CN106841376A - A kind of micro-fluidic on-line preconcentration MALDI TOF detection means and method - Google Patents
A kind of micro-fluidic on-line preconcentration MALDI TOF detection means and method Download PDFInfo
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- CN106841376A CN106841376A CN201611259831.6A CN201611259831A CN106841376A CN 106841376 A CN106841376 A CN 106841376A CN 201611259831 A CN201611259831 A CN 201611259831A CN 106841376 A CN106841376 A CN 106841376A
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- tof
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- G01N27/64—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode using wave or particle radiation to ionise a gas, e.g. in an ionisation chamber
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/041—Connecting closures to device or container
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
Abstract
The present invention relates to a kind of micro-fluidic on-line preconcentration MALDI TOF detection means and method, the device includes：The MALDI TOF target plates of some target spots are provided with, the target spot is provided with enrichment material layer；It is provided with microfluidic channel, its geomery and MALDI TOF the target plates micro-fluidic cover plate for coordinating and the micro fluidic device that mobile phase is enriched with for loading and control.The present invention carries out on-line sample pretreatment by micro-fluidic mode, can effectively save the MALDI TOF example enrichments of a large amount of very complicateds, elutes, the cumbersome pretreatment operation such as desalination, effectively time-consuming, improves detection efficiency and Detection accuracy.
Description
Technical field
The invention belongs to field of mass spectrometry, and in particular to a kind of micro-fluidic on-line preconcentration MALDI-TOF detection means and method.
Background technology
In complex biological sample, many low abundance materials carry a large amount of disease biomarkers, therefore also become in recent years
Carry out the focus of attention in the field such as biological, chemical.MALDI-TOF is a kind of high-efficiency and high-resolution of component detection in complex biological sample
Rate technology, full name is Matrix Ass i sted Laser Desorpt ion Ioni zat ion Time of Fl ight
Mass Spectrometry, i.e. matrix solid-dispersion flight time mass spectrum.This technology this year is fast to develop
Speed, overall technology means are very ripe, have been widely used for the multiple fields such as drug development, bioanalysis, chemical detection, together
When also further popularized to other every field.
For the detection of the MALDI-TOF of complex biological sample, first have to pre-process complex biological sample, it is rich through material
Collection, rinses, and by sample elution, then by eluent point target, adds matrix, sample is formed cocrystallization with matrix, then target plate is put
Enter in sample room, into vacuum environment, laser irradiation, sample occurs electric charge transfer, parses a series of processes such as attached with matrix, makes
Ionized sample molecule, enters into separation detection in flight time mass spectrum.As can be seen here, the pretreatment before vacuum environment is being entered
Step is very cumbersome, is not easy to quick detection, and the extensive examination of high flux.While the MALDI-TOF targets of commercialization at present
Plate is mainly used reuses target plate, and is generally cleaning by hand, further increases workload, takes and tend to have residual.
In view of drawbacks described above, the present invention is by research and innovation, it is intended that development and design one kind effectively convenient can carry out complexity
The process of biological sample pretreatment, saves enrichment, elutes, the troublesome operation such as desalination.Detection time can be effectively saved, inspection is improved
Efficiency is surveyed, strengthens detection accuracy.
The content of the invention
To solve drawbacks described above, the invention provides a kind of micro-fluidic on-line preconcentration MALDI-TOF detection means and method,
Simplify pretreatment process, save the troublesome operations such as enrichment, wash-out, desalination, save detection time, improve detection efficiency.
A kind of micro-fluidic on-line preconcentration MALDI-TOF detection means that the present invention is provided includes：
MALDI-TOF target plates, the MALDI-TOF target plates are provided with some target spots, and the target spot is provided with enrichment material
Layer；
Micro-fluidic cover plate, its front side is provided with some microfluidic channels and the collecting with the microfluidic channel insertion
Chamber, the host cavity coordinates the annular seal space that mobile phase is housed for being formed with the target spot；Stream is provided with the micro-fluidic cover plate
It is dynamic mutually to export, for waste liquid after exclusion enrichment；
Microfluidic channel insertion on micro fluidic device, with the micro-fluidic cover plate is used for loading and enrichment mobile phase.
In the present invention, the MALDI-TOF target plates agree with according to sample room situation controllable adjustment, its shape with sample room；
The enrichment material composition is according to specific detection object controlled adjustment；
Micro-fluidic cover plate, leaves the cover plate of microfluidic channel, and its geomery agrees with MALDI-TOF target plates；
Micro fluidic device, for loading and on-line preconcentration flowing phase control.
In the present invention, it can be any conductive material, preferably conductive plastics or stainless steel that the target plate prepares material.
In the present invention, target spot size is 1mm~5mm on the target plate.
In the present invention, target spot quantity is 3~200 on the target plate.
In the present invention, enrichment material is functional porous material, and surface-functionalized group can be received for different enrichment targets
Control adjustment.
In the present invention, enrichment material material overall dimensions are more than microfluidic channel.
In the present invention, micro-fluidic slab passageway size is 0.5 μm~500 μm.
In the present invention, end is provided with mobile phase outlet after micro-fluidic cover plate enrichment, for waste liquid after discharge enrichment.
In the present invention, micro fluidic device is engaged with micro-fluidic cover plate, and for applied sample amount, the parameter such as flow rate of mobile phase can
Control.
In the present invention, micro fluidic device can realize the micro-fluidic effects device, preferably micro perseverance for any in form
Stream syringe pump.
Detection method of the invention is specific as follows：The micro-fluidic cover plate is placed on MALDI-TOF target plates correspondence target
Point position, through micro fluidic device loading, object is directly adsorbed in enrichment material surface, is rinsed through mobile phase, discharges waste liquid；Take
Under micro-fluidic cover plate, on target spot directly be added dropwise Mass Spectrometer Method matrix, formed cocrystallization film；Mass spectrum sample room is put into, directly
Detection.
Compared with prior art, the device have the advantages that being：
1st, with reference to micro-fluidic and MALDI-TOF technologies, on-line preconcentration is carried out, improves detection efficiency；
2nd, specific enrichment is carried out to object using enrichment material, improves detection resolution, clear part is disturbed；
3rd, the cumbersome pre-treatment step such as elution samples and desalination is saved, simplifies experimental procedure, improve efficiency；
4th, unartificial loading, using micro-fluidic operation, is easy to repeated experiment, improves detection repeatability.
Brief description of the drawings
Fig. 1 is micro-fluidic on-line preconcentration MALDI-TOF structure of the detecting device schematic diagrames of the invention.
Fig. 2 is micro-fluidic cover plate schematic diagram of the invention.
Component label instructions
MALDI-TOF target plates 1
Enrichment material layer 2
Micro-fluidic cover plate 3
Microfluidic channel 4
Mobile phase outlet 5
Micro fluidic device 6
Specific embodiment
Embodiments of the present invention are illustrated below by way of specific instantiation, those skilled in the art can be by this specification
Disclosed content understands other advantages of the invention and effect easily.The present invention can also be by specific realities different in addition
The mode of applying is embodied or practiced, the various details in this specification can also based on different viewpoints with application, without departing from
Various modifications or alterations are carried out under spirit of the invention.
Below explanation describes embodiment of the invention in detail and instructs how those skilled in the art realize this
The reproduction of invention.In order to instruct technical scheme, some the conventional aspects for having simplified or having omitted.Those skilled in the art
It should be appreciated that deformation from these implementation methods or will within the scope of the invention.Those skilled in the art should understand that following
Feature can combine to form multiple deformations of the invention in a variety of ways.Thus, the invention is not limited in the optional reality of subordinate
Mode is applied, and only claim and their equivalent is limited.
Refer to shown in Fig. 1 and Fig. 2, the present invention provides a kind of micro-fluidic on-line preconcentration MALDI-TOF detection means, its bag
Include：MALDI-TOF target plates 1, the MALDI-TOF target plates are provided with some target spots, and the target spot is provided with enrichment material layer 2；
Micro-fluidic cover plate 3, its front side is provided with some microfluidic channels 4 and the receipts with the microfluidic channel insertion
Cavity volume, the host cavity coordinates the annular seal space that mobile phase is housed for being formed with the target spot；It is provided with the micro-fluidic cover plate 3
Mobile phase outlet 5, for waste liquid after exclusion enrichment；
Microfluidic channel insertion on micro fluidic device 6, with the micro-fluidic cover plate is used for loading and enrichment mobile phase.
The micro-fluidic on-line preconcentration MALDI-TOF target plates that the present invention is provided include：MALDI-TOF target plates are according to sample room feelings
Condition controllable adjustment, its shape is agreed with sample room；
The enrichment material composition of enrichment material layer is according to specific detection object controlled adjustment；
Micro-fluidic cover plate, leaves the cover plate in micro-fluidic duct, and its geomery agrees with MALDI-TOF target plates；
Micro fluidic device, for loading and on-line preconcentration flowing phase control.
In the present invention, it can be any conductive material, preferably conductive plastics or stainless steel that the target plate prepares material.Target plate
Upper target spot size is 1mm~5mm.Target spot quantity is 3~200.
In the present invention, enrichment material is functional porous material, and surface-functionalized group can be received for different enrichment targets
Control adjustment.Enrichment material material overall dimensions are more than microfluidic channel.
In the present invention, micro-fluidic slab passageway size is 0.5 μm~500 μm.End is provided with mobile phase outlet after enrichment, uses
In waste liquid after discharge discharge enrichment.
In the present invention, micro fluidic device is engaged with micro-fluidic cover plate, and for applied sample amount, the parameter such as flow rate of mobile phase can
Control.It is any to realize that the micro-fluidic effects device is all considered as viable means in form, preferably micro constant current syringe pump.
Present invention additionally comprises a kind of detection method of micro-fluidic on-line preconcentration MALDI-TOF, the method is comprised the following steps：
1) the micro-fluidic cover plate is placed on MALDI-TOF target plates, host cavity and target spot on the micro-fluidic cover plate
Position correspondence, the host cavity coordinates the annular seal space that mobile phase is housed for being formed with the target spot；
2) micro fluidic device loading is started, the object in sample is directly adsorbed in the enrichment material layer surface on target spot,
Starting micro fluidic device simultaneously carries out mobile phase flushing；
3) waste liquid after the mobile phase outlet discharge enrichment；
4) micro-fluidic cover plate is removed, Mass Spectrometer Method matrix is directly added dropwise on target spot, form cocrystallization film；
5) by step 4) after formed device be put into mass spectrum sample room detection.
Specifically, referring to shown in Fig. 1 to Fig. 2, micro-fluidic on-line preconcentration MALDI-TOF target plates of the invention include：
MALDI-TOF target plates 1, a series of controllable target spots of uniform arrangement number size on target plate, with filling on the target spot
Specific richness material 2.Micro-fluidic target plate 3 cover target spot number as needed parallel test number determine, by micro fluidic device 6
Drive after micro-fluidic target plate upper channel 4 enters through example enrichment, waste liquid is discharged from outlet 5.After being enriched with through material selectivity
Target plate cover plate is removed, addition matrix forms film of receiving a crime report altogether, is put into MALDI-TOF mass spectrum samples room, carries out sample detection.
So, the present invention effectively overcomes various shortcoming of the prior art and has high industrial utilization.
The above-described embodiments merely illustrate the principles and effects of the present invention, not for the limitation present invention.It is any ripe
The personage for knowing this technology all can carry out modifications and changes under without prejudice to spirit and scope of the invention to above-described embodiment.Cause
This, those of ordinary skill in the art is complete with institute under technological thought without departing from disclosed spirit such as
Into all equivalent modifications or change, should be covered by claim of the invention.
Claims (9)
1. a kind of micro-fluidic on-line preconcentration MALDI-TOF detection means, it is characterised in that：The detection means includes：
MALDI-TOF target plates, the MALDI-TOF target plates are provided with some target spots, and the target spot is provided with enrichment material layer；
Micro-fluidic cover plate, its front side is provided with some microfluidic channels and the host cavity with the microfluidic channel insertion,
The host cavity coordinates the annular seal space that mobile phase is housed for being formed with the target spot；Mobile phase is provided with the micro-fluidic cover plate
Outlet, for waste liquid after exclusion enrichment；
Microfluidic channel insertion on micro fluidic device, with the micro-fluidic cover plate is used for loading and enrichment mobile phase.
2. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：It is described
MALDI-TOF target plates are conductive plastics or stainless steel.
3. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：The target spot
It is circle, its a diameter of 1mm~5mm.
4. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：The target spot
Quantity is 3~200.
5. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：The enrichment
Material layer is functional porous material.
6. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：The enrichment
Scantling is more than microfluidic channel.
7. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：The miniflow
Control channel diameter is 0.5 μm~500 μm.
8. micro-fluidic on-line preconcentration MALDI-TOF detection means according to claim 1, it is characterised in that：The miniflow
Control device is micro constant current syringe pump.
9. the method that the device described in a kind of use claim 1 to 8 any one is detected, it is characterised in that：The method
Comprise the following steps：
1) the micro-fluidic cover plate is placed on MALDI-TOF target plates, host cavity and target position on the micro-fluidic cover plate
Correspondence, the host cavity coordinates the annular seal space that mobile phase is housed for being formed with the target spot；
2) micro fluidic device loading is started, the object in sample is directly adsorbed in the enrichment material layer surface on target spot, while
Starting micro fluidic device carries out mobile phase flushing；
3) waste liquid after the mobile phase outlet discharge enrichment；
4) micro-fluidic cover plate is removed, Mass Spectrometer Method matrix is directly added dropwise on target spot, form cocrystallization film；
5) by step 4) after formed device be put into mass spectrum sample room detection.
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| CN201611259831.6A CN106841376A (en) | 2016-12-30 | 2016-12-30 | A kind of micro-fluidic on-line preconcentration MALDI TOF detection means and method |
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| CN201611259831.6A CN106841376A (en) | 2016-12-30 | 2016-12-30 | A kind of micro-fluidic on-line preconcentration MALDI TOF detection means and method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109490536A (en) * | 2018-12-27 | 2019-03-19 | 南京大学 | A kind of affine in immunity micro-fluidic chip-mass spectrometry device and its application in Residue of Antibiotics in Milk analysis |
| CN111330655A (en) * | 2018-12-18 | 2020-06-26 | 深圳先进技术研究院 | Microfluidic chip for lipid compound detection and lipid compound detection method |
| CN113466320A (en) * | 2021-08-06 | 2021-10-01 | 河北省食品检验研究院 | MALDI-TOF detection method for bovine-derived components in milk powder |
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| US20040245102A1 (en) * | 2002-09-09 | 2004-12-09 | Gilbert John R. | Implementation of microfluidic components, including molecular fractionation devices, in a microfluidic system |
| CN1629640A (en) * | 2003-12-18 | 2005-06-22 | 中国科学院大连化学物理研究所 | Micro flow control chip with integrated dialysis function and preparing process thereof |
| CN101008594A (en) * | 2007-01-25 | 2007-08-01 | 重庆大学 | Micro-fluidic chip containing sample pre-treatment film and production method therefor |
| CN101067621A (en) * | 2007-06-05 | 2007-11-07 | 中国科学院上海微系统与信息技术研究所 | Production of micro-solid phase extraction chip based on micro-processing method and using method |
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| US20040099310A1 (en) * | 2001-01-05 | 2004-05-27 | Per Andersson | Microfluidic device |
| US20040245102A1 (en) * | 2002-09-09 | 2004-12-09 | Gilbert John R. | Implementation of microfluidic components, including molecular fractionation devices, in a microfluidic system |
| CN1629640A (en) * | 2003-12-18 | 2005-06-22 | 中国科学院大连化学物理研究所 | Micro flow control chip with integrated dialysis function and preparing process thereof |
| CN101008594A (en) * | 2007-01-25 | 2007-08-01 | 重庆大学 | Micro-fluidic chip containing sample pre-treatment film and production method therefor |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111330655A (en) * | 2018-12-18 | 2020-06-26 | 深圳先进技术研究院 | Microfluidic chip for lipid compound detection and lipid compound detection method |
| CN109490536A (en) * | 2018-12-27 | 2019-03-19 | 南京大学 | A kind of affine in immunity micro-fluidic chip-mass spectrometry device and its application in Residue of Antibiotics in Milk analysis |
| CN113466320A (en) * | 2021-08-06 | 2021-10-01 | 河北省食品检验研究院 | MALDI-TOF detection method for bovine-derived components in milk powder |
| CN113466320B (en) * | 2021-08-06 | 2022-10-14 | 河北省食品检验研究院 | MALDI-TOF detection method for bovine-derived components in milk powder |
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Application publication date: 20170613 |