JPS59220645A - Chromatogram display apparatus - Google Patents

Abstract

PURPOSE:To obtain display easy to observe by enhancing the separation degree of components, by using the holding time and the expanse corresponding to a column different from the column used in analysis of chromatography and the peak height obtained by a comparator means. CONSTITUTION:An analogue like or digital like chromatogram signal is inputted from an input means and compared with the function curve signal corresponding to a specific component supplied from a memory means 12 by a comparator means to be made approximate thereto. The function curve made approximate by the comparator means 13 is synthesized by a chromatogram generating means 14 to be displayed by a display means 15 and a new chromatogram is displayed. The generation of each component wave form relating to each component is performed by the chromatogram generating means and the result thereof is supplied to the display means 15 to be displayed. As a result, a chromatogram can be fabricated and displayed in a high separation degree similar to that of the chromatogram obtained through a long column and highly precise measurement can be performed.

Description

DETAILED DESCRIPTION OF THE INVENTION This invention relates to an apparatus for displaying a chromatogram obtained by analyzing a sample containing at least two components using a chromatogram.

Background> Conventionally, in a dichromatodarum display device, a sample separated by a column is supplied to a flow cell, where it is converted into a signal using absorbance, refractive index, fluorescence intensity, etc. Some displayed the signal directly, others displayed the logarithm value of the signal from the flow cell, and some displayed the signal after removing or smoothing the (m) minute peak from the flow cell.

In the previous direct display, we were able to display the most linear information from the flow cell and correctly, but the inspection innovation 1
Noise on the means itself, noise generated during operation of the detection means, noise generated during signal transmission, etc., are included in the display and become an impediment to the observation of the desired peak of the waveform. Even when these noises are small, it is difficult to visually observe the characteristics of each peak due to the overlap of adjacent peaks, or to directly measure peak values such as the peak area, peak height, and standard deviation of the waveform. It was difficult to calculate.

Last night, in the 4-number display (d, signal from the flow cell), the signals that were widely separated in 2 were compressed logarithmically, and were often difficult to separate visually. A device that removes or smoothes minute peaks was sometimes used in conjunction with the above-mentioned direct display, shunting, or logarithmic display to remove or smooth noise, but it is important to distinguish between noise and peak I by component. She is a very troubled sister,
In some cases, it was removed or smoothed out during the beak adjustment, which was the intended purpose.

As described above, all conventional methods have had many problems in visually characterizing and displaying chromatographs.

<Summary of the Invention> An object of the present invention is to provide a chromatogram display device that makes it possible to correctly observe peaks for each component without being affected by noise.

According to this invention, function curves corresponding to individual chromatograms of a plurality of specific components are stored in the storage means. The long-term chromatodalam signal and the function curve signal generated by reading out the storage means are compared with each other in a graphical way.
The chromatogram generator generates a new curved chromatogram signal corresponding to a specific component in the input chromatogram signal based on the comparison result. S1 occurs. The new taromadogram signal is displayed on the display means. When a new chromatogram signal is generated by the chromatogram generation means, the retention time (richin/contime) and spread (dispersion) corresponding to a column different from the chromatography column used for the analysis, and the comparison means described above can be used to generate a new chromatogram signal. By using the peak height, it is possible to improve the degree of separation of components in the input chromatogram signal and obtain a display that is easy to observe.

Here, "corresponding to a specific component" means corresponding to a chromatogram curve obtained by analyzing only a specific component. In this invention, the known function curve has extremely important significance in that the unique components correspond to the known function curve as its support.

The function curve or line corresponding to a specific component can often be a curve that can be expressed by a mathematical formula.

For example, statistical distribution function curves such as Gaussian distribution curves, binomial distribution curves, chi-square distribution curves, F distribution curves, etc.
Alternatively, there are function curves such as trigonometric function curves, logarithmic function curves, and exponential function curves, and function curves created by combinations of these function curves. For example, the function curve corresponding to a constant component may be one that is included in the sample in advance.
The chromatogram peak curve is known as a result of analyzing only one component.

As means for detecting this chromatogram, for example, there are methods using ultraviolet absorption, refraction, fluorescence, coloration, electrical conductivity, etc. Ultraviolet absorption is particularly inexpensive and easy to implement. Furthermore, samples that contain various components such as biological fluids such as serum and urine can be processed using gel permeation chromatography, ion exchange chromatography, partition chromatography, adsorption chromatography, size exclusion chromatography, etc. When separating by chromatography such as t9 layer chromatography or gas chromatography, it is more useful if it easily forms unseparated peaks on the chromatogram. .

<Embodiment> FIG. 1 shows an embodiment of a chromatogram display device according to the present invention, in which an analog or digital chromatogram signal is inputted from an input terminal 11, and a chromatogram signal corresponding to a specific component supplied from a storage means 12 is input. The comparison means 13 compares and approximates the function curve signal. In this case, a method may be adopted in which the function curve signals are supplied one after another from the storage means 12 and the signal that most closely approximates the chromatograph is selected, or after supplying the basic function curve signal, this basic signal is
You may also use a method of approximating by changing the coefficients of Gin.

Next, the function curves approximated by the comparing means 13 are synthesized by the chromatogram generating means 14 and displayed on the display means 15, so that a new chromatogram is displayed.

For example, a 200 cm long (25 cm x 8) GPC (
Suppose that the chromatogram obtained by introducing serum into a gel permeation chromatography column is as shown in Figure 2. Peak 16 in this chromatodarum
, 1.7, I8 is Igλ (immunoglobulin A),
Contrasts with each component of IgG (immunocloprin G) and T f (+-transferrin) are shown. In this chromatogram, it is difficult to visually observe the characteristics of the peaks of these three components. Therefore, in this embodiment, processing is performed as follows. A normal distribution was assumed as the approximate function curve corresponding to each component. Normal distribution f (t) f(tl--exp (-'(
”-')2)2 σ h .

It is shown by the formula. In this X, 7 is the peak height, T, 1'
+ is retention time, σ is standard deviation, L
is the time. This normal distribution f (t) is stored in the storage means 12
It is stored as a function curve according to JJy, min and 6j. Although the retention time T1 spread (standard deviation) σ in the chromatogram of each component in this trial cut 1 is known in advance, its amount, that is, the peak height h is unknown. From the value of the chromatogram f (t) at the retention time of each component, each j
You can know the peak height of the component. Using the predetermined retention time and standard deviation corresponding to each component, the peak height corresponding to each component was determined when the chromatogram was approximated as the sum of normal distributions corresponding to these three components. By adding these normal distribution curves together, a composite curve, that is, a new romatogram, was created and displayed. This is shown, for example, in FIG.

Peaks 16', 17', and 18' in FIG. 3 correspond to components IgA, igG, and Tf, respectively. In this way, the valley peak becomes clear, and it is possible to display a chromatogram in a form that makes it easy to visually observe the peak characteristics of each component.

The relationship between the column length and the retention time T during which each component appears is as shown in FIG. 4, and the column length is linearly proportional to the retention time T. On the other hand, the relationship between the length L of the force column and the standard deviation σ of the peak is non-linear, as shown in the vJ5 diagram, as the length of the column increases. σ tends to become saturated.

In any case, once the length of the column is determined, the retention time T and standard deviation σ of each component are automatically determined by being calculated in advance.

Next, the operation for obtaining the new romatogram display shown in FIG. 3 will be explained with reference to the flowchart in FIG. 6.

The length Li of the chromatography column whose sample was analyzed in step S1 and the length LO of the column corresponding to the chromatography column displayed on the display means 15 are input. The retention time Tj (Li) and standard deviation σj (Li) (j-IgG, IgA, or Tf) of each component IgG, IgA, and Tf are determined. -1p The relationships shown in FIGS. 4 and 5 are stored in the notation means 12, and the column length L
Tj and σJ of each component are read out using l.

In step S3, the input chromatogram (E No. f (t
).

Next, in step S4, a normal distribution curve f(t) is read out from the storage means 12 as a function curve corresponding to the specific component,
In step S5, TIgo(L
i), σrg(+(Li), determined in step S3 (
hl, o/σIgG)Li is read in step S5.
A waveform fIgo(t) corresponding to the component IgG is created by substituting T, σ, and (h/σ) of (t), respectively. The comparison means 13 compares fIg(J(L) with step S6
is compared with the input chromatogram signal f(t) to obtain a subtracted waveform f''(1).This waveform f'(t) is 8
In the waveform f (t) in Figure 2, the waveform fIg of the component IgG
o(t) is removed, and this waveform fI, o
The influence of the tail portion of (t) on the peaks of components IgA and Tf is removed.

Therefore, in step S7, Tj (Li
), the component IgA of the input chromatogram f (t),
Tf peak height reduction, /σxgA)Li + (h
Tf/'Determine Tf offender 1. Next, in step S8 in the chromatogram generating means 14, Tj (Li
), σj (Li), and (hj/σJ)Li are substituted into the normal function curve f(t), and these waveforms are simultaneously created and synthesized. The synthesis and ♀ ノ I / ♀ no I / Step S9, output to the table / j <hand F 15, and display the brit. This reduced/reduced chromatogram has peak waveforms 16'', 17', and 18' corresponding to a chromatochromatogram containing only the components IgA, IgG, and Tf, and the characteristics of each of these component waveforms can be clearly observed.

Next, a case will be described in which a new chromatogram corresponding to a column different from the column used for the input chromatogram f(t) is created. Assume that the input chromatogram f (t) is as shown in FIG. This has a length Li of 50 on
It was obtained by introducing serum into a (25on x 2') GPC column.
], , 22 and 23 are component IgA, respectively.

Compatible with IgG and Tf. For this chromatogram, the length Lo of the column corresponding to the new chromatogram to be displayed is set to 200c7n. In step S1 in FIG. 6, unlike the previous case, Li=50 cm'
, L O = 200 cm, and in step S2, not only Tj (Ll) and σj (Li) but also T J (L
o) +σj (Lo) is also determined. After that, in steps s3 to S7, the same thing as in the previous case is performed. Step S: l, the component 1 obtained in S7. gG, IgA.

Each peak of Tf? From r G (h /σ)L,j and σj(Li) obtained in step S1, step S10
hIgG.

11□ Find h, , f. In step Sll, the chromatogram generating means generates a waveform for each component by simultaneously substituting these hj and Tj (Lo) and σj (Lo) obtained in step S1 into the normal distribution curve f (t). 14, and the result is sent to step S9.
is supplied to the display means 15 for display. As a result, as shown in Fig. 8, a new low-q togerum corresponding to the 200 crnO column is displayed, and its peak 21'
, 22', and 23' correspond to the components IgA, IgG, and Tf, respectively, when passed through a 200 cmO column. In this way, the q
From the obtained input chromatogram f (t), it is possible to create and display a chromatogram with a high degree of resolution similar to that obtained through a long column, and measurement can be performed with a correspondingly higher precision.

<Effect> As described above, this emission has the effect of displaying a chromatogram in a form that makes it easy to visually observe the peak characteristics of each component (ψ).

That is, there is an effect that peak values such as peak height, peak area, half-value, standard deviation, retention time, waveform, etc. can be easily read from the chromatogram. I praise you.
This has the effect of being able to display chromatograms excluding unintended peaks and peaks caused by miscellaneous axes. ′? It has the effect of being able to predict or phantomly display the chromatogram that will be analyzed using a column of any length. It has the advantage that even peaks that are close to each other and overlap can be displayed as a single component.

[Brief explanation of drawings]

FIG. 1 shows a chroma I crumb display device according to the present invention.
Figure 3 is a diagram showing an example of the chromatodaram obtained by analysis using a GPC column of the present invention.
Figure 4 shows the relationship between IgA and IgG as a function of column length.
, FIG. 5 is a diagram showing the relationship between the retention times of IgA, IgG, and Tf with respect to column length. FIG. 6 is a flowchart showing an example of the operation of the present invention. FIG. 7 Figure 8 shows an example of a curved chromatogram obtained by phase-separating serum with a 50-on GPC column. It is a figure showing an example. 11: input terminal, 12: storage means, ]3: ratio E1 means, 14: chromatogram generation means, 15: display means. Patent Applicant Asahi Kasei Corporation Representative Takui Kusano 1 Takui 2 Zu+0LILJll+1JIJjLILILJjl)LI
U(9)7173 Figure 1'71''4 Procedural amendment (voluntary) Commissioner of the Japan Patent Office ■, Indication of small matter Patent application 1987-967542, Famous invention ＼ Chromatogram display device 8, Person making the amendment Relationship with 3 cases per month Patent applicant Asahi Kasei Kogyo Co., Ltd. 4, Agent 4-2-21 Shinjuku, Shinjuku-ku, Tokyo
Soigoku Building 5, subject of amendment 8 IJ Specification, Detailed Description of the Invention Column 6, Contents of Amendment (1) After "Gas Chromatography" on page 5, lines 16-17 of the specification, V F l" fractionation).

Claims (1)

[Claims] (1) Function curve corresponding to each 1β chromatogram of multiple specific components! a storage means for storing a chromatogram signal, a comparison means for comparing an input chromatogram signal with an increment number code generated by reading out the storage means, and a comparison result of the comparison means also includes an input chromatogram signal. 6. A chromatogram generating means comprising: a chromatogram generating means for generating a force or a romatogram for a specific component in a vision, and a display means for displaying a romatogram for a particular component of the force.