JPS59218951A - Calculating device for peak value of chromatogram - Google Patents

Abstract

PURPOSE:To enable exact calculation of the peak value such as height, spread, retention time, area, trough and inclination of one crest of a chromatogram by providing a storage means, comparing means and peak value calculating means. CONSTITUTION:An analog or digital chromatogram signal is inputted from an input terminal 11 and is compared with the signal indicating the function curve corresponding to the specific component supplied from a storage means 12 by a comparing means 13. The constants necessary for generating the function curves corresponding to various specific components are stored in the means 12 and various function curve signals are generated from said constants or the function curve signals are directly stored therein. The peak value of the approximate function curve obtd. from the result of the comparison by the means 13 is calculated by a peak value calculating means 14. The peak value such as height, spread, retention time, area, trough and inclination of one crest of the chromatogram is exactly calculated.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an apparatus for calculating the first peak of a chromatogram obtained by analyzing a sample containing at least two components by chromatography.

<Background> Some conventional devices set arbitrary linear boundaries on a chromatogram to determine peak values.

However, in the device of , the rise point of a peak or the valley between two peaks was used to determine a straight boundary.

Therefore, it is often impossible to find the valley of a peak in the case of small peaks that depend on the reach of a large peak (f≧15) or in the case of closely spaced peaks. There were times when I couldn't make a decision. Further, there was a problem in that the valley of the peak moved due to an order of magnitude change in the content of a certain component in the sample, and the boundary of this peak also changed L, causing the peak value, where the content did not change, to change and be read out.

<Objective of the invention> The object of the invention is to accurately calculate peak values such as the height, spread, retention time (richin/contime), area, valley, slope (differential value), etc. of one peak in a chromatogram. An object of the present invention is to provide a chromatogram peak value calculation device that can calculate peak values of chromatograms.

SUMMARY OF THE INVENTION The present invention is an apparatus for calculating the peak value of a chromatogram obtained by analyzing a sample containing at least two components by chromatography, and which calculates the peak value of a chromatogram obtained by analyzing a sample containing at least two components by chromatography. is stored in the storage means,
The input chromatogram signal is compared graphically or numerically with the function curve signal read out and generated by the two-key distribution means, and the peak of the approximate function curve obtained from the comparison result is The value is determined by the peak value calculation means.

Here, "corresponding to a specific component" means corresponding to a chromatogram curve obtained by analyzing a specific component.

Conventionally, curves obtained as experimental results have been approximated using known function curves based on their shapes, but in this invention, the chromatograms of each component are known, and the chromatograms of each component are approximated. These corresponding to are approximated using known function curves. In other words, the known function curve used in this invention has important significance in that unique components correspond to it. The function curve corresponding to this time-varying component is generally a curve that can be expressed by a mathematical formula, but for example, it is a chromatogram peak curve that is known as a result of pre-analysis of one component contained in a sample and is generally known. It may be something that cannot be expressed using a simple mathematical formula. Function curves that can be expressed using the above formulas include, for example, the Callas (normal) 1" curve, the binomial distribution area, the chi-square distribution curve, the L distribution curve, the tr" distribution curve, and the statistical distribution function curve. 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.

Chromatogram detection can be carried out using ultraviolet absorption, photorefraction, fluorescence, color, electrical conductivity, etc., but it can be carried out at low cost by using Shiyu Ikkatsu absorption. Chromatograms (obtained by gel permeation chromatography, ion exchange chromatography, partition chromatography, adsorption chromatography, size exclusion chromatography, Wf layer chromatography, gas chromatography, etc.) to which this invention applies It is.

<Embodiment> FIG. 1 shows an example of a device according to the present invention. Input terminal 1
A digital chromatogram signal is inputted from the alarm log, and is compared with a signal representing a function curve corresponding to a specific component supplied from the marking means 12 by the comparing means 13. Ki 1. In the means 12, constants necessary for generating function curves corresponding to various specific components are stored, and various function curve signals are generated from the constants, or function curve signals are directly stored therein.

1. Determining a function curve that approximates the input chromatogram signal by comparison by the comparison means 13. In this case, the function curve signals are read out one after another from the storage means 12 and the signal that most closely approximates the chromatogram is selected. may be approximated by supplying a basic function curve signal and then changing the coefficients of this basic signal.

Here, the main components included in the target region 1'+ are known in advance, and the spread of the chromatogram and retention time for each component are known, and each component in the manual chromatogram signal is The case where the height of each is calculated will be specifically explained below. The storage means 12 stores the I-i fundamental wave, in this example a normal distribution curve, g(Ll-11r exp (-' (PJ2L)
2) 2 σ1 Remember, where t is time, hi is the height of one peak in the chromatogram, σl is the spread (standard deviation) of 111, and Rtj is the holding surface. The input chromatogram signal f (tl is a chromatogram obtained by introducing 7 hours of blood into a GPC (gel permeation chromatography) filled with polyvinyl alcohol gel, for example, as shown by the curve in Figure 2. The components in this chromatogram are 1gM (immunoglobulin M), Ig
A (immunoglobulin A), 1, gG (immunoglobulin G
), Tf (transferrin) and Atb (albumin) as shown in FIG.
It is approximated by 16117+18 and 19. In other words, each of these components is IgM.

Retention times in chromatograms of IgA, IgG, Tf and Atb "trgM+"t+gA+ "trgo+
Rtrf and Rtazb are known in advance, and the spread of each of these components 'rgM''Igktsu'rga (
7, , , r and 'AAb are also known 0, i.e. Rti and σl(r-I g M +
IgA + IgQ, Tf or A2b) is known, and by determining the V:JShi of each component based on a manual chromatogram, the value of each component can be known.

The determination of each peak value is performed, for example, as follows.9 In this example, Ig+V starting from the input chromatogram f(t)
Each component of l+ 1gCx + A l b is extracted and subtracted from the input chromatogram f[tl, and the peaks 1111 of each component of IgA and Tf are determined from the remaining components.

In this way, the influence of overlapping 14 tangent components is avoided. As shown in FIG.
Before and after each retention time of IgM, IgG, and Atb known in advance in 1, time 1 in the input chromatogram f (t); = 1920-1965 seconds, t = 2790-28
30'i'J' and the respective maximum values at t = 3300 to 3500 seconds, Igfvl, IgG, At
Each peak height h i (i = I g M,
IgG or Atb). Then I g+N, I
The retention time "tI" of each single J component sample phase of gG and Atb was determined by analyzing the chromatogram f(t) of the sample shown in Figure 2 in advance using the same apparatus (column).
gM+ RLzgar RtAlb and A standard) difference'r
gM+σ1. Step s for each part of g Q +σAtb
2 is done manually.

Next, in step S, 3, 1i12':', V;', means to normal function g (tl to a'rl:, and) component 1 grvi, I gG, Atb Step S1
By substituting hi obtained in step s2, Rti and σl input in step s2, gj(tl is obtained by equation 11-).In this way, each component 1gM, IgG+Atb component is calculated by calculating the approximate function gi(tl). The comparison means takes the difference from the input chromatogram f (tl. That is, in step s5 'i
f(t)-(g,, Jt) +g1. Jul+
gA nine phrases) are performed. This remaining waveform f”(t
The component IgA therein.

't''f is unaffected by the component 1gG.

At step S6, the time t=26 of this remaining waveform f"
20 to 2δ70 seconds and t=2940 to 2970 seconds, respectively, and the peak heights of the components I gA and i″f are determined.

In step S7, the retention time RL□gA+1 of the components IgA and Tf is determined in advance using the same method as that determined in step s2.
尤L1. Input the f1 standard deviation σIgAlσ, 1, and f, and in step S8, calculate these and the /
By substituting this peak height into g ft), the approximation function g
(tl, gl, f(L)IgA are generated respectively. In step 89, the peak height hi or waveform area for each JJy and minute obtained in steps Sl and S6 and the human charomatogram are generated. f(υ
The difference between the sum of the approximate functions gi(tl) of each component, that is, the waveform (digital 11σ) shown by the broken line in FIG. 14.Other family approximation functions gi (tl's half width, variance, average, etc., peaks 11α are calculated as necessary.

Since the retention time Rti of each component is known in advance, to obtain the peak height hi in steps Sl and S6, the retention time 1 likely L1 of each component is substituted for the time of the input chromatogram f (t). You can ask for it. If the function curve source stored in the storage means cannot be represented by a mathematical formula, 1-1, create a plurality of function curves whose center and spread are constant and whose peak heights are sequentially trough;
About the ingredients 1. I'll keep it. 1 tally is I & minute Ig
If such a curve is used as a one-period curve of M, A'y1 is, for example, as shown in A'y 51%I, when the component IgM is held at step Szo 141: J-, -Ij-
Value of (t) 1] 1gM
Load. Increase J by 1 in step Sll, and step S1
2nd 1. Stream the peak height of 1g1Vl's revised number of songs that I remember or January's rJ3, and compare that hj and htgh+ (Step 513), if they do not match, step S
Returning to II, if there is a match, there are more A in step S14, and II! Let hj of i be the beep height, and output the function song as an approximate function song. Thereafter, other components are obtained in the same manner, and the deviation of adjacent waveforms can also be removed by the same processing as step S5 in FIG. 4.

<Effects> As described above, according to the device i'lf of the present invention, since approximation is made by a function curve for a specific component for each rfi: minute, the shadow envelope of the lμr tangential component is Therefore, when determining the peak 11'1 from a sample chromatogram containing two or more components, it is not necessary to completely determine the boundaries of the peak, and the error associated with this determination results. This has the effect of eliminating errors in peak values. The weather in the valley of the peak is such that it won't freeze.j1. This method also has the effect of being able to separate peaks that are mixed together. This method has the advantage that the peak value can be determined without being affected by the movement of the valley of the peak due to the change in a content of a certain component.

Therefore, the present invention is more effective for chromatography using, for example, a GI) C column packed with a polyvinyl alcohol gel, which tends to form unseparated peaks on a chromatogram.

[Brief explanation of the drawing]

FIG. 1 is a block diagram showing an example of a chromatogram peak value calculation device according to the present invention, and FIG. 2 is a block diagram showing an example of a chromatogram peak value calculation device according to the present invention.
Figures 2B and 3 are diagrams showing the chromatogram obtained by analysis with the C column, and curves approximated to each component of the analysis waveform obtained by the apparatus of the present invention, and the deviations of these and chromatography. Figure 4 5 is a flowchart showing one operation of the apparatus of the present invention, and FIG. 5 is a θ1 flowchart showing 15 variations of the operation example. Patent applicant Takuo Akino, agent for Asahi Kasei Industries Co., Ltd. 1 Figure 3 Figure ↑ 5 Damage to amendment of figure procedure (voluntary) September 2, 1982 Commissioner of the Japan Patent Office 1, Case indication 4 Toki I4 1982 946202 Name of the invention Chromatogram peak value calculation device 3, relationship with tenant who makes the correction Patent issued by 11 people Asahi Kasei Kogyo Co., Ltd. 4, Agent 4-2-2' Shinjuku, Urashuku-ku, Tokyo 1
Contents of Sagami Bill 6 Amendment (1) Add "1'' F I Pa (Field Flow Fractional Yon/Yon) to you in "Cass Chromatography" on page 4, lines 15-16 of the specification.

Claims (1)

[Claims] (1) A storage means for storing function curves corresponding to individual chromatograms of a plurality of specific components, and comparing a manually inputted chromatogram signal with a function curve signal generated by the storage means. A chromatogram peak value calculation device comprising a ratio soft means and a peak value calculation means for calculating a peak value of an approximate function curve based on the comparison result.