JP4680761B2 - Liquid chromatography condition determination support apparatus, liquid chromatograph, and liquid chromatography condition determination support program - Google Patents

Description

  The present invention relates to liquid chromatography using a liquid solvent (eluent) as a mobile phase. Specifically, the present invention relates to examination of conditions for performing liquid chromatography.

  In liquid chromatography, an eluent corresponding to a mobile phase composed of a sample having a plurality of components and a plurality of solvents mixed at a predetermined mixing ratio flows through a separation column packed with a stationary phase. At this time, the sample flowing into the column together with the eluent moves along with the flow of the eluent while adsorbing to the stationary phase packed in the column, and is discharged from the column after a predetermined time.

  Here, the time required for discharging each component contained in the sample depends on the affinity with the eluent, the interaction between the stationary phase of the column and each component, etc., and differs for each component. Specifically, those having weak affinity with the eluent and those having strong interaction with the stationary phase remain in the column for a long time. On the other hand, those having a strong affinity with the eluent and those having a weak interaction with the stationary phase are quickly discharged. As a result, the sample placed on the column is separated for each component and eluted.

A liquid chromatograph for performing such liquid chromatography is disclosed in, for example, Patent Document 1.
JP 2003-240765 A

  In the above liquid chromatography, the size and load of the separation column can be determined simply by experience and intuition, for example, in a column with an inner diameter of 2 cm, the load should be around 1 gram. There were many. In addition, the column size and load amount were simply determined without considering the affinity of each component with the eluent and the difference (selectivity) in the interaction between each component and the stationary phase, and therefore, the prediction was not correct. There were many things. Therefore, analysis and fractionation by liquid chromatography has frequently failed, and unnecessary long time has occurred frequently, reducing the efficiency of chromatography.

Means and effects for solving the problems

  The problems to be solved by the present invention are as described above. Next, means for solving the problems and the effects thereof will be described.

According to a first aspect of the present invention, there is provided a liquid chromatography condition determination support device configured as follows. That is, for a plurality of components, the mobility R _f and the actual measurement value input means for inputting a measured value of the mobility R _f values of each constituent in the thin layer chromatography, which is input by the actual value input means Load amount calculating means for obtaining the load amount of the sample on the column of the liquid chromatography in terms of value, and load amount output means for outputting the load amount obtained by the load amount calculating means.

With this configuration, it is possible to calculate an appropriate load amount from the mobility result (R _f value) obtained by thin layer chromatography and present it to the user, and the user side determines the conditions while referring to this. Thus, work efficiency such as separation can be improved.

The liquid chromatography condition determination support apparatus is preferably configured as follows. In other words, the load amount calculating means obtains the number of theoretical plates necessary to make the plurality of components separable with a predetermined degree of separation from the mobility R _f value input by the measured value input means. Based on the relationship stored in the storage means, the storage means for storing in advance the relationship between the means, the theoretical plate number, and the amount of the sample loaded on the column of the liquid chromatography. 2nd calculating means for calculating | requiring load amount from the theoretical plate | plate number calculated | required by (2).

  With this configuration, a load amount for realizing an appropriate degree of separation can be presented to the user, and a good chromatographic result can be easily obtained.

The liquid chromatography condition determination support apparatus is preferably configured as follows. In other words, the load amount calculation means obtains the load amount for each type of column for a plurality of types of columns of liquid chromatography from the mobility R _f value. The load amount output means is configured to output a load amount for each type of the column.

  With this configuration, for example, when a load amount of a sample to be separated is determined, it is possible to support the user's condition determination process so that a column capable of suitably separating the sample can be appropriately selected from a plurality of types.

The liquid chromatography condition determination support apparatus is preferably configured as follows. That is, the actual measurement value input means includes a pointing device capable of designating an arbitrary point on the display image of the display device. The measured value of the mobility R _f value can be input by designating a point on the image of the thin layer plate drawn on the display device with the pointing device.

With this configuration, it is possible to provide an intuitive and excellent operability as if the state of the thin layer plate in the thin layer chromatography performed in advance is input as it is, and the mobility R _f value can be accurately determined by an easy operation. You can enter.

According to the second aspect of the present invention, there is provided a liquid chromatography condition determination support program configured as follows. That is, the measured value input means for inputting the measured value of the mobility _Rf value of each component when thin-layer chromatography is performed on a plurality of components, the mobility input by this measured value input means The R _f value functions as load amount calculation means for obtaining the load amount of the sample to the column of liquid chromatography, and load amount output means for outputting the load amount obtained by the load amount calculation means.

With this configuration, an appropriate load can be calculated and presented to the user from the result of the mobility R _f value obtained by thin layer chromatography, and the user can determine the conditions while referring to this. , Work efficiency such as separation can be improved.

The liquid chromatography condition determination support program is preferably configured as follows. In other words, the load amount calculating means obtains the number of theoretical plates necessary to make the plurality of components separable with a predetermined degree of separation from the mobility R _f value input by the measured value input means. Based on the relationship stored in the storage means, the storage means for storing in advance the relationship between the means, the theoretical plate number, and the amount of the sample loaded on the column of the liquid chromatography. 2nd calculating means for calculating | requiring load amount from the theoretical plate | plate number calculated | required by (2).

  With this configuration, a load amount for realizing an appropriate degree of separation can be presented to the user, and a good chromatographic result can be easily obtained.

The liquid chromatography condition determination support program is preferably configured as follows. In other words, the load amount calculation means obtains the load amount for each type of column for a plurality of types of columns of liquid chromatography from the mobility R _f value. The load amount output means is configured to output a load amount for each type of the column.

  With this configuration, for example, when a load amount of a sample to be separated is determined, it is possible to support a user's condition determination process so that a column capable of suitably separating the sample can be appropriately selected from a plurality of types.

In the liquid chromatography condition determination support program, in the actual measurement value input means, the actual measurement value of the mobility _Rf value is obtained by pointing a point on the image of the thin layer plate drawn on the display device of the computer with a pointing device. It is preferable to be able to input by instructing.

With this configuration, it is possible to provide an intuitive and excellent operability as if the state of the thin layer plate in the thin layer chromatography performed in advance is input as it is, and the mobility R _f value can be accurately determined by an easy operation. You can enter.

  According to another aspect of the present invention, there is provided a liquid chromatograph including the liquid chromatography condition determination support device having the above-described configuration.

Next, embodiments of the invention will be described. FIG. 1 is an explanatory diagram showing a method of actually measuring the mobility R _f value of each component by thin layer chromatography.

First, with reference to FIG. 1, the thin layer chromatography (TLC) as a process before the liquid chromatography is demonstrated. This thin layer chromatography is performed in order to actually measure the mobilities R _f1 and R _f2 of the components 1 and 2 of the sample to be separated.

  The method of thin layer chromatography will be described. First, silica gel powder as a stationary phase is thinly and uniformly applied on the surface of an elongated flat plate (for example, a glass plate) to prepare a silica gel thin layer plate 11. Then, a drop of the sample to be separated is dropped on the silica gel on the silica gel thin layer plate 11 to form a sample spot 12. The sample to be dropped is a mixture of at least two components (component 1 and component 2).

  Next, the lower part of the silica gel thin layer plate 11 is immersed in the eluent 14 stored in the container 13 as shown on the left side of FIG. Then, the eluent 14 is sucked up into the silica gel thin layer on the thin layer plate 11 by capillary action. Along with this, the sample spot 12 also moves upward, but the speed at which each component contained in the sample rises depends on the affinity with the eluent 14, the interaction with silica gel as the stationary phase, and the like. Different for each. Therefore, when a predetermined time elapses, as shown on the right side of FIG. 1, the component 1 contained in the sample spot 12 moves to the position 12a, and the component 2 moves to the position 12b. In this way, separation of the two components is realized.

In the above state, the distance L ₀ from the position of the sample spot 12 before immersing the thin layer plate 11 in the eluent 14 to the upper end position of the sucked eluent 14 is measured, and the movement of each component of the sample is measured. The distances L _r1 and L _r2 are measured together. Then, the mobility R _f1 and R _f2 of each component is obtained according to the formulas R _f1 = L _r1 / L ₀ and R _f2 = L _r2 / L ₀ . The obtained value is appropriately recorded for input to a personal computer of a liquid chromatograph described later.

  Next, a liquid chromatograph will be described with reference to FIGS. FIG. 2 is a schematic diagram and block diagram of a liquid chromatograph, and FIG. 3 is a block diagram showing functions realized by a condition determination support program in a personal computer for controlling the liquid chromatograph.

  The liquid chromatograph 21 stores two containers 31 and 32, an electromagnetic valve 33 provided at an interconnection point of a liquid feeding path extending from the containers 31 and 32, a pump 36, and an eluent 34, respectively. The mixer 35, the pump 37, the injector 38, the column 39, the detector 40, and the fraction collector 41 are arranged in the above order according to the liquid feeding direction. The liquid chromatograph 21 includes a personal computer 51. The personal computer 51 functions as a control device that controls each of the above devices according to desired chromatography conditions, and when the user determines the chromatography conditions. It has a function as a condition determination support device that supports it.

  The container 31 stores the solvent A, and the container 32 stores the solvent B. In addition, the solvent used is not limited to two types of A and B, You may increase a number according to a use condition and the objective. In general, as the solvent A and the solvent B, a combination of a nonpolar molecule and a polar molecule is used.

  The pump 36 is configured to pump up the solvents A and B through the electromagnetic valve 33. The electromagnetic valve 33 selects the solvent to be pumped up from the solvent A or the solvent B based on the control signal from the personal computer 51 and adjusts the ratio of the selection time, whereby the eluent 34 having a predetermined mixing ratio. And The eluent 34 is the same type as the eluent 14 used in the above-mentioned thin layer chromatography (FIG. 1).

  The injector 38 holds the sample, and the eluent 34 sucked and discharged from the mixer 35 by the pump 37 passes through the injector 38 so that the sample is sent out.

  The column 39 is a tubular container filled with a stationary phase, and liquid chromatography is performed by passing a sample through the stationary phase together with the eluent 34. As this stationary phase, the same one as that laminated on the thin layer plate 11 in the thin layer chromatography (FIG. 1) is used, and silica gel is used in this embodiment.

  The column 39 is provided with a plurality of types having different column dimensions (column diameter and column length), the type of the stationary phase, and the like. It is configured so that it can be set and used.

  The detector 40 is disposed on the downstream side of the column 39 and detects the result of liquid chromatography performed in the column 39. The detection result is transmitted to the personal computer 51. Further, the fraction collector 41 includes a plurality of test tubes, and is configured so that each component contained in the sample can be sorted into each test tube based on the analysis result of the detector 40.

  The personal computer 51 uses a general-purpose computer. As shown in FIG. 2, a CPU 52, a memory 53, a hard disk 54, an external interface 55, a keyboard 56, a mouse (pointing device) 57, and the like. And a display device 58.

Then, the personal computer 51, an actual measurement value input unit 61 of the mobility R _f1, R _f2 of each component when performing the thin layer chromatography, found, the mobility R _f1 input by the input means 61 R _The load amount calculating means 62 for calculating the load amount of the sample on the column 39 of the liquid chromatography at _f2 and the load amount output means 63 for outputting the load amount obtained by the load amount calculating means 62 are provided. .

Further, the load amount calculation means 62 has a theoretical plate number N required to make it possible to separate the two components with a predetermined degree of separation Rs from the mobility R _f1 and R _f2 inputted by the actual value input means 61. Based on the first calculation means 64 for obtaining the relationship, the storage means 65 capable of storing the relationship between the theoretical plate number N and the load amount of the sample to the column 39, and the relationship stored in the storage means 65, Second calculation means 66 for obtaining the load amount from the theoretical stage number N obtained by the first calculation means 64 is provided.

  Specifically, various types of software including a liquid chromatography control program and a condition determination support program are stored in the hard disk 54 of the personal computer 51. And said each means 61-66 shown in FIG. 2 is constructed | assembled by combining the said hardware and software.

  Note that the above software can be stored in the hard disk 54 of the personal computer 51 by installing it using a recording medium such as a CD-ROM, FD, or MO in which the program is recorded. Alternatively, the installation can be performed via a network such as the Internet.

  Liquid chromatography conditions are set using the personal computer 51 described above, and these conditions include, for example, the mixing ratio of the eluent 34 and the flow rate of the eluent 34 flowing into the column 39. Based on these conditions, the personal computer 51 controls the electromagnetic valve 33, the pumps 36 and 37, etc., and performs liquid chromatography. Further, the personal computer 51 can analyze the detection signal from the detector 40, draw an outflow curve on the display device 58, and store the result in the hard disk 54 or the like.

  Here, the conditions of the liquid chromatography include which column of the plurality of types of columns 39 is selected and used, and the amount of sample (load amount) set in the injector 38. In the present embodiment, the determination process is supported by the above condition determination support program so that the user can appropriately select the type of column 39 and the load amount.

  Hereinafter, this condition determination support program will be described. FIG. 4 is a diagram showing a window displayed on the display by the condition determination support program, FIG. 5 is a diagram showing a state in which a calculation result of a load amount for each size column is displayed, and FIG. 6 is a case where an L size column is used. FIG. 5 is a diagram showing the relationship between the load amount and the number of theoretical plates necessary to realize the degree of separation Rs = 1.

When this condition determination support program is executed, a window 71 having a GUI (graphical user interface) as shown in FIG. 4 is displayed on the display device 58, for example. In this window 71, text boxes 72 and 73 for inputting the numerical values of the mobility R _f1 and R _f2 measured by the thin layer chromatography and the spot positions on the silica gel thin layer plate 11 of the thin layer chromatography are displayed. A spot indication area 74 for specifying the mobility R _f1 and R _f2 by instructing above, a result display area 75 for displaying the result of the calculated load amount, and an OK button 76 for instructing the calculation of the load amount And a cancel button 77 for closing the window 71 and ending the execution of the condition determination support program.

With the screen configuration described above, the user operates and clicks the mouse 57 so that the mouse pointer 80 is aligned with the text boxes 72 and 73 of the window 71, and sets the values of the mobility R _f1 and R _f2 to “0.5”. Or “0.3” or the like is input from the keyboard 56. After inputting the mobility R _f1 and R _f2 in both the text boxes 72 and 73, the OK button 76 is clicked. Then, the load amount of the sample is calculated according to the procedure described later, and the calculation result is displayed in the result display area 75.

  The result display area 75 is in a table format, and for each type of the plurality of columns 39, the numerical value of the corresponding load amount can be output in a list format. In the present embodiment, there are seven types of columns 39 in the “size” column (total of seven types of columns 39 having different sizes from S, M, L, 2L, 3L, 4L, and 5L in the liquid chromatograph 21 of the present embodiment). In the column “g”, how many grams of the sample should be set (load amount) when the column 39 of the size is used is displayed for each size. For example, as shown in FIG. 5, “0.1” is displayed on the “M” line, and “1.0” is displayed on the “L” line.

  From the display of FIG. 5, the user can obtain information that, for example, if a 1-gram sample is to be sampled, an excellent result will be obtained if the L-size column 39 is used. Therefore, the user can select an appropriate column 39 size with reference to this information, so that liquid chromatography does not take an unnecessarily long time, and separation does not succeed and the separation is not repeated. Work can be remarkably streamlined.

The mobility R _f1 and R _f2 can be input by operating the mouse 57 using the spot indicating area 74 instead of directly inputting from the keyboard 56 using the text boxes 72 and 73. In this spot indicating area 74, a figure 81 imitating the thin layer plate 11 in the thin layer chromatography is drawn, and in this figure 81, circular shapes corresponding to the spot positions 12a and 12b of the component 1 and component 2 are drawn. The two spot figures 82a and 82b are drawn. The long horizontal line at the bottom of the spot indicating area 74 corresponds to the position of the sample spot 12 before movement, and the long horizontal line at the top corresponds to the position of the sucked eluent 14. That is, the distance between the upper and lower long horizontal lines corresponds to the distance L ₀ shown in FIG.

In the display example of FIG. 4, the upper spot graphic 82a is drawn at the position of R _f1 = 0.5, and the lower spot graphic 82b is drawn at the position of R _f2 = 0.3. In order to change this, for example, the mouse button 80 is depressed while the mouse pointer 80 is aligned with the lower spot graphic 82b, the mouse pointer is moved upward in this state, and the mouse button is released at a desired position. Then, the spot figure 82b can be moved to the upper side, for example, to the position of R _f2 = 0.4 (programmed as such). Simultaneously with this movement, the numerical value input in the text box 73 is automatically changed to a numerical value (0.4) corresponding to the position of the new spot graphic 82b. Also, the mobility R _f1 of the component 1 can be easily set by dragging the upper spot figure 82a in the same manner as described above.

As described above, the mobility R _f1 and R _f2 of each component can be designated as if the spot positions 12a and 12b of the thin layer chromatography result of FIG. 1 are designated on the screen of the display device 58 as they are, and intuitive and excellent operability. Is realized. In addition, in order to further easily indicate the mobility R _f1 and R _{f2 by} the mouse 57, the spot indicating area 74 is provided with a scale 78 that equally divides the distance L ₀ at an appropriate interval. An explanatory note 79 explaining the operation method is displayed at a position beside the instruction area 74.

When the mobility values R _f1 and R _f2 are directly input from the text boxes 72 and 73 as numerical values, the spot graphics 82a and 82b in the spot indicating area 74 automatically move to positions corresponding to the numerical values. ing. As a result, the user can easily check the input values of the mobility R _f1 and R _f2 , and input errors are prevented.

なお、上記の式において、Ｎは理論段数、αは選択性因子、ｋ’は保持比である。 Next, the calculation of the load amount will be described in detail below. In general, in liquid chromatography of a sample, there is a resolution Rs as an index (peak and band proximity in an outflow curve) indicating the degree of separation between component 1 and component 2, which are components of the sample. It is known that it can be expressed as equation (1).

In the above formula, N is the number of theoretical plates, α is a selectivity factor, and k ′ is a retention ratio.

Here, the retention ratio k ′ is t ₀ when the sample starts to flow into the column 39 together with the eluent 34 and the eluent 34 flows out of the column 39 (so-called void time). When the time from beginning to flow into the column 39 until the effluent from the column 39 and t _r, it is expressed by the following equation (2).

Then, the liquid chromatograph 21 of the present embodiment, as the peak time t _r of the target substance is almost 4 times the value of the void time t _0, configured to appropriately automatically set chromatographic conditions in the personal computer 51 (T _r = 4 × t ₀ ). When this condition is applied to the above equation (2), k ′ = 3.

Further, in the liquid chromatograph 21 of the present embodiment, Rs = 1 is set as a specific value of the degree of separation necessary and sufficient for the fraction collector 41 to appropriately sort into each test tube. Yes. Therefore, by substituting Rs = 1 and k ′ = 3 into the above equation (1) and further transforming it into the form of N =, the equation of the number of theoretical plates N necessary to realize the separation degree Rs = 1 is obtained. The following equation (3) can be derived.

The selectivity factor α is specifically represented by the following formula (4).

In the above equation (4), k ₁ ′ and k ₂ ′ are individual holding ratios for the component 1 and the component 2. That is, when the time from the start of liquid chromatography until the peak of the component 1 appears is t _r1 and the time until the peak of the component 2 appears is _{tr 2} , k ₁ ′ and k ₂ ′ are expressed by the following formula (5 ).

上記の式は、薄層クロマトグラフィと液体クロマトグラフィとを具体的に関係付ける重要な式として知られている。 Here, the mobility R _f1 and R _f2 of each component in the above-described thin layer chromatography can be regarded as a ratio of the moving speed of each component in the stationary phase to the eluent moving speed. On the other hand, in the liquid chromatography, the ratio of the moving speed of each component in the stationary phase to the eluent moving speed is the above-described void time t ₀ , the time t _r1 required for each component to move in the column 39, It is obtained by dividing each by _tr2 . Therefore, for each component, the ratio of the mobility rate in thin layer chromatography (mobility R _f1 , R _f2 ) and the ratio of the retention time in liquid chromatography (t ₀ / t _r1 , t ₀ / t _r2 ) are equal. By doing so, the following equation (6) can be derived.

The above formula is known as an important formula that specifically relates thin layer chromatography and liquid chromatography.

Then, by modifying the above equation (5), k ₁ ′ and k ₂ ′ can be expressed as the following equation (7) using the mobility R _f1 and R _f2 .

By substituting the relationship of equation (7) into equation (4) and further substituting the obtained α into equation (3), the following equation (8) is derived.

Here, 256 / 9≈28, and by _setting a = 1 / R _f1 , the above equation (8) can be transformed into the following equation (9).

As described above, the relationship between the number of theoretical plates N necessary for realizing the degree of separation Rs = 1 and the mobilities R _f1 and R _f2 is obtained. That is, in the above condition determination support program, when the above OK button 76 is clicked, from the mobility R _f1 and R _f2 input in the text boxes 72 and 73, using the above equation (9), The number N of theoretical plates necessary for realizing Rs = 1 is obtained. As described above, the function of the first calculation means 64 shown in FIG. 3 is realized. For example, if R _f1 = 0.5 and R _f2 = 0.3 are designated as shown in the input example in the window of FIG. 4, N≈86 is obtained from equation (9).

  After obtaining the above theoretical plate number N, the load amount corresponding to the theoretical plate number N is obtained for each column 39 of each size described above. That is, the hard disk 54 stores, for example, the relationship between the theoretical plate number N and the load amount as shown in FIG. 6 (function of the storage means 65 shown in FIG. 3). In the graph illustrated in FIG. 6, the horizontal axis represents the load amount, and the vertical axis represents the theoretical plate number N on a logarithmic scale. As shown in this graph, when the load amount is less than the predetermined load amount, the theoretical plate number N is constant at a large value, but when the load amount exceeds the predetermined load amount, the theoretical plate number N is a logarithm as the load amount increases. It shows a tendency to decrease linearly (linear function) on the scale.

  In the present embodiment, the condition determination support program stored in the hard disk 54 includes a predetermined relational expression representing the relationship between the theoretical plate number N and the load amount as illustrated in FIG. This relational expression is obtained based on a prior experiment.

  Further, in FIG. 6, the relationship with the L-size column 39 is shown as a representative, but the relationship between the theoretical plate number N and the load amount is also tested for other sizes of the column 39 such as the S size and the M size. The condition determination support program includes a formula obtained in advance and expressed as a relational expression. In this way, the function of the storage means 65 shown in FIG. 3 is realized.

  Next, the load amount corresponding to the theoretical plate number N obtained by the above-described calculation is obtained using the relational expression. For example, in the above example, the theoretical plate number N was calculated to be 86, but assuming that the L-size column 39 is used, it can be seen that the load corresponding to the theoretical plate number N = 86 is 1 gram from the graph of FIG. . As described above, the function of the second calculation means 66 shown in FIG. 3 is realized. The load amount thus obtained is displayed in the result display area 75 of the window 71 as described above. In light of the above example, the calculation result “1.0” is displayed in the “L” row of the result display area 75 as shown in FIG.

  Although only a part of the numerical values of the calculation results is shown in FIG. 5, the calculation of the corresponding load amount is performed for all sizes of S, M, L,. For example, if the relational expression in the case of the M-size column 39 is used and the load corresponding to N = 86 is calculated to be 0.1 gram, “0. The calculation result “1” is displayed.

As described above, the personal computer 51 provided in the liquid chromatograph 21 of the present embodiment inputs the actual measured values of the mobility R _f1 and R _f2 of each component when thin layer chromatography is performed for a plurality of components. An actual value input means 61 for calculating the load amount of the sample on the liquid chromatography column 39 from the mobility R _f1 , R _f2 input by the actual value input means 61, Load amount output means 63 for outputting the load amount obtained by the load amount calculation means 62.

Therefore, an appropriate load can be calculated and presented to the user from the results of the mobility R _f1 and R _f2 obtained by thin layer chromatography, and the user can determine the conditions while referring to this. , Work efficiency such as separation can be improved.

Since the load amount is calculated based on the mobility R _f1 and R _f2 of the components 1 and 2 as described above, the load amount can be calculated rationally. That is, in liquid chromatography, when the difference in affinity between the eluent and the interaction with the stationary phase is small between component 1 and component 2, in order to achieve better resolution than when it is large A large number of theoretical plates (small load) is required. In this regard, in this embodiment, mobility R _f1 and R _f2 , which are parameters indicating the affinity of the components 1 and 2 with the eluent and the strength of the interaction with the stationary phase, are used in thin layer chromatography. Since the load amount is calculated, the load amount can be calculated while appropriately considering the above circumstances, and as a result, a highly reliable load amount can be presented to the user.

Further, the load amount calculating means 62 can separate the plurality of components with a predetermined degree of separation (Rs = 1 in this embodiment) from the mobility R _f1 and R _f2 input by the actual value input means 61. First calculation means 64 for obtaining the number N of theoretical plates necessary for performing, the relationship (the relationship illustrated in FIG. 6) between the theoretical plate number N and the amount of sample loaded on the column 39 of liquid chromatography is stored in advance. Storage means 65 for storing, and second calculation means 66 for determining the load amount from the theoretical plate number N determined by the first calculation means 64 based on the relationship stored in the storage means 65. .

  Therefore, conditions for realizing an appropriate resolution can be presented to the user, and a good chromatographic result can be easily obtained.

Further, the load amount calculating means 62 calculates the load amount for each type of the column 39 from the mobility R _f1 , R _{f2 for} a plurality of types of columns 39 of S, M, L,. Looking for. As shown in FIG. 5, the load amount output means 63 is configured to output a load amount for each type (S, M, L,...) Of the column 39.

  Therefore, for example, when the number of grams of the sample to be separated is determined, the user's condition determination process so that the column 39 having a size that can be suitably separated can be appropriately selected from S, M, L,. Can support.

Further, the actual measurement value input means 61 includes a mouse 57 capable of designating an arbitrary point of the display image of the display device 58, and the actual measurement values of the mobility R _f1 and R _f2 are obtained from the display device 58. It is possible to input by instructing a point on the thin layer board figure 81 drawn in the spot instruction area 74 by a drag operation of the mouse 57.

Therefore, it is possible to provide intuitive and excellent operability as if the state of the thin layer plate 11 in the thin layer chromatography performed in advance is input as it is, and the mobility R _f1 , R _f2 accurately with easy operation. Can be entered.

  Although the preferred embodiment of the present invention has been described above, the above is an example, and can be modified as follows, for example.

  In the above embodiment, the case where the column 39 having an appropriate size is selected from a plurality of types of columns having different sizes has been described. However, the present invention is not limited thereto, and for example, an appropriate column is selected from a plurality of types of columns having different internal stationary phases. Also in this case, the above support program can be applied.

  The condition determination support program and the control program can be linked to each other. For example, double-clicking the “L” row in the result display area 75 of FIG. 5 closes the window 71 and terminates the condition determination support program, and uses the L-size column 39 in the liquid chromatography control program. In such a case, a suitable condition parameter is automatically set immediately.

Further, in the calculation of the above-mentioned load, the above _{Rs = 1, t r = 4} × t 0 such conditions can be appropriately changed in consideration of the purpose and the like of the analysis.

As shown in FIG. 4, in the initial state in which the window 71 is displayed, a numerical value “0.5” (so-called default value) is input to the text box 72 of the mobility R _f1 from the beginning. This is because when R _f1 = 0.5, a = 2 in the above equation (9), the calculation is simplified, and the processing is convenient. However, the default value is not limited to 0.5, and other values may be adopted.

Explanatory drawing which shows the method of actually measuring the mobility of each component by thin layer chromatography. The schematic diagram and block diagram of a liquid chromatograph. The block diagram which shows the function which a condition determination assistance program implement | achieves in the personal computer for control of a liquid chromatograph. The figure which shows the window which a condition determination assistance program displays on a display. The figure which shows a mode that the condition determination assistance program displayed the calculation result of the load amount with respect to the column of each size. The figure which shows the relationship between a load amount at the time of using the column of L size, and the theoretical plate number required in order to implement | achieve the separation degree Rs = 1.

Explanation of symbols

21 liquid chromatograph 39 column 51 personal computer 71 window 72, 73 text box 74 spot indication area 75 result display area

Claims (9)

An actual value input means for inputting an actual value of the mobility R _f value of each component when thin layer chromatography is performed for a plurality of components;
A load amount calculating means for determining the load amount of the sample to the column of the liquid chromatography at the mobility R _f value input by the actual value input means;
Load amount output means for outputting the load amount obtained by the load amount calculation means;
A condition determination support apparatus for liquid chromatography, comprising: The condition determination support apparatus for liquid chromatography according to claim 1,
The load amount calculating means includes
First computing means for obtaining the number of theoretical plates necessary for separating the plurality of components with a predetermined degree of separation from the mobility R _f value input by the actual value input means;
Storage means for storing in advance the relationship between the number of theoretical plates and the amount of sample loaded on the column of liquid chromatography;
Based on the relationship stored in the storage means, a second calculation means for determining the load amount from the number of theoretical plates determined by the first calculation means;
A condition determination support apparatus for liquid chromatography, comprising: The condition determination support apparatus for liquid chromatography according to claim 1 or 2,
The load amount calculation means obtains the load amount for each type of the column for a plurality of types of columns of liquid chromatography from the mobility R _f value,
The load amount output unit is configured to output a load amount for each type of the column, and is a liquid chromatography condition determination support device. The condition determination support apparatus for liquid chromatography according to any one of claims 1 to 3,
The actual measurement value input means includes a pointing device capable of instructing an arbitrary point of the display image of the display device,
The actual measurement value of the mobility R _f value can be input by instructing a point on the image of the thin-layer plate drawn on the display device by the pointing device. Support device.   A liquid chromatograph comprising the liquid chromatography condition determination support device according to any one of claims 1 to 4. An actual value input means for inputting an actual value of the mobility R _f value of each component when thin layer chromatography is performed for a plurality of components;
A load amount calculating means for determining a load amount of the sample to the column of the liquid chromatography in the mobility R _f value input by the actual value input means; and
Load amount output means for outputting the load amount obtained by the load amount calculation means,
A liquid chromatography condition determination support program characterized by causing a computer to function as A condition determination support program for liquid chromatography according to claim 6,
The load amount calculating means includes
First computing means for obtaining the number of theoretical plates necessary for separating the plurality of components with a predetermined degree of separation from the mobility R _f value input by the actual value input means;
Storage means for storing in advance the relationship between the number of theoretical plates and the amount of sample loaded on the column of liquid chromatography;
Based on the relationship stored in the storage means, a second calculation means for determining the load amount from the number of theoretical plates determined by the first calculation means;
A condition determination support program for liquid chromatography, comprising: A condition determination support program for liquid chromatography according to claim 6 or 7,
The load amount calculation means obtains the load amount for each type of the column for a plurality of types of columns of liquid chromatography from the mobility R _f value,
The load amount output means is configured to output a load amount for each type of the column, and is a liquid chromatography condition determination support program. A condition determination support program for liquid chromatography according to any one of claims 6 to 8,
In the actual measurement value input means, the actual measurement value of the mobility R _f value can be input by instructing a point on the image of the thin layer plate drawn on the display device of the computer with a pointing device. A program for determining conditions for liquid chromatography.

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