JP5966007B2 - Liquid chromatograph - Google Patents

Description

  The present invention relates to a liquid chromatograph, and more particularly, to an increase in the speed and separation of an analytical method.

In recent years, higher speed and higher separation of analysis methods for liquid chromatograph devices (hereinafter referred to as LC) have been further demanded. One of the methods for examining the high-speed performance and high-separation performance of analytical methods related to LC is kinetic plot analysis (hereinafter referred to as KPA). Non-Patent Document 1 describes this technique. KPA uses the relationship of the theoretical plate height H (hereinafter also referred to as H (also referred to as u ₀ )) at each linear velocity u ₀ (m / s) as input data. The linear velocity u ₀ (m / s) is proportional to the flow rate F (ml / min) and the column cross-sectional area considering the porosity ε as a coefficient. In general, this data is called a H-u curve or a van Demeter curve (FIG. 2). This data is unique to the column and is ideally considered a unique property of the packing material. This data is usually shown as a graph or a table. Here, the theoretical plate height H (u ₀ ) indicates the column length required per number of theoretical plates N (H (u ₀ ) = L / N), and one theoretical plate with a shorter column length. It means that the filler to be achieved has a higher separation capacity. KPA is not only a Hu plot but also column permeability Kv (m ² ) showing the liquid permeability or fluidity inherent to the column, the viscosity η (Pa · s) of the mobile phase used for the analysis, and the use conditions Or the maximum analysis pressure (system pressure) ΔPmax (MPa) limited by the apparatus system to be used is required as input data.

In KPA, a t _o -N plot is obtained based on these input data (Hu plot, Kv, η, ΔPmax) (FIG. 3). Here, t _o (s) indicates a void time which is a movement time of the non-retained component in the column. The t _o -N plot is a very useful graph when considering the high speed or high separation performance of a method. The method is a condition parameter of the analysis method composed of the column length L and the linear velocity u ₀ .

FIG. 4 shows the case where the number of theoretical plates N = 20,000 is obtained, and it can be seen that the void time t _o = 10 s is the fastest column and method.
FIG. 5 shows a case where a void time t _o = 100 s is obtained, and it can be seen that the column and the method can realize high separation with the theoretical plate number N = 60,000.
Parameters relating to liquid flow (Kv, η, ΔPmax) act simultaneously with the column length L and limit the linear velocity u ₀ . These parameters are necessary for uniquely determining the analytical method conditions in the conversion to the above t _o -N plot.
Hereinafter, the concept of the conversion method from the Hu plot to the t _o -N plot will be described. The linear velocity u ₀ and the column length L are inversely proportional. The inversely proportional constant is referred to as a constant of flow Cf (m ² / s). The flow constant Cf is uniquely determined from Equation 1 from the input parameters (Kv, η, ΔPmax) relating to liquid flow.

カラム長さＬは理論段数Ｎと理論段高さＨ（ｕ₀）とで表され、数式２を得る。

The column length L is expressed by the theoretical plate number N and the theoretical plate height H (u ₀ ), and Equation 2 is obtained.

一方、カラム長さＬは線速度とボイド時間ｔ_oとによっても表され、数式３を得る。

On the other hand, the column length L is also expressed by the linear velocity and the void time t _o to obtain Equation 3.

Ｈ−ｕプロットから、複数の（ｕ₀, Ｈ）セットを得る。次に複数の（ｕ₀, Ｈ）セットを入力として数式２と数式３からそれぞれ複数の（ｔ₀, Ｎ）セットを求める。これをグラフに表現したものがｔ₀−Ｎプロットである。

A plurality of (u ₀ , H) sets are obtained from the Hu plot. Next, a plurality of (t ₀ , H) sets are input, and a plurality of (t ₀ , N) sets are obtained from Equation 2 and Equation 3, respectively. A graph representing this is a t ₀ -N plot.

In these processing operations, the column length L is uniquely calculated for each linear velocity u ₀ using Equation 1 under the restriction condition of the given flow constant Cf. That is, the column length L at which the maximum pressure loss (hereinafter referred to as ΔPmax) is obtained at the linear velocity u ₀ is obtained.

In the linear velocity u _0, Once the column length L is determined, for H (u ₀₎ is uniquely determined corresponding to the linear velocity u _0, from Equation 2 theoretical plates N is automatically Desired. Similarly, once the column length L is determined at the linear velocity u ₀ , the void time t _o is automatically obtained from Equation 3.
KPA converts the Hu plot into a t _o -N plot by the above calculation process.

  In addition, Patent Document 1 considers that when a method is converted from a general-purpose liquid chromatograph measurement system to a measurement system under high-pressure and high-speed conditions, due to the great influence of changes in analysis conditions, A method of reflecting the volume value is described.

JP 2009-281897 A

Desmet et al., 2005, Anal. Chem., 77, 4058-4070

However, in the method of simply obtaining the column length L that becomes ΔPmax for each linear velocity u ₀ by the above processing in the KPA under the restriction condition of the given flow constant Cf, the optimum column length L is not always obtained. It has been found by the inventors' investigation.
That is, even if the required high speed performance (void time t ₀ ) or high separation performance (theoretical plate number N) is obtained at a pressure value (system pressure) lower than ΔPmax, the maximum ΔPmax Is unambiguously set as an asymptotic limit value, and a situation may occur in which the pressure is unnecessarily increased.
An unnecessary increase in pressure leads to a burden on the LC hard member and wear of parts, leading to a problem of shortening the product life.
The present invention has been made in view of the above problems, and provides a device capable of obtaining a realistic and optimum column length L and flow rate F (linear velocity u ₀ ), and a method using the device. That is.

  As one aspect for solving the above-described problems, the present invention has the following features. A liquid feeding section for feeding a mobile phase, a sample injection section for injecting a sample into the fed mobile phase flow path, a column for separating the injected sample, and detecting the separated sample A detection unit and a control unit for processing the detected result, wherein the control unit is an index related to the analysis time based on the data on the flow rate dependency of the column efficiency and the data on the fluidity And whether or not the obtained index exceeds a predetermined allowable range, and based on the result of the determination, set the analysis conditions for the column length and the mobile phase flow rate. And a method using the device.

According to the above-described aspect, it is not necessary to raise the pressure excessively when considering the high speed and high separation of the LC. Therefore, more realistic and optimum column length L and flow rate F (linear velocity u ₀ ) are set. Can be sought. In addition, since the burden on the hard member can be reduced, the life of the entire apparatus can be extended.

FIG. 3 is a flowchart of speeding up processing showing Embodiment 1 of the present invention. FIG. The graph which shows the example of Hu plot. Graph showing an example of t ₀ -N plot of various columns. Graph showing an example of t ₀ -N plot of various columns (N fixed). Graph showing an example of t ₀ -N plot of various columns (t ₀ fixed). Shows the layer structure of t ₀ -N plot of Example 1 of the present invention. The separation process flowchart which shows Example 2 of this invention. 1 is a system configuration diagram showing an embodiment of the present invention. The system flow-path figure which shows the Example of this invention. The figure which shows the display screen of the input device 8 which shows the Example of this invention. The figure which shows the display screen of the output device 9 which shows the Example of this invention.

Hereinafter, a layered kinetic plot analysis method (hereinafter referred to as LKPA method) according to an embodiment of the present invention will be described with reference to the drawings.
As a premise, a method for uniquely determining the column length L for each linear velocity u ₀ using (Formula 1) using the conventional KPA method under the restriction condition of the given flow constant Cf, It will be explained that the method of obtaining the column length L that provides the maximum pressure loss ΔPmax at the speed u ₀ is not sufficient to obtain the optimum column length L in practice.

As an example, when using a filler column having a particle diameter of 5 μM, in KPA, the calculation is performed under the condition that the pressure loss ΔPmax is increased to 100 MPa. However, in reality, it is not necessary to increase the pressure to 100 MPa, and the analysis is often performed under a pressure condition lower than this value (for example, about 20 MPa).
The reason is as follows. Normally, in the region near the theoretical plate number N = 10,000, when a packing column with a particle size of 5 μM is used as in this example, the pressure loss ΔPmax is increased by about ΔPmax = 20 MPa. However, even if the column length L is increased, the void time t _o is not shortened. Further, no matter how much the pressure loss ΔPmax is larger than the above value, only the flow constant Cf, the column length L, and the linear velocity u ₀ (flow rate) increase, and there is no performance advantage in LC analysis. Because there is no.

The above phenomenon is a useless repetitive operation and is a vicious circle. That is, when the linear velocity u ₀ is increased for speeding up, it is necessary to consider the aforementioned Hu curve. At a particle size of 5 μM, when the linear velocity u ₀ is increased even if it exceeds the minimum value of the theoretical plate height H, the theoretical plate height H increases linearly with respect to the linear velocity u ₀ . In this linear increase region, even if the linear velocity u ₀ is increased, the theoretical plate number N deteriorates and the void time t ₀ cannot be shortened efficiently. In other words, increasing the linear velocity u ₀ by 10% increases the theoretical plate height H by nearly 10%. In this case, in order to maintain the theoretical plate number N, the column length L must be increased by about 10% according to Equation 2, and the void time t ₀ eventually returns to the original value according to Equation 3. .

KPA has no problem as a methodology because it is intended to determine the reach performance of the particle size packing column even in unrealistic or extreme assumptions. However, when KPA is used, it is impossible to determine how much mobile phase of linear velocity u ₀ (flow rate) should actually be sent to a column of length L. The present invention provides a methodology for providing realistic column length L and linear velocity u ₀ solutions, and t _o -N plots, and provides the necessary setup variables.

To summarize the above contents, the input data in KPA (H-u plot, Kv, eta,? Pmax) when outputting the t _o -N plot from also intermediately method consisting column length L and the linear velocity u ₀ calculate. At this time, if it is accompanied by useless repetitive operation, the condition set (u _o , L) calculated incidentally may be limitless in reality.
The LKPA according to the present invention provides an actual solution of the conditions (u _o , L) assuming actual analysis. Specific embodiments thereof will be described below with reference to the drawings.

  FIG. 9 is a flow chart of the liquid chromatograph system according to the embodiment of the present invention.

In FIG. 9, a pump 11A for transporting the eluent (mobile phase) 10A and a pump 11B for transporting the eluent (mobile phase) 10B are provided, and the eluent 10A and the eluent 10B sent by each pump are mixers. 12 is mixed. Gradient liquid feeding is also possible with the pump 11A and the pump 11B. The eluent mixed by the mixer 12 is sent to the analysis column 15 together with the sample injected by the autosampler 13. The analytical column 15 includes a column oven 14 that adjusts the temperature of the column.
In the analysis column 15, the sample is separated for each component and sent to the detector 16. The cell 17 of the detector 16 is irradiated with light, and a chromatographic waveform is obtained from the signal intensity. The sample and eluent are then sent to the waste liquid tank 18.

  FIG. 8 shows a configuration diagram of the liquid chromatograph system according to the embodiment of the present invention. The liquid chromatograph apparatus 1 corresponds to the configuration described above with reference to FIG. 9 and includes a pump 2, an autosampler 3, a column oven 4, and a detector 5. Each module in the liquid chromatograph apparatus 1 is connected to the data processing apparatus 6 to exchange data. The processing result in the data processing device 6 is displayed on the output device 9. The data processing device 6 exchanges data with the method conversion processing computer 7. The method conversion processing computer 7 can perform processing based on the conditions input from the input device 8. The data processing device 6 and the method conversion processing computer 7 may perform processing by the same computer. In the method conversion processing computer 7, processing using LKPA according to the present invention is executed.

  FIG. 1 shows a flowchart of acceleration processing (acquisition of acceleration parameters) according to an embodiment of the present invention. Here, the pressure loss (system pressure) ΔP is originally described as an example of the scanning parameter, but another scanning parameter includes a flow constant Cf that is an inversely proportional constant. Scanning will be described by taking an example of processing discretely for each predetermined width. Here, the pressure loss ΔPmax is set as an initial value, and subtraction processing is performed in steps of, for example, 10 MPa. In FIG. 1, the step is indicated as s = 10 MPa, but it means a step width of ΔP, and can be expressed as ΔP = 10 MPa. That is, when the pressure loss ΔP is gradually decreased and it is determined that the above-described useless repetition has been resolved, the decrease in the pressure loss ΔP is stopped.

The index for determining the elimination of the useless repetitive operation was the void time t _o that is an index related to the analysis time. For example, plate time (t _o / N), impedance time (t _o / N ² ), etc. can be adopted as the determination index, but in the case of FIG. 1, the number of theoretical plates N _target to be acquired is fixed. Similar processing can be performed when these are used.

In this embodiment, the step of the pressure loss ΔP is set to 10 MPa, and a plane graph of a t _o -N plot is generated for each ΔP. By laminating them one after another, a layered structure is formed.

A specific process is demonstrated along the flowchart of FIG. First, when the speed-up process is started (S101), input settings for various parameters are performed (S102). As necessary information, in addition to the H-u plot, Kv, η, and ΔPmax, N _target to be set as a target value in this processing, a descending step width s of ΔP, and an allowable rate are input. Each input information will be described below.
Kv, η, and ΔPmax are parameters that are required as input variables from the beginning when KPA is used, and the flow constant Cf is uniquely determined by Equation 1 using these values.
The Hu plot is used for conversion to a t ₀ -N plot.
N _target is a value required when interpreting the t ₀ -N plot obtained by KPA, and is a fixed value for each process. For example, as described above, taking as an example the case shown in FIG. 4, as a void time corresponding to N _target = 20,000 was set as a target value, that t _o = 10 s is the fastest Recognize.
In addition to the information described above, a descending step width s and an allowable rate% are required as specific input variables when the LKPA according to the present invention is used. The allowable rate is a fixed value for each process. The step width s (that is, ΔP width) corresponds to the vertical interval of the layers in FIG.
Next, the value of pressure loss ΔP is initially set as ΔPmax (S103), and a t ₀ -N plot at ΔPmax is generated, and among t ₀ required to obtain N _target at ΔPmax, T indicating the minimum time is obtained (S104).
Thereafter, a step-down process from ΔPmax is performed using the set step width s (S105).
A t ₀ -N plot at each ΔP is generated, and t ₀ @N _target at ΔP is obtained from the interpolation curve (S106).

The allowable rate% is an allowable range related to the ratio of t ₀ to T obtained by the descending step process, with T indicating the minimum time as 100% of the void time t ₀ obtained when ΔP = ΔPmax.
That is, here, it is determined whether or not a value obtained by multiplying T by the allowable rate% is larger than t ₀ (S107). When this value becomes larger than t ₀ , the descending step process of S105 is repeated. On the other hand, if it does not exceed t ₀ , the descent step process is stopped at ΔP at this time (S108).
Here, S107 will be described using a specific example using Table 1.
For example, starting from ΔPmax = 100 MPa, T = 5.0 s and subtracting every ΔP = 10 MPa as the step width, as shown in Table 1, N _target is acquired even when ΔP = 90 MPa. The time required for this is t ₀ = 5.0 s, which is the same as when ΔPmax.
As shown in Table 1, the allowable rate becomes 110% or more only when the pressure is lowered to ΔP = 60 MPa.
As in the above example, there is a ratio that divides (1) the upper region of pressure loss where useless repetition occurs and (2) the boundary where high pressure loss is beneficial for speeding up. Permissible rate is%.

これは、Ｔ（s）が既知である場合には、許容率％の代わりにオペレータが目標ボイド時間ｔ_{o target}（s）を入力設定することもできる。
通常のフローはΔＰの減算処理だが、目標ボイド時間ｔ_{o target}（s）を入力する設定方法の場合は、１０， ２０， …ＭＰａとΔＰを加算する処理方法もありうる。この場合は無益な繰返し操作を行う前に処理を停止することとなる。
加算する処理方法の短所は圧力損失ΔＰを引き上げで達成可能な限界値を知ることができない点である。ＬＫＰＡにより圧力損失ΔＰを上昇・下降させることは、実験的な操作ではなく計算処理なので、ボイド時間ｔ_oの限界値を知ることができる減算処理のほうが望ましい。

In this case, when T (s) is known, the operator can input and set the target void time t _{o target} (s) instead of the allowable rate%.
The normal flow is subtraction processing of ΔP, but in the setting method of inputting the target void time t _{o target} (s), there can be a processing method of adding 10, 20,... MPa and ΔP. In this case, the processing is stopped before the useless repetition operation is performed.
The disadvantage of the processing method to be added is that the limit value achievable by increasing the pressure loss ΔP cannot be known. Increasing / decreasing the pressure loss ΔP by LKPA is not an experimental operation but a calculation process, so a subtraction process that can know the limit value of the void time t _o is preferable.

Thereafter, the determined parameter group, that is, the pressure loss ΔP at the time of stopping the process, the void time t _o , the column length L and the linear velocity u ₀ for realizing it are output (S109), and the process is terminated.

The operator procures a column having a length L as an output result and sends it at a flow rate F derived from the linear velocity u ₀ to realize a high-speed analysis method. The pressure loss ΔP and the void time t _o are indexes obtained as a result of measurement under these analysis conditions, and are measured and confirmed as necessary.
In the above embodiment, the case where ΔP is discretely processed has been described for convenience, but mathematically ΔP can be expanded to a dense real number. In the case of a real number, the t _o -N curve on the plane exhibits a curved surface that hangs down like a curtain (film) in a three-dimensional space stretched by three axes of N, t _o , and ΔP.
FIG. 10 shows a display screen of the input device 8 in the above processing. As shown in this drawing, necessary parameters and processing conditions for each column to be evaluated are input, and a processing start button 1001 is pressed to start processing.
FIG. 11 shows a display screen of the output device 9 in the above processing. As shown in the figure, the operator can recognize the column length L and the linear velocity u ₀ which are finally required analysis conditions.

In the first embodiment, the high-speed processing has been described. In the present embodiment, the high-speed processing will be described. Here, the liquid chromatograph apparatus to which this process is applied is the same as that described with reference to FIGS.
A specific process is demonstrated along the flowchart of FIG. First, when high separation processing is started (S701), input settings of various parameters are performed (S702). As necessary information, in addition to the H-u plot, Kv, η, and ΔPmax, t _{0 target} to be set as a target value in this processing, a descending step width s of ΔP, and an allowable rate are input. Each input information will be described below.
Kv, η, and ΔPmax are parameters that are required as input variables from the beginning when KPA is used, and the flow constant Cf is uniquely determined by Equation 1 using these values.
The Hu plot is used for conversion to a t ₀ -N plot.
t _{0 target} is a value required when interpreting the t ₀ -N plot obtained by KPA, and is a fixed value for each process. For example, as described above, taking the case shown in FIG. 5 as an example, N = 60,000 may be the maximum resolution as the number of theoretical plates corresponding to t _{0 target} = 100 s set as the target value. Recognize.
In addition to the information described above, a descending step width s and an allowable rate% are required as specific input variables when the LKPA according to the present invention is used. The allowable rate is a fixed value for each process. The step width s (that is, ΔP width) corresponds to the vertical interval of the layers in FIG.
Next, the value of the pressure loss ΔP is initially set as ΔPmax (S703), and a t ₀ -N plot at ΔPmax is generated to generate a t _{0 target} at ΔPmax. Nmax indicating the highest resolution is obtained (S704).
Thereafter, a step-down process from ΔPmax is performed using the set step width s (S705).
A t ₀ -N plot at each ΔP is generated, and N @ t _{0 target} at ΔP is obtained from the interpolation curve (S706).
The permissible rate% refers to a permissible range regarding the ratio of N to Nmax obtained by the descending step process, where Nmax indicating the highest resolution among the theoretical plate numbers N obtained when ΔP = ΔPmax is 100%.
That is, here, it is determined whether or not a value obtained by multiplying Nmax by the allowable rate% is smaller than N (S707). If this value is not smaller than N, the descent step process of S705 is repeated. On the other hand, if it is smaller than N, the descent step process is stopped at ΔP at this time (S708).
Here, S707 will be described using a specific example using Table 2.
When useless repetition occurs, for example, starting from ΔPmax = 100 MPa, N = 20,000, and subtracting every ΔP = 10 MPa as the step width, as shown in Table 2, ΔP = 90 MPa The number of theoretical plates required to obtain t ₀ is 20,000, which is the same as ΔPmax.
Then, as shown in Table 2, the allowable rate becomes 90% or less only when it is lowered to ΔP = 60 MPa.

上記のように、高分離化処理の場合は、目標ボイド時間ｔ_{o target}を基準に圧力損失ΔＰを減算しながら、圧力損失ΔＰごとに理論段数Ｎを計算する。この場合、Ｓ７０７において、許容率を乗じた理論段数Ｎｍａｘ以下になるところで、圧力損失ΔＰの減算処理を停止する。減算処理が終了すると計算生成物として、処理停止時の圧力損失ΔＰ、ボイド時間ｔ_o、およびそれを実現するカラム長さＬ、線速度ｕ₀が決定される。
その後、決定されたパラメータ群、すなわち、処理停止時の圧力損失ΔＰ、理論段数Ｎ、およびそれを実現するカラム長さＬ、線速度ｕ₀を出力し(Ｓ７０９)、処理を終了する。

As described above, in the case of high separation process, while subtracting the pressure loss [Delta] P based on the target void time t _{o target,} to calculate the number of theoretical plates N for each pressure loss [Delta] P. In this case, in S707, the subtraction process of the pressure loss ΔP is stopped when the theoretical plate number Nmax or less multiplied by the allowable rate is reached. When the subtraction process is completed, a pressure loss ΔP, a void time t _o , a column length L, and a linear velocity u ₀ when the process is stopped are determined as calculation products.
Thereafter, the determined parameter group, that is, the pressure loss ΔP when the process is stopped, the theoretical plate number N, the column length L that realizes it, and the linear velocity u ₀ are output (S709), and the process ends.

The operator procures a column of length L obtained as a solution as a method, and sends the solution at a flow rate F derived from the linear velocity u _0, thereby realizing a high speed and high separation analysis method. The pressure loss ΔP and the void time t _o are indexes obtained as a result of measurement under these analysis conditions, and are measured and confirmed as necessary.
As described above, with respect to the column length L and the flow rate F (linear velocity u ₀ ), which are indispensable indexes for optimizing the LC analysis method, the present embodiment can obtain a practically optimal solution. The possible LKPA method has been described.
According to this method, even in the case where the conventional KPA brings an extreme solution to the column length L and the flow rate F, it becomes possible to perform more stable analysis without imposing a burden on the hard member.
The present invention can be applied to method optimization software, method transfer programs, and UHPLC systems.

DESCRIPTION OF SYMBOLS 1 Liquid chromatograph apparatus 2 Pump 3 Autosampler 4 Column oven 5 Detector 6 Data processing apparatus 7 Method conversion processing computer 8 Input apparatus 9 Output apparatus 10A Eluent 10B Eluent 11A Pump 11B Pump 12 Mixer 13 Autosampler 14 Column oven 15 Analysis column 16 Detector 17 Cell 18 Waste liquid tank 1001 Process start button

Claims (9)

A liquid feeding part for feeding the mobile phase;
A sample injection portion for injecting a sample into the liquid phase mobile phase flow path,
A column for separating the injected sample;
A detection unit for detecting the separated sample;
A data processing unit for processing the detected result;
In a liquid chromatograph apparatus having a method conversion processing unit connected to the data processing unit ,
The method conversion processing unit
Based on the data on the flow rate dependence of the column efficiency and the data on fluidity,
Find metrics on analysis time,
The calculated index is
Determine whether it exceeds the predetermined tolerance,
Based on the result of the determination,
The liquid chromatograph apparatus is characterized by setting analysis conditions relating to a length of the column and a flow rate of the mobile phase. In the liquid chromatograph apparatus according to claim 1,
The method conversion processing unit
The calculated indicator is
If it exceeds the predetermined tolerance,
Based on the indicator in the liquidity data when the range is exceeded,
The liquid chromatograph apparatus is characterized by setting analysis conditions relating to a length of the column and a flow rate of the mobile phase. In the liquid chromatograph apparatus according to claim 1,
The indicator related to the analysis time is
A liquid chromatograph apparatus characterized by a time required for acquiring a predetermined target theoretical plate number. In the liquid chromatograph apparatus according to claim 1,
The fluidity data is a pressure condition value,
The method conversion processing unit
The pressure condition value is sequentially changed at predetermined intervals,
Based on the data on the flow rate dependence of the column efficiency at the changed pressure condition value,
A liquid chromatograph apparatus for obtaining an index relating to the analysis time. In the liquid chromatograph device according to any one of claims 1 to 4,
The liquid chromatograph apparatus, wherein the data processing unit and the method conversion processing unit are provided in the same computer. An optimization method for liquid chromatographic analysis,
A first step of inputting data relating to flow dependency of column efficiency and data relating to fluidity;
Based on the data on the flow dependency of the column efficiency and the data on the flowability,
A second step for obtaining an index relating to analysis time;
The calculated index is
A third step for determining whether or not a predetermined allowable range is exceeded;
Based on the result of the determination,
A fourth step of setting analysis conditions relating to the length of the column and the flow rate of the mobile phase, and a method for optimizing a liquid chromatograph analysis. A method for optimizing liquid chromatographic analysis according to claim 6, comprising:
In the fourth step,
The calculated index is
If it exceeds the predetermined tolerance,
Based on the indicator in the liquidity data when the range is exceeded,
A method for optimizing a liquid chromatograph analysis, wherein analysis conditions relating to a length of the column and a flow rate of the mobile phase are set. A method for optimizing liquid chromatographic analysis according to claim 6, comprising:
The indicator related to the analysis time is
A method for optimizing a liquid chromatograph analysis, characterized in that it is the time required to acquire a predetermined target theoretical plate number. A method for optimizing liquid chromatographic analysis according to claim 6, comprising:
In the second step,
The fluidity data is a pressure condition value,
The pressure condition value is sequentially changed at predetermined intervals,
Based on the data on the flow rate dependence of the column efficiency at the changed pressure condition value,
A method for optimizing a liquid chromatograph, characterized in that an index relating to the analysis time is obtained.

Images