JPS6332132B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for measuring the degree of chyle, degree of jaundice, and degree of hemolysis in biochemical samples, particularly serum.

It is widely known that chromogens such as chyle, jaundice, and hemolysis are substances that interfere with the measurement of general test items by automatic analyzers in the field of biochemical analysis. In particular, serum color (jaundice, hemolysis, etc.) and turbidity (chyle, etc.) can cause errors in accuracy in measurements, especially in endpoint measurements. Therefore, consideration has been given to the influence of chromogen on the measured values of general test items.
The test results are displayed, for example, on the test slip, based on the inspector's visual observation, indicating the amount that is thought to be contained in four levels of symbols, eg, +, -. However, with this method, there are large individual differences among inspectors, and not only is it not possible to quantitatively determine the amount of interfering chromogen, but in recent years, inspections have been automated,
As the number of processed specimens increases, this type of external observation becomes difficult. In Japanese Patent Application Laid-open No. 116283/1983,
In consideration of the above points, an analytical method for quantitatively determining chromogen is disclosed. This invention is based on “Chyle,
A sample that is affected by chromogen due to at least one of hemolysis and jaundice is irradiated with light, and chromogen is analyzed based on the optical characteristics of the sample, and at least two of chylosis, hemolysis, and jaundice are affected. Items are measured, preferably three items. Of these, chylosis is an appropriate value in the long wavelength range of the visible light wavelength range, which is affected by chyle but not by hemolysis and jaundice.
It is determined by measuring the difference in absorbance at two wavelengths. The degree of hemolysis is determined based on the value of the absorbance difference at two appropriate wavelengths in the medium wavelength range, which is not affected by jaundice, in the visible light wavelength range, and the measured value in the long wavelength range. The degree of jaundice is determined based on the absorbance difference value at two appropriate wavelengths in the short wavelength range affected by hemolysis in the visible light wavelength range and the measured value in the medium wavelength range. ” is its content. However, this analysis method requires at least six different wavelengths to be used, and furthermore, the degree of chyle, degree of jaundice, and degree of hemolysis must be determined in order based on each measurement result, which is cumbersome. There is. In addition, the invention disclosed in JP-A No. 54-84781 uses "multi-wavelength photometry technology to measure the absorbance of a sample liquid at least four or more wavelengths, and The concentration, chyle, hemolysis, and jaundice of the test substance in the sample are determined by solving simultaneous equations using the reference spectra of the test substance, chyle, hemolysis, and jaundice, and then the wavelength near the absorption center of the test substance is determined. The content is to find the ratio of the absorbance at each wavelength to the absorbance at each other wavelength. However, in the analysis method according to the present invention, standard absorption spectra for chyle, jaundice, and hemolysis must be memorized in advance, which requires storage capacity. There is a difference between the standard absorption spectrum for hemolysis and that of actual serum, and there is a drawback that accurate results cannot always be expected. In addition, for jaundice and hemolysis, there are standard methods such as bilirubin and hemoglobin, respectively, but for chyle, which is the most basic in the above analysis method,
There is no standard one, and even if glass powder were to be used as a substitute, the particle size would be different and would be inappropriate.

The purpose of the present invention is to eliminate the various drawbacks mentioned above, to easily detect abnormal absorption samples due to chyle, jaundice, and hemolysis in serum, and to store the degree of absorption abnormality and absorption spectra of chyle, jaundice, and hemolysis samples. The purpose of the present invention is to provide a method for measuring the degree of chylosity, degree of jaundice, and degree of hemolysis in serum, which can be easily determined without having to do so.

The present invention primarily provides a method for measuring the content of chyle, jaundice, and hemolysis in a biochemical sample, particularly serum, based on spectral characteristics indicating the wavelength and absorbance of light irradiated to serum. is the chyle,
Absorbance is measured using a short wavelength in the visible range that shows absorption in all cases of jaundice and hemolysis, and for serum with absorbance above a certain value,
It is determined that the abnormal serum is due to at least one of chyle, jaundice, and hemolysis, and then, secondarily, the degree of chyle, jaundice, and hemolysis of the abnormal serum is obtained as a result of measurement using at least four wavelengths. By comparing and contrasting the numerical value obtained by calculating the absorbance with several preset standards,
This method is characterized by the fact that chyle, jaundice, and hemolysis can be determined without using standard samples for each.

The present invention will be described in detail below with reference to the drawings.

Figure 1 shows the experimental results of examining the spectral absorption spectra of chyle, jaundice, and hemolysis samples.The horizontal axis shows wavelength, especially wavelength in the visible range, and the vertical axis shows absorbance. It shows the relationship between wavelength and absorbance for a sample. The following became clear from this figure.

(1) The chyle sample curve 1 shown by the solid line descends while drawing a gentle curve over the entire visible wavelength region from the short wavelength region to the long wavelength region. Now, each point 1a, 1b on the chyle sample curve 1 at wavelengths of 410nm, 480nm and 570nm
Focusing on the absorbance at 1c and 1c, the wavelength
Absorbance difference between 410nm and wavelength 480nm Δ1E410
-480 and the absorbance difference Δ1E480-570 between wavelengths of 480 nm and 570 nm are almost equal.

(2) Jaundice sample curve 2 shown by the dotted line has a short wavelength.
It exhibits the highest absorbance approximately midway between 410 nm and 480 nm, and then decreases to a wavelength of around 570 nm. Similarly to (1) above, focusing on the absorbance at each point 2a, 2b, and 2c on the jaundice sample curve 2 at wavelengths of 410nm, 480nm, and 570nm, the absorbance at wavelengths of 410nm and 480nm is almost equal; The difference in absorption with the wavelength of 570 nm is extremely large.

(3) Hemolysis sample curve 3 shown by the chain line has a short wavelength.
After showing maximum absorbance around 410 nm, it drops rapidly, and after that it traces a slightly undulating trajectory.
Overall, the wavelength gradually decreases to around 620 nm. As in cases (1) and (2) above, the wavelength
Focusing on the absorbance at each point 3a, 3b, and 3c on the hemolysis sample curve 3 at 410nm, 480nm, and 570nm, we can see that the wavelength 480nm and the wavelength
The absorbance at 570 nm is almost equal, and the difference in absorbance between wavelengths of 410 nm and 480 nm is extremely large.

(4) In a long wavelength range, for example, a wavelength of 660 nm or more, the absorbance according to the jaundice sample curve 2 and the hemolysis sample curve 3 is almost zero.

(5) In a short wavelength region, for example, around a wavelength of 410 nm, all of the curves of chyle sample curve 1, jaundice sample curve 2, and hemolysis sample curve 3 show high absorbance.

As mentioned above, various facts have been revealed from the experimental results shown in FIG. The method for measuring the degree of chyle, jaundice, and hemolysis in serum according to the present invention makes it possible to easily determine the degree of chyle, jaundice, and hemolysis by effectively and appropriately utilizing the above-mentioned relationships. This is what I did. In the following, an example of its use will be given as an example of the method of the present invention.

(1) The absorbance of serum or diluted serum is measured at a short wavelength in the visible range, for example 410 nm, and if it exceeds a certain value, it is determined that the sample is abnormal, such as chyle, jaundice, or hemolysis.

(2) Next, measure the degree of each abnormal sample as follows.

Measurement of chyle - Absorbance at a long wavelength in the visible range, for example 660 nm, which is almost unaffected by absorption due to jaundice and hemolysis, is measured, and the degree of chyle is determined by the magnitude of the absorbance.

Measurement of jaundice degree - Use a wavelength near the wavelength that shows specific absorption for jaundice, for example, 480 nm, and a wavelength that shows almost the same absorbance as 480 nm, for example, and a wavelength that has low absorbance in the jaundice sample, for example 570 nm. Ratio of absorbance difference Δ2E410−480 to absorbance difference Δ2E480−570 between wavelengths 480 nm and 570 nm K2 = Δ2E410−
Find 480/Δ2E480−570. Then, if K2 is a small value, for example, 1 or less, it is assumed that the patient has jaundice, and the degree of jaundice is determined from the magnitude of the difference in absorbance between wavelengths of 480 nm and 570 nm.

Measurement of the degree of hemolysis - one near the wavelength that exhibits specific absorption for hemolysis, e.g. 410 nm, and two wavelengths at which the hemolyzed sample has low and almost equal absorption, e.g.
480nm and 570nm are used, and wavelength 410nm and
Absorbance difference Δ3E410-480 and wavelength with 480nm
Find the ratio K3=Δ3E410−480/Δ3E480−570 of the absorbance difference Δ3E480−570 between 480 nm and 570 nm. If K3 is a large value, for example 2 or more, it is considered to be hemolysis, and the wavelength is 410 nm.
Determine the degree of hemolysis from the difference in absorbance from 480 nm. In this way, in the present invention,
By simply using light of four wavelengths, 410 nm, 480 nm, 570 nm, and 660 nm, the presence or absence of chyle, jaundice, and hemolysis can be qualitatively detected, and the degree of chyle, jaundice, and hemolysis can be determined quantitatively. That is, in the present invention, the short wavelength (410 nm) has high absorption for all of chyle, jaundice, and hemolysis, the long wavelength (660 nm) has almost no absorption for jaundice and hemolysis, and the long wavelength (660 nm) has almost no absorption for jaundice. Using four wavelengths, the first medium wavelength (570 nm) and the second medium wavelength (480 nm), which has a large absorption due to jaundice and a small absorption approximately equal to the first medium wavelength against hemolysis, are used to detect breast cancer. frequency,
The degree of hemolysis and jaundice can be measured accurately. However, if the degree of hemolysis is significant,
Absorbance at wavelength 410nm is the normal measurement range
Since OD.2 may be exceeded, the degree of hemolysis should be determined using wavelengths other than 410 nm, such as 570 nm, where the specific absorption of the hemolyzed sample appears.
Absorbance at even less absorbing wavelengths, such as 600 nm, may also be used.

Note that the present invention is not limited to the above-mentioned examples, and can be used in various ways within the scope of the present invention. The wavelengths listed above are also merely examples.

As explained above, according to the measurement method of the present invention, by fully understanding the relationship between the wavelength and absorbance shown by each sample curve for chyle, jaundice, and hemolysis, serum Since it is possible to detect chyle, jaundice, and hemolysis, and to measure the degree of these, there is no need to use standard substances for chyle, jaundice, and hemolysis, which are particularly difficult to obtain, as in conventional methods. This method has the advantage that samples with abnormal appearance in serum can be detected without relying on the absorption spectrum of chyle.

[Brief explanation of the drawing]

FIG. 1 is a diagram showing the relationship between wavelength and absorbance for each sample of chyle, jaundice, and hemolysis in serum. 1... Chyle sample curve, 2... Jaundice sample curve, 3... Hemolysis sample curve.

Claims (1)

[Scope of Claims] 1. A method for measuring the content of chyle, jaundice, and hemolysis in a biochemical sample, especially serum, based on spectral characteristics indicating the wavelength and absorbance of light irradiated to the serum, comprising: Next, the absorbance is measured using a short wavelength in the visible range that shows absorption in all of the chyle, jaundice, and hemolysis, and if the absorbance is above a certain value, the chyle in the serum is , the serum is determined to be due to at least one of jaundice and hemolysis, and then secondarily, the serum is determined to be chyle due to the magnitude of absorbance at the first wavelength λ ₁ , where there is almost no absorption due to both jaundice and hemolysis. A second wavelength λ ₂ with high absorption due to jaundice and hemolysis, a third wavelength λ ₃ with high absorption due to jaundice but low absorption due to hemolysis, and a fourth wavelength with low absorption due to both jaundice and hemolysis. The absorbance at λ ₄ is determined _, and the difference ΔE ₃ - ₂ between the absorbance at the third wavelength λ _{3 and the absorbance at the second wavelength λ 2} and the absorbance at the third wavelength λ ₃ and the absorbance at the fourth wavelength λ ₄ are determined. and the difference ΔE ₃ - ₄ is determined, and the degree of jaundice is measured from the value of the difference ΔE ₃ - ₄ in the absorbance.
A method for measuring chyle, jaundice, and hemolysis in serum, characterized in that the degree of hemolysis is measured from the value of the difference in absorbance ΔE ₃ - ₂ , thereby measuring the degree of hemolysis using absorbance at at least four wavelengths.