JPH04326058A - Apparatus for sampling component by liquid chromatograph - Google Patents

Abstract

PURPOSE:To certainly sample an objective component regardless of a condition such as a holding time or peak sequence by preliminarily storing the spectrum data of a component to be sampled and comparing the same with the spectrum data of an analyzed outflow component. CONSTITUTION:After a mobile phase is sent to a column 1 and a sample is injected in a sample injection part 8, the absorption spectrum data of the eluate of the column 1 is read in a buffer memory MB. A comparing operation part C performs the comparing operation of said data and the adsorption spectrum data of an objective component stored in a reference data memory MS. The calculated value of the degree of coincidence is compared with the judging level stored in a judging level memory part J and, based on whether said value is larger than the judging level, a three-way valve 3 is changed over to a sampling container 4 or a drainage receiver 5. As mentioned above, since the outflow component of a chromatograph and the objected component are identified and the outflow component regarded as a substance of the same kind is sampled, the objective component can be certainly sampled even when a holding time is shifted or the peak of impurity appears on the way.

Description

[Detailed description of the invention]

0001

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a component separation device for separating sample components using a liquid chromatograph and collecting designated components.

0002

[Prior Art] In order to separate sample components using a liquid chromatograph, it is necessary to set the separation time of the target component in advance depending on the retention time of the component to be fractionated, or to place the target component on the chromatogram. Depending on whether it is a peak, a method is used in which peak detection is performed and components exhibiting peaks of that order are fractionated.

Among these conventional methods, the time-based method changes the retention time of each component due to slight differences in temperature, mobile phase, or sample injection volume, so the peak of the target component may deviate from the peak. There is a risk that the preparative operation will be performed at a different location, and if the order is based on the order of the peaks, the order may be changed due to the introduction of unexpected impurity peaks and a completely different component may be separated. In any case, it was difficult to carry out accurate fractionation.

0004

[Problems to be Solved by the Invention] The present invention provides a fractionation device that can reliably fractionate target components without relying on indicators that may change depending on conditions such as retention time or peak order as described above. This is what we are trying to provide.

[0005]

[Means for solving the problem] The spectral data of the component to be separated is stored, the liquid chromatograph effluent is analyzed and the spectral data of the effluent component is collected, and the spectral data of the effluent component and the above-mentioned pre-stored The degree of agreement with certain spectral data was determined, and if the degree of agreement was greater than a certain degree, the component exhibiting that peak was fractionated.

[0006] As a method for identifying substances, a method of comparing spectra or mass spectra is used. When the spectra of two substances match well, the two components can be considered to be the same type of substance. The present invention uses this spectral comparison to identify the chromatographic effluent component and the target substance, and separates the effluent component that can be considered to be the same substance. The target component can be fractionated without error.

[0007]

DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 shows an apparatus according to an embodiment of the present invention. 1 is a liquid chromatography column, 2 is a flow cell through which the effluent from column 1 flows, 3 is a component separation three-way valve, 4 is a separation container, and 5 is a drain receiver. 6 is a mobile phase liquid, 7 is a pump that sends the mobile phase to the column 1, and 8 is a sample injection part. Across the flow cell 2, 9 is a light source, 10 is a spectrometer, and 11
is a photodiode array placed on the spectral image plane formed by the spectrometer, and the light source, spectrometer, and photodiode array constitute a multiwavelength spectrophotometer.
Reference numeral 12 denotes a control device that takes in the output signal of the photodiode head array 11, processes the data, and operates the three-way valve 3.

As shown in FIG. 2, the control device 12 includes a buffer memory MB for temporarily storing output data of the photodiode array 11, and a standard data memory MS for storing spectral data of component substances to be separated. ,
Comparison calculation section C, judgment level storage section J, and comparison calculation section C
The output of the comparison calculation section C is compared with the judgment level, and when the output of the comparison calculation section C is equal to or higher than the judgment level, the three-way valve 3 is switched to the separation container 4 side, and in other cases, the three-way valve 3 is switched to the drain receiver 5 side. It has each functional part of the three-way valve control section V.

FIG. 3 is a flowchart of the operation of the control section 12 described above. A mobile phase is sent to column 1,
After the sample is injected into the sample injection section 8, the operation is started. Spectral data is read from the photodiode array 11 into the buffer memory MB (a). As is clear from FIG. 1, this spectrum data is absorption spectrum data of the column effluent. Standard data memory M
Perform a comparison calculation with the absorption spectrum data of the target component substance stored in S (b), and check whether the calculated degree of coincidence value is greater than the judgment level stored in the judgment level storage unit (h) ), if it is large (YES), switch the three-way valve 3 to the preparative container side (d); if NO, switch the three-way valve 3 to the drain receiver side (e), and continue this operation for a certain period of time. ,
The operation ends after that time has elapsed. This fixed time is set at a time when all the components of the injected sample have finished flowing out and the next sample can be injected into the column.

[0010] An example of comparison calculation between the spectra of the column effluent components and the spectra of the target substance will be described. Spectral spectrum data is composed of photometric outputs at several wavelengths λ1, λ2,...λn. This set of n values can be considered as one vector. If the vector of spectral data of the column outflow component is represented by S, and the vector of spectral data of the target component is represented by U, then when both vectors are parallel, both spectral data match, and the length of the vector simply represents the concentration ratio. It only represents. In reality, the component flowing out of the column is a solution of the mobile phase, and the absorption spectrum of the mobile phase overlaps, and it is rare that it completely matches the standard spectroscopic spectrum of the target component, so both spectra are parallel to some extent. When they are close, the two substances are considered the same. The calculation of the degree of coincidence is the calculation of the parallelism of two vectors, and when the two vectors are parallel, the normalized vector whose component is the value obtained by dividing the vector component by the vector length for each vector, The scalar product becomes 1. Therefore, when this scalar product exceeds a certain determination level close to 1, it is determined that both substances are the same. To write this calculation concretely, let each component of the vector S of the spectrum of the column outflow component be Si (i = 1, 2,...n), and each component of the vector U of the spectrum of the target substance be Ui. , the components of each normalized vector are for the column effluent components,

[Math 1] About target ingredients

[Math 2] The parallel condition is

[Math 3] The above formula is expressed in Si and Ui.

[Math 4] becomes.

[0011] In the above-mentioned embodiments, the determination is made based on the degree of coincidence of spectroscopic data, but this is not limited to spectra, and mass spectra can also be used.

0012

According to the present invention, even if the retention time or peak order of the component to be fractionated changes, the target component can be fractionated without fail, and the reliability of sample fractionation work is improved.

[Brief explanation of drawings]

FIG. 1 is a block diagram showing the overall configuration of an apparatus according to an embodiment of the present invention;

[Figure 2] Functional block diagram of the control device of the same embodiment

[Figure 3] Flowchart of the operation of the above control device

[Explanation of symbols]

1 Column 2 Flow cell 3 Three-way valve 4 Separation container 5 Drainage receiver 6 Mobile phase

Claims (1)

[Claims] A means for storing spectral data of a component to be fractionated, a means for collecting spectral data of a liquid chromatograph effluent, and a spectral data obtained by the spectral data collecting means and stored in the storage means. Component separation using a liquid chromatograph, comprising means for calculating the degree of coincidence with spectral data, and switching means for guiding the liquid chromatograph effluent to a preparative container when the calculated degree of coincidence is equal to or higher than a predetermined value. removal device.