JPS61159160A - Quantitative determination in chromatography nd data processing unit therefor - Google Patents

Abstract

PURPOSE:To make possible the quantitative determination by regarding the chromatograms of a standard product and unknown sample as respective functions, regarding the chromatogram of the unknown sample as the primary association of the standard product and determining the coefft. thereof by a method of least squares. CONSTITUTION:A program 0 is a program to move a data processing unit body. Said program samples the respective peaks of the chromatogram obtd. by high-speed liquid chromatography and stores the resulted signal levels as a peak file into a RAM2. A program 1 calls the peak file from the RAM2, takes out the area value of each segment necessary for the subsequent calculation or the area value of each peak and writes out the same on a floppy disk 3 by reorganization to the data file. A program 2 is a program to correct the file written out on the disk 3 formed by the program 1 by manual input. A program 3 calls the data file of the segment area or peak area formed by the program 1 or 2 and determines the primary association coefft. The content of each component is thus quantitatively determined.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to a quantitative determination method in chromatography of a sample in which target components are composed of a plurality of chemical species, and a data processing device for the same, and particularly relates to a method for quantifying a sample in which a target component is composed of a plurality of chemical species, and a data processing device for the same.
), etc., where the sample is a surfactant that is made up of many chemical species, such as a detergent, and the content of each interface is measured rather than the content of each individual chemical species. The present invention relates to a chromatographic quantitative method based on a completely new perspective that can be effectively applied when analysis to determine the content of an activator is required, and a data processing device for quantitative analysis. A particularly suitable industrial field is the quality control department in the manufacturing field, where final products are made by mixing surfactants, I-rimers, crude drug extracts, etc.

Conventional technology Chromatography such as HPLC is originally a means of separating a sample into its components for quantitative analysis, but when the target component itself is a mixture of many chemical species, such as the above-mentioned surfactant, Since the components are also separated into their respective chemical species, quantitative calculations have been performed using processing methods such as time grouping. However, in many cases, it is extremely rare for the many chemical species that make up the components of these mixtures to be separated in a chromatogram without interfering with each other, and in such cases, time grooving, etc. Treatment methods cannot be applied, and quantitative analysis by HPLC or the like has been considered impossible.

In other words, in conventional quantitative methods (and data processing equipment R for quantitative determination), the objective is achieved by making the chemical amount correspond to the peak area (or height) K of the separated chemical species. However, as mentioned above, quantitative analysis cannot be carried out unless the peaks of each type of cursive character are clearly separated due to mutual interference.

Problems to be Solved by the Invention Accordingly, the purpose of the present invention is to provide a completely separate new perspective that enables quantitative analysis of a target sample even in cases where the conventional quantitative methods described above are difficult to apply in principle. The purpose of this patent is to provide a quantitative determination method in chromatography based on chromatography and a data processing device therefor.

Means for Solving the Constituent Problems of the Invention In order to achieve the above object, the quantitative method in chromatography of the present invention includes the steps of storing a chromatogram measured by high performance liquid chromatography or the like in a memory as a peak file; Retrieving the peak file from memory, extracting the area value of each section in the case of division processing, and reorganizing it as a data file in the case of peak waveform processing; The method is characterized by the step of calculating and obtaining each coefficient according to the method of least squares when 2m is regarded as a linear combination of the chromatograms of each sample.

Further, the data processing device for quantitative analysis in chromatography of the present invention comprises means for implementing each step of the above quantitative method, specifically, an icrocomputer including a fixed memory (RAM) and a floppy disk. The system is characterized by being composed of hardware such as a computer and software such as a program that causes the hardware to perform operations corresponding to each of the above steps.

EXAMPLES Hereinafter, examples of the present invention will be explained in detail with reference to the drawings, but first, the principle of the quantitative method according to the present invention will be briefly explained.

The quantitative method of the present invention differs from conventional methods in that it starts from a completely new perspective of understanding the chromatogram as a function. tb, and the chromatogram obtained by analyzing the unit amount of component l is ft(t), then the chromatogram F of an unknown sample containing components 1 y n in amounts C1 to Cn, respectively.
(t) is expressed as a -order combination of f l (t) as follows: F(t) - ΣC1ft(t) Here, F(t) and ft(t) are the unknown sample and each It is obtained by measuring a standard sample with a chromatograph such as HPLC, and the quantitative operation is nothing but determining the coefficient C1 of the above-mentioned -order bond.

This can be obtained as the value of C1 that minimizes the sum of squared residuals expressed as R=〒(F x C*fJ)2 by applying the principle of least squares method to the above equation. Note that yj, 11 is a function of each chromatogram obtained discretely.

FIG. 1 is a conceptual block diagram showing the configuration of a data processing apparatus according to the present invention, and the method and apparatus of the present invention will be explained below with reference to the same figure.

First, the data processing device is mounted on the data stage of the Chromadog 2F, such as an HPLC, and the key and interface (
(None of these are shown) as well as RAM for program storage.
1, RAM for storage of peak files 2 (R
A floppy disk 3j for storing the reorganized data files
and an output device 4 for outputting calculation results.

Program 0 is a program 2 that operates the main body of the data processing device stored in RAM 1, and each peak of a chromatogram obtained by measurement using HPLC etc.
．． Sampling is performed every 1 s@e, and the resulting signal level is stored in RAM Z as a peak file.

Program l calls the above-mentioned peak file of the chromatogram from RAM 2, and from there the area value of each division if the division processing necessary for later calculations has been performed, or the area of each peak if the peak waveform processing has been performed. This is a program that extracts values, reorganizes them into a data file, and writes them onto the floppy disk 3.

Program 2 is a program 2 that is used when manually modifying a data file created using program 1 and written onto floppy disk 3, or when manually inputting data without using program 1. The data files written to the floppy disk 3 may be automatically reorganized from the peak file by program 1, or may be manually reorganized from the display of the peak file by program 2. .

Program 3 is the main calculation program, which calls the data file of the section area or peak area created by the above program 1 or 2, executes the following calculation, and obtains the coefficient Ci of the linear combination described above: Here, 11 Product: -8tdl (A: AV-AA) # Unknown 9: (AV, -A:) sta2 [AVA
・Mu Meng).

However, M1 is the actual value of Category 1. Stdm (A〒whip...cod).

Content of each component C1...C determined by the above calculation
n is output to the output device 4. As a result, it is possible to determine the content of each component, which could not be measured individually by conventional methods because the components interfere with each other.

Next, the results of a model experiment to which the present invention was applied are shown in the following Examples and Figures *2 and 3.

(Example) Polystyrene oligomers with different degrees of polymerization as Odel samples ``HIGH'' (average molecular weight approximately 800) and ``LOW''
" (average molecular weight of about 200). Note that these two components have conventionally been impossible to quantitatively analyze individually because their chromatograms overlap.

The above @HIGH" and "1,01iV'" are regarded as standard products, and these are respectively (10:90), (25 near 5), (
50:50).

(75:25) and (90:10), and this was used as an unknown sample to be quantified. Standard and unknown samples were analyzed by injection of 10 pL each under the following HPLC conditions, and the results were monitored and analyzed using the data processor of the present invention.

Column: Ftn@pak GWL 101 mobile phase;
Chloroform flow rate: 0.5 mt/min As a result, the chromatograms shown in Figures 2 (&) and 2 (b) for the @HIGH and IOW samples, and the 3rd chromatogram for unknown samples of each mixing ratio. The chromatograms and quantitative results shown in Figures (1) to 2 (•) were obtained. In other words, the accuracy of the quantitative value is at most 1.131 even in the case where the error is the widest (50:50). In all other cases, the deviation is within l-, and the standard deviation of the quantitative value in each case is (b) are all within 1%. These results are fully satisfactory as a HPLC quantitative method.

Effects of the Invention As described above, according to the present invention, the chromatograms of the standard and unknown samples are each regarded as a function, the chromatogram (function) of the unknown sample is regarded as a linear combination of the standard, and its coefficients are minimized. Quantitative analysis is performed from a completely new perspective by applying the method of squares, making it possible to quantify things that could not be individually quantified using conventional methods. Therefore, the present invention provides remarkable effects in the field of quantitative analysis of mixtures consisting of many chemical species that interfere with each other and cannot be separated individually, such as detergent surfactants.

[Brief explanation of drawings]

Fig. 1 is a conceptual block diagram of a data processing device that implements the quantitative method of the present invention, Figs. 2m and 2b are diagrams showing polystyrene oligomer "HIGH" and HI@LOW' glue quadrograms, and Fig. 3 & ~3・The figure is @HIGH” and @
LOW' (10:90), (25 near 5), (50:
50), (75:25), (90:10). FIG. 3 is a diagram showing a chromatogram and quantitative values of each sample mixed at a ratio of . 1... RAM for storing programs, 2-... RAM for storing peak files, 3... Floppy disk for storing data files, 4-... Output device.

Claims (2)

[Claims] (1) The step of storing a chromatogram measured by high-performance liquid chromatography, etc. as a peak file in memory, loading the above-mentioned peak file from memory, and in the case of segmentation processing, the area value of each segment, and in the case of peak waveform processing, each The step of extracting the area values of the peaks and reorganizing them as a data file, and calling the above data file and calculating each coefficient using the method of least squares when the chromatogram of the unknown sample is regarded as a linear combination of the chromatograms of each standard. A method for quantitative determination in chromatography, characterized by comprising the step of determining. (2) Means for storing a chromatogram measured by high-performance liquid chromatography, etc. in a memory as a peak file, and loading the above-mentioned peak file from the memory. Means for extracting peak area values and reorganizing them as a data file, calling the above data file, and calculating each coefficient according to the method of least squares when the chromatogram of the unknown sample is regarded as a linear combination of the chromatograms of each standard. A data processing device for performing quantitative analysis in chromatography, characterized in that it includes means for determining.