an acidic compound if the said ratio is more than a threshold value, and will classify as a neutral and a basic compound if less than a threshold value. That is, the parameter used for determining the pretreatment of the acidic compound is different from the parameter used for determining the pretreatment of the neutral / basic compound.

Here, an example will be described in which an experimentally obtained threshold value = 1.4 is used when a retention index calculated based on a measurement method described later is used. That is, the “acidic compound” in the present embodiment refers to a compound that satisfies the following conditions, and the “neutral compound” and “basic compound” refer to compounds that do not satisfy the following conditions.
(Retention index of acidic condition) / (Retention index of basic condition) ≧ 1.4
Hereinafter, (Retention index under acidic conditions) / (Retention index under basic conditions) is abbreviated as an acid / base ratio.

  As described above, the pretreatment DB 120 is a database that stores data indicating the established pretreatment method and the parameters calculated for the compound for the compound for which the pretreatment method has been established. Since the parameter can be calculated from the Retention index, the Retention index may be stored instead of or together with the parameter.

〔Measuring method〕
A method for measuring the retention index will be described with reference to FIG. FIG. 11 is a diagram illustrating an example of the retention index measurement conditions. The column used is BEH C18 2.1 × 50 mm, 1.7 μm (Waters), and is a reverse phase column as in the above embodiment. The mobile phase is a mixed solution of solution A and solution B (acetonitrile). Solution A is 0.1% formic acid for acidic conditions (first condition), 0.1% ammonium hydroxide for basic conditions (second condition), and neutral conditions (third condition). In the case of (condition), it is 10 mM ammonium acetate.

  As shown in the figure, the flow rate was 0.7 mL / min, and the gradient was increased from 2% to 98% in 3 minutes from 0.0 minutes to 3.0 minutes. The ratio of B was maintained at 98% for 0.5 minutes, and thereafter the ratio of B was reduced to 2% again in 0.1 minutes, and measurement was performed up to 4.5 minutes. The “curve” in the figure is a parameter indicating how to apply a gradient (inclination).

[Measured compounds]
Retention index was measured for 27 types of compounds under acidic, neutral, and basic conditions. The results are shown in FIG. 12 and FIG. For example, the salicylic acid in FIG. 12 has a retention index of about 1.2 under acidic conditions and a retention index of about 0.5 under basic conditions, and therefore satisfies the condition of acid / base ratio ≧ 1.4. Categorized as acidic compounds. On the other hand, clemastine has a retention index of about 1.7 under acidic conditions and a retention index of about 2.7 under basic conditions, so it does not satisfy the above conditions and is classified as a neutral / basic compound. Is done.

  As will be described below, suitable pretreatments were examined for each of these 27 types of compounds, and the correspondence between parameters based on the Retention index and suitable pretreatments was examined.

[Pretreatment (solid phase extraction)]
Each compound whose Retention index was measured as described above was pretreated using three types of cartridges for solid phase extraction (Oasis HLB, Oasis MCX, Oasis MAX). These cartridges are the same as those used in the first embodiment. The HLB is a cartridge filled with a reverse phase filler having both a hydrophilic group and a lipophilic group. MCX is a reversed-phase / cation exchange mixed-mode (mixed) solid phase and is generally suitable for selective extraction of basic compounds. MAX is a mixed-phase solid phase of reversed-phase / weak anion exchange, and is generally suitable for selective extraction of acidic compounds.

  More specifically, the pretreatment with MAX is performed by loading a compound sample on a cartridge, washing with 5% ammonium hydroxide, then eluting with a neutral eluent (100% methanol), and further using an acidic eluent. Elute with (2% formic acid methanol solution). In MCX pretreatment, the compound sample was loaded on the cartridge, washed with 2% formic acid, eluted with neutral eluent (100% methanol), and then basic eluent (5% water). Elution with ammonium oxide methanol solution). Thus, in the pretreatment with these two types of cartridges, two types of eluates were obtained for each type of cartridge. On the other hand, in the pretreatment with HLB, a compound sample was loaded onto a cartridge, washed with a 5% aqueous methanol solution, and then eluted with a neutral eluent (100% methanol). That is, there is only one type of eluate obtained by pretreatment with HLB.

[Pretreatment (liquid-liquid extraction)]
In addition, for each of the compounds for which the retention index was measured as described above, in three types of solutions (diethyl ether, ethyl acetate, diethyl ether and isopropyl alcohol (IPA) 8: 2 (v / v) mixed solution). Liquid-liquid extraction was performed. Diethyl ether is a low polarity solvent, and ethyl acetate is a medium to high polarity solvent. Extraction with each solution was performed under acidic and alkaline conditions.

[Evaluation by score]
In order to identify a suitable pretreatment, the recovery rate of the compound by each of the above pretreatments was evaluated by a score. A part of the result is shown in FIG. FIG. 14 is a diagram showing the correspondence between pretreatment and score for each compound. The recovery rate was calculated as follows.
(1) Human plasma (Recovery QC) to which a mixed standard solution is added and human plasma (Blank) to which a mixed standard solution is not added and the above three solid-phase extraction conditions (HLB / MCX / MAX) and liquid-liquid extraction Pretreatment is performed under 6 conditions (diethyl ether / ethyl acetate / diethyl ether-IPA extraction under acidic and basic conditions). For Blank, a mixed standard solution was added to the solution before evaporation to dryness (Contorol QC).
(2) About the injection sample of Recovery QC and Controlol QC, it measured using LC and a tandem mass spectrometer, and the recovery rate was computed from the measurement result.

  A score of 7 was given when the recovery rate was 70% or more, 3 when it was 40% or more and less than 70%, 1 when it was less than 40%, and 0 when no peak was detected by LC. In addition, the sum of the scores for the six conditions of liquid-liquid extraction was calculated as the LLE score, and the total of the scores for the five conditions of solid phase extraction was calculated as the SPE score.

  FIG. 14 also shows the peak detection sensitivity in LC. The sensitivity is evaluated in three stages, and “◯”, “Δ”, and “×” are evaluated in the descending order of sensitivity. More specifically, among Recovery QCs, “○” indicates that sensitivity was obtained at both high concentration (High) and low concentration (Low) (a peak was detected), and “△” indicates that only high concentration was detected. “A” indicates that it was difficult to detect a peak at any concentration. In this example, the peak detection sensitivity is not reflected in the score. However, the sensitivity may be reflected numerically and added to the score. FIG. 14 shows only 12 of the 27 types of compounds, and the evaluation results of the remaining 15 types of compounds are omitted.

[Correlation between LLE score and retention index for neutral and basic compounds]
FIG. 15 shows the coordinates of the LLE score calculated as described above for the neutral / basic compound and the total value of the retention index of the acidic, neutral and basic conditions (hereinafter abbreviated as the total value as appropriate). It is the figure plotted on the plane. Further, in the same figure, the optimum pretreatment types are shown by the shape of the points plotted (diamonds are liquid-liquid extraction with diethyl ether, squares are liquid-liquid extraction with diethyl ether / IPA mixture, triangles are MCX By solid phase extraction).

  As shown in the figure, the neutral / basic compound having a high total value has a high LLE score. More specifically, for neutral / basic compounds having a total value of 4 or more, the optimal pretreatment is liquid-liquid extraction with diethyl ether or a mixture of diethyl ether / IPA, and the total value is less than 4. For basic compounds, the optimal pretreatment was solid phase extraction with MCX.

  FIG. 16 shows the values of (Retention index of basic condition) / (Retention index of neutral condition) and the total value of acidic, neutral and basic retention index for neutral / basic compounds. It is the figure plotted on the coordinate plane. As shown in this figure, for neutral / basic compounds, liquid-liquid extraction is a preferred pretreatment method when the total value is 4 or more, and solid-phase extraction is preferred when the total value is less than 4.

  As described above, with respect to the neutral / basic compound, it is possible to specify whether the preferable pretreatment method is solid phase extraction or liquid-liquid extraction by using the total value as a parameter. That is, in the pretreatment determining apparatus 100, a total value is obtained for each of a plurality of neutral / basic compounds for which it is known whether the preferred pretreatment method is solid phase extraction or liquid-liquid extraction. A value and a preferred preprocessing method are associated with each other and stored in the preprocessing DB 120 in advance. Then, the boundary determination unit 102 refers to the pretreatment DB 120 to determine the boundary values for neutral / basic compounds for determining whether the preferred pretreatment method is solid phase extraction or liquid-liquid extraction. obtain.

[Correlation between LLE score and retention index for acidic compounds]
FIG. 17 is a diagram in which the LLE score calculated as described above and the value of the acid / base ratio are plotted on a coordinate plane for an acidic compound. Also, in the figure, the optimum pretreatment types are indicated by the shape of the points plotted (diamonds are liquid-liquid extraction with diethyl ether and solid phase extraction with MAX, open squares are with diethyl ether and ethyl acetate. Liquid-liquid extraction, MCX (neutral extract), solid phase extraction with HLB, triangles for solid phase extraction with HLB, circles for solid phase extraction with MAX, solid squares for MAX, MCX (neutral extract), HLB By solid phase extraction).

  As shown in the figure, the acidic compound having a high acid / base ratio has a high LLE score. More specifically, for acidic compounds with an acid / base ratio value of 1.7 or more, the optimal pretreatment includes liquid-liquid extraction with diethyl ether or ethyl acetate, and for acidic compounds of less than 1.7 The optimal pretreatment was solid phase extraction.

  FIG. 18 is a diagram in which an acid / base ratio value and a total value of acidic, neutral, and basic retention indexes are plotted on a coordinate plane for an acidic compound. As shown in this figure, for acidic compounds, liquid-liquid extraction is the preferred pretreatment method when the acid / base ratio is 1.7 or more, and liquid-liquid extraction is less than 1.7. ing.

Thus, for acidic compounds, it is possible to specify whether the preferred pretreatment method is solid phase extraction or liquid-liquid extraction by using the acid / base ratio as a parameter. In addition, since the acidic compound in this example has an acid / base ratio of 1.4 or more, the pretreatment method can also be specified under the following conditions. In other words, when the compound to be specified in the pretreatment method is specialized in an acidic compound, only the retention index of the acidic condition and the basic condition needs to be obtained, and the retention index of the neutral condition can be omitted. .
Conditions for determining solid phase extraction: 1.4 ≦ (acid / base ratio) <1.7
Conditions for determining liquid-liquid extraction: 1.7 ≦ (acid / base ratio)
That is, in the pretreatment determining apparatus 100, the acid / base ratio is obtained for each of a plurality of acidic compounds for which it is known whether the preferred pretreatment method is solid phase extraction or liquid-liquid extraction, and the obtained acid / base is obtained. The ratio and the preferred preprocessing method are associated with each other and stored in the preprocessing DB 120 in advance. Then, the boundary determination unit 102 refers to the pretreatment DB 120 to obtain a boundary value for an acidic compound for determining whether the preferred pretreatment method is solid phase extraction or liquid-liquid extraction.

[Preprocessing decision processing]
Next, the preprocess determination process (preprocess determination method) executed by the preprocess determination apparatus 100 will be described with reference to FIG. FIG. 19 is a flowchart illustrating an example of a pre-processing determination process for determining the pre-processing of the target compound from the retention time of the target compound to be determined for pre-processing.

First, when the input unit 2 receives an input of the retention time of the target compound (S20, data acquisition step), the data acquisition unit 10 outputs this data to the classification unit 103, and the classification unit 103 that has received the data receives the retention time. The target compounds are classified based on the ratio (S21). The retention time is a value obtained by conducting LC under the same conditions as those for obtaining the retention time in the data of FIGS. 12 and 13 (acidic, neutral, basic) for the compound for which pretreatment is to be determined. Retention index in each condition is used. Further, the classification based on the retention time ratio is, as described above, the acid / base ratio ≧ 1.4.
The target compound that satisfies the above condition is classified as an acidic compound, and the compound that does not satisfy the condition is classified as a neutral / basic compound.

  Here, when the target compound is classified as an acidic compound (YES in S22), the classification unit 103 transmits the retention time received from the data acquisition unit 10 to the preprocessing determination unit 101 and notifies the classification result. To do. Then, the pretreatment determining unit 101 obtains a predetermined threshold value for acidic compounds (boundary value: 1.7 in the examples of FIGS. 17 and 18) from the boundary determining unit 102, and the retention time ratio (acid / base ratio). ) Is greater than or equal to the threshold value (S23).

  If the result of this determination is greater than or equal to the threshold value (YES in S23), the pretreatment determination unit 101 determines that the pretreatment method suitable for the target compound is liquid-liquid extraction (S24, pretreatment determination step). On the other hand, if it is less than the threshold value (NO in S23), the pretreatment determining unit 101 determines that the pretreatment method suitable for the target compound is solid phase extraction (S25, pretreatment determining step). In other words, the pretreatment determining unit 101 performs correlation information indicating a correlation between a predetermined parameter (acid / base ratio) and the pretreatment method (liquid-liquid extraction if the acid / base ratio is equal to or greater than the threshold value). If it is less than that, information indicating solid phase extraction) is stored in advance, and the pretreatment method is determined based on this correlation information. Then, the preprocessing determination unit 101 notifies the output unit 4 of the preprocessing method determined in S24 or S25 and outputs it (S26), and ends the preprocessing determination process.

  Here, when the target compound is classified as a neutral / basic compound in S21, the classification unit 103 determines NO in S22, and the retention time received from the data acquisition unit 10 to the preprocessing determination unit 101 Is sent, and the classification result is notified. Then, the preprocessing determination unit 101 obtains a predetermined threshold value (boundary value: 4.0 in the examples of FIGS. 15 and 16) for the neutral / basic compound from the boundary determination unit 102, and the total value of the retention times It is determined whether (the total value of each Retention index of acidic, neutral, and basic conditions) is equal to or greater than the threshold value (S27).

  If the result of this determination is greater than or equal to the threshold value (YES in S27), the pretreatment determination unit 101 determines that the pretreatment method suitable for the target compound is liquid-liquid extraction (S24, pretreatment determination step). On the other hand, if it is less than the threshold (NO in S27), the pretreatment determining unit 101 determines that the pretreatment method suitable for the target compound is solid phase extraction (S28, pretreatment determining step). In other words, the preprocessing determination unit 101 performs correlation information indicating a correlation between a predetermined parameter (total value) and the preprocessing method (liquid-liquid extraction if the total value is greater than or equal to a threshold value, and if less than the threshold value) Information indicating solid phase extraction) is stored in advance, and the pretreatment method is determined based on the correlation information. Then, the preprocessing determining unit 101 notifies the output unit 4 of the preprocessing method determined in S24 or S28 and outputs it (S26), and ends the preprocessing determining process.

  In the above example, after classification in S21, a parameter (acid / base ratio or total value) is compared with a threshold value in S23 or S27, and a pretreatment method is determined in S25, S24, or S28. ing. However, these treatments can be performed together by using a combination of the conditions for classifying the compound and the conditions for determining the pretreatment method. In other words, solid phase extraction may be determined if 1.7> acid / base ratio ≧ 1.4, and liquid-liquid extraction may be determined if 1.7 ≦ acid / base ratio. If the acid / base ratio <1.4 and the total value ≧ 4, the liquid-liquid extraction is determined. If the acid / base ratio <1.4 and the total value <4, the solid-phase extraction is determined. You may decide. In this case, the correlation information of the acidic compound indicates that the acid / base ratio is solid phase extraction if the acid / base ratio is not less than a predetermined lower limit value and less than the predetermined upper limit value, and is liquid-liquid extraction if the acid / base ratio is not less than the predetermined upper limit value. It becomes information to show. The correlation information of the neutral / basic compound is liquid-liquid extraction when the acid / base ratio is less than a predetermined lower limit and the total value is equal to or higher than the predetermined upper limit, and the acid / base ratio is predetermined. If it is less than the lower limit value and the total value is less than the predetermined upper limit value, it is information indicating that the extraction is solid phase. In this case, the classification unit 103 can be omitted.

[Modification]
In the above example, the Retention index in each of acidic, neutral, and basic conditions was measured by changing only the type of substance used in the mobile phase (solution A in the example of FIG. 11). The conditions may be changed. However, the conditions for measuring the retention index of the target compound and the conditions for measuring the retention index of the compound for which pretreatment suitable for pretreatment DB 120 is stored need to be the same.

  In the above example, the total value of the retention index and the acid / base ratio in each of the acidic, neutral, and basic conditions are used as parameters, but the parameter based on the retention index is not limited to these examples. . For example, instead of the total value of Retention indexes under acidic, neutral, and basic conditions, the average value of these Retention indexes may be used.

  Furthermore, in the above example, the preprocessing method is determined using one parameter and one boundary value, but the preprocessing method may be determined using a plurality of parameters. In this case, for example, as in the example of the second embodiment, the preprocessing method may be determined from the plot position of the target compound on the coordinate plane with each parameter as the coordinate axis.

  In the above example, solid phase extraction or liquid-liquid extraction is determined as a pretreatment method. However, liquid-liquid extraction with, for example, diethyl ether is preferable by changing parameters and boundary values to be used. It is also possible to determine a more detailed preprocessing method.

[Demonstration of effect]
In order to demonstrate the effect of the pretreatment determining apparatus 100, the pretreatment of the compound used in the first embodiment was determined by the pretreatment determining apparatus 100. This will be described with reference to FIG. FIG. 20 is a diagram showing parameters obtained for each compound as a target of the effect verification. As shown in the figure, the obtained parameters are the acidic, basic, and neutral retention indexes, the acid / base ratio calculated from these values, and the total value (the sum of the acidic, basic, and neutral retention indexes. Value).

  In addition, the “pre-processing method” in the figure describes the pre-processing method determined to be suitable in the first embodiment. Note that methotrexate was not measurable in FIG. 4, but this was due to the influence of the contaminating peak, which could be detected by changing the LC conditions and separating the contaminating peak. Since the preferred pretreatment method for methotrexate was solid phase extraction with MCX, in FIG. 20, the “pretreatment method” for methotrexate is MCX. In the figure, “ether” is liquid-liquid extraction with diethyl ether.

  As described above, the pretreatment determining apparatus 100 classifies target compounds that satisfy the condition of “acid / base ratio ≧ 1.4” as acidic compounds, and classifies those compounds that do not satisfy the conditions as neutral / basic compounds. Therefore, among the compounds of FIG. 20, four types of compounds from fluvastatin to probenecid are classified as acidic compounds, and the remaining seven types are classified as neutral / basic compounds.

  The pretreatment determining apparatus 100 determines that a suitable pretreatment method is liquid-liquid extraction if the condition of “acid / base ratio ≧ 1.7” is satisfied for an acidic compound, and is preferable if the condition is not satisfied. Determine that the pretreatment method is solid phase extraction. Thus, among the four acidic compounds, the preferred pretreatment method for fluvastatin, methotrexate, and chlorzoxazone is determined to be solid phase extraction, and the preferred pretreatment method for probenecid is liquid-liquid extraction. It is determined.

  Here, as shown in the figure, the “pretreatment method” of fluvastatin, methotrexate, and chlorzoxazone is all solid phase extraction by MCX. That is, for these three types of compounds, the pretreatment determining apparatus 100 was able to determine the correct pretreatment method. The “pretreatment method” of probenecid was solid phase extraction by MCX, and the pretreatment determination apparatus 100 could not determine the correct pretreatment method for this compound.

  Further, the pretreatment determining device 100 determines that a suitable pretreatment method is liquid-liquid extraction if the condition of “total value ≧ 4” is satisfied for neutral / basic compounds, and is preferable if not satisfied. Determine that the pretreatment method is solid phase extraction. Thus, the preferred pretreatment method is determined to be liquid-liquid extraction for pitavastatin, cerivastatin, dextromethorphan, and desipramine, and the preferred pretreatment method is determined to be solid phase extraction for omeprazole and lansoprazole. .

  Here, as shown in the figure, the “pretreatment methods” for pitavastatin, cerivastatin, dextromethorphan, and desipramine are all liquid-liquid extraction with “ether”, ie, diethyl ether. That is, the pretreatment determining apparatus 100 was able to determine the correct pretreatment method for these four types of compounds. On the other hand, for omeprazole and lansoprazole, the pretreatment determination device 100 could not determine a correct pretreatment method.

  As described above, according to the pretreatment determining apparatus 100, it was proved that a correct pretreatment method can be determined for the compound used in the first embodiment. Of the compounds for which the correct pretreatment method has not been determined, the acid / base ratio of probenecid is 1.8, which is close to the threshold value (1.7). The total value of lansoprazole is 3.91, which is also close to the threshold value (4). Such misjudgment near the threshold value can be reduced by accumulating data of a larger number of compounds in the preprocessing DB 120 and updating the threshold value. In addition, it may be possible to reduce misjudgment by optimizing the LC condition for obtaining the retention index.

[Embodiment 4]
Hereinafter, another embodiment of the present invention will be described in detail with reference to FIGS. 21 and 22. In the present embodiment, as in the third embodiment, the present invention is applied to the target compound based on the predetermined parameter calculated using the Retention index and the correlation information indicating the correlation between the parameter and the pretreatment method. Determine the power pre-treatment method. Since the difference from the third embodiment is only the parameter to be used and the correlation information, this difference will be mainly described here.

[Parameters to be used]
In the present embodiment, in addition to the acid / base ratio that is the parameter used in the third embodiment, the parameters of the acidic K ′ / neutral K ′ ratio, the basic K ′ / neutral K ′ ratio, and the total value K ′ are used. Is used to determine the pretreatment method.

The acidity K ′ is a value obtained by correcting the retention index of the acidic condition with t0. Similarly, the neutral K ′ is the value obtained by correcting the retention index of the neutral condition with t0, and the basic K ′ is that of the basic condition. This is a value obtained by correcting the Retention index with t0. The acidic K ′ / neutral K ′ ratio is expressed by the following mathematical formula.
(Acid K ′ / neutral K ′ ratio) = {(Retention index of acidic conditions−t0) / t0} / {(Retention index of neutral conditions−t0) / t0}
The basic K ′ / neutral K ′ ratio is expressed by the following mathematical formula.
(Basic K ′ / neutral K ′ ratio) = {(Retention index of basic condition−t0) / t0} / {(Retention index of neutral condition−t0) / t0}
The total value K ′ is expressed by the following mathematical formula.
(Total value K ′) = acidic K ′ + basic K ′ + neutral K ′
Note that t0 in the above formula is the retention time of components not retained in the column. The time when the solvent peak in the LC is detected may be t0, or t0 may be calculated by using the formula “t0 = column volume / LC flow rate”. Regardless of which method is used, the value of t0 is theoretically the same. Since the value of t0 is uniquely determined by the LC condition, it may be stored, for example, in the storage unit 5 or the like so that the preprocessing determination apparatus 100 can refer to it.

  By adding t0, the Retention index is a value that more clearly reflects how strongly the compound is retained in the column (affinity between the compound and the column). This is because the retention index can change depending on factors such as the performance of the column and the degree of deterioration, but such a change can be offset by calculating the ratio to t0.

[Correlation between parameters and pretreatment method]
Next, the correlation between the above three parameters and the preprocessing method will be described with reference to FIGS. FIG. 21 is a diagram showing a suitable pretreatment method and parameters for a compound classified as an acidic compound by the method of the present embodiment (details will be described later), and FIG. 22 is a diagram of the method of the present embodiment. It is the figure which showed the suitable pre-processing method and parameter about the compound classified into the neutral and basic compound in (it mentions later for details). In these figures, “ether” refers to liquid-liquid extraction with diethyl ether, “ethyl acetate” refers to liquid-liquid extraction with ethyl acetate, and “E / IPA” refers to diethyl ether and isopropyl alcohol (IPA). 8: Refers to liquid-liquid extraction with 2 (v / v) mixed solution.

  In the present embodiment, acidic compounds and neutral / basic compounds are classified using parameters of acid / base ratio and acidic K ′ / neutral K ′ ratio. Specifically, the classification unit 103 classifies a compound that satisfies both the condition of the acid / base ratio ≧ 1.4 and the condition of the acidic K ′ / neutral K ′ ratio ≧ 1 as an acidic compound, and at least one of them is classified. Compounds that do not satisfy the above conditions are classified as neutral / basic compounds. Since it can be said that a compound having a larger value of the acidic K ′ / neutral K ′ ratio has a stronger tendency to be an acidic compound, by using the acidic K ′ / neutral K ′ ratio for classification, an acidic compound and a neutral / base The physical properties of a compound can be more accurately reflected in the classification of a chemical compound. Of the compounds used in Embodiment 3 and the compounds used in Embodiment 1, FIG. 21 shows compounds classified as acidic compounds based on such criteria, and FIG. 22 shows such compounds based on such criteria. Compounds classified as neutral and basic compounds are shown.

  For acidic compounds, as in Embodiment 3, for compounds that satisfy the condition of acid / base ratio ≧ 1.7, a pretreatment method for liquid-liquid extraction is associated, and for compounds that do not satisfy this condition, solid phase is used. An extraction pre-processing method is associated. Based on this association, the correct pretreatment method can be determined for 11 types of compounds excluding probenecid among the 12 types of compounds shown in FIG.

  On the other hand, the neutral / basic compound is further classified by whether or not the classification unit 103 satisfies the condition of the basic K ′ / neutral K ′ ratio ≧ 1. A compound that satisfies this condition is classified as a basic compound because it has a higher affinity with the column under basic conditions than under neutral conditions, and a compound that does not satisfy these conditions is classified as neutral. To do. It can be said that a compound having a larger basic K ′ / neutral K ′ ratio has a stronger tendency to be a basic compound.

  For compounds that satisfy the condition of the total value K ′ ≧ 10.38 among the neutral compounds, the pretreatment method for liquid-liquid extraction is associated, and for the compounds that do not satisfy this condition, the pretreatment method for solid phase extraction. Are associated. Based on this association, a correct pretreatment method can be determined for 8 types of compounds other than lovastatin among 10 types of neutral compounds shown in FIG.

  That is, the boundary determination unit 102 determines the threshold value (10.38) in the above condition based on the data stored in the preprocessing DB 120. Further, the preprocessing determination unit 102 stores correlation information indicating that liquid-liquid extraction is performed when the total value K ′ is equal to or greater than a threshold value, and solid-phase extraction is performed when the total value K ′ is less than the threshold value. Thereby, the preprocessing determination unit 102 can acquire the threshold value from the boundary determination unit 102 and determine a preprocessing method according to the comparison result between the threshold value and the total value K ′.

  The threshold value “10.38” is the highest total K ′ value (10.32 of labetalol) among the compounds effective for solid-phase extraction and the compound effective for liquid-liquid extraction with diethyl ether. It is an intermediate value between the smallest total value K ′ (10.44 of omeprazole). Needless to say, the threshold value is not limited to this example as long as it can distinguish between a compound effective for solid-phase extraction and a compound effective for liquid-liquid extraction with diethyl ether.

  Here, since lovastatin has the same Retention index under any of acidic, neutral, and basic conditions, it is considered that the retention time is a compound that is hardly affected by the pH of the LC mobile phase. That is, lovastatin is considered to be a compound that is not suitable for the method of this embodiment in which the pretreatment method is determined by using the change in retention time due to the difference in pH of the mobile phase based on the physical properties of the compound. Therefore, for compounds such as lovastatin that have a retention time that is not easily affected by the pH of the LC mobile phase (compounds that have the same or close retention time in the mobile phase at each pH), determine the pretreatment method. It may be excluded.

  As for basic compounds, 14 out of 16 types shown in FIG. 22 are effective for solid phase extraction with MCX, and 11 types are effective for liquid-liquid extraction with diethyl ether. Therefore, for a basic compound, a pretreatment method for solid phase extraction with MCX and a pretreatment method for liquid-liquid extraction with diethyl ether are associated. Based on this association, the correct pretreatment method can be determined for all of the 16 types of basic compounds shown in FIG.

  That is, the pretreatment determining unit 102 stores the correlation information indicating that the basic compound is a solid phase extraction pretreatment method using MCX and liquid-liquid extraction using diethyl ether. Thereby, the pretreatment determination unit 102 can determine the pretreatment method of the compound classified as the basic compound by the classification unit 103 as the pretreatment method of solid phase extraction using MCX and the liquid-liquid extraction using diethyl ether. . Any one of these may be determined as the pretreatment method.

  Similarly to the third embodiment, the compound satisfying the condition “total value ≧ 4” is associated with the pretreatment method for liquid-liquid extraction, and the compound not satisfying this condition is associated with the pretreatment method for solid phase extraction. May be. In this case, only procainamide does not satisfy the above conditions, the pretreatment method is determined as solid phase extraction, and the others are determined as liquid-liquid extraction. Even in this case, the correct pretreatment method can be determined for all of the 16 types of basic compounds shown in FIG.

  In addition, the pretreatment method may be determined using a combination of the compound classification condition and the pretreatment method determination condition. That is, if 1.7> acid / base ratio ≧ 1.4 and acidic K ′ / neutral K ′ ratio ≧ 1, solid phase extraction is determined, and 1.7 ≦ acid / base ratio and acidic K ′ / medium. If the property K ′ ratio ≧ 1, liquid-liquid extraction may be determined. Further, at least one of the conditions of acid / base ratio ≧ 1.4 and acid K ′ / neutral K ′ ratio ≧ 1 is not satisfied, and basic K ′ / neutral K ′ ratio <1, and the total If the value K ′ ≧ 10.38, it is determined as liquid-liquid extraction, the acid / base ratio ≧ 1.4 and the acidic K ′ / neutral K ′ ratio ≧ 1 are not satisfied, and the base If the ratio of sex K ′ / neutral K ′ <1 and the total value K ′ <10.38, solid phase extraction may be determined. If the acid / base ratio ≧ 1.4 and the acidic K ′ / neutral K ′ ratio ≧ 1 are not satisfied, and if the basic K ′ / neutral K ′ ratio ≧ 1, then MCX The pretreatment method for solid phase extraction and liquid-liquid extraction with diethyl ether may be determined. In these cases, the classification unit 103 can be omitted.

  As described above, the correlation information in the present embodiment is information indicating the following (1) to (3). The following (1) is an acidic compound, (2) is a basic compound, and (3) is correlation information for determining a pretreatment method for a neutral compound. In addition, the pretreatment determining apparatus 100 may determine the pretreatment method of the target compound using all of the correlation information, or may determine the pretreatment method of the target compound using only one or two of them. May be. That is, the preprocessing determination device 100 may be a preprocessing determination device specialized for one or two of acidic compounds, basic compounds, and neutral compounds.

  (1) The acid / base ratio is less than a predetermined upper limit value (1.7 in the above example) and not less than a predetermined lower limit value (1.4 in the above example), and further an acidic K ′ / neutral K ′ ratio If ≧ 1, the pretreatment method is solid-phase extraction, the acid / base ratio is not less than a predetermined upper limit (1.7 in the above example), and the acidic K ′ / neutral K ′ ratio ≧ 1 If present, the pretreatment method is liquid-liquid extraction.

  (2) Acid / base ratio ≧ predetermined lower limit value (1.4 in the above example) and acidic K ′ / neutral K ′ ratio ≧ 1 when basic K ′ / medium is not satisfied If the property K ′ ratio> 1, the pretreatment method is a solid phase extraction with a reversed phase-cation exchange mixed solid phase (MCX) and a liquid-liquid extraction with diethyl ether. In addition, if it is a solid phase of the mixed system of reverse phase-cation exchange, an effective pretreatment is possible in the same manner as MCX.

  (3) Acid / base ratio ≧ predetermined lower limit (1.4 in the above example) and at least one of acidic K ′ / neutral K ′ ratio ≧ 1 and basic K ′ / neutral When K ′ ratio ≦ 1, if the total value K ′ ≧ predetermined lower limit (10.38 in the above example), the pretreatment method is liquid-liquid extraction, and the total value K ′ <predetermined If it is the lower limit (10.38 in the above example), the pretreatment method is solid phase extraction.

  In addition, the value of said upper limit and lower limit is an example, and is not restricted to these examples. By accumulating data of a larger number of compounds in the pretreatment DB 120 and updating the above upper limit value and lower limit value, it is possible to increase the estimation accuracy of the pretreatment method. In addition, in order to increase the estimation accuracy, the LC condition for obtaining the Retention index may be optimized. In this case, an upper limit value and a lower limit value are set according to the optimized LC condition.

  As described above, in this embodiment, the values (acidic K ′, neutral K ′, basic K ′) calculated from the numerical values of the retention index itself are used instead of the numerical values of the retention index itself. Then, the preprocessing method is determined using various parameters obtained from these values. This makes it possible to determine the correct pretreatment method for the compounds (omeprazole and lansoprazole) for which the correct pretreatment method could not be determined in the third embodiment.

  In addition, the parameter used for determination of a pre-processing method should just be a parameter calculated using Retention index, and is not restricted to said example. The same applies to the correlation information, and the correlation information is not limited to the above example as long as the information can determine the preprocessing method from the parameter calculated using the Retention index. By setting appropriate parameters and correlation information, it is possible to further improve the estimation accuracy of the preprocessing method.

  Appropriate parameters and correlation information are stored in the pretreatment DB 120 for the retention indexes and suitable pretreatment methods of a large number of compounds, and the correlation between the value of the parameter calculated using the retention index and the suitable pretreatment method, It can be determined by finding the parameters, such as by plotting them on a coordinate plane.

[Combination of Embodiments]
The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in different embodiments. Is also included in the technical scope of the present invention.

[Example of software implementation]
Finally, each block of the preprocessing determination devices 1, 50, 100, in particular, the control unit 3 may be realized in hardware by a logic circuit formed on an integrated circuit (IC chip), or may be a CPU (Central It may be realized by software using a Processing Unit).

  In the latter case, the preprocessing determination devices 1, 50, and 100 include a CPU that executes instructions of a program that realizes each function, a ROM (Read Only Memory) that stores the program, and a RAM (Random Access Memory) that expands the program. ), A storage device (recording medium) such as a memory for storing the program and various data. The object of the present invention is to enable the computer to read the program code (execution format program, intermediate code program, source program) of the control program of the preprocessing determination devices 1, 50, 100, which is software that implements the functions described above. This can also be achieved by supplying the recorded recording medium to the preprocessing determining devices 1, 50, 100, and the computer (or CPU or MPU) reading and executing the program code recorded on the recording medium.

  Examples of the recording medium include non-transitory tangible medium, such as magnetic tape and cassette tape, magnetic disk such as floppy (registered trademark) disk / hard disk, and CD-ROM / MO. Discs including optical discs such as / MD / DVD / CD-R, cards such as IC cards (including memory cards) / optical cards, semiconductor memories such as mask ROM / EPROM / EEPROM (registered trademark) / flash ROM Alternatively, logic circuits such as PLD (Programmable Logic Device) and FPGA (Field Programmable Gate Array) can be used.

  Further, the preprocessing determination devices 1, 50, 100 may be configured to be connectable to a communication network, and the program code may be supplied via the communication network. The communication network is not particularly limited as long as it can transmit the program code. For example, the Internet, intranet, extranet, LAN, ISDN, VAN, CATV communication network, virtual private network, telephone line network, mobile communication network, satellite communication network, and the like can be used. The transmission medium constituting the communication network may be any medium that can transmit the program code, and is not limited to a specific configuration or type. For example, even with wired lines such as IEEE 1394, USB, power line carrier, cable TV line, telephone line, ADSL (Asymmetric Digital Subscriber Line) line, infrared rays such as IrDA and remote control, Bluetooth (registered trademark), IEEE 802.11 wireless, HDR ( It can also be used by radio such as High Data Rate (NFC), Near Field Communication (NFC), Digital Living Network Alliance (DLNA), mobile phone network, satellite line, and digital terrestrial network. The present invention can also be realized in the form of a computer data signal embedded in a carrier wave in which the program code is embodied by electronic transmission.

[Summary]
The pretreatment determination device of the present invention is a pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed, and the target compound is chromatographed under a predetermined condition. Data acquisition means for obtaining the retention time when the compound is subjected to the pretreatment method, and the retention time when the compound is subjected to chromatography under the above conditions for each of the plurality of compounds for which the pretreatment method has been established, and the physical properties of the compound Is associated with a compound having a retention time within a predetermined range from the retention time acquired by the data acquisition means, based on database data recorded in association with the physical property value indicating A pretreatment determining means for determining a pretreatment method as a pretreatment method for the target compound and outputting the pretreatment method together with physical property values associated with the pretreatment method. It is characterized in.

  Further, in order to solve the above-described problem, the pretreatment method determination method of the present invention is a pretreatment by a pretreatment determination device that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. A method for determining a method, the data acquisition step for acquiring a retention time when the target compound is subjected to chromatography under a predetermined condition, and each of a plurality of compounds for which a pretreatment method has been established, Based on the database data recorded in association with the retention time when subjected to chromatography under conditions, the physical property value indicating the physical property of the compound, and the established pretreatment method, the data was acquired in the data acquisition step. A pretreatment method associated with a compound having a retention time within a predetermined range from the retention time is determined as a pretreatment method for the target compound, and the pretreatment method is supported. It is characterized in that it comprises a pretreatment determination step of outputting together with the physical properties being eclipsed.

  Which pretreatment method is effective is highly correlated with the physical properties of the target compound, and the retention time in an analyzer such as chromatography varies depending on the physical properties, etc. It can be a factor for determining the processing method.

  In fact, as a result of experiments by the inventor of the present application, when plotting retention times of liquid chromatography and physical property values on a coordinate plane, the plots of substances with common pretreatment methods are within a certain region. It was found that they gathered together and appeared.

  Therefore, based on the data in the database recorded by associating the retention time and the pretreatment method for a plurality of compounds, the pretreatment method associated with the compound having a retention time within a predetermined range from the retention time of the target compound It can be determined as a pretreatment method of the target compound.

  Moreover, according to said structure, since a physical-property value is output with a pre-processing method, the user can select the pre-processing method according to the physical-property value of the object compound. For example, when the pretreatment determination apparatus determines a plurality of pretreatment methods, a pretreatment method for a compound having a physical property value close to the physical property value of the target compound can be selected.

  That is, according to the above configuration, an effective pretreatment method can be determined only from the retention time when the target compound is subjected to chromatography under predetermined conditions. This makes it possible to quickly determine an appropriate pretreatment method regardless of knowledge or experience. Furthermore, according to the above configuration, since only the retention time has to be input, the pretreatment method can be determined even when the structure information of the target compound is completely unknown. And even if it is a case where the physical property value of a target compound is unknown, since the pre-processing method is output by the input of only retention time, examination of a pre-processing method can be advanced with reference to this.

  Another pretreatment determining apparatus of the present invention is a preprocessing determining apparatus for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed in order to solve the above-described problems. The data acquisition means for acquiring the retention time when the target compound is subjected to chromatography under predetermined conditions and the plurality of compounds for which pretreatment methods have been established, are subjected to chromatography under the above conditions. When plotting on the coordinate plane the retention time when provided and the physical property values indicating the physical properties of the compound, the line segment dividing the plurality of compounds for each established pretreatment method is plotted on the coordinate plane. Preprocessing determination that determines a pretreatment method to be applied to the target compound for each range of the physical property value from the retention time acquired by the data acquisition unit and the mathematical expression. It is characterized in that it comprises a stage.

  In order to solve the above-described problem, another pretreatment method determination method of the present invention is based on a pretreatment determination device that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. A pretreatment method determination method, wherein a data acquisition step for obtaining a retention time when the target compound is subjected to chromatography under a predetermined condition, and each of a plurality of compounds for which a pretreatment method has been established, When the retention time when subjected to chromatography under the above conditions and the physical property values indicating the physical properties of the compound are plotted on a coordinate plane, the pretreatment in which the plurality of compounds are established on the coordinate plane A mathematical expression indicating a line segment to be divided for each method is acquired, and the pretreatment method to be applied to the target compound is determined from the retention time acquired in the data acquisition step and the mathematical expression. It is characterized in that it comprises a pretreatment determination step of determining for each range of values.

  As a result of experiments by the inventors of the present application, when chromatographic retention time and physical property values are plotted on a coordinate plane, a region where a plot of a substance with a suitable pretreatment method appears is separated by a line segment. It has been found possible.

  Therefore, according to the above configuration, on the coordinate plane, a mathematical expression indicating a line segment for dividing a plurality of compounds for each established pretreatment method is acquired, and applied to the target compound from the acquired mathematical expression and the retention time. The power pretreatment method is determined for each range of physical property values.

  Therefore, according to the above configuration, an effective pretreatment method can be determined only from the retention time when the target compound is subjected to chromatography under a predetermined condition, and it can be promptly determined regardless of knowledge or experience. It is possible to determine an appropriate pretreatment method. Furthermore, according to the above configuration, the pretreatment method can be determined even when the structural information of the target compound is completely unknown.

  The physical property value is a partition coefficient between water and octanol, and the column used for the chromatography is preferably a reverse phase column.

  As a result of experiments by the inventors of the present application, by using the above physical property values and using the retention time by the above column, a pretreatment method by liquid-liquid extraction and a reverse phase system or ion exchange system cartridge are obtained. It has been found that the solid phase extraction used can be distinguished from suitable compounds.

  For this reason, according to said structure, it becomes possible to determine an appropriate pre-processing method as liquid-liquid extraction or solid-phase extraction.

  Another pretreatment determining apparatus of the present invention is a preprocessing determining apparatus for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed in order to solve the above-described problems. The target compound is subjected to liquid chromatography under a second condition in which the retention time when the target compound is subjected to liquid chromatography under the first condition and the pH of the mobile phase is different from the first condition. A data acquisition means for acquiring at least the holding time when the data is acquired, a predetermined parameter calculated using at least each holding time obtained under the first and second conditions acquired by the data acquisition means, and the parameter And a pretreatment determining means for determining a pretreatment method to be applied to the target compound based on correlation information indicating a correlation between the pretreatment method and the pretreatment method.

  In order to solve the above-described problem, another pretreatment determination method of the present invention uses a pretreatment determination apparatus that determines a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed. A method for determining a treatment method, wherein the target compound is subjected to liquid chromatography under the first condition and the target compound is subjected to a second condition in which the pH of the mobile phase is different from the first condition. The data acquisition step for obtaining at least the retention time when the sample was subjected to liquid chromatography, and each retention time obtained under the first and second conditions obtained in the data acquisition step was calculated using at least A preprocessing determining step for determining a preprocessing method to be applied to the target compound based on a predetermined parameter and correlation information indicating a correlation between the parameter and the preprocessing method; It is characterized in Mukoto.

  In general, the state of a compound in a solution often varies depending on the pH of the solution, and therefore the retention time when the compound is subjected to LC often varies depending on the pH of the mobile phase. That is, characteristics (physical properties) corresponding to the pH of the compound are reflected in the retention time when the compound is subjected to LC with a mobile phase having a different pH.

  Therefore, the inventor of the present application calculates various parameters from each retention time obtained by subjecting the compound to LC with a mobile phase having a different pH, and the relationship between the calculated parameter and a pretreatment method suitable for the compound. An experiment was conducted to investigate. And it discovered that the parameter obtained from each retention time obtained by subjecting a compound to LC with a mobile phase having a different pH and a pretreatment method suitable for the compound had a correlation.

  Based on this, according to said structure, each holding time obtained on the 1st and 2nd conditions from which pH of a mobile phase differs is acquired. Then, based on the parameters calculated using at least these retention times and the correlation information indicating the correlation between the parameters and the pretreatment method, the pretreatment method to be applied to the target compound is determined.

  According to this configuration, it is possible to determine a pretreatment method that is considered effective for a target compound from only the retention time when the target compound is subjected to chromatography under a predetermined condition. Regardless, it is possible to quickly determine an appropriate pretreatment method.

  The mobile phase under the first condition is acidic, the mobile phase under the second condition is basic, and the parameter is the retention time under the second condition of the retention time under the first condition. The correlation information indicates that the pretreatment method is solid-phase extraction if the ratio is less than a predetermined upper limit value, and the pretreatment method is liquid-liquid extraction if the ratio is equal to or greater than the predetermined upper limit value. It is preferable that the information indicates.

  The inventor of the present invention uses the ratio of the retention time in acidic conditions to the retention time in basic conditions as a parameter, so that the pretreatment method to be applied is a solid phase extraction and the pretreatment method to be applied is It has been found that a compound that is liquid-liquid extraction can be distinguished. Therefore, the pretreatment determining means determines the pretreatment method to be applied by comparing the ratio calculated from the retention times under the two conditions of acid and basic acquired by the data acquisition means with the upper limit value. Whether it is phase extraction or liquid-liquid extraction can be determined.

  In addition, the mobile phase of the first condition is acidic, the mobile phase of the second condition is basic, and the data acquisition unit is configured to perform the acquisition of the target compound under the third condition where the mobile phase is neutral. The retention time when the sample is subjected to liquid chromatography is further obtained, and the ratio of the retention time in the first condition to the retention time in the second condition is a predetermined value for determining a parameter used for determination of pretreatment Determining means for determining whether the ratio is less than a predetermined threshold, and the parameters used when the determining means determines that the ratio is less than a predetermined threshold are the holding time in the first condition and the second Is the sum of the retention time under the above condition and the retention time under the above third condition, and the correlation information used when the determination means determines that the ratio is less than a predetermined threshold, Pretreatment if numerical values in total is less than the predetermined upper limit value is solid phase extraction, pretreatment methods if at least a predetermined upper limit liquid - is preferably information indicating the liquid extraction.

  When the ratio of the retention time in the acidic condition to the retention time in the basic condition is less than a predetermined threshold, the inventor of the present invention uses the sum of the retention times in the acidic, basic, and neutral mobile phases as a parameter. Thus, it has been found that a compound in which the pretreatment method to be applied is solid-phase extraction and a compound in which the pretreatment method to be applied is liquid-liquid extraction can be distinguished. Therefore, the preprocessing determining means compares the sum of the retention times calculated from the results of measuring the retention times under the three conditions of acid, basic, and neutral acquired by the data acquisition means with the upper limit value. It is possible to determine whether the pretreatment method to be applied is solid phase extraction or liquid-liquid extraction.

  In addition, the mobile phase of the first condition is acidic, the mobile phase of the second condition is basic, and the data acquisition unit is configured to perform the acquisition of the target compound under the third condition where the mobile phase is neutral. Is further obtained for the liquid chromatography, the retention time of the component not retained in the column used for the liquid chromatography is t0, the retention time in the first condition is t1, and the second condition is When the holding time is t2, the holding time in the third condition is t3, and (t1-t0) / t0 = K1, (t3-t0) / t0 = K3, the parameters are t1 / t2 and K1 / K3, and the correlation information indicates that t1 / t2 is less than a predetermined upper limit value and greater than or equal to a predetermined lower limit value, and if K1 / K3 is 1 or more, the pretreatment method is solid phase extraction, and t1 / t2 Is prescribed Not less than the upper limit value, and pre-treatment if K1 / K3 is 1 or more liquid - is preferably information indicating the liquid extraction.

  The inventor of the present invention applies a compound whose pretreatment method to be applied is solid-phase extraction by using the ratio of the retention time in the acidic condition to the retention time in the basic condition and K1 / K3 as parameters. It has been found that the power pretreatment method can be distinguished from compounds that are liquid-liquid extraction. Therefore, according to the correlation information, the preprocessing determining means performs a preprocessing method corresponding to each parameter calculated from the result of measuring the retention time under the three conditions of acid, basic, and neutral acquired by the data acquiring means. Can be determined. The retention time t0 of the component not retained on the column may be, for example, the time when the peak corresponding to the solvent when the target compound is subjected to LC is detected, or the value of (column volume / LC flow rate) Also good. Since the value of t0 is uniquely determined by the LC condition, it may be stored so that the preprocessing determination device can refer to it.

  In addition, the mobile phase of the first condition is acidic, the mobile phase of the second condition is basic, and the data acquisition unit is configured to perform the acquisition of the target compound under the third condition where the mobile phase is neutral. Is further obtained for the liquid chromatography, the retention time of the component not retained in the column used for the liquid chromatography is t0, the retention time in the first condition is t1, and the second condition is When the holding time is t2, the holding time in the third condition is t3, and (t1-t0) / t0 = K1, (t2-t0) / t0 = K2, (t3-t0) / t0 = K3, The parameters are t1 / t2, K1 / K3, and K2 / K3, and the correlation information includes at least a condition that t1 / t2 is equal to or greater than a predetermined lower limit value and a condition that K1 / K3 is equal to or greater than 1. what If the condition that K2 / K3 is greater than 1 is satisfied, the pretreatment method is a pretreatment method for solid phase extraction using a solid phase of a reverse phase-cation exchange mixed system and a liquid solution using diethyl ether. The information is preferably information indicating at least one of liquid extraction.

  The inventor of the present invention uses the ratio of the retention time in the acidic condition to the retention time in the basic condition, and K1 / K2 and K2 / K3 as parameters, so that the pretreatment method to be applied is reversed-phase-positive. It has been found that a compound that is at least one of a pretreatment method of solid-phase extraction by a solid phase of an ion exchange mixed system and a liquid-liquid extraction by diethyl ether can be identified. Therefore, according to the correlation information, the preprocessing determining means performs a preprocessing method corresponding to each parameter calculated from the result of measuring the retention time under the three conditions of acid, basic, and neutral acquired by the data acquiring means. Can be determined.

  In addition, the mobile phase of the first condition is acidic, the mobile phase of the second condition is basic, and the data acquisition unit is configured to perform the acquisition of the target compound under the third condition where the mobile phase is neutral. Is further obtained for the liquid chromatography, the retention time of the component not retained in the column used for the liquid chromatography is t0, the retention time in the first condition is t1, and the second condition is When the holding time is t2, the holding time in the third condition is t3, and (t1-t0) / t0 = K1, (t2-t0) / t0 = K2, (t3-t0) / t0 = K3, The above parameters are t1 / t2, K1 / K3, K2 / K3, and K1 + K2 + K3, and the correlation information includes the condition that t1 / t2 is equal to or greater than a predetermined lower limit and K1 / K3 is equal to or greater than 1. If the condition that at least one of the above conditions is not satisfied and K2 / K3 is 1 or less and K1 + K2 + K3 is equal to or greater than a predetermined lower limit value, the pretreatment method is liquid-liquid extraction, and t1 / t2 Does not satisfy at least one of the condition that K1 / K3 is 1 or more, and satisfies the condition that K2 / K3 is 1 or less, and K1 + K2 + K3 is a predetermined lower limit. If it is less than the value, it is preferable that the pretreatment method is information indicating that it is solid phase extraction.

  The inventor of the present invention uses the ratio of the retention time in acidic conditions to the retention time in basic conditions, and K1 / K3, K2 / K3, and K1 + K2 + K3 as parameters to determine the pretreatment method to be applied. It was found that it was possible to distinguish between solid phase extraction and liquid-liquid extraction. Therefore, according to the correlation information, the preprocessing determining means performs a preprocessing method corresponding to each parameter calculated from the result of measuring the retention time under the three conditions of acid, basic, and neutral acquired by the data acquiring means. Can be determined.

  The preprocessing determination device may be realized by a computer. In this case, the computer is operated as each unit of the preprocessing determination device, thereby controlling the preprocessing determination device by the computer. A program and a computer-readable recording medium on which the program is recorded also fall within the scope of the present invention.

  The present invention can be used to determine the pretreatment method of a compound.

1 Preprocessing Determination Device 10 Data Acquisition Unit (Data Acquisition Unit)
11 Preprocessing determining unit (Preprocessing determining means)
20 Pre-processing DB (database)
50 Preprocessing Determination Device 51 Preprocessing Determination Unit (Preprocessing Determination Unit)
100 Preprocessing Determination Device 101 Preprocessing Determination Unit (Preprocessing Determination Unit)
103 Classification part (determination means)

Claims (14)

A pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed,
Data acquisition means for acquiring a retention time when the target compound is subjected to chromatography under predetermined conditions;
For each of a plurality of compounds for which a pretreatment method has been established, the retention time when the compound is subjected to chromatography under the above conditions, the physical property value indicating the physical property of the compound, and the established pretreatment method Based on the data stored in the database, a pretreatment method associated with a compound having a retention time within a predetermined range from the retention time acquired by the data acquisition means is determined as a pretreatment method for the target compound. And preprocessing determining means for outputting together with the physical property values associated with the preprocessing method ,
The preprocessing method determined by the preprocessing determining means is any one of solid phase extraction using a predetermined stationary phase and liquid-liquid extraction using a predetermined solvent. . A pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed,
Data acquisition means for acquiring a retention time when the target compound is subjected to chromatography under predetermined conditions;
For each of a plurality of compounds for which a pretreatment method has been established, when the retention time when the compound is subjected to chromatography under the above conditions and the physical property values indicating the physical properties of the compound are plotted on a coordinate plane, On the coordinate plane, a mathematical expression indicating a line segment for dividing the plurality of compounds for each established pretreatment method is acquired, and applied to the target compound from the retention time acquired by the data acquisition means and the mathematical expression. Pre-processing determining means for determining a power pre-processing method for each range of the physical property values ,
The preprocessing method determined by the preprocessing determining means is any one of solid phase extraction using a predetermined stationary phase and liquid-liquid extraction using a predetermined solvent. .   The pretreatment determination device according to claim 1, wherein the physical property value is a partition coefficient between water and octanol, and the column used for the chromatography is a reverse phase column. A pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed,
When the target compound is subjected to liquid chromatography under a second condition in which the target compound is subjected to liquid chromatography under the first condition and the second condition in which the pH of the mobile phase is different from the first condition. Data acquisition means for acquiring at least the holding time of
Predetermined parameters calculated using at least each retention time obtained under the first and second conditions acquired by the data acquisition means, and a plurality of compounds for which a pretreatment method has been established, It should be applied to the target compound based on the correlation information indicating the correlation between the parameter and the pretreatment method obtained based on each retention time when subjected to liquid chromatography under each second condition Preprocessing determining means for determining a preprocessing method ,
The preprocessing method determined by the preprocessing determining means is any one of solid phase extraction using a predetermined stationary phase and liquid-liquid extraction using a predetermined solvent. . The mobile phase of the first condition is acidic, the mobile phase of the second condition is basic,
The parameter is a ratio of the holding time in the first condition to the holding time in the second condition,
The correlation information is information indicating that the pretreatment method is solid-phase extraction when the ratio is less than a predetermined upper limit value, and the pretreatment method is liquid-liquid extraction when the ratio is equal to or greater than the predetermined upper limit value. The preprocessing determination device according to claim 4, wherein The mobile phase of the first condition is acidic, the mobile phase of the second condition is basic,
The data acquisition means further acquires a retention time when the target compound is subjected to liquid chromatography under a third condition where the mobile phase is neutral,
Determining means for determining whether the ratio of the holding time in the first condition to the holding time in the second condition is less than a predetermined threshold for determining a parameter used for determination of preprocessing;
The parameters used when the determination unit determines that the ratio is less than a predetermined threshold are the retention time in the first condition, the retention time in the second condition, and the third condition. The sum of the retention time and
The correlation information used when the determination means determines that the ratio is less than a predetermined threshold is that the pretreatment method is solid-phase extraction if the summed numerical value is less than a predetermined upper limit, and the predetermined upper limit 6. The pretreatment determining apparatus according to claim 4, wherein the pretreatment method is information indicating that the pretreatment method is liquid-liquid extraction if the value is equal to or greater than the value. The mobile phase of the first condition is acidic, the mobile phase of the second condition is basic,
The data acquisition means further acquires a retention time when the target compound is subjected to liquid chromatography under a third condition where the mobile phase is neutral,
The retention time of the component not retained in the column used for the liquid chromatography is t0, the retention time in the first condition is t1, the retention time in the second condition is t2, and the retention time in the third condition is t3. , (T1-t0) / t0 = K1, and (t3-t0) / t0 = K3,
The above parameters are t1 / t2 and K1 / K3,
In the above correlation information, if t1 / t2 is less than a predetermined upper limit value and not less than a predetermined lower limit value, and if K1 / K3 is 1 or more, the pretreatment method is solid phase extraction, and t1 / t2 is a predetermined upper limit value. 5. The preprocessing determination device according to claim 4, wherein the preprocessing method is information indicating that the preprocessing method is liquid-liquid extraction when K1 / K3 is 1 or more. The mobile phase of the first condition is acidic, the mobile phase of the second condition is basic,
The data acquisition means further acquires a retention time when the target compound is subjected to liquid chromatography under a third condition where the mobile phase is neutral,
The retention time of the component not retained in the column used for the liquid chromatography is t0, the retention time in the first condition is t1, the retention time in the second condition is t2, and the retention time in the third condition is t3. , (T1-t0) / t0 = K1, (t2-t0) / t0 = K2, and (t3-t0) / t0 = K3,
The above parameters are t1 / t2, K1 / K3, and K2 / K3,
The correlation information does not satisfy at least one of the condition that t1 / t2 is equal to or greater than a predetermined lower limit and the condition that K1 / K3 is equal to or greater than 1, and satisfies the condition that K2 / K3 is greater than 1. For example, the pretreatment method is information indicating that it is at least one of a pretreatment method of solid phase extraction by a solid phase of a reversed phase-cation exchange mixed phase and a liquid-liquid extraction by diethyl ether. The preprocessing determination device according to claim 4. The mobile phase of the first condition is acidic, the mobile phase of the second condition is basic,
The data acquisition means further acquires a retention time when the target compound is subjected to liquid chromatography under a third condition where the mobile phase is neutral,
The retention time of the component not retained in the column used for the liquid chromatography is t0, the retention time in the first condition is t1, the retention time in the second condition is t2, and the retention time in the third condition is t3. , (T1-t0) / t0 = K1, (t2-t0) / t0 = K2, and (t3-t0) / t0 = K3,
The above parameters are t1 / t2, K1 / K3, K2 / K3, and K1 + K2 + K3,
The correlation information satisfies the condition that t1 / t2 is not less than a predetermined lower limit value and / or K1 / K3 is not less than 1 and K2 / K3 is not more than 1. If K1 + K2 + K3 is equal to or greater than the predetermined lower limit, the pretreatment method is liquid-liquid extraction, and the condition that t1 / t2 is equal to or greater than the predetermined lower limit and the condition that K1 / K3 is equal to or greater than 1 Information indicating that the pretreatment method is solid-phase extraction if at least one of the conditions is satisfied and K2 / K3 is 1 or less and K1 + K2 + K3 is less than the predetermined lower limit. The preprocessing determination device according to claim 4. A pretreatment method determination method by a pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed,
A data acquisition step for acquiring a retention time when the target compound is subjected to chromatography under predetermined conditions;
For each of a plurality of compounds for which a pretreatment method has been established, the retention time when the compound is subjected to chromatography under the above conditions, the physical property value indicating the physical property of the compound, and the established pretreatment method Based on the data recorded in the database, the pretreatment method associated with the compound having the retention time within the predetermined range from the retention time acquired in the data acquisition step is determined as the pretreatment method of the target compound. Te, I saw including a pre-processing determining step of outputting together with the physical property values associated with the pretreatment method,
The pretreatment method determined in the pretreatment determination step is any one of solid phase extraction using a predetermined stationary phase and liquid-liquid extraction using a predetermined solvent. Method. A pretreatment method determination method by a pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed,
A data acquisition step for acquiring a retention time when the target compound is subjected to chromatography under predetermined conditions;
For each of a plurality of compounds for which a pretreatment method has been established, when the retention time when the compound is subjected to chromatography under the above conditions and the physical property values indicating the physical properties of the compound are plotted on a coordinate plane, On the coordinate plane, a mathematical expression indicating a line segment for dividing the plurality of compounds for each established pretreatment method is acquired, and applied to the target compound from the retention time acquired in the data acquisition step and the mathematical expression. the to pretreatment methods saw including a pre-processing determining step of determining for each range of the physical property value,
The pretreatment method determined in the pretreatment determination step is any one of solid phase extraction using a predetermined stationary phase and liquid-liquid extraction using a predetermined solvent. Method. A pretreatment method determination method by a pretreatment determination device for determining a pretreatment method for removing impurities from a sample solution containing a target compound to be analyzed,
When the target compound is subjected to liquid chromatography under a second condition in which the target compound is subjected to liquid chromatography under the first condition and the second condition in which the pH of the mobile phase is different from the first condition. A data acquisition step for acquiring at least a holding time of
Predetermined parameters calculated using at least the retention times obtained under the first and second conditions obtained in the data acquisition step and each of the plurality of compounds for which pretreatment methods have been established are And applied to the target compound based on the correlation information indicating the correlation between the parameter and the pretreatment method obtained based on each retention time when subjected to liquid chromatography under each of the second conditions. look including a pre-processing determining step of determining a pre-treatment method should,
Pretreatment methods for determining the above pretreatment determining step, solid-phase extraction using a predetermined stationary phase, and a liquid using a predetermined solvent - either the liquid extraction, determination pretreatment method characterized by Method. A control program for causing a computer to execute each step of the preprocessing method determination method according to any one of claims 10 to 12 .   A computer-readable recording medium on which the control program according to claim 13 is recorded.

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