JPS61213743A - Automatic specimen introducing apparatus - Google Patents

Abstract

PURPOSE:To introduce a liquid specimen and a reaction liquid while accurately weighing the same to sufficiently mix both of them, by opening and closing first and second valve members synchronously at every minute time interval and alternately feeding the liquid specimen and the reaction liquid to hold both of them in a hollow holding coil. CONSTITUTION:A six-way valve 8 and a three-way valve 9 are brought to a solid line state and solenoid valves 3, 6 are alternately opened and closed at every minute time interval in such a state that a weighing cylinder 10 is driven to suck a liquid specimen 2 and a reaction liquid 5 into a holding coil 7 to hold the same therein. The liquid specimen 2 and the reaction liquid 5 are alternately arranged in the holding coil 7 at every predetermined length and held in a stationary state until sufficient chemical reaction is finished and, thereafter, the six-way valve 8 and the three-way valve 9 are changed over to introduce the reaction product into an analytical column 15. When the introducing operation of the specimen is finished, containers 1, 5 are substituted with containers of a washing solution to wash the holding coil 7.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention involves introducing and arranging a sample solution and a reaction solution alternately in minute amounts into a holding coil, and introducing the resulting reaction products into an analytical column. This invention relates to an automatic sample introduction device.

Conventional technology In high-performance liquid chromatography, some kind of chemical reaction occurs between a specific analyte in a sample solution and a reaction solution, and the resulting reaction product is introduced into an analytical column. An analytical method is used to separate and detect the reaction products (Brecolumn derivatization method). Examples of this pre-column reaction include a reaction using 9-anthryldiazomethane (ADAM) as a reagent when a fatty acid is the analyte, and a reaction using 0-phthalate when the analyte is an aminoglycoside antibiotic. Reaction using aldehyde (OPA),
When an amino acid is used as an analyte, a reaction using dinitrofluorobenzene may be used.

When carrying out the above analysis method, a microsyringe is inserted into a sample liquid container into which a sample liquid has been injected, a predetermined amount of sample liquid is aspirated, and this is discharged into an empty reaction vessel, and then the reaction liquid is injected. A predetermined amount of reaction liquid is aspirated from the reaction liquid container by a similar operation, this is discharged into the reaction container, and after a predetermined period of time, the reaction product produced in the reaction container is injected into an analytical column. It is common practice to perform a series of operations manually.

On the other hand, a device is used that automatically performs a series of operations in which a sample solution and a reaction solution are introduced into a precolumn, and the resulting reaction product is introduced into an analytical column together with a mobile phase. The number is very small.

An automatic sample introduction device equipped with such a pre-column reaction mechanism is described, for example, in "Journal of Chromat
ogrαphy, 305 (1984) P, 3
46 J.

The operation will be briefly explained with reference to FIG. 4, which shows this in a simplified manner. When the hexagonal valve 47 forms the flow path shown by the solid line, the aminoglycoside antibiotic is injected from the sample liquid injection part 43. The container 41 is filled with serum containing serum and the pump 42 is driven.
It is introduced into the reaction column 48 together with the sample liquid 40 inside. Next, the pump 46 is driven, and the 0-7 taraldehyde (OPA) solution in the container 45 is introduced into the reaction column 48 as a reaction liquid. After a predetermined period of time, the hexagonal valve 47 is switched to form a flow path shown by the dotted line, and the OPA derivative of the aminoglycoside antibiotic produced in the reaction column 48 is pumped together with the mobile phase in the container 50 by driving the pump 51. The sample is introduced into an analytical column 52.

Problems to be Solved by the Invention In the manual operation described above, the sample solution or reaction solution is measured and aspirated while visually checking the scale lines on the microsyringe, so there is no possibility that the measurement will be inaccurate. Therefore, the analysis results will be inaccurate, and if the microsyringe is inserted into the sample liquid container, the sample liquid will adhere to the surrounding area, and if the microsyringe is inserted into the reaction liquid container in this state, the sample liquid will adhere to the surrounding area of the microsyringe. The sample solution mixed with the reaction solution, and the microsyringe coated with the reaction solution,
When inserted into a sample liquid container, the reaction liquid mixes with the sample liquid, causing cross contamination. Furthermore, it is necessary to carry out analysis at a constant time from the start of the reaction until the solution containing the reaction product is introduced into the analytical column, resulting in a complicated operation. This operation is performed after a predetermined period of time has elapsed from the end of the second analysis, especially in the gradient analysis method in which analysis is performed by constantly changing the composition of the mobile phase passing through the analytical power system.
In addition to the requirement that the test liquid must be introduced into the column, the pre-column reaction must be started to match this introduction time, making the operation even more complicated.

In addition, the following problems occur in the automatic sample introduction device that is equipped with a reaction column in addition to an analysis column.

The first problem is that a sample liquid containing an analyte and a transfer liquid for transporting the sample liquid to the reaction column are first introduced into the reaction column. In this state, the analyte must first be adsorbed onto the reaction column packing material. Next, a reaction solution is introduced into the reaction column. A chemical reaction occurs here, and the reaction products must be adsorbed on the reaction column packing material while the reaction liquid is flowing until the reaction occurs completely. In the next step, an analytical mobile phase is introduced into the reaction column, and at this time the reaction products must be quickly desorbed from the reaction column packing material. The processes of adsorption, adsorption, and desorption in the reaction column described above are controlled by the properties of the packing material packed in the reaction column and the reaction liquid introduced into the column. It is difficult to select the type. For these reasons, an automatic sample introduction device equipped with a reaction column cannot use conventional techniques to form a membrane and use a large number of analytes in combination with the core solution.

The second problem is that the sample liquid introduced into the reaction column contains various compounds.

Some of these compounds strongly adsorb to the packing material of the reaction column, so a large amount of washing liquid is required to desorb and wash them.

For the above reasons, the widespread use of automatic sample introduction devices equipped with a pre-column reaction mechanism has been hindered, and manual operation has instead been used.

Therefore, in order to solve the above-mentioned drawbacks of the prior art, the present invention enables accurate metering and introduction of a sample liquid and a reaction liquid, sufficiently mixes the two liquids, and facilitates cleaning operations. The purpose is to provide an automatic sample introduction device.

Means for Solving the Problems The present invention causes a chemical reaction between a sample liquid and a reaction liquid, and introduces the resulting mixture into an analytical column together with a mobile phase, in which the sample liquid is transferred by opening and closing at minute intervals. A first valve member, a second valve member that feeds the υ reaction liquid by closing and opening synchronized with the opening and closing of the first valve member, and a first valve member introduced through the first and second valve members. A hollow holding coil that holds sample liquid and reaction liquid alternately arranged at predetermined lengths, and a first stripe and a second stripe.
This automatic sample introduction device includes a suction means for introducing a predetermined amount of sample liquid and reaction liquid into a holding coil through a valve member.

In conjunction with the suction operation of the action suction means, the first and 20th valve members are synchronously and alternately opened and closed, and the holding coil alternately arranges and holds the sample liquid and the reaction liquid at predetermined lengths. A reaction product produced by the reaction is introduced into an analytical column together with a mobile phase.

Example An embodiment of the present invention will be described below.

FIG. 1 shows a configuration diagram of the device of this embodiment, and shows 1
3 is a solenoid valve as a first valve member, 4 is a reaction liquid container in which a reaction liquid 5 is injected, and 6 is a solenoid valve as a second valve member. It is. The solenoid valves 3 and 6 are alternately energized by an unillustrated energizing device at the same time as the metering syringe starts to operate, which will be described later. When the solenoid valve 3 is opened and closed at intervals of, for example, 1 second, the solenoid valves are energized in synchronization with this. 6 is closed and opened at 1 second intervals, and the sample liquid 2 and reaction liquid 5 are alternately discharged at 1 second intervals. 7 is a hollow holding coil made of metal or Teflon, etc., and not filled with a filler, and is used as a reed or conduit! , through conduit 11 or! , to the solenoid valves 3 and 6. Then, the sample liquid 2 and the reaction liquid 5 are alternately introduced and held in the holding coil 7 by the suction operation of the metering syringe and the alternating opening and closing operations of the electromagnetic valves 3 and 6, which will be described later. Reference numeral 8 denotes a large-way valve as a flow path switching means, which has a first switching mode having a flow path shown by a solid line and a second switching mode having a flow path shown by a dotted line. Reference numeral 9 is a three-way valve, which forms a flow path shown by a solid line and a flow path shown by a dotted line as many times as K. 1
0 is a measuring syringe as a suction means, and 11 is a handle of the metering syringe, which is driven by a drive device (not shown) to accurately measure and aspirate the sample liquid 2 and the reaction liquid 5. As the suction means, the capacity of the holding coil 7 may be increased and a measuring means such as a microsyringe may be used, or the capacity of the holding coil 7 may be fixed at 50 plK, for example, and a well-known Verisflick pump or the like may be used as the suction means. A liquid larger than the volume of the holding coil 7 is sucked, and the holding coil i; 71! There is a way to make it function as a measuring means.

12 is a mobile phase container into which the mobile phase 13 is injected, and 14 is a bonder, which together form a mobile phase supply means. 15 is
This is an analytical column.

Here, to explain the formation of the first and 20th flow path systems by the hexagonal valve 8, when the hexagonal valve 8 is switched to the first switching mode and the flow path shown by the solid line is formed, the solenoid valve 3 or 6, Conduit 11 or! , conduit! , and the flow path P of the hexagonal valve 8
1, the holding coil 7, the flow path P of the six-way valve 8, and the conduit 1
4, a three-way valve 9 forming a flow path shown by a solid line, and a conduit l!
and the metering syringe 1° are connected to form a tenth channel system. Next, when the six-way valve 8 is switched to the second switching mode to form the flow path shown by the dotted line, the mobile phase supply means, the conduit l, the flow path P4 of the six-way valve 8, and the holding coil 7, The flow path P6 of the six-way valve 8, the conduit E, and the analytical column 1
5 are communicated with each other to form a second flow path system.

Next, the operation of the device of this embodiment will be explained. Now Ogata dialect 8
is switched to the first switching mode and the three-way valve 9 is switched to the flow path shown by the solid line, the solenoid valve 3 or 6, the holding coil 7, and the metering syringe 10 are switched to the first switching mode, as described above.
are communicated with each other to form a first channel system. The measuring syringe 1o is a driving device (not shown)! Driven by tK, handle 1
At the same time as the valve 1 is moved to the right, the solenoid valves 3 and 6 of the valve 1 are alternately biased by a biasing device (not shown) to open and close.

When the solenoid valve 3 opens, the solenoid valve 6 is closed, and the sample liquid 2
is passed through the conduits l, and l, and when the solenoid valve 3 is closed, the solenoid valve 6 is opened, and the reaction liquid 5 is introduced into the inlet pipe l, and! , and through the flow path PI of the L-way valve 8, the sample liquid 2 and the reaction liquid 5 having a volume corresponding to the opening time of the solenoid valve 3 or 6 are alternately attracted and held in the holding coil 7. . The metering syringe IO is driven for a predetermined stroke, and the drive is stopped when a predetermined amount of suction has been completed. In the meantime, due to the alternate opening and closing action of solenoid valves 3 and 6,
The sample liquid 2 and the reaction liquid 5 are alternately attracted to the holding coil 7, and are alternately arranged and held every predetermined length.

Here, the sample liquid 2 and reaction liquid 5 sucked into the holding coil 7
The arrangement of the holding coil 7 will be explained in detail with reference to FIG. 2, which shows a part of the holding coil 7. Now, the opening and closing time of the solenoid valves 3 and 6 is 1 second, and the suction time of one time of the metering syringe 10 is 10 seconds.
Assuming that 50 μl of liquid is aspirated in 0 seconds, since electromagnetic valves 3 and 6 are alternately opened and closed for 1 second each, 25 μl of sample liquid 2 and reaction liquid 5 are each sucked over 50 seconds.
It will be gradually absorbed. Therefore, the sample liquid 2 and the reaction liquid 5 are each sucked for 1 second at a rate of 25μ1150 seconds=0°5μ2/second. Here, if the inner diameter of the holding coil 7 is 0.8-, its cross-sectional area is approximately 0.5 wax
”The sample solution 2 and reaction solution 5 with a volume of 0.5μ! are aspirated for 1 second each, and the holding coil 7 has a length of 11
1Ill liquid columns are alternately formed. In this way, a large number of sample liquids 2 and reaction liquids 5 are alternately arranged and held in the holding coil 7, so that both liquids are sufficiently mixed and a chemical reaction can be carried out quickly. In addition,
- Although the description has been given as an example in which equivalent amounts of the reaction liquid and sample liquid are aspirated, the present invention is not limited to aspirating equivalent amounts of the reaction liquid and sample liquid. In order to cause the chemical reaction in the holding coil 7 to occur more quickly, it is necessary to provide a heating device for heating the holding coil 7.

Note that even if the viscosity of the sample liquid and reaction liquid differs due to the suction operation of the measuring syringe 10 and the opening/closing operation of the solenoid valves 3 and 6, for example, when the sample liquid is serum and the reaction liquid is a methanol solution of ADAM, Also, the sample liquid and reaction liquid can be accurately measured and introduced into the holding coil 7.

Now, the sample liquid 2 and the reaction liquid 5 arranged and held in the holding coil 7 are kept stationary and held in that state until a sufficient chemical reaction is completed. On the other hand, the mobile phase 13 is transferred to the conduit by driving the pump 14! , the flow path P of the six-way valve 8, and the conduit 1. The liquid is sent to the analytical column 15 via.

After a predetermined period of time has passed, the six-way valve 8 is switched to the second switching mode, and the three-way valve 9 is switched to the flow path indicated by the dotted line.
Mobile phase supply means and conduit! , the flow path P4 of the six-way valve 8, the holding coil 7, the flow path P of the six-way valve 8, the conduit 18, and the analytical column 15 are communicated to form a second flow path system. Together with the mobile phase 13, the handle 1 of the metering syringe 10 is driven by the reaction generated in the holding coil 7.
1 to the left, the flow path of the three-way valve 9 shown by the dotted line in the conduit 21, and the conduit 1. is discharged to the outside through the

This completes one sample introduction operation for analysis.

When cleaning the holding coil 7, the sample liquid container 1 and the reaction liquid container 5 are replaced with containers filled with cleaning liquid, the six-way valve 8 is switched to the first switching mode, and the three-way valve 9 is switched to the solid line. Switch to the flow path shown by and drive the metering syringe 10 for suction, then switch the three-way valve 9 to the flow path shown by the dotted line, discharge the cleaning liquid sucked into the metering syringe 10, and repeat this to hold. Clean the coil 10. As mentioned above, the holding coil 10 is not filled with a filler and is in a hollow state, so the filler does not retain the analyte, etc., and requires a large amount of cleaning liquid and a long cleaning time. It is not necessary and cleaning work is extremely easy.

Next, two modified examples of the above-mentioned valve member will be explained with reference to FIG.

In FIG. 3(A) showing a modified example of the valve member, a solid cylindrical body 31 that penetrates perpendicularly to the conduit through which the sample liquid 2 flows and the conduit E through which the reaction liquid 5 flows is provided with a phase difference. 90' different flow holes 31α and 31b are bored. Backings 32 and 33 for preventing liquid leakage are provided at the portions of the conduits l□ and 4 through which the cylindrical body 31 passes. When the cylindrical body 31 is rotated 90'' in the direction of the arrow from the illustrated state, the flow hole 31α coincides with the axial direction of the conduit l, allowing the sample liquid 2 to flow, and the flow hole 31b is aligned with the axial direction of the conduit l. orthogonal,
The flow of reaction solution 5 is shut off.

Still another modification is shown in FIG. 3(B), in which a cylindrical body 34 that is perpendicular to and penetrates the conduits 11 and l has a conduit E1! , communication holes 34α and 34b are bored in a direction coinciding with the axial direction of . One end of the cylindrical body 34 is a cam 35
A coil spring 37 is loaded between the other end of the cylindrical body 34 and the chassis 36, and biases the cylindrical body 34 in the right direction. The cylindrical body 34 passes through the conduit 11 and 1.
Similarly, backings 32 and 3 for preventing liquid leakage are placed in the part 4.
3 are provided. In the situation shown, conduit l
, is in a blocked state and conduit l is in a flowing state. However, when the cam 35 is rotated 180 degrees, the cylindrical body 34 is moved to the right by the bias of the coil spring 37, and the flow hole 34.6 is in a blocked state. is the conduit j and moves to the outside, and the flow hole 34α is the conduit 1
Located within m, conduit! , is in circulation and the conduit l.

is in a cut-off state.

The modified example of the valve member described above is capable of communicating and blocking the flow between the sample liquid 2 and the reaction liquid 5 in 6-times of operation.

DETAILED DESCRIPTION OF THE INVENTION According to the present invention, in a system in which a sample liquid and a reaction liquid are chemically reacted and introduced into an analytical column together with a mobile phase, the first and second
Since the valve members of the holding coil are opened and closed alternately in synchronization at minute time intervals, the retention coil The sample liquid and the reaction liquid are arranged and held alternately at predetermined lengths, so that the two liquids are sufficiently mixed and reaction products are easily generated. Even if there is a difference in viscosity between the reaction liquid and the reaction liquid, both liquids can be accurately measured and introduced into the holding coil. Furthermore, since the holding coil is a hollow body that does not contain a filler, reaction products are not adsorbed or desorbed to the filler as in conventional automatic sample introduction devices, and therefore the combination of the analyte and reagent is prevented. This makes it possible to perform analysis without being restricted by the above conditions, and since a large amount of washing water is not required, washing becomes easy, and it becomes possible to popularize analytical devices using pre-column reaction devices.

[Brief explanation of the drawing]

FIG. 1 is a configuration diagram of an embodiment of the present invention, FIG. 2 is an explanatory diagram for explaining the arrangement state of the sample liquid and reaction solution held in the holding coil of the embodiment shown in FIG. 1, and FIG. FIG. 4A is a top view of a modification of the valve member, FIG. 4B is a top view of another modification of the valve member, and FIG. 4 is a configuration diagram of a conventional device. 1 is a sample liquid container, 2 is a sample liquid, 4 is a reaction liquid container, 5 is a reaction liquid, 3 and 6 are electromagnetic valves, 7 is a holding coil, 8 is a hexagonal valve,
9 is a three-way valve, 10 is a metering syringe, 11 is a handle, 12 is a mobile phase container, 13 is a mobile phase, 14 is a pump, 15 is an analytical column,! 8~! , denotes a conduit, and P, to P6 denote a flow path of the six-way valve 8. Figure 1 Figure 2

Claims (2)

[Claims] (1) A first valve member that transports a sample liquid by opening and closing at minute time intervals, and a second valve that transports a reaction liquid by opening and closing in synchronization with the opening and closing of the first valve member. a hollow holding coil that holds the sample liquid and reaction liquid introduced through the first and second valve members, arranged alternately at predetermined lengths, and the first and second valve members; An automatic sample introduction device comprising suction means for introducing a predetermined amount of sample liquid and reaction liquid into a holding coil via a valve member. (2) The automatic sample introduction device according to claim (1), further comprising heating means for heating the holding coil.