CN113252763A - Multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis - Google Patents

Multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis Download PDF

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Publication number
CN113252763A
CN113252763A CN202110429173.5A CN202110429173A CN113252763A CN 113252763 A CN113252763 A CN 113252763A CN 202110429173 A CN202110429173 A CN 202110429173A CN 113252763 A CN113252763 A CN 113252763A
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capillary
sampling probe
sample introduction
sample
fluorescence analysis
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CN202110429173.5A
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陈进
孙雨
王凤超
韩睿祎
庞建鑫
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Shanghai Institute of Technology
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Shanghai Institute of Technology
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416Systems
    • G01N27/447Systems using electrophoresis
    • G01N27/44704Details; Accessories
    • G01N27/44717Arrangements for investigating the separated zones, e.g. localising zones
    • G01N27/44721Arrangements for investigating the separated zones, e.g. localising zones by optical means
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416Systems
    • G01N27/447Systems using electrophoresis
    • G01N27/44704Details; Accessories
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416Systems
    • G01N27/447Systems using electrophoresis
    • G01N27/44756Apparatus specially adapted therefor
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N2001/002Devices for supplying or distributing samples to an analysing apparatus

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Electrochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

The invention relates to a multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis, which comprises a retractable and rotatable stepping motor, a sample introduction disc in transmission connection with the stepping motor, a plurality of hollow capillary sampling probes, a power supply, a plurality of detection windows and a waste liquid pool, wherein a plurality of working areas are arranged on the sample introduction disc along the circumferential direction, three replaceable centrifugal tubes which are respectively used as a sample pool, a liquid storage pool and a water purification pool are inserted into each working area, the three centrifugal tubes are arranged along the circumferential direction according to the sequence of the sample pool, the liquid storage pool and the water purification pool, the sampling ends of the capillary sampling probes can movably extend into the centrifugal tubes, the detection window is arranged on the capillary sampling probe, the outlet end of the capillary sampling probe is inserted into the waste liquid pool, one pole of the power supply is electrically connected with the inlet end of the capillary sampling probe, and the other pole of the power supply is electrically connected with the outlet end of the capillary sampling probe to form a loop. Compared with the prior art, the method can realize multi-channel simultaneous detection, improve the detection rate and have application prospect.

Description

Multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis
Technical Field
The invention belongs to the technical field of analytical instruments, and particularly relates to a multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis.
Background
With the development of science and technology, people have higher and higher requirements on the efficiency of analysis and test, so that the automatic continuous sample feeding is very important for high-throughput analysis. In electrophoretic analysis, the introduction and replacement of samples is often performed manually in many cases. The typical sample changing mode adopted by the chip electrophoresis analysis specifically comprises the following steps: cutting off the high-voltage power supply, removing the microchip from the detection device, emptying and cleaning the sample liquid storage tank for multiple times, then re-injecting new sample solution, resetting the microchip, inserting the electrode into the liquid storage tank, adjusting the detection point, and turning on the high-voltage power supply for secondary detection. This approach is time consuming and greatly reduces the throughput of the analysis of the entire system.
The analytical testing of samples is currently moving towards lower cost, faster analysis and more compact devices. After a traditional capillary array electrophoresis is performed for one analysis, the analysis needs to be stopped, the power supply needs to be cut off, a new sample tray needs to be replaced, or the sample tray needs to be cleaned and then the sample is injected into the sample tray, so that the next analysis can be performed. The sample introduction mode greatly reduces the analysis flux of the whole system, the separation channel of the short capillary electrophoresis analysis is short, and the analysis speed is high, so the introduction speed of the sample and the length of the sample band in the sample introduction can have great influence on the separation speed and the separation effect of the sample.
Patent CN101398436B discloses an automatic introducing device of rotation type micro-sample, including integration sample dish, capillary sampling probe, step motor, high voltage power supply, electrode, liquid storage tube, its characterized in that: the integrated sample disc is in a gear shape, namely liquid storage tanks are uniformly distributed on the outer edge of the circular disc body, and each liquid storage tank is provided with a pore; the outer wall of the sample inlet end of the capillary sampling probe is plated with one or more metal layers, the sample inlet end of the capillary sampling probe can be inserted into a pore at the bottom of a liquid storage tank of the integrated sample disc, and the capillary sampling probe does not contact with the disc body of the integrated sample disc when the integrated sample disc rotates. However, this patent can not detect different materials simultaneously, and when detecting same material, because capillary sampling probe only has one, can not accomplish to detect and just in time wash, can cause the sample pollution, influences the detection effect.
Disclosure of Invention
The invention aims to provide a multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis, which adopts multi-channel automatic sample introduction, can realize multi-channel simultaneous detection, greatly improves the detection speed, and has good application prospect as a sample introduction device of a capillary electrophoresis detector.
The purpose of the invention is realized by the following technical scheme:
the utility model provides a multichannel autoinjection device for capillary electrophoresis fluorescence analysis, the device is including scalable and rotatable step motor, advance the sample dish of being connected with the step motor transmission, a plurality of hollow capillary sampling probe, power, a plurality of detection window and waste liquid pond, it is equipped with a plurality of workspace to advance along circumference on the sample dish, all inserts in every workspace and is equipped with three and regard as sample cell, liquid storage tank and clean water reservoir and removable centrifuging tube respectively, three centrifuging tube arrange along circumference according to the order of sample cell, liquid storage tank and clean water reservoir, capillary sampling probe sets up along the axis direction that is on a parallel with advances the sample dish, the portable centrifuge tube that stretches into of sample end of capillary sampling probe, detection window sets up on capillary sampling probe, capillary sampling probe's exit end inserts in the waste liquid pond, an utmost point of power and capillary sampling probe's entrance point electricity are connected, the other pole is electrically connected with the outlet end of the capillary sampling probe to form a loop.
The stepping motor intermittently rotates at a speed of not more than 10rad/s, the intermittent time is controlled by a microcomputer or a singlechip in the stepping motor and is matched with the stretching speed, stretching is carried out in the rotating intermittent, and the specific rotating speed is adjusted according to actual operation.
The sample feeding disc is circular, and a shaft of the stepping motor and the sample feeding disc are concentrically arranged.
The centrifuging tube passes through the elastic component to be fixed on advancing the sample dish, the elastic component can be the rubber circle, the size of elastic component can be: the thickness is 0.8-1 mm, the bore is 0.2-1 mm, and the length is cut according to the size of the centrifuge tube.
The centrifugal tube is in an inverted cone shape.
The volume of the centrifugal tube is 30-150 mu L.
And a pore is reserved at the head part of the centrifugal tube and used for inserting a capillary sampling probe.
Be equipped with the platinum electrode on the entrance point lateral wall of capillary sampling probe, when the device carries out work, fill liquid in the centrifuging tube, capillary sampling probe and platinum electrode all stretch into in the liquid, capillary sampling probe and platinum electrode all keep away from the interior pipe wall of centrifuging tube, avoid polluting. The platinum electrode can be directly adhered with the capillary sampling probe through glue.
And an electrode is also arranged at the outlet end of the capillary sampling probe, and the electrode positioned at the outlet end is directly placed in the waste liquid pool.
The capillary sampling probe adopts a fused quartz capillary or a high molecular polymer capillary, the capillary has high strength and elasticity, and the separation speed can be accelerated by the smooth inner surface like a mirror.
The inner diameter of the capillary sampling probe is 5-250 mu m.
The axial length of the capillary sampling probe is 20-100 mm.
The power supply is driven by a voltage-adjustable direct-current electric field, the voltage range is 0-30kV, and a common high-voltage power supply on the market is adopted.
The detection window is electrically connected with an external data processing center through an optical fiber, and data is output for processing. The number of detection windows is consistent with the number of capillary sampling probes. The optical fiber detection technology is adopted to detect data, so that the detection speed is high, multiple groups of data can be detected simultaneously, multi-channel simultaneous sample injection detection is realized, and data are output.
The number of the working areas is 3-10.
The device still includes the fixed plate that is on a parallel with advance kind dish and interval set up, be equipped with a plurality of through-holes that supply capillary sampling probe to run through on the fixed plate, can avoid capillary sampling probe slope.
When the capillary tube sampling probe works, one pole of a high-voltage power supply is connected with an outlet end electrode of the capillary tube sampling probe through a lead, the other pole of the high-voltage power supply is connected with an inlet end electrode, and the connection modes (namely positive and negative voltage poles) of the two poles are switched according to the charge of a sample to be analyzed. The step motor rotates and stretches according to the working process, the capillary sampling probe is used as a separation channel, and a sample marked by fluorescent substances is separated in the capillary sampling probe under the action of an electric field.
Compared with the prior art, the invention has the following advantages:
1. the invention adopts the replaceable centrifuge tube to replace the original liquid storage tank, thereby avoiding the pollution of different samples;
2. the invention adopts multi-channel simultaneous detection, which can greatly improve the detection speed;
3. the invention can realize automatic sample introduction and capillary electrophoresis technology, can realize the effects of low consumption and high flux, and has good application prospect.
Drawings
FIG. 1 is a schematic structural diagram of a multi-channel automatic sample injection device;
FIG. 2 is a schematic structural view of the connection between the sample plate and the stepping motor;
FIG. 3 is an isometric view of a sample tray;
FIG. 4 is a top view of a sample tray;
FIG. 5 is a schematic view of a centrifuge tube;
FIG. 6 is a schematic diagram of a capillary sampling probe;
fig. 7 is an enlarged view of a portion a in fig. 6.
In the figure: 1-a sample tray; 11-a working area; 111-a sample cell; 112-a liquid storage tank; 113-a clean water tank; 2-a step motor; 3-capillary sampling probe; 31-platinum electrode; 32-a fixed plate; 4-a power supply; 5-detection window; 6-waste liquid pool.
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments.
Examples
As shown in fig. 1, 2, 3, 4, 5, 6, 7, a multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis comprises a retractable and rotatable stepping motor 2, a sample introduction disc 1 in transmission connection with the stepping motor 2, a plurality of hollow capillary sampling probes 3, a power supply 4, a plurality of detection windows 5 and a waste liquid pool 6, wherein the sample introduction disc 1 is circumferentially provided with a plurality of working areas 11, each working area 11 is inserted with three replaceable centrifuge tubes which are respectively used as a sample pool 111, a liquid pool 112 and a water purification pool 113, the three centrifuge tubes are circumferentially arranged according to the sequence of the sample pool 111, the liquid pool 112 and the water purification pool 113, the centrifuge tubes are fixed on the sample introduction disc 1 through elastic members (which can be rubber rings and are omitted in the figure), the centrifuge tubes are in an inverted cone shape (as shown in fig. 5), the capillary sampling probes 3 are arranged along the axial direction parallel to the sample introduction disc 1, the centrifuging tube is stretched into to capillary sampling probe 3's sample end is movably, and detection window 5 sets up on capillary sampling probe 3, carries out the electricity through optic fibre and external data processing center and connects, and capillary sampling probe 3's exit end inserts in waste liquid pond 6, and one utmost point of power 4 and capillary sampling probe 3's entrance point electricity are connected, and another utmost point and capillary sampling probe 3's exit end electricity is connected and is formed the return circuit. The sample plate 1 is circular, and the shaft of the stepping motor 2 is arranged concentrically with the sample plate 1 (as shown in fig. 4, a circle at the center is a region connected with the shaft of the stepping motor 2). Be equipped with platinum electrode 31 (as shown in fig. 6, 7) on the entrance point lateral wall of capillary sampling probe 3, when the device worked, contain liquid in the centrifuging tube, capillary sampling probe 3 and platinum electrode 31 all stretch into liquid, and capillary sampling probe 3 and platinum electrode 31 all keep away from the interior pipe wall of centrifuging tube. The position of the multi-channel automatic sampling device during working is selected according to actual conditions.
Wherein, capillary sampling probe 3 adopts fused silica capillary or high molecular polymer capillary, and power 4 adopts the power by the drive of 0-30 kV's voltage direct current electric field, and the workspace 11 sets up 3 ~ 10 altogether, sets up to three in this embodiment, and the volume of centrifuging tube is 30 ~ 150 mu L, and capillary sampling probe 3's internal diameter is 5 ~ 250 mu m, and capillary sampling probe 3's axial length is 20 ~ 100 mm.
The stepping motor rotates intermittently at a speed not more than 10rad/s, the intermittent time is controlled by a microcomputer or a singlechip in the stepping motor and is matched with the stretching speed, and stretching is carried out in the rotating intermittent.
The device also comprises a fixing plate 32 which is parallel to the sample injection disc 1 and is arranged at intervals, and a plurality of through holes for the capillary sampling probes 3 to penetrate through are arranged on the fixing plate 32.
Before sample introduction, after all components are assembled in the sequence shown in the figure, samples, buffer solution and cleaning liquid are sequentially and respectively filled in three centrifuge tubes (comprising a sample cell 111, a liquid storage cell 112 and a clear water cell 113) in a sample introduction disc 1, then a stepping motor 2 and a power supply 4 are started, the sample introduction disc 1 is driven by the stepping motor 2 to rotate at a fixed angular speed, stops after rotating to a certain angle and extends out towards the direction of a capillary tube sampling probe 3, the inlet end of the capillary tube sampling probe 3 enters the sample cell 111 to be in contact with the samples in the sample cell 111, a certain voltage is applied to two ends of the capillary tube sampling probe 3 through the power supply 4, the samples are introduced under the action of an electric field, after sample introduction is completed, simultaneously, the capillary tube sampling probe 3 is aligned to the liquid storage cell 112 after the stepping motor 2 retracts and rotates at the fixed angular speed, and then extends out again, make the entrance point of capillary sampling probe 3 and the contact of liquid storage tank 112 accomplish and advance the appearance, the sample that adopts the fluorescent substance mark separates in capillary sampling probe 3 under the electric field effect, through detection window 5, adopt the optical fiber detection technique to accomplish the detection, the analysis data, the exit end intercommunication of capillary sampling probe 3 has waste liquid pool 6, the sample waste liquid etc. of accomplishing the detection are arranged into waste liquid pool 6 in, then step motor 2 passes through the flexible rotation of third wheel, the entrance point and the water purification pond 113 contact of capillary sampling probe 3, wash, the waste liquid of having washd also arranges into waste liquid pool 6.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Claims (10)

1. The utility model provides a multichannel autoinjection device for capillary electrophoresis fluorescence analysis, a serial communication port, the device is including scalable and rotatable step motor (2), with step motor (2) transmission connection advance sample dish (1), a plurality of hollow capillary sampling probe (3), power (4), a plurality of detection window (5) and waste liquid pond (6), it is equipped with a plurality of workspace (11) along circumference to advance sample dish (1), all inserts in every workspace (11) and is equipped with three and regard as sample cell (111), liquid reserve pool (112) and water purification pool (113) and removable centrifuging tube respectively, and three centrifuging tube are arranged along circumference according to the order of sample cell (111), liquid reserve pool (112) and water purification pool (113), capillary sampling probe (3) set up along the axis direction that is on a parallel with advance sample dish (1), the sample end of capillary sampling probe (3) is movably stretched into the centrifuging tube, detection window (5) set up on capillary sampling probe (3), the exit end of capillary sampling probe (3) is inserted in waste liquid pond (6), the entrance point electricity of a utmost point and capillary sampling probe (3) of power (4) is connected, and the exit end electricity of another utmost point and capillary sampling probe (3) is connected and is formed the return circuit.
2. The multi-channel autosampler device for capillary electrophoresis fluorescence analysis according to claim 1, characterized in that the stepper motor (2) rotates intermittently at a speed not larger than 10 rad/s.
3. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, wherein the sample introduction disk (1) is circular, and the shaft of the stepping motor (2) is concentrically arranged with the sample introduction disk (1).
4. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, wherein the centrifuge tube is fixed on the sample introduction disc (1) through an elastic member;
the centrifugal tube is in an inverted cone shape.
5. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, wherein a platinum electrode (31) is disposed on an outer side wall of an inlet end of the capillary sampling probe (3), when the device is in operation, the centrifuge tube contains a liquid, the capillary sampling probe (3) and the platinum electrode (31) both extend into the liquid, and the capillary sampling probe (3) and the platinum electrode (31) both are far away from an inner tube wall of the centrifuge tube.
6. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, wherein the capillary sampling probe (3) is a fused silica capillary or a high molecular polymer capillary.
7. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, wherein the capillary sampling probe (3) has an inner diameter of 5-250 μm and an axial length of 20-100 mm.
8. The multi-channel autosampler device for capillary electrophoresis fluorescence analysis according to claim 1, characterized in that the power supply (4) is driven by an adjustable voltage dc electric field of 0-30 kV.
9. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, wherein the detection window (5) is electrically connected with an external data processing center through an optical fiber.
10. The multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis according to claim 1, further comprising a fixing plate (32) parallel to the sample introduction plate (1) and arranged at intervals, wherein a plurality of through holes for the capillary sampling probes (3) to pass through are formed on the fixing plate (32).
CN202110429173.5A 2021-04-21 2021-04-21 Multi-channel automatic sample introduction device for capillary electrophoresis fluorescence analysis Pending CN113252763A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114280254A (en) * 2021-12-29 2022-04-05 河北香宇肉类制品有限公司 Edible oil detection device and method
CN115106141A (en) * 2022-05-30 2022-09-27 上海应用技术大学 Novel space-configuration space continuous flow microfluidic chip

Citations (5)

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Publication number Priority date Publication date Assignee Title
US20040112749A1 (en) * 2001-03-29 2004-06-17 Takehiko Shibasaki Electrophoresis device
US20070163882A1 (en) * 2006-01-16 2007-07-19 Hitachi High-Technologies Capillary electrophoresis apparatus and electrophoresis method
CN101165472A (en) * 2006-10-20 2008-04-23 中国科学院大连化学物理研究所 Multiple-pass mode capillary tube electrophoresis device
CN101398436A (en) * 2008-09-27 2009-04-01 东北大学 Rotating micro-example auto-introducing device
CN103018310A (en) * 2012-12-14 2013-04-03 凯晶生物科技(苏州)有限公司 Capillary chip electrophoresis structure for implementing multiple sample injection and implementation method thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040112749A1 (en) * 2001-03-29 2004-06-17 Takehiko Shibasaki Electrophoresis device
US20070163882A1 (en) * 2006-01-16 2007-07-19 Hitachi High-Technologies Capillary electrophoresis apparatus and electrophoresis method
CN101165472A (en) * 2006-10-20 2008-04-23 中国科学院大连化学物理研究所 Multiple-pass mode capillary tube electrophoresis device
CN101398436A (en) * 2008-09-27 2009-04-01 东北大学 Rotating micro-example auto-introducing device
CN103018310A (en) * 2012-12-14 2013-04-03 凯晶生物科技(苏州)有限公司 Capillary chip electrophoresis structure for implementing multiple sample injection and implementation method thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114280254A (en) * 2021-12-29 2022-04-05 河北香宇肉类制品有限公司 Edible oil detection device and method
CN114280254B (en) * 2021-12-29 2024-04-30 河北香宇肉类制品有限公司 Edible oil detection device and method
CN115106141A (en) * 2022-05-30 2022-09-27 上海应用技术大学 Novel space-configuration space continuous flow microfluidic chip

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