CN113391010B - Sample introduction method of clinical automatic analysis instrument - Google Patents
Sample introduction method of clinical automatic analysis instrument Download PDFInfo
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- CN113391010B CN113391010B CN202110775691.2A CN202110775691A CN113391010B CN 113391010 B CN113391010 B CN 113391010B CN 202110775691 A CN202110775691 A CN 202110775691A CN 113391010 B CN113391010 B CN 113391010B
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- 238000000034 method Methods 0.000 title claims abstract description 19
- 238000004458 analytical method Methods 0.000 title abstract description 6
- 239000007788 liquid Substances 0.000 claims abstract description 53
- 238000001514 detection method Methods 0.000 claims abstract description 33
- 238000002347 injection Methods 0.000 claims abstract description 30
- 239000007924 injection Substances 0.000 claims abstract description 30
- 238000005406 washing Methods 0.000 claims abstract description 19
- 238000005086 pumping Methods 0.000 claims abstract description 4
- 239000000523 sample Substances 0.000 claims description 86
- 238000011534 incubation Methods 0.000 claims description 22
- 238000004140 cleaning Methods 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 3
- 239000012723 sample buffer Substances 0.000 claims description 2
- 239000011324 bead Substances 0.000 abstract description 12
- 239000012535 impurity Substances 0.000 abstract description 4
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 238000005070 sampling Methods 0.000 description 2
- 229910000838 Al alloy Inorganic materials 0.000 description 1
- 229910001069 Ti alloy Inorganic materials 0.000 description 1
- 229910000963 austenitic stainless steel Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- 230000005415 magnetization Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/24—Automatic injection systems
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
The invention discloses a sample introduction method of a clinical automatic analysis instrument, which comprises the following steps: 1) Buffer storage and sample introduction: closing the on-off valve, adjusting the three-way valve to enable the liquid discharge pipe to be communicated with the second branch pipe, starting the magnetic suction filter, starting the injection pump, and sucking the sample into the buffer quantitative ring through the liquid transfer needle; 2) Injecting a sample into the detection unit: adjusting the three-way valve to enable the first branch pipe and the second branch pipe to be communicated, and pumping the sample in the buffer quantitative ring into the detection unit through the injection pump; 3) Needle washing: and adjusting the three-way valve to enable the liquid discharge pipe to be communicated with the second branch pipe, opening the on-off valve, and starting the suction pump to clean the second branch pipe, the buffer quantitative ring, the liquid discharge pipe, the magnetic suction filter and the inner cavity of the liquid transfer needle. The invention has accurate sample introduction and convenient operation, relieves the contradiction that the sample outlet frequency of the pretreatment unit is not matched with the processing speed of the detection unit, and reduces the damage probability of the sample containing impurities and residual magnetic beads on the detection equipment.
Description
Technical Field
The invention relates to the technical field of mass spectrometry detection equipment, in particular to a sample introduction method of a clinical automatic analyzer.
Background
The liquid chromatography tandem mass spectrometry (LC-MS/MS) can increase additional analysis capability, and can accurately identify and quantify trace compounds in complex sample matrixes such as cell and tissue lysate, blood, plasma, urine, oral fluid and the like, so that the method is increasingly applied to clinical application. The sample needs to be pretreated before LC-MS/MS detection. The magnetic bead method is a novel LC-MS/MS pretreatment method, and comprises the steps of adding a reagent containing magnetic beads into a sample for incubation, separating a target substance in the sample by the magnetic beads, and then sampling. However, the magnetic beads in the sample are inevitably left after the separation of the magnetic beads, and a certain amount of magnetic beads are left in the sample, which shortens the life of the liquid phase column for subsequent detection. In addition, the sampling frequency of the pretreatment unit and the processing frequency of the LC-MS/MS detection unit are difficult to be consistent, and the two have the flux matching problem, so that the normal work of the detection unit is influenced to a certain extent.
Disclosure of Invention
In order to solve the problems, the invention provides a sample introduction method of a clinical automatic analyzer with compact structure, safety and high efficiency, which can specifically adopt the following technical scheme:
the sample introduction method of the clinical automatic analyzer of the invention carries out sample caching, filtering, sample introduction and needle washing through the sample introduction unit arranged between the front processing unit and the detection unit,
the sample introduction unit comprises:
the incubation buffer disk is used for loading the sample cup sent out by the pretreatment unit;
the liquid transferring needle is arranged on one side of the incubation buffer disk and is provided with an inlet end and an outlet end, the inlet end is used for extracting a sample in the reaction cup, and the outlet end is communicated with a liquid discharge pipe;
the magnetic suction filter is arranged on the liquid discharge pipe;
the three-way valve is provided with a first port, a second port and a third port, the first port is connected with an outlet of the liquid discharge pipe, the second port is communicated with a first branch pipe, the first branch pipe is communicated with a sample inlet of the detection unit, and the third port is communicated with a second branch pipe;
the injection pump is arranged at the outlet end of the second branch pipe and is provided with an accommodating cavity;
the buffer quantitative ring is arranged on the second branch pipe and is positioned between the three-way valve and the injection pump;
the needle washing tube is communicated with the accommodating cavity, an on-off valve is arranged on the needle washing tube, and a suction pump for sucking cleaning liquid is arranged at the other end of the needle washing tube;
the sample injection method comprises the following steps:
1) Buffer storage and sample introduction: closing the on-off valve, adjusting the three-way valve to enable the liquid discharge pipe to be communicated with the second branch pipe, starting the magnetic suction filter, starting the injection pump, and sucking the sample into the buffer quantitative ring through the liquid transfer needle; 2) Injecting a sample into the detection unit: adjusting the three-way valve to enable the first branch pipe and the second branch pipe to be communicated, and pumping the sample in the buffer quantitative ring into the detection unit through the injection pump; 3) Needle washing: and adjusting the three-way valve to enable the liquid discharge pipe to be communicated with the second branch pipe, opening the on-off valve, and starting the suction pump to complete cleaning of the second branch pipe, the cache quantitative ring, the liquid discharge pipe, the magnetic suction filter and the inner cavity of the liquid transfer needle.
A reaction cup manipulator is arranged between the incubation buffer disk and the pretreatment unit.
Incubation buffer disk including the incubation dish of fixed setting with be located its top around center pivoted and hold the dish, it puts the position to be provided with the sample cup on the dish to hold, the play appearance position department of incubation dish is provided with first magnetism and inhales the piece.
The magnetic suction filter comprises a barrel body communicated with a liquid discharge pipe, the barrel body is of a split structure, a filter sieve plate is arranged at the inlet end of the barrel body, and a second magnetic suction piece is arranged on the side face of the barrel body.
The cylinder and the filtering sieve plate are made of demagnetizing materials, and the second magnetic attraction piece is a sucker type electromagnet.
And a cleaning liquid tank is arranged below the suction pump.
And a cleaning and liquid draining station is arranged on one side of the liquid transferring needle.
And the sample inlet of the detection unit is communicated with a plurality of sample feeding channels.
The sample introduction method of the clinical automatic analyzer provided by the invention has the advantages that the sample introduction is accurate, the operation is convenient, the sample introduction unit is internally provided with the incubation buffer disk, the liquid transferring needle, the injection pump and the detection unit which are connected with the three-way valve, and the needle washing tube which is connected with the injection pump, so that the integration of sample caching, filtering, sample introduction and needle washing is realized, the contradiction that the sample outlet frequency of the pretreatment unit is not matched with the processing speed of the detection unit is relieved, and the damage probability of the sample containing impurities and residual magnetic beads to the detection equipment is reduced.
Drawings
Fig. 1 is a schematic structural view of an automatic clinical analyzer according to the present invention.
FIG. 2 is a schematic diagram of the structure of the incubation buffer tray in FIG. 1.
Fig. 3 is a schematic structural view of the magnetic filter in fig. 1.
FIG. 4 is a liquid flow diagram of the sample buffer state of the sample injection unit in FIG. 1.
FIG. 5 is a liquid flow diagram showing the state of the sample introduction unit in FIG. 1 introducing the sample into the detection unit.
FIG. 6 is a liquid flow diagram of the needle washing state of the sample injection unit in FIG. 1.
Detailed Description
The following describes embodiments of the present invention in detail with reference to the drawings, which are implemented on the premise of the technical solution of the present invention, and give detailed implementation manners and specific working procedures, but the scope of the present invention is not limited to the following embodiments.
As shown in fig. 1 to 6, the sample injection method of the clinical automatic analyzer according to the present invention performs sample buffering, filtering, sample injection, and needle washing through the sample injection unit 300 disposed between the front processing unit 100 and the detection unit 200.
The detection unit 200 is generally composed of a preceding liquid phase analysis module (LC) and a following Mass Spectrometer (MS), wherein a sample inlet of the liquid phase analysis module (LC) is communicated with a plurality of sample introduction channels.
The sample injection unit 300 comprises an incubation buffer tray 301, a cuvette manipulator 302, a pipette 303, a three-way valve 304, an injection pump 305, a syringe 306, and the like. The incubation buffer tray 301 comprises an annular bearing tray 301a and an incubation tray 301b fixedly arranged below the annular bearing tray, wherein the bearing tray 301a is connected with a stepping motor and can rotate around the center in a forward or reverse direction, and a plurality of sample cup placing positions are arranged on the bearing tray 301 a; the annular incubation disc 301b is provided with a constant temperature device for providing a constant temperature environment of 0-100 ℃ which is beneficial to sample incubation, a reaction cup manipulator 302 is arranged on one side of a sample inlet position, and a liquid transferring needle 303 is arranged on one side of a sample outlet position; the cuvette manipulator 302 is used to transfer cuvettes at the outlet of the pretreatment unit 100 onto the tray 301a, and when the cuvettes are rotated to the sample-out position, the sample in the cuvettes is extracted by the inlet end of the pipetting needle 303 and enters the drain pipe 307 whose outlet end is communicated with the pipetting needle 303.
The drain 307 is connected to the detection unit 200 and the syringe pump 305 through the three-way valve 304. The first port of the three-way valve 304 is connected to the outlet of the drain 307, the second port of the three-way valve 304 is connected to the detection unit 200 through a first branch 308, and the third port of the three-way valve 304 is connected to the syringe pump 305 through a second branch 309. The syringe pump 305 has a receiving chamber, and a piston is installed at one end of the receiving chamber, and the volume of the receiving chamber can be controlled by pushing and pulling the piston by external force. Further, for the purpose of precise sample injection, a buffer quantitative ring 310 is installed on the second branch pipe 309. When the conducting drain pipe 307 and the second branch pipe 309 are conducted, the suction and temporary storage of the sample (or the washing of the needle) can be realized; when the second branch tube 309 and the first branch tube 308 are conducted, the sample can be temporarily stored in the detection unit 200. The change of the pipeline conducting state can be realized by adjusting the valve core direction of the three-way valve 304.
The holding cavity of the syringe pump 305 is also communicated with a syringe 306, the syringe 306 is provided with an on-off valve 311, and the other end of the syringe 306 is connected with the liquid supply mechanism. The liquid supply mechanism is generally composed of a cleaning liquid tank 312 and a suction pump 313, wherein the suction pump 313 is mounted on the syringe 306.
In order to reduce the damage of the sample containing impurities and residual magnetic beads to the detection device, a filtering mechanism is further disposed in the sample introduction unit, and includes a first magnetic attraction element 314 mounted at the sample outlet position of the incubation tray 301b and a magnetic attraction filter 315 mounted on the drain pipe 307. The first magnetic attraction piece 314 is only required to be a magnet, and can adsorb magnetic beads in a sample before the liquid transferring needle 303 samples the sample, so that the magnetic beads are prevented from entering the subsequent detection unit 200; the magnetic filter 315 includes a cylinder 315a connected to the drain pipe 307, the cylinder 315a is a split structure connected by bolts, a filter screen plate 315b with a pore size of 200nm is installed at an inlet end of the cylinder 315a for filtering impurity particles with a diameter of 200nm or more in the sample, a suction cup type electromagnet is installed at a side surface of the cylinder 315a as a second magnetic member 315c, and under an electrified condition, the magnetic filter can further adsorb residual magnetic beads in the sample. The cylinder 315a and the filtering screen plate 315b are made of demagnetizing materials (such as aluminum alloy, titanium alloy, austenitic stainless steel, etc.), so as to avoid magnetization.
In order to discharge the waste needle washing liquid, a washing and draining station 316 is arranged on one side of the liquid transferring needle 303.
The sample injection working process of the invention is divided into three stages: 1) Buffer storage and sample introduction: closing the on-off valve 311, adjusting the three-way valve 304 to conduct the liquid discharge pipe 307 and the second branch pipe 309, starting the magnetic suction filter 315, starting the injection pump 305, and sucking the sample into the buffer quantitative ring 310 through the liquid transfer needle 303; 2) Injecting a sample into the detection unit: adjusting the three-way valve 304 to conduct the first branch pipe 308 and the second branch pipe 309, and driving the sample in the buffer quantitative ring 310 into the detection unit 200 through the injection pump 305; 3) Needle washing: the three-way valve 304 is adjusted to conduct the liquid discharge pipe 307 and the second branch pipe 309, the on-off valve 311 is opened, the suction pump 313 is started, and the cleaning of the inner cavity of the second branch pipe 309, the buffer quantitative ring 310, the liquid discharge pipe 307, the magnetic suction filter 315 and the liquid transfer needle 303 is completed.
The method comprises the following specific steps:
1) Buffer sample introduction
a. The second magnetic member 315c is energized, the on-off valve 311 is closed, and the three-way valve 304 is adjusted to conduct the drain pipe 307 and the second branch pipe 309;
b. inserting the inlet end of the pipetting needle 303 into a sample cup at the sample outlet position of the carrier tray 301a, starting the injection pump 305 to suck, and enabling the sample to enter the buffer quantitative ring 310 after passing through the magnetic suction filter 315;
c. the syringe pump 305 is turned off;
2) Introduction of sample into the detection unit
a. Adjusting the three-way valve 304 to make the first branch pipe 308 and the second branch pipe 309 conduct;
b. starting the injection pump 305, and pumping the sample in the buffer quantitative ring 310 into the detection unit 200;
c. the syringe pump 305 is turned off;
3) Needle washing device
a. Opening the on-off valve 311 to cut off the second magnetic attraction piece 315c, rotating the inlet end of the pipetting needle 303 to the position above the cleaning and draining station 316, and adjusting the three-way valve 304 to conduct the draining pipe 307 and the second branch pipe 309;
b. starting a suction pump 313 to suck the cleaning liquid in the cleaning liquid tank 312, and completing cleaning of the inner cavities of the second branch pipe 309, the buffer quantitative ring 310, the liquid discharge pipe 307, the magnetic suction filter 315 and the liquid transfer needle 303;
c. and (5) closing the suction pump 313 to complete all sample feeding work.
It should be noted that in the description of the present invention, terms of orientation or positional relationship such as "front", "rear", "left", "right", "vertical", "horizontal", "inner", "outer", etc., are based on the orientation or positional relationship shown in the drawings, and are only for convenience of description and simplification of description, but do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.
Claims (4)
1. The utility model provides a kind method of advancing of clinical autoanalyzer, advances the appearance unit through setting up between preceding processing unit and detecting element and carries out sample buffer memory, filtration, advance kind and wash the needle, its characterized in that:
the sample introduction unit comprises:
the incubation buffer disk is used for loading the sample cup sent out by the pretreatment unit;
the liquid transferring needle is arranged on one side of the incubation buffer disk and is provided with an inlet end and an outlet end, the inlet end is used for extracting a sample in the reaction cup, and the outlet end is communicated with a liquid discharge pipe;
the magnetic suction filter is arranged on the liquid discharge pipe;
the three-way valve is provided with a first port, a second port and a third port, the first port is connected with an outlet of the liquid discharge pipe, the second port is communicated with a first branch pipe, the first branch pipe is communicated with a sample inlet of the detection unit, and the third port is communicated with a second branch pipe;
the injection pump is arranged at the outlet end of the second branch pipe and is provided with an accommodating cavity;
the buffer quantitative ring is arranged on the second branch pipe and is positioned between the three-way valve and the injection pump;
the needle washing tube is communicated with the accommodating cavity, an on-off valve is arranged on the needle washing tube, and a suction pump for sucking cleaning liquid is arranged at the other end of the needle washing tube;
the sample injection method comprises the following steps:
1) Buffer storage and sample introduction: closing the on-off valve, adjusting the three-way valve to enable the liquid discharge pipe to be communicated with the second branch pipe, starting the magnetic suction filter, starting the injection pump, and sucking the sample into the buffer quantitative ring through the liquid transfer needle; 2) Injecting a sample into the detection unit: adjusting the three-way valve to enable the first branch pipe and the second branch pipe to be communicated, and pumping the sample in the buffer quantitative ring into the detection unit through the injection pump; 3) Needle washing: adjusting the three-way valve to enable the liquid discharge pipe to be communicated with the second branch pipe, opening the on-off valve, and starting the suction pump to complete cleaning of the second branch pipe, the cache quantifying ring, the liquid discharge pipe, the magnetic suction filter and the inner cavity of the liquid transfer needle;
the incubation buffer storage disc comprises an incubation disc which is fixedly arranged and a bearing disc which is positioned above the incubation disc and rotates around the center, a sample cup placing position is arranged on the bearing disc, and a first magnetic suction piece is arranged at a sample outlet position of the incubation disc;
the magnetic filter comprises a cylinder body communicated with the liquid discharge pipe, the cylinder body is of a split structure, a filter sieve plate is arranged at the inlet end of the cylinder body, and a second magnetic part is arranged on the side surface of the cylinder body;
the cylinder body and the filtering sieve plate are made of demagnetizing materials, and the second magnetic attraction piece is a sucker type electromagnet;
the sample inlet of the detection unit is communicated with a plurality of sample feeding channels.
2. The sample injection method for clinical automatic analyzer according to claim 1, wherein: a reaction cup manipulator is arranged between the incubation buffer disk and the pretreatment unit.
3. The sample injection method of the clinical automatic analyzer according to claim 1, wherein: and a cleaning liquid tank is arranged below the suction pump.
4. The sample injection method for clinical automatic analyzer according to claim 1, wherein: and a cleaning and liquid draining station is arranged on one side of the liquid transferring needle.
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| CN202110775691.2A CN113391010B (en) | 2021-07-07 | 2021-07-07 | Sample introduction method of clinical automatic analysis instrument |
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| CN202110775691.2A CN113391010B (en) | 2021-07-07 | 2021-07-07 | Sample introduction method of clinical automatic analysis instrument |
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| CN116116472A (en) * | 2023-02-03 | 2023-05-16 | 中国科学技术大学 | Liquid drop size self-adjusting sample injector |
| CN119985802A (en) * | 2025-04-14 | 2025-05-13 | 凯莱谱科技股份有限公司 | Chromatographic device and chromatographic analysis method |
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| US5200084A (en) * | 1990-09-26 | 1993-04-06 | Immunicon Corporation | Apparatus and methods for magnetic separation |
| CA2183638C (en) * | 1995-03-20 | 2001-03-20 | Tajima Hideji | Liquid processing method making use of pipette device and apparatus for same |
| US20040021073A1 (en) * | 2002-04-12 | 2004-02-05 | California Institute Of Technology | Apparatus and method for magnetic-based manipulation of microscopic particles |
| EP3490694A4 (en) * | 2016-07-31 | 2020-04-15 | Ancera Corp. | Multilayer disposable cartridge for ferrofluid-based assays and method of use |
| WO2018234115A1 (en) * | 2017-06-19 | 2018-12-27 | Lab-On-A-Bead Ab | Combinatory separation |
| WO2019014306A2 (en) * | 2017-07-13 | 2019-01-17 | Argos Therapeutics, Inc. | Automated processing systems, apparatus, methods and related devices |
| CN111033265B (en) * | 2017-09-20 | 2024-05-17 | 深圳迈瑞生物医疗电子股份有限公司 | Automatic analysis device and working method thereof |
| CN108414737B (en) * | 2018-06-13 | 2024-06-04 | 安徽为臻生物工程技术有限公司 | Full-automatic chemiluminescence immunoassay instrument |
| CN109100458B (en) * | 2018-10-16 | 2020-10-23 | 安徽皖仪科技股份有限公司 | Automatic chromatographic sample injector and automatic sample injection method |
| CN109959742B (en) * | 2019-04-25 | 2024-02-20 | 天津海关动植物与食品检测中心 | Online analysis pretreatment and offline analysis integrated liquid chromatograph and use method thereof |
| CN109959741B (en) * | 2019-04-25 | 2024-02-23 | 天津海关动植物与食品检测中心 | Dual-channel online analysis pretreatment and offline analysis integrated liquid chromatograph and application method thereof |
| CN209821138U (en) * | 2019-04-29 | 2019-12-20 | 安徽皖仪科技股份有限公司 | Sample injection needle cleaning system of liquid chromatogram autosampler |
| CN111999512A (en) * | 2019-05-11 | 2020-11-27 | 南京岚煜生物科技有限公司 | Chemiluminescence immunity analyzer |
| US20210008566A1 (en) * | 2019-07-14 | 2021-01-14 | Zhuhai Sanmed Biotech Ltd | System and processes for isolation and enrichment by magnetic separation |
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