CN110018264A - The method that multichannel integrates liquid chromatogram column device and carries out liquid phase preparation - Google Patents

Abstract

The invention discloses a multi-channel integrated liquid chromatography column device, comprising a box body with an open front end, an electric heating tube is coiled around the rear wall of the box body, and a plurality of analytical high-efficiency liquid chromatography columns are arranged in parallel and spaced in the box body, namely For the analysis column, the side wall of the box is hinged with a box door at the opening, and the top of each analysis column is connected with a liquid outlet pipe; the bottom end of each analysis column is connected to a liquid inlet pipe through a liquid separation mechanism, and the liquid inlet pipe extends out of the box. The side wall of the body is connected with the delivery pump. The invention also discloses a method for liquid phase preparation using the above multi-channel integrated liquid chromatography column device. The invention combines the advantages of good separation effect of the analytical column and large injection volume of the preparation column, and overcomes the defects of the small injection volume of the analytical column and the poor separation effect of the preparation type high performance liquid chromatography column, that is, the preparation column. Multiple separations, efficient and convenient, can separate a variety of desired compounds at one time, saving the cost of separation of complex compounds.

Description

Multi-channel integrated liquid chromatography column device and method for liquid phase preparation

technical field

The invention relates to the technical field of high performance liquid phase preparative chromatography.

Background technique

The method of preparative liquid chromatography has become an important separation and analysis technology in the fields of chemistry, biochemistry, medicine, industry, agriculture, environmental protection, commodity inspection and legal inspection, and it is indispensable in the hands of analytical chemistry, biochemistry and environmental chemists. tool. Taking medicine as an example, in the Pharmacopoeia of the People's Republic of China, there are more than about 600 kinds of chemically synthesized drugs and more than about 400 kinds of traditional Chinese medicines using liquid chromatography for quality control. However, due to the sample throughput and sample separation efficiency of traditional preparative chromatography. The contradiction between them has not been resolved. In order to obtain a better separation and purification effect, it is necessary to reduce the sample processing volume, which makes the separation cumbersome and the cost increases, which in turn makes some liquid-phase preparation related drugs on the market very expensive.

At present, high-performance liquid chromatographs are only equipped with analytical columns (ie analytical high-performance liquid chromatography columns), semi-preparative columns or preparative columns. When the preparative column separates some complex compounds, the separation effect often fails to achieve the expected separation effect. The analytical column can achieve a good separation effect, but due to its small injection volume, it takes a long time for preparation, and the flow is relatively large, so its application in the preparation of liquid phase is limited. For those skilled in the art, it is a conventional technical understanding that an analytical column is used for analytical testing, a preparative column is used for liquid phase preparation, and that an analytical column and a preparative column cannot be used universally.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a multi-channel integrated liquid chromatography column device, which not only has good separation effect, but also has the advantages of large injection volume and high preparation capacity.

In order to achieve the above purpose, the multi-channel integrated liquid chromatography column device of the present invention includes a box body with an open front end, an electric heating tube is coiled around the rear wall of the box body, and an electric fan is arranged in the box in front of the electric heating tube. A number of analysis columns are arranged in parallel and spaced in the front box, the side wall of the box is hinged with a box door at the opening, and the box door includes a door frame and a double-layer transparent material embedded in the door frame, and there is a cavity between the double-layer transparent materials;

The top of each analysis column is connected with a liquid outlet pipe, and each liquid outlet pipe is collected into a liquid outlet pipe bundle. The liquid outlet pipe bundle leads out of the box. One liquid outlet pipe in the liquid outlet pipe bundle is used to connect an external liquid chromatography detection. The other liquid outlet pipes in the liquid outlet pipe bundle lead to the collection container outside the box;

The bottom ends of each analysis column are respectively connected to the liquid inlet pipe through the liquid separation mechanism, and the liquid inlet pipe extends out of the side wall of the box body and is connected with the transport pump for transporting the mobile phase;

The electric heating tube and the electric fan are connected with an electric control device, the electric control device is connected with a display screen and a temperature sensor, the electric control device and the display screen are both located on the outer wall of the box, and the temperature sensor is located inside the box body.

A metal orifice plate is arranged in the box between the analysis column and the electric fan, a plurality of small holes are evenly distributed on the metal orifice plate, and two are symmetrically arranged up and down on the metal orifice plate.

A plurality of analytical columns are arranged side by side at intervals, and the liquid separation mechanism adopts any one of the following two structures:

a. The liquid separation mechanism includes a multi-port valve. The multi-port valve has a liquid inlet and several liquid outlets. The liquid outlet of the multi-port valve is connected with the analysis column one by one; the liquid inlet of the multi-port valve is connected with the liquid inlet manifold. connected;

b. The liquid separation mechanism includes several three-way valves, and each three-way valve includes one liquid inlet and two liquid outlets; with the liquid flow direction as the downstream direction, the liquid inlet and the liquid inlet of the three-way valve located at the most upstream In the three-way valves connected in the upstream and downstream directions, the two liquid outlets of the one upstream three-way valve are connected with the liquid inlets of the two downstream three-way valves in one-to-one correspondence; The total number of the liquid outlets of each three-way valve is the same as the number of the analytical columns and is in one-to-one correspondence.

The invention also discloses a method for liquid phase preparation using the above multi-channel integrated liquid chromatography column device. The process of preparing the liquid phase is as follows: the sample enters each analysis column uniformly with the mobile phase through the liquid inlet manifold and the liquid separation mechanism, and each separation group With the mobile phase, it flows out of each analytical column successively through the liquid outlet pipe, and finally enters the collection container, and the separated components that flow out at different times are collected in different collection containers by replacing the collection container;

In the above-mentioned process of preparing the liquid phase, the temperature in the box is maintained at a predetermined temperature range by a constant temperature method, and the constant temperature method is:

The electric control device turns on the electric fan, and the electric fan forms an air circulation in the box, and the air flow makes the temperature of all parts of the box more uniform, so that the heat generated by the electric heating tube evenly heats each analysis column;

The target temperature TM°C is stored in the electronic control device; the electronic control device receives the temperature signal detected by the temperature sensor, and the actual temperature inside the box detected by the temperature sensor is TS°C; when TS<TM-2, the electronic control device controls The electric heating tube is continuously energized and heated; when (TM-2) ≤ TS < (TM-1), the electric control device controls the electric heating tube to stop for 1 second every 2 seconds; when (TM-1) ≤ TS < TM When TS = TM, the electric control device controls the electric heating tube to stop for 3-5 seconds every 2 seconds; when (TM+1)>TS>TM, the electric heating The control device keeps the working state of the electric heating tube unchanged; when (TM+1)≤TS, the electric control device turns off the electric heating tube.

The multi-channel integrated liquid chromatographic column device of the present invention connects a plurality of analytical columns in parallel through structures such as multi-port valves, thereby improving the total sample processing capacity, and combining the advantages of good separation effect of the analytical column and large injection volume of the preparation column It overcomes the defects of small injection volume of the analytical column and poor separation effect of the preparative column. One separation can be equivalent to the original multiple separation operations, which is efficient and convenient for the use of the chromatographic column, and can separate a variety of desired compounds at one time. Saving the cost of separation of complex compounds is of great significance for the detection of national departments, drug preparation (reducing drug preparation costs and drug prices), and small living environments.

There is an air interlayer between the double layers of transparent materials, which not only improves the thermal insulation performance of the box door, but also facilitates the operator to observe the working conditions of each analytical column through the box door.

On the one hand, the metal orifice plate plays the role of facilitating the fixing of the chromatographic column, and on the other hand, the metal orifice plate plays the role of uniform heat dissipation. The good thermal conductivity of the metal makes the temperature of all parts of the box more uniform, and the porous structure is conducive to the circulation of air. The air flow caused by the electric fan flows through the small holes and the space outside the two metal orifice plates, and the heat generated by the electric heating tube is evenly distributed to all parts of the inner space of the box.

The liquid dispensing mechanism has a simple structure and uniform liquid distribution. The components used, such as pipelines, multi-way valves and three-way valves, are commonly used in the laboratory, and maintenance and replacement are very convenient.

By adopting the liquid phase preparation method of the present invention, when the actual temperature inside the box body is far from the target temperature, the electric heating tube is operated at full speed to rapidly increase the temperature inside the box body; the actual temperature inside the box body rises to the target temperature When nearby, gradually reduce the opening time of the electric heating tube to reduce the temperature rise rate in the box, prevent the temperature rise too fast and make the temperature in the box too high, on the one hand to avoid energy waste, on the other hand to make the temperature inside the box better controlled around the target temperature. When the actual temperature in the box is the same as the target temperature, the opening time of the electric heating tube is shorter, and it is easier to match the speed of heat dissipation in the box, the temperature in the box changes more slowly, and the constant temperature effect is good.

When the actual temperature in the box is more than 1°C higher than the target temperature, the electric heating tube is completely turned off. At this time, the residual heat of the electric heating tube and the heat in the box higher than the target temperature can keep the temperature in the box at a long time. The target temperature is near, so it is not necessary to turn on the electric heating tube before the actual temperature TS in the box drops to TM.

In a word, the method for liquid phase preparation of the present invention can make the air temperature in the box change slowly when it is near the target temperature, while the air temperature in the box changes rapidly when it is far from the target temperature (rapid heating or rapid cooling), reducing energy Waste, improve the constant temperature effect.

The present invention can change the type of the analytical column according to the different chemical properties of the substances to be separated, so as to achieve the purpose of separating products; in order to meet the requirements of different preparation amounts, the number of analytical columns can be increased; the present invention has a simple structure and is helpful for Mass production and extensive application. The multi-channel integrated liquid chromatography column device has an important influence on the liquid phase preparation in the laboratory and scientific research field.

Description of drawings

Fig. 1 is the structural representation of the multi-channel integrated liquid chromatography column device in the present invention after removing the box door;

Fig. 2 is the left side schematic diagram of the multi-channel integrated liquid chromatography column device after removing the electronic control device and the display screen;

Fig. 3 is the electric control principle diagram of the present invention;

Fig. 4 is the separation effect diagram of trifluorouracil nucleotide solution during preparation;

Fig. 5 is the analysis effect diagram of the purified product obtained by each analytical column of the multi-channel integrated liquid chromatography column device; each waveform line from top to bottom in the figure is the analysis effect curve of the purified product obtained by No. 1-4 analytical columns;

FIG. 6 is an analysis and comparison diagram of the purified product obtained from each analytical column of the multi-channel integrated liquid chromatography column device and the preparation liquid of the preparative column. The four wavy lines from top to bottom in the figure are the analysis effect curves of purified products obtained from analytical columns 1-4 respectively; the bottom wavy line in the figure (that is, the fifth line from top to bottom) is the purified product obtained from the preparative column analysis effect curve.

Detailed ways

As shown in FIG. 1 to FIG. 6 , the multi-channel integrated liquid chromatography column device of the present invention includes a box body 1 with an open front end, an electric heating tube 2 is wound around the rear wall of the box body 1 , and the box body in front of the electric heating tube 2 1 is provided with an electric fan 3, a number of analysis columns 4 are arranged in parallel and spaced in the box body 1 in front of the electric fan 3, the side wall of the box body 1 is hinged with a box door at the opening, and the box door includes a door frame 5 and a box embedded in the door frame 5. The inner double layer transparent material 6 has a cavity 7 between the double layer transparent materials 6; the transparent material is glass, preferably plexiglass.

The top of each analysis column 4 is connected with a liquid outlet pipe 8, and each liquid outlet pipe is collected into a liquid outlet pipe bundle 9. The liquid outlet pipe bundle 9 leads out of the box body, and one liquid outlet pipe in the liquid outlet pipe bundle 9 is used to connect an external outlet. The liquid chromatographic detector in the liquid chromatographic detector, the other liquid outlet branch pipes in the liquid outlet pipe bundle 9 lead to the collection container 10 outside the box 1; the collection container 10 is preferably a collection bottle, and the liquid chromatography detector is a conventional device, not shown in the figure.

The bottom ends of each analysis column 4 are respectively connected to the liquid inlet pipe 11 through the liquid separation mechanism, and the liquid inlet pipe 11 extends out of the side wall of the box 1 and is connected with the transfer pump for transferring the mobile phase; the transfer pump is a conventional technology, not shown in the figure.

The electric heating tube 2 and the electric fan 3 are both connected to an electric control device 12. The electric control device 12 is connected with a display screen 13 and a temperature sensor 14. The electric control device 12 and the display screen 13 are both located on the outer wall of the box body 1. The temperature sensor 14 is located inside the box body 1 . In order to make the temperature signal more accurate, multiple temperature sensors can be evenly arranged in the box, and the electric control device takes the average value of each temperature sensor as the value of TS. TS is a real number.

There is a cavity 7 between the double layers of transparent materials 6, which not only improves the thermal insulation performance of the box door, but also facilitates the operator to observe the working conditions of each analysis column 4 through the box door.

The electronic control device 12 is an 86 computer or a single-chip microcomputer, and a PLC can also be used.

A metal orifice plate 15 is arranged in the box 1 between the analysis column 4 and the electric fan 3 . The metal orifice plate 15 is evenly distributed with a plurality of small holes 16 , and two metal orifice plates 15 are symmetrically arranged up and down. The left and right ends of the metal orifice plate 15 are fixedly connected to the side wall of the box body 1 . On the one hand, the small holes 16 are convenient for fixing screws, and the analysis column 4 is fixed by the screws, and on the other hand, the air convection in the box body 1 is convenient.

A plurality of analytical columns are arranged side by side at intervals, and the liquid separation mechanism adopts any one of the following two structures:

a. The liquid separation mechanism includes a multi-port valve. The multi-port valve has a liquid inlet and several liquid outlets. The liquid outlet of the multi-port valve is connected with the analysis column one by one; the liquid inlet of the multi-port valve is connected with the liquid inlet manifold. connected;

b. The liquid separation mechanism includes several three-way valves, and each three-way valve includes one liquid inlet and two liquid outlets; with the liquid flow direction as the downstream direction, the liquid inlet and the liquid inlet of the three-way valve located at the most upstream In the three-way valves connected in the upstream and downstream directions, the two liquid outlets of the one upstream three-way valve are connected with the liquid inlets of the two downstream three-way valves in one-to-one correspondence; The total number of the liquid outlets of each three-way valve is the same as the number of the analytical columns and is in one-to-one correspondence.

The first type is simple and compact in structure, but requires the use of a special multi-port valve; the second type has a large number of structural components and the connection is relatively complicated, but because the laboratory already has a three-way valve, it is convenient for local materials and on-site connection and production. The advantages.

In this embodiment, four analysis columns 4 are arranged side by side at intervals, and the liquid separation mechanism adopts the second structure, including a total three-way valve 17, the liquid inlet of the total three-way valve 17 is connected to the liquid inlet manifold 11, and the total three The two liquid outlets of the through valve 17 are respectively connected to the liquid inlet of a liquid distribution three-way valve 18, each liquid distribution three-way valve 18 has two liquid outlets, and the four liquid distribution three-way valves 18 have four outlets. The liquid ports are connected with the liquid inlets of the four analysis columns 4 through the liquid inlet branch pipes 19 in one-to-one correspondence;

The liquid dispensing mechanism has a simple structure and uniform liquid distribution. The components used, such as pipelines and three-way valves, are common parts in the laboratory, and maintenance and replacement are very convenient.

The invention also discloses a method for liquid phase preparation using the above multi-channel integrated liquid chromatography column device.

The process of preparing the liquid phase is as follows: the sample enters each analysis column 4 evenly with the mobile phase through the liquid inlet manifold 11 and the liquid separation mechanism, and each separated component flows out of each analysis column 4 through the liquid outlet branch pipe 8 successively with the mobile phase, and finally enters the collection container. In 10, the separated components flowing out at different times are collected in different collection containers 10 by changing the collection container 10;

In the above-mentioned process of preparing the liquid phase, the temperature in the box 1 is maintained in a predetermined temperature range by a constant temperature method, and the constant temperature method is:

The electric control device 12 turns on the electric fan 3, and the electric fan 3 forms an air circulation in the box body 1, and the air flow makes the temperature in the box body 1 more uniform, so that the heat generated by the electric heating tube 2 evenly heats each analysis column 4;

The target temperature TM° C. is stored in the electronic control device 12 , and TM is a real number. The electronic control device 12 receives the temperature signal detected by the temperature sensor 14, and the actual temperature inside the box 1 detected by the temperature sensor 14 is TS°C; when TS<TM-2, the electronic control device 12 controls the electric heating tube 2 to continue energizing Heating; when (TM-2)≤TS＜(TM-1), the electric control device 12 controls the electric heating tube 2 to stop for 1 second every 2 seconds; when (TM-1)≤TS＜TM, every Work for 2 seconds and stop for 2 seconds; when TS=TM, the electric control device 12 controls the electric heating tube 2 to stop for 3-5 seconds every 2 seconds of operation (the specific value depends on the overall heat dissipation speed of the box 1, the box The faster the body 1 dissipates heat, the shorter the stop time of the electric heating tube 2); when (TM+1)>TS>TM, the electronic control device 12 keeps the working state of the electric heating tube 2 unchanged (in this temperature range, When the whole is in the temperature rising period, the electric heating tube 2 stops working for 3-5 seconds every 2 seconds; when the whole is in the temperature falling period, the electric heating tube 2 stops working until TS=TM); when (TM+1)≤TS , the electric control device 12 turns off the electric heating tube 2 .

Using the constant temperature method of the present invention, when the actual temperature inside the box 1 is far from the target temperature, the electric heating tube 2 is operated at full speed to rapidly increase the temperature inside the box 1; the actual temperature inside the box 1 rises to the target temperature When the temperature is near, gradually reduce the opening time of the electric heating tube 2 to reduce the temperature rise rate in the box 1, prevent the temperature rise too fast and make the temperature in the box 1 too high, on the one hand to avoid energy waste, on the other hand The temperature in the box 1 is better controlled around the target temperature. When the actual temperature in the box 1 is the same as the target temperature, the electric heating tube 2 is turned on for a shorter time, which is easier to match with the rate of heat dissipated by the box 1, the temperature in the box 1 changes more slowly, and the constant temperature effect is good.

When the actual temperature in the box 1 is higher than the target temperature by more than 1°C, the electric heating tube 2 is completely turned off. At this time, the residual heat of the electric heating tube 2 and the heat in the box 1 higher than the target temperature can make the The temperature remains near the target temperature for a long time, so it is not necessary to turn on the electric heating tube 2 until the actual temperature TS in the tank 1 drops to TM.

In a word, the method for liquid phase preparation of the present invention can make the air temperature in the box 1 change slowly when it is near the target temperature, while the air temperature in the box 1 changes rapidly when it is far from the target temperature (rapid heating or rapid cooling) , reducing energy waste and improving the constant temperature effect.

To verify the effectiveness of the multi-channel integrated liquid chromatography column device, the inventors conducted experiments.

Experimental program:

(1) Configure the trifluorouracil nucleotide solution to be separated, use the multi-channel integrated liquid chromatography column device of the present invention to separate the solution, and then use the analytical column to identify the separation effect of the purified product obtained from each analytical column in the present invention.

(2) Use a preparative column to separate the same amount of trifluorouracil nucleotide solution, and use an analytical column to identify the separation effect of the preparative solution.

Figure 4 shows the separation effect of the trifluorouracil nucleotide solution during preparation.

The four analytical columns 4 of the multi-channel integrated liquid chromatography column device are respectively numbered as No. 1-4 analytical columns. The analysis diagrams of analytical columns 1-4 are shown in Figure 5.

By analyzing the purified products obtained from the multi-channel integrated liquid chromatography column device, we can find that each analytical column 4 has a good separation effect and the separation effect is basically the same (as shown in Figure 5). By comparing the separation effect of the multi-channel integrated liquid chromatography column device with the separation effect of the preparation column, we can find that under the condition of a small injection volume, the method for liquid phase preparation of the multi-channel integrated liquid chromatography column device of the present invention can The same preparative liquid phase effect as the preparative column is achieved, which shows the feasibility of the device in the preparative liquid phase.

The above embodiments are only used to illustrate rather than limit the technical solutions of the present invention. Although the present invention has been described in detail with reference to the above embodiments, those of ordinary skill in the art should understand that the present invention can still be modified or equivalently replaced without departing from the Any modification or partial replacement of the spirit and scope of the present invention should be included in the scope of the claims of the present invention.

Claims (4)

1. The multi-channel integrated liquid chromatography column device is characterized in that: it comprises a box body with an open front end, an electric heating pipe is wound around the rear wall of the box body, and an electric fan is arranged in the box in front of the electric heating pipe, and the front of the electric fan is provided with an electric fan. A number of analysis columns are arranged in parallel and spaced apart in the box, the side wall of the box is hinged with a box door at the opening, the box door includes a door frame and a double-layer transparent material embedded in the door frame, and there is a cavity between the double-layer transparent materials; The top of each analysis column is connected with a liquid outlet pipe, and each liquid outlet pipe is collected into a liquid outlet pipe bundle. The liquid outlet pipe bundle leads out of the box. One liquid outlet pipe in the liquid outlet pipe bundle is used to connect an external liquid chromatography detection. The other liquid outlet pipes in the liquid outlet pipe bundle lead to the collection container outside the box; The bottom ends of each analysis column are respectively connected to the liquid inlet pipe through the liquid separation mechanism, and the liquid inlet pipe extends out of the side wall of the box body and is connected with the transport pump for transporting the mobile phase; The electric heating tube and the electric fan are connected with an electric control device, the electric control device is connected with a display screen and a temperature sensor, the electric control device and the display screen are both located on the outer wall of the box, and the temperature sensor is located inside the box body. 2 . The multi-channel integrated liquid chromatography column device according to claim 1 , wherein a metal orifice plate is arranged in the box between the analysis column and the electric fan, and a plurality of metal orifice plates are evenly distributed on the metal orifice plate. 3 . There are two small holes on the metal orifice plate symmetrically up and down. 3. The multi-channel integrated liquid chromatographic column device according to claim 1 is characterized in that: the analytical column is provided with a plurality of side-by-side intervals, and the liquid-separating mechanism adopts any one of the following two structures: a. The liquid separation mechanism includes a multi-port valve. The multi-port valve has a liquid inlet and several liquid outlets. The liquid outlet of the multi-port valve is connected with the analysis column one by one; the liquid inlet of the multi-port valve is connected with the liquid inlet manifold. connected; b. The liquid separation mechanism includes several three-way valves, and each three-way valve includes one liquid inlet and two liquid outlets; with the liquid flow direction as the downstream direction, the liquid inlet and the liquid inlet of the three-way valve located at the most upstream In the three-way valves connected in the upstream and downstream directions, the two liquid outlets of the one upstream three-way valve are connected with the liquid inlets of the two downstream three-way valves in one-to-one correspondence; The total number of the liquid outlets of each three-way valve is the same as the number of the analytical columns and is in one-to-one correspondence. 4. utilize the multi-channel integrated liquid chromatographic column device described in claim 2 to carry out the method for liquid phase preparation, it is characterized in that: The process of preparing the liquid phase is as follows: the sample enters into each analytical column evenly with the mobile phase through the liquid inlet manifold and the liquid separation mechanism, and each separated component flows out of each analytical column through the liquid outlet branch pipe with the mobile phase, and finally enters the collection container. The collection container collects the separated components that flow out at different times in different collection containers; In the above-mentioned process of preparing the liquid phase, the temperature in the box is maintained at a predetermined temperature range by a constant temperature method, and the constant temperature method is: The electric control device turns on the electric fan, and the electric fan forms an air circulation in the box, and the air flow makes the temperature of all parts of the box more uniform, so that the heat generated by the electric heating tube evenly heats each analysis column; The target temperature TM°C is stored in the electronic control device; the electronic control device receives the temperature signal detected by the temperature sensor, and the actual temperature inside the box detected by the temperature sensor is TS°C; when TS<TM-2, the electronic control device controls The electric heating tube is continuously energized and heated; when (TM-2) ≤ TS < (TM-1), the electric control device controls the electric heating tube to stop for 1 second every 2 seconds; when (TM-1) ≤ TS < TM When TS = TM, the electric control device controls the electric heating tube to stop for 3-5 seconds every 2 seconds; when (TM+1)>TS>TM, the electric heating The control device keeps the working state of the electric heating tube unchanged; when (TM+1)≤TS, the electric control device turns off the electric heating tube.