JPS62249067A - Multi-component simultaneous quantitative flow injection analyzing method - Google Patents

Abstract

PURPOSE:To simultaneously execute a quantitative analysis of a multi-component in a sample, by delaying in order each reaction liquid which has passed through a spare reaction passage, respectively, feeding it to a main reaction passage without mixing them, respectively, and calculating an obtained peak height. CONSTITUTION:Carriers 1-1-1-n containing each reaction reagent are fed by a liquid feeding pump 2, respectively, and each constant temperature bath 7 is set to a selected temperature and set to a stationary state. Subsequently, from a sample injector 3, samples containing the first - the n-th components to be determined are led simultaneously into each spare reaction passage through each sample leading-in part 4. These samples are fed in order to a main reaction passage 101 without being mixed to each other by a delay coil 8, and a peak height is detected by a detector 102. Subsequently, from an obtained peak height, an arithmetic operation is executed by making each standard liquid containing a component to be determined of a known concn. pass through the spare reaction passage and the main reaction passage, and using an inclination of a measuring straight line which is detected, by which an unknown concn. is calculated.

Description

Detailed Description of the Invention (a) Industrial Application Field This invention relates to a flow injection analysis method, and particularly to a flow injection analysis method (hereinafter abbreviated as FIA method) that can simultaneously analyze multiple components contained in a sample. Ru.

(b) Conventional technology In the conventional FIA method, as shown in Fig. 5, a sample is injected under pressure from a sample injector (8a), and the target component in the sample is sent by a liquid pump (2a). A carrier solution containing a reaction reagent (la) and a reaction coil (4a)
), the reaction product is reacted with a reagent, and the reaction product is detected by a detector (5a) and quantified. In addition, efforts have been made to pre-process the sample by providing two or more channels, or to reduce the amount of reagent by marling. However, in any case, only one component is quantitatively defined. Attempts have also been made to quantify multiple components simultaneously; for example, an example of analysis of Mg/Sr system or Ca/Sr system using two or more flow paths that cause specific reactions [H. Kagenou and H.
Jensen, Analytica Shimi-Riki Acta, (H@K
agenow and AsJensen, Ana
l, Chim, Acta) 114, 229° 1980)
Examples of Hg/Zn-based analysis using two or more detectors that can specifically detect
Deci, Analytical Chemistry (D, Hool)
ey and R, Dessy.

Anal, Chem, )t 55.81J 1988
), but all of these utilize specific effects with high selectivity for each sample, and it is essential that the independence of reaction or detection for each sample is maintained. However, in reality, for example, a reaction that should be specific and independent to component A may also have some effect on component B, or conversely, a reaction that should be specific and independent to component B may also act slightly on component B. It often happens that FIA has a slight effect on FIA components, and as a result, the possibility of simultaneous determination of components using the FIA method is severely restricted.
This method was developed to improve the above-mentioned problems in the IA method, and is an F method that allows multiple components in a sample to be analyzed simultaneously.
The purpose is to provide IA law.

B) Means and operation for solving the problems This invention is constructed by sequentially connecting a liquid feeding section for a carrier containing a reaction reagent, an introduction section for samples containing the first to nth quantified components, and a preliminary reaction section of a reaction coil. 1st! The fP pre-reaction flow path and the second to nth pre-reaction flow paths, each formed by having the same configuration and further connecting a liquid sending delay coil to the outlet side thereof, are arranged in parallel and merged at the outlet, and the merged portion is Using an analyzer in which a main reaction channel is constructed by sequentially connecting a reaction coil and a detector, samples are simultaneously pressurized into the sample introduction portions of the first to nth preliminary reaction channels, and these Each reaction solution that has passed through the channel is
The samples are introduced into the main reaction channel without mixing with each other in the order of ~n, and are introduced into the main reaction channel without mixing with each other, while the samples pressurized into each preliminary reaction channel pass through each preliminary reaction section and the following main reaction section. The reaction liquids that are subjected to different reactions and generated are sent to the detector in order, and the peak height P (1) of the reaction liquids that have passed through each of the first to n preliminary reaction channels and the main reaction section is ~P
(n) is detected, and the following simultaneous equation: % formula % However, 01 to Cn indicate the unknown concentration of the 1st to nth neglected component in the sample, and X, for example, X12 is the first chaotic component of known concentration. The slope of a test straight line indicating the relationship between the peak height detected by passing the 100% standard solution through the second preleprosy reaction channel and the main repulsion channel and the concentration of the first analyte component, which is measured in advance. .

Solve and calculate the concentrations C1 to C of the first to nth test components in the sample.
This invention provides a multi-component simultaneous flow-in fraction analysis method consisting of calculating Examples include a mixed liquid of wind ions.

According to this invention, the sample is simultaneously pressurized into each pre-reaction channel from each sample introduction section, but the sample introduction section includes a loop for holding a certain amount of sample for each pre-reaction channel. An example of this is a known device that consists of 60,000 valves and a shaft common to these valves, and that rotates the shaft to simultaneously introduce the samples in the loop into the respective preliminary reaction channels.

In addition, each reaction solution that has passed through the first to nth preliminary reaction channels is delayed in the order of the first to nth preliminary reaction channels and sent to the main reaction channel without being mixed. Each of the reactors is provided with a delay coil, but this delayed liquid feeding can also be performed by adjusting the flow rate of each mixed reaction liquid.

In addition, in this invention, the sample passing through each preliminary reaction channel and the main reaction section following it is subjected to different reactions while passing through each preliminary reaction section and the main reaction section (with the same polarity). This reaction is mainly carried out by a chemical reaction using a reaction reagent contained in a carrier, but heating means may be provided in the preliminary reaction section and the main reaction section. In addition to a chemical reaction, a photochemical reaction by irradiation with light, radiation, or electromagnetic waves, or an electrochemical oxidation or reduction reaction may be used.

In addition, the above heating means is constant temperature! Well-known ones such as No. 3 can be used.

An appropriate detector is selected depending on the reaction product of the sample, and examples include absorption, fluorescence, and custom-made electrochemical detectors.

Moreover, the simultaneous equations can be solved automatically and quickly using a suitable calculator. .

(e) Example The analytical method of this defense will be explained using -1 (first doctor) of the analyzer used in this analytical method.

In Figure 1 (l-1, 1-2, 1-8...
, 1-n) is a carrier containing a reaction reagent, (2-1,2-
2, 2-8..., 2-n) is a pump for feeding a carrier containing a reaction reagent, (3) is a sample presser, (4-
1, 4-2, 4-8..., 4-n) is the delayed sample interrogator, (5-1, 5-2, 5-8=, 5-n
311151~n'P equipped reaction section, (6-1, 6-2,
6-8=・-6-n, 9) is the reaction coil, (7-i
, 7-2.7-8=T-n.

LOon is fit, (8-2, 8-8, 8-4
・! ...8-n) is a delay coil, (9-1, 9-2
, 9-8・beat・, 9-n) are the first to nth preliminary reaction channels, (
100) is the main reaction section, (101) is the main reaction flow path, (10
2) is a detector.

Next, an example of an analysis method of the present invention performed using this apparatus will be explained.

The reaction reagent for each preliminary reaction channel and the temperature of each constant temperature bath are selected so that different reactions are imparted to each sample as it passes through each preliminary reaction channel and the main reaction channel. Carrier containing reaction reagent (1-111-2, 1-8...
. . . , 1-n) are connected to the liquid feeding pumps (2-1, 2-n), respectively.
-2, 2-3, song, 2-n), and each constant temperature bath is set at the selected temperature to keep it in a steady state.

Next, samples containing the first to nth analyte components are simultaneously introduced from the sample injector (3) into each pre-reaction channel through each sample introduction section. First, the sample press-fitted into the first preliminary reaction channel (9-1) passes through this and first enters the main reaction channel (10-1).
1) sent to -10,000th 2.・6. ...The samples pressurized into the pre-reaction channels pass through each pre-reaction channel and are fed to each delay coil to perform the main reaction in the order of No. 2... No. 1 without mixing with each other. It is sent to the flow path (101). Each sample is subjected to a different reaction while passing through each preliminary reaction channel and the main reaction channel. Then, the peak heights P(L), P(2), P(3), . . . are sent to the detector (102) and detected in order.
P (n) is obtained and discharged.

The peak heights P(1), p(2), P(3) obtained here are
)......, P(n), the daytime is, for example, at the peak of P 0.1, in addition to the contribution of C1 in the line between the first preliminary reaction channel and the main reaction channel, C2 , C8...Cn, and at the peak of P(2), the contribution of CI in the line between the second preliminary reaction channel and the main reaction channel is the sum of the contributions of NC2. Order, C
a...Contribution of Cn is totaled, and the same applies hereafter. As a result, the following simultaneous equations are established.

P (11=Xt t C10X 12C2+---
-=+X1 nCnP (2) = X21 C1+
X22 C2+-=-・→-X2nCnP (n) =
Xnl cm + Xn2 cg +・−−−−−+
xnncnIn addition, in each line of the first to n-th preliminary reaction channels and the main reaction channel, different reactions or reactions of different degrees are given to the same reaction, and the coefficients Xtt to Xnn in the above equation are each unique. For example, X1
2 is a calibration line showing the relationship between the peak height detected by passing a standard solution containing the first quantified component with a known concentration through the second preliminary reaction channel and the main reaction channel and the first quantified component Yumaro; is the slope of , and is measured in advance.

By substituting the actually measured P(1) to P(n) into this simultaneous equation, the unknown concentration C1xCn is calculated.

EXPERIMENTAL EXAMPLE The results of analyzing both components of glucose and maltose-containing water (m#L) using an example of the analyzer used in the present invention shown in FIG. 2 are shown below.

In Figure 2, A1 is boric acid 0.1M, sodium borate Na2B40
Aqueous solution B1 of 70.05M and 0.1% arginine is a 0.01M aqueous solution of perchloric acid, A2 and 82 are liquid pumps for A1 and B1, respectively, and A8 and 88 are sample introductions to preliminary reaction channels A and B. A4 and 84 C-J are the pre-reaction sections of the pre-reaction channels A and B respectively, C is the main reaction section, A5, B5 and C1 are the reaction coils, A6, B6 and C2 are the constant temperature chambers, D is the detector.

Note that the preliminary reaction section B4 and main reaction section C of the preliminary reaction channel B are maintained at 140°C by their respective constant temperature baths, but the temperature of the preliminary reaction section A4 of the preliminary reaction channel B is not raised and remains in the palm-knit state. .

Note that this experimental example uses boric acid ig containing arginine as the reducing sugar.
Fluorophores are produced in IhJa and are stable (Hirohisa Mikami and Yasuo Ishida 9 Analytical Chemistry, 32°E207, 19
(see 1988) and utilizes the fact that maltose is hydrolyzed by acid to produce glucose. That is, a sample of a mixed solution of maltose and glucose is press-injected into the above-mentioned apparatus and fluoresced, and the maltose in the sample press-filled into the pre-reaction channel B is oxidized with perchloric acid in the pre-reaction section B4. Glucose is produced, and then in the main reaction section C, both components are made to fluoresce by arginine from the flow path, and each is detected by a detector.

(1) Creating a test line a) Creating a test line between glucose error and peak height Al liquid at a flow rate of 0.6111/cm, BL liquid at a flow rate of 0.6111/cm.
Send at 4ml/sa and use as steady state. In addition, 0.2% glucose aqueous solution was injected in the direction of the arrow from the six-way valves A8 and B8 set as shown in the solid line, and the loops AL and BL attached to each valve were inserted into the same place 7) [(10
μl) of the aqueous solution is retained. Next, by switching the A8 and B3 valves at the same time as shown in the dotted line, the AL
and the standard samples in BL are introduced into the AM path and the B flow path, respectively. First, the reaction liquid that has passed through the preliminary reaction section A4 and the main reaction section C of the A channel is detected by a fluorescence detector, and the peak height of al shown in Fig. 8 is obtained, and then the preliminary reaction of the B channel is detected. 86(1-
The fluorescence when excited by the light source was detected by the fluorescence detector, and the peak height of bl (ff) in Figure 8 was obtained (
All measurements were taken twice. same as below〕.

Next, in the same manner, Q, 4, Q, cs: $i and 0.8% glucose aqueous dispersion were press-fitted, respectively, as shown in FIG. 8a21.
b2ya8, B8. and B4 and B4 peak values were obtained.

Next, the height on the chart of peaks a1, B2, B8 and B4 is ff1l) and the glucose quality is 0.2.0.4.0.
．． The fourth test line G-A showing the relationship with 6 and 0.8%
Shown in the figure. Similarly, peaks bl, B2. b3$ and B4
High g (ff) and glucose concentration on the chart of 0.2
．． The standard line GB showing the relationship with 0.4, 0.6 and 0.8% is shown as the fourth factor.

Next, beaks a5 and b5 shown in FIG. 8 are prepared in the same manner as above except that glucose is changed to maltose. a6, b6
．． A7, b7, a8, and b8 were obtained, and calibration curves M-A and M-B corresponding to G-A and G-B were obtained from these as shown in FIG. The slope of each calibration curve was as follows.

G-A229ff/% sugar G B 184.5 u M-A40.5u M-B 63,5u (2) Analysis of aqueous solution containing glucose and maltose Water bath liquid containing 0.85% glucose and 0.66% maltose Add 10 μl of each synthetic sample to A in the same manner as above.
The beaks a and b shown in FIG. 3 were obtained by simultaneously press-fitting into the line and B line (returning was repeated 4 times).

Create the following simultaneous equations from the above data and calculate A peak height (at) = G - A (7) slope (229B/%) x
CG (glucose level 9%) + M-A slope (40.5 Ja/%) XCG (maltose concentration 9%) B-beak height Jlml) = G-H slope (184,
4, wI/%)XCG (same as above) +MB slope C68, 5JII,'%)

Content'M (gradient content (to) Relative error %) Glucose 0.85 0.826 0.7 Maltose
0.65 0.646 0.6 The calculation results and the content show good agreement.

(e) Effect of explanation According to the present invention, it is possible to simultaneously analyze the hypothetical constituents of a sample containing a large number of neglected constituents.

[Brief explanation of drawings]

Fig. 1 is an explanatory diagram of the configuration of an example of the apparatus used in this invention, Fig. 2 is an explanatory diagram of the configuration of the apparatus used in the experimental example, the eighth factor is a beak chart diagram obtained in the experimental example, and Fig. 4 is the diagram of the configuration of the apparatus used in the experimental example. FIG. 5 is a graph showing a line to be detected based on the data obtained from the eight factors, and is an explanatory diagram of the configuration of an example of a conventional FIA method apparatus. (1-1,1-2,1-8=z 1-n )=reaction #A
A certain containing carrier, (2-1, 2-2, 2-8..., 2-n)...
Liquid feed pump, (3)...sample injector, (4-1, 4-2, 4-8...~4-n)...sample introduction section, (5-1, 5-2 ,5-3...--5-n
)...preliminary reaction section, (6-1, 6-2, 6-3...
・～6-9)・・Reaction coil, (7-1, 7-2, 7
-8-----17-n, 10) = constant temperature bath, (8-2,
8-8,...,8-n)...delay coil, (9
-1,9-2,9-8...9-n)...Preliminary reaction channel, (100)...Main reaction section, (101)...
...Main reaction path, (102)...detector.婉1 字 2 fig. 3 11 11 1 1 藺 (minutes)

Claims (1)

[Scope of Claims] 1. A first preliminary reaction flow path formed by sequentially connecting a liquid feeding section for a carrier containing a reaction reagent, an introduction section for samples containing the first to nth analyte to be quantified, and a preliminary reaction section of a reaction coil; , with the same configuration, the second to nth preliminary reaction channels, each formed by connecting a liquid sending delay coil to the outlet side, are arranged in parallel and merged at the outlet, and the reaction coil and the detection channel are connected in parallel at the outlet. Using an analyzer in which main reaction channels are connected in sequence, samples are simultaneously pressurized into the sample introduction portions of the first to nth preliminary reaction channels, and each sample passing through these channels is The reaction solutions are introduced into the main reaction channel in the order of numbers 1 to n without being mixed with each other, and the sample press-fitted into each preliminary reaction channel is transferred to each preliminary reaction section and the following main reaction section. The reaction liquids that are subjected to different reactions while passing through are sent to the detector in order, and the peak height of the reaction liquid that has passed through each of the first to nth preliminary reaction channels and the main reaction section is calculated. SaP(1)~P(
n) is detected and the following simultaneous equations are created: P(1)=X_1_1C_1+X_1_2C_2+...
...+X_1_nC_nP(2)=X_2_1C_
1+X_2_2C_2+...+X_2_nC_
nP(n)=X_n_1C_1+X_n_2C_2+・
... + This is the slope of a calibration straight line that indicates the relationship between the peak height detected by passing through the main reaction channel and the concentration of the first quantified component, and is measured in advance. Solve to find the concentration C_1~ of the first to nth quantified components in the sample.
A multicomponent simultaneous quantitative flow injection analysis method consisting of calculating C_n.