CN205484232U - Micro -fluidic chip and little extraction system of online automatic chip magnetism solid phase - Google Patents
Micro -fluidic chip and little extraction system of online automatic chip magnetism solid phase Download PDFInfo
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- CN205484232U CN205484232U CN201620228337.2U CN201620228337U CN205484232U CN 205484232 U CN205484232 U CN 205484232U CN 201620228337 U CN201620228337 U CN 201620228337U CN 205484232 U CN205484232 U CN 205484232U
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Abstract
The utility model provides a micro -fluidic chip and online automatic chip magnetism solid phase micro -extraction liquid chromatogram inductively coupled plasma mass spectrometry system, this system contains the constant current syringe pump, aseptic syringe, the polyethylene pump line, the quartz capillary, high performance liquid chromatography, the chromatography post, the inductively coupled plasma mass spectrum, the control of self -control chip pneumatic valve, micro -fluidic chip, magnet, the six -way valve is constituteed with the ration ring, promote aseptic syringe through the polyethylene pump line with sample and the leading -in micro -fluidic chip of reagent by the constant current syringe pump, accomplish the sample treatment after the quartz capillary on the chip, the six -way valve detects with the leading -in liquid chromatogram inductively coupled plasma mass spectrum of ration ring, and use the control of self -control chip pneumatic valve to control the processing procedure on the chip. This system has low reagent / sample consumption, the repeatability is good, the height integrates and the automatic characteristics, very is applicable to the research of trace element morphological analysis among the cell sample.
Description
Technical field
This utility model belongs to analytical chemistry field, relates to a kind of micro-fluidic chip and a kind of on-line automaticization chip magnetic solid-phase microextraction system.
Background technology
Elemental Speciation Analysis is typically realized with the combination of high-sensitive element specific detection technology by efficient isolation technics.In many element specific detection devices, inductivity coupled plasma mass spectrometry (ICP-MS) has a prominent advantage, the most highly sensitive, detection limit is low, range of linearity width, but also is provided that isotope relevant information.It is the conventional means of Elemental Speciation Analysis by itself and chromatographic technique (such as HPLC, GC, CE) combination.Wherein, the analytical technology that HPLC Yu ICP-MS is combined is most study in mercury morphological analysis, be most widely used.But, use conventional HPLC-ICP-MS that the mercury shape in cell directly detects or also exists some problems: (1) cell sample volume is less, it is impossible to match with conventional H PLC sampling volume；(2) cellular matrix can cause matrix interference and Polyatomic ion；(3) needs of the Elemental Speciation Analysis of ultra trace during the sensitivity of instrument is insufficient for a small amount of cell.Small bore columns (small-bore columns) HPLC uses column dimension and the less chromatographic column of fixing phase particle diameter, this small bore columns is had sampling volume is little, chromatographic isolation degree high, back pressure is high and flow rate of liquid is lower feature.Accordingly, it is capable to preferably the volume with cell sample matches, reduce the impact that ICP source stabilization and Ionization Efficiency can be caused by the introducing of organic facies and salt simultaneously.As a example by micro column, 40 μ L min-1Flow rate of mobile phase decrease and introduce ionogenic substrate and greatly reduce the usage amount of reagent and salt and the introduction volume of organic solvent, be more beneficial for stablizing of plasma.But, latter two problems to be solved, then need to be aided with, before microHPLC-ICP-MS detects, removal and the enrichment of target analytes that suitable sample-pretreating method carries out substrate to it.
Chip magnetic solid-phase microextraction (MSPME) is the novel solid phase micro extraction system technically grown up by magnetic Solid-Phase Extraction (MSPE) and micro-total analysis system (μ TAS).Micro-total analysis system is one of Disciplinary Frontiers of Modern Analytical Chemistry, and its target is to realize the chemical analysis system overall miniaturization, automatization, integrated and portability from sample treatment to detection by analytical chemistry, micro electronmechanical processing, computer, electronics, material science and the intersection of biology, medical science.At present, it has been widely used among chemistry and biological study, particularly cell operation and cell analysis.Micro-fluidic chip, as a kind of chemical analysis instrument, has an advantage of many: low biological sample and the usage amount (μ L or nL level) of reagent, high flux, quickly, high sensitivity and spatial resolution etc..Therefore, micro-fluidic chip is to solve the powerful measure of miniaturization issues in cell analysis.All kinds of manipulations that magnetic nano-particle is widely used on micro-fluidic chip and analysis system.Chip is that magnetic nano-particle provides superior time and spatial control platform, simultaneously in chip system, the closely advantage brought due to miniaturization also makes the Miniature magnetic parts controlling magnetic Nano ion show higher magnetic and the higher maneuvering capability of the thing followed.Being based on these advantages, the magnetic manipulation technology on micro-fluidic chip is applied to the analysis system of many.
In the magnetic region passage that magnetic nano-particle is self-assembled on micro-fluidic chip by Hu etc., to form Solid-Phase Extraction microtrabeculae, thus establish on micro-fluidic chip the solid-phase microextraction platform for cell sample.In this system, due to the micron-sized size characteristics of micro-fluidic chip, number of cells needed for sample greatly reduces for detecting compared to regular growth, in view of the existence that cell is heterogeneous, the testing result of this system the most just more can embody element and the truth of form thereof in cell than the result of regular growth detection.Chen etc. utilize sulfonic group modified nano-magnetic ion to be filled in chip channel, set up micro-fluidic chip and the new method of Se form in liquid chromatograph-inductivity coupled plasma mass spectrometry binding analysis yeast cells, have opened up new approaches for Se form analysis in cell.But in this work, five kinds little molecule Se form (SeCys, MeSeCys, SeMet, SeGlu and SeEt) extraction efficiency equal less than 80%, also needing to ultrasonic wave added in desorption phase carries out eluting simultaneously, and due to the operation needed for the operator scheme of off-line, the method or relatively complicated, there is also the risk that sample is contaminated after the extraction simultaneously, based on this, develop online automatization's HPLC-ICP-MS detection platform and seem the most meaningful.
Utility model content
In order to overcome the deficiencies in the prior art, this utility model devises a kind of micro-fluidic chip, establishes a kind of magnetic Solid-Phase Extraction and micro column high efficiency liquid phase chromatograph-inductivity coupled plasma mass spectrometry on-line automaticization combination system simultaneously.This system has low reagent and sample consumption, good repeatability, the highly integrated and feature of automatization, is very suitable for the research of the trace biological sample trace element forms such as cell.
The technical solution of the utility model is specific as follows:
A kind of micro-fluidic chip, including symmetrical two microchannels, magnetic nano-particle and Magnet；Described microchannel includes being positioned at the serpentine channel of head end and being positioned at the rupture of membranes straight channel of tail end；Article two, microchannel is respectively provided with four micro-fluidic chip entrances, and described micro-fluidic chip entrance includes cell sample holes, rupture of membranes liquid sample holes, strippant sample holes and magnetic nano-particle sample holes；Article two, the tail end of microchannel is respectively provided with a waste outlet, and two microchannel tail ends are connected with the outlet of same micro-fluidic chip；
The head end of serpentine channel draws two short passages, connects with cell sample holes and rupture of membranes liquid sample holes respectively, and the tail end of serpentine channel is connected with rupture of membranes straight channel head end by short passage；
The head end of rupture of membranes straight channel passes sequentially through short passage and connects with tail end, strippant sample holes and the magnetic nano-particle sample holes of serpentine channel；
The tail end of rupture of membranes straight channel draws two short passages, and one connects with waste outlet, and one and micro-fluidic chip outlet, the short passage connected with waste outlet is provided with control air valve；
It is filled with magnetic nano-particle in described rupture of membranes straight channel, and rupture of membranes straight channel both sides are equipped with Magnet.
Described Magnet is permanent magnet, and described magnetic nano-particle is the Fe that γ-mercaptopropyl trimethoxysilane is modified3O4Nanoparticle.
A kind of on-line automaticization chip magnetic solid-phase microextraction system, including above-mentioned micro-fluidic chip, constant current syringe pump and syringe, six-way valve injector, quantitative loop, liquid chromatography pump, liquid-phase chromatographic column, is connected by pipeline between each parts；Described constant current syringe pump and syringe are connected with micro-fluidic chip entrance；The described valve opening on six-way valve injector is by being divided into the first valve opening, the second valve opening, the 3rd valve opening, the 4th valve opening, the 5th valve opening, the 6th valve opening clockwise, first valve opening is connected with micro-fluidic chip outlet, second valve opening is connected with waste liquid pool, 3rd valve opening and the 6th valve opening are connected with quantitative loop, 4th valve opening is connected with liquid chromatography pump, and the 5th valve opening is connected with liquid-phase chromatographic column sample introduction end；When handle is positioned at sample position, the first valve opening and the connection of the 6th valve opening, the second valve opening and the connection of the 3rd valve opening, the 4th valve opening and the connection of the 5th valve opening；When handle is positioned at sample introduction position, the first valve opening and the connection of the second valve opening, the 3rd valve opening and the connection of the 4th valve opening, the 5th valve opening and the connection of the 6th valve opening.
Described liquid-phase chromatographic column, its sample outlet end is connected with inductivity coupled plasma mass spectrometry.
This utility model has the following advantages and beneficial effect:
This utility model can extract inorganic mercury, methyl mercury, ethyl hydrargyrum and phenyl mercury simultaneously, is used for analyzing inorganic mercury in cell sample, methyl mercury, ethyl hydrargyrum and phenyl mercury form；There is device on-line analysis, analyze and change degree height, favorable reproducibility, the sample substrate detergent power advantage such as by force fast and automatically, be applicable to inorganic mercury in trace biological sample, methyl mercury, ethyl hydrargyrum and the analysis of phenyl mercury form.
Accompanying drawing explanation
Fig. 1 is the structural representation of micro-fluidic chip；Wherein, 1,2 is microchannel, and 1-1,2-1 are cell sample holes, and 1-2,2-2 are rupture of membranes liquid sample holes, 1-3,2-3 are strippant sample holes, 1-4,2-4 are magnetic nano-particle sample holes, and 1-5,2-5 are waste outlet, and 3-1 is Magnet one, 3-2 is Magnet two, 3-3 is Magnet three, and 4 export for micro-fluidic chip, and 5-1,5-2,5-3,5-4 are for controlling air valve.
nullFig. 2 is that on-line automaticization chip magnetic solid-phase microextraction liquid chromatograph inductivity coupled plasma mass spectrometry combined system analyzes view，Wherein: 6 is micro-fluidic chip，7 is inductivity coupled plasma mass spectrometry，8 is liquid-phase chromatographic column，9 is liquid chromatography pump，10 is quantitative loop，11-1 is strippant constant flow pump and syringe，11-2 is cell sample constant flow pump and syringe，11-3 is cell rupture of membranes liquid constant flow pump and syringe，11-4 is cell rupture of membranes liquid constant flow pump and syringe，11-5 is cell sample constant flow pump and syringe，11-6 is strippant constant flow pump and syringe，11-7 is that magnetic ball is filled and phosphate buffer constant flow pump and syringe，11-8 is that magnetic ball is filled and phosphate buffer constant flow pump and syringe，12 is waste liquid pool，13-1、13-2、13-3、13-4、13-5、13-6 respectively is the first valve opening of six-way valve、Second valve opening、3rd valve opening、4th valve opening、5th valve opening、6th valve opening.
Fig. 3 is that on-line automaticization chip magnetic solid-phase microextraction liquid chromatograph inductivity coupled plasma mass spectrometry combined system extracts view.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, this utility model done further detailed description, but embodiment of the present utility model is not limited to this.
Embodiment 1
As it is shown in figure 1, a kind of micro-fluidic chip, including symmetrical two microchannels, magnetic nano-particle and Magnet；Described microchannel includes being positioned at the serpentine channel of head end and being positioned at the rupture of membranes straight channel of tail end；Article two, microchannel is respectively provided with four micro-fluidic chip entrances, and described micro-fluidic chip entrance includes cell sample holes, rupture of membranes liquid sample holes, strippant sample holes and magnetic nano-particle sample holes；Article two, the tail end of microchannel is respectively provided with a waste outlet, and two microchannel tail ends are connected with the outlet of same micro-fluidic chip；
The head end of serpentine channel draws two short passages, connects with cell sample holes and rupture of membranes liquid sample holes respectively, and the tail end of serpentine channel is connected with rupture of membranes straight channel head end by short passage；
The head end of rupture of membranes straight channel passes sequentially through short passage and connects with tail end, strippant sample holes and the magnetic nano-particle sample holes of serpentine channel；
The tail end of rupture of membranes straight channel draws two short passages, and one connects with waste outlet, and one and micro-fluidic chip outlet, the short passage connected with waste outlet is provided with control air valve；
It is filled with magnetic nano-particle in described rupture of membranes straight channel, and rupture of membranes straight channel both sides are equipped with Magnet.
Described Magnet is permanent magnet, and described magnetic nano-particle is the Fe that γ-mercaptopropyl trimethoxysilane is modified3O4Nanoparticle.
The design of PDMS micro-fluidic chip: being integrated with symmetrical two microchannels 1 and 2 on this chip, four control air valve 5-1,5-2,5-3,5-4 and magnetic linkage growth, cell rupture of membranes and the big module of target analytes desorbing three.The height of microchannel is 50 μm, and width is 400 μm, and the height of air valve passage is 50 μm, and width is 600 μm.The curve that wriggles in Fig. 1 is cell rupture of membranes district, and sinuous passage contributes to the mixing between cell sample and rupture of membranes liquid, accelerates cell cracking process, and four entrances 1-1,1-2,2-1,2-2 are introduced two-way cell sample and two-way rupture of membranes liquid by constant current syringe pump respectively.Be connected with straight channel from top to bottom in Fig. 1 is followed successively by strippant sample holes, magnetic nano-particle sample holes, in extraction process, magnetic nano-particle sample holes also serves as buffering the entrance of cleanout fluid, fixes three pieces of permanent magnets (1.0*0.5*0.2cm) the most respectively at two parallel passages.
The processing of PDMS micro-fluidic chip: the making of silicon template uses soft lithography process, uses AZ-50XT photoresist.Fluid passage makes: component A (performed polymer) and the B component (firming agent) of GE RTV 615 (PDMS) are mixed with mass ratio 10:1, stir, it is placed in vacuum desiccator use oil pump evacuation 10min, static after taking-up treats bubble collapse.PDMS colloidal sol is cast on silicon masterplate, the permanent magnet of model-aided is positioned at the 3-1 Magnet one shown in Fig. 2,3-2 Magnet two, 3-3 Magnet one, 75 DEG C of solidification 3h, peel off the PDMS of solidification from silicon masterplate, punch at feeder connection and the port of export.
Control passage to make: component A and the B component of GE RTV 615 (PDMS) are mixed with mass ratio 15:1, stir, be placed in vacuum desiccator use oil pump evacuation 10min, stand after taking-up and treat bubble collapse.Being placed on sol evenning machine pallet by silicon formpiston, pour into a mould PDMS colloidal sol, open sol evenning machine, (forward 600rpm rotates 15s to arrange rotary speed parameter；After turn 1200rpm, rotate 30s), utilize sol evenning machine is rotated in silicon male mold surfaces one layer of PDMS film of coating, is then placed in baking oven, 75 DEG C of solidification 30min.The making of double-deck PDMS chip: the silicon formpiston by the PDMS containing fluid passage processed with containing the PDMS film controlling passage is placed in plasma cleaner, 1min is processed with oxygen plasma, immediately the two is fitted after taking-up, make surface bond, and guarantee that fluid passage is vertical corresponding with control channel position.It is placed in 75 DEG C of solidification 30min in baking oven, makes bonding face aging.Then this bilayer PDMS is peeled off from formpiston, controlling the punching of feeder connection end.Itself and slide being positioned in plasma cleaner, process 1min with oxygen plasma, be bonded rapidly after taking-up, then 75 DEG C of solidification 10min make bonding face the most aging.
The method using co-precipitation prepares magnetic Solid-Phase Extraction magnetism of material nanoparticle Fe3O4, the method is prior art, specifically comprises the following steps that and weighs 11.7g iron chloride FeCl3, it is dissolved in 150mL high purity water.Add 4.3g ferrous chloride FeCl2, add 50mL high purity water.After being completely dissolved, at N2The lower agitating heating of protection is back to 85 DEG C, rapidly joins 40mL strong aqua ammonia and by mixing speed and protection gas N2Becoming big, solution colour is become black from Chinese red.After half an hour, it is cooled to room temperature, gained magnetic nano-particle Fe3O4Use magnetism separate method to use high purity water and washing with alcohol for several times respectively, be stored in ethanol stand-by.The method using base catalysis prepares magnetic Nano silicon ball Fe3O4@SiO2.Magnetic nano-particle Fe prepared by upper step3O4Take half in dry beaker, add 100mL isopropanol, after ultrasonic disperse 10min, add to the there-necked flask of 250mL, add 12mL distilled water, after dropping 7mL strong aqua ammonia, drip 8mL tetraethoxysilane TEOS, N2Room temperature reaction 12h is stirred, respectively with high purity water and washing with alcohol magnetic Nano silicon ball Fe under protective condition3O4@SiO2For several times, it is stored in ethanol stand-by.The preparation of γ-MPTS modified magnetic nano silicon spheres: magnetic Nano silicon ball Fe prepared by upper step3O4@SiO2Take half in dry beaker, add 200mL ethanol, 5mL strong aqua ammonia, after 2mL γ-mercaptopropyl trimethoxysilane (γ-MPTS), ultrasonic disperse 10min, add to the there-necked flask of 500mL, N2Room temperature reaction 12h is stirred, respectively with high purity water and washing with alcohol magnetic Nano silicon ball Fe under protective condition3O4@SiO2@SH for several times, is stored in ethanol stand-by.0.5mol L is used before using-1After nitric acid cleans twice under ultrasound condition, use ammonium acetate to be washed till neutrality, finally clean three times with high purity water, stand-by.Magnetic nano-particle is formulated as 25mg mL-1Suspension and supersound process 15 minutes, afterwards under the effect of outer magnet, with 4mL min in the case of chip controls air valve 5-2 and 5-3 closes-1Flow velocity be filled in straight type passage.
Embodiment 2
As in figure 2 it is shown, a kind of on-line automaticization chip magnetic solid-phase microextraction system, including above-mentioned micro-fluidic chip, constant current syringe pump, six-way valve injector, quantitative loop, liquid chromatography pump, liquid-phase chromatographic column, connected by pipeline between each parts；Described constant current syringe pump is connected with micro-fluidic chip entrance；The described valve opening on six-way valve injector is by being divided into the first valve opening, the second valve opening, the 3rd valve opening, the 4th valve opening, the 5th valve opening, the 6th valve opening clockwise, first valve opening is connected with micro-fluidic chip outlet, second valve opening is connected with waste liquid pool, 3rd valve opening and the 6th valve opening are connected with quantitative loop, 4th valve opening is connected with liquid chromatography pump, and the 5th valve opening is connected with liquid-phase chromatographic column sample introduction end；When handle is positioned at sample position, the first valve opening and the connection of the 6th valve opening, the second valve opening and the connection of the 3rd valve opening, the 4th valve opening and the connection of the 5th valve opening；When handle is positioned at sample introduction position, the first valve opening and the connection of the second valve opening, the 3rd valve opening and the connection of the 4th valve opening, the 5th valve opening and the connection of the 6th valve opening.
Above-mentioned on-line automaticization chip magnetic solid-phase microextraction system, it is also possible to the sample outlet end of liquid-phase chromatographic column is connected with inductivity coupled plasma mass spectrometry.
Wherein: constant current syringe pump is TS2-60 type constant current syringe pump (Baoding LanGe constant flow pump Co., Ltd, China), asepsis injector is 1mL (Shanghai Jinta Medical Equipment Co., Ltd., China).High performance liquid chromatography is Ultimate 3000 type (Dionex, Germerring, Germany), chromatography post be RP-C18micro chromatographic column (Dionex, C18Acclaim, 3 μm,1.0mm i.d.), inductivity coupled plasma mass spectrometry is Xseries type ICP-MS (Thermo, USA).Chromatographic isolation gradient is: 1-8min, 2% methanol-98% flowing phase；8-10min, is changed to 55% methanol-45% flowing phase；10-12min, keeps 55% methanol-45% flowing phase；12-20min, is changed to 2% methanol-98% flowing phase.Chip air valve control system is laboratory self-control, by computer control.
Cell sample is imported micro-fluidic chip entrance 1-1 and 2-1 by polyethylene pump line from syringe, cell rupture of membranes liquid is imported micro-fluidic chip entrance 1-2 and 2-2 by polyethylene pump line from syringe, strippant is imported micro-fluidic chip entrance 1-3 and 2-3 by polyethylene pump line from syringe, and phosphate buffer is imported micro-fluidic chip entrance 1-4 and 2-4 by polyethylene pump line from syringe.Serpentine channel in chip completes cell rupture of membranes, completing solid-phase microextraction process at straight type passage, waste liquid imports quartz capillary from chip waste outlet 1-5 and 2-5, stripping liquid from outlet 4, and enter quantitative loop, eventually enter into microHPLC-ICP-MS and be analyzed detection.Its detailed process is as follows: 0-10 minute, a cell sample imports from chip entrance 1-1, cell rupture of membranes liquid imports from chip entrance 1-2 simultaneously, enter serpentine channel and complete cell rupture of membranes, sample enters straight type passage afterwards, and the target analytes in sample is adsorbed by magnetic Nano material, and now air valve 5-1 opens, 5-2,5-3,5-4 close, and waste liquid is discharged by waste outlet 3-1；10-20 minute, strippant imports from chip entrance 1-3, target analytes is eluted from magnetic Nano material, now air valve 5-2 opens, 3-1 closes, eluent is by exporting 4 importing quartz capillaries and entering quantitative loop, another part of cell sample imports from chip entrance 2-1 simultaneously, cell rupture of membranes liquid imports from chip entrance 2-2 simultaneously, entering serpentine channel and complete cell rupture of membranes, sample enters straight type passage afterwards, and the target analytes in sample is adsorbed by magnetic Nano material, now air valve 5-3 closes, and 5-4 opens waste liquid and discharged by waste outlet 3-2；When 20 minutes, system is analysis state persistently 10 minutes by extracting State Transferring, is converted to extraction state in 30 minutes time；20-30 minute, left channel was introduced phosphate buffer by chip entrance 1-4 and is carried out, and now air valve 5-1 opens, and 5-2 closes；30-40 minute, 3rd part of cell sample imports from chip entrance 1-1, cell rupture of membranes liquid imports from chip entrance 1-2 simultaneously, enter serpentine channel and complete cell rupture of membranes, sample enters straight type passage afterwards, target analytes in sample is adsorbed by magnetic Nano material, now air valve 5-1 opens, 5-2 closes, strippant imports from chip entrance 2-3, is eluted by target analytes from magnetic Nano material, and now air valve 5-3 opens, 5-4 closes, and eluent is by exporting 4 importing quartz capillaries and entering quantitative loop.Gone up process back and forth to carry out, due to liquid chromatograph inductivity coupled plasma mass spectrometry analyze 20 minutes used times, system sample throughput be 3 per hour.The extraction of cell sample, Air Valve Control and six-way valve switching all can be by computer controls, it is achieved that the integrated and automatization of sample extraction, desorbing and analysis.
Its programmed instruction is as shown in Table 1 below:
Table 1 on-line automaticization chip magnetic solid-phase microextraction liquid chromatograph inductivity coupled plasma mass spectrometry combined system programmed instruction
Under conditions of optimum, the analytical performance of body series, as shown in table 2, Hg are investigated2+, MeHg+, EtHg+And PhHg+Detection limit be respectively as follows: 18.8,12.8,17.4 and 41.8 ng L-1, this method is respectively as follows: 9.5,9.9,9.4 and 9.6 times to the enrichment times of four kinds of target analytes
The analytical performance of table 2 on-line automaticization chip magnetic solid-phase microextraction liquid chromatograph inductivity coupled plasma mass spectrometry combined system
A: in sample solution, four kinds of mercury shape concentration are is 0.25 ng mL-1
Investigate chip magnetic packed column to four kinds of goal analysis forms Hg2+、MeHg+、EtHg+And PhHg+The every passage of adsorption capacity be respectively 0.80,0.73,0.65 and 0.67 μ g.
Investigate different chip channel (each one passage of any selection on seven pieces of different chips) Hg under optimum experiment condition2+、MeHg+、EtHg+And PhHg+Repeatability, by calculate, the relative standard deviation of extraction is respectively 6.1%, 4.8%, 6.4% and 8.5% (CHg 2+,MeHg+,EtHg+,PhHg+=2ng mL-1, n=7), illustrate that the chip magnetic solid phase packed column that this experiment uses magnetic nano-particle self assembly stacking method to prepare has and preferably prepare repeatability.
The memory effect of nano magnetic silicon ball solid phase column and service life are also the important indicators evaluating chip magnetic solid phase packed column performance.
In experiment use single channel in optimal conditions four kinds of target morphologies of mesh are extracted, result show its after single extraction completes, its four kinds of target analytes Hg2+、MeHg+、EtHg+And PhHg+Memory effect be respectively as follows: 5.8%, 3.6%, 1.9%, 1.1%；After chip nano magnetic silicon ball solid phase packed column reuses 10 times, its extraction efficiency remains at 85-115%.
With 10,000 HepG2 cell suspending liquid is sample substrate, has investigated the mark-on recovering state of method, is 1.0ng mL at spiked levels-1Time, Hg2+、MeHg+、EtHg+And PhHg+Recovery result be respectively as follows: 106.5 ± 6.0%, 103.2 ± 6.4%, 98.3 ± 7.4% and 99.8 ± 9.8%.
Finally select 10,100 and 500 μ g L-1The MeHg of three kinds of variable concentrations+/Hg2+HepG2 cell is hatched, after hatching 12,18 and 24 hours, cell sample is taken out, use the chip-based onlinemicroHPLC-ICP-MS in this work that the mercury shape in cell is analyzed, every part of sample comprises 10000 cells.Its result is as shown in table 3.
Table 3 inorganic mercury and methyl mercury hatch this analysis system anlysis result of HepG2 cell
A: cannot be quantitative
The cell that b: methyl mercury is hatched
The cell that c: inorganic mercury is hatched
The Sample Pretreatment Technique of miniaturization is combined by the on-line automaticization chip magnetic solid-phase microextraction liquid chromatograph inductivity coupled plasma mass spectrometry combined system set up with the form fractionation detection technique of miniaturization, it is achieved that to the analysis of element morphology in micro-example.The method has low sample/reagent consumption, highly integrated automatization, highly sensitive, selectivity good, the advantage of favorable reproducibility, and the Elemental Speciation Analysis of a small amount of cell is had the biggest potentiality.
Above-described embodiment is this utility model preferably embodiment; but embodiment of the present utility model is also not restricted to the described embodiments; other any without departing from the change made under spirit of the present utility model and principle, modify, substitute, combine, simplify; all should be the substitute mode of equivalence, within being included in protection domain of the present utility model.
Claims (4)
1. a micro-fluidic chip, including symmetrical two microchannels, magnetic nano-particle and Magnet；Described microchannel
Including being positioned at the serpentine channel of head end and being positioned at the rupture of membranes straight channel of tail end；Article two, microchannel is respectively provided with four micro-fluidic chip entrances,
Described micro-fluidic chip entrance includes cell sample holes, rupture of membranes liquid sample holes, strippant sample holes and magnetic nano-particle sample introduction
Hole；Article two, the tail end of microchannel is respectively provided with a waste outlet, and two microchannel tail ends export phase with same micro-fluidic chip
Even；
The head end of serpentine channel draws two short passages, connects with cell sample holes and rupture of membranes liquid sample holes respectively, serpentine channel
Tail end is connected with rupture of membranes straight channel head end by short passage；
The head end of rupture of membranes straight channel passes sequentially through tail end, strippant sample holes and the magnetic nano-particle of short passage and serpentine channel and enters
Sample hole connects；
The tail end of rupture of membranes straight channel draws two short passages, and one connects with waste outlet, one and micro-fluidic chip outlet,
The short passage connected with waste outlet is provided with control air valve；
It is filled with magnetic nano-particle in described rupture of membranes straight channel, and rupture of membranes straight channel both sides are equipped with Magnet.
Micro-fluidic chip the most according to claim 1, it is characterised in that: described Magnet is permanent magnet, described magnetic
Nanoparticle is the Fe that γ-mercaptopropyl trimethoxysilane is modified3O4Nanoparticle.
3. an on-line automaticization chip magnetic solid-phase microextraction system, it is characterised in that: include described in claim 1 or 2 is micro-
Fluidic chip, constant current syringe pump and syringe, six-way valve injector, quantitative loop, liquid chromatography pump, liquid-phase chromatographic column, each portion
Connected by pipeline between part；Described constant current syringe pump and syringe are connected with micro-fluidic chip entrance；Described six-way valve enters
Valve opening on sample device is by being divided into the first valve opening, the second valve opening, the 3rd valve opening, the 4th valve opening, the 5th valve opening, the 6th valve clockwise
Hole, the first valve opening is connected with micro-fluidic chip outlet, and the second valve opening is connected with waste liquid pool, and the 3rd valve opening and the 6th valve opening are with quantitative
Ring is connected, and the 4th valve opening is connected with liquid chromatography pump, and the 5th valve opening is connected with liquid-phase chromatographic column sample introduction end；Handle is positioned at sampling position
When putting, the first valve opening and the connection of the 6th valve opening, the second valve opening and the connection of the 3rd valve opening, the 4th valve opening and the connection of the 5th valve opening；Hands
When handle is positioned at sample introduction position, the first valve opening and the connection of the second valve opening, the 3rd valve opening and the connection of the 4th valve opening, the 5th valve opening and the 6th
Valve opening connects.
On-line automaticization chip magnetic solid-phase microextraction system the most according to claim 3, it is characterised in that: described liquid
Phase chromatographic column, its sample outlet end is connected with inductivity coupled plasma mass spectrometry.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107607608A (en) * | 2017-09-08 | 2018-01-19 | 武汉大学 | A kind of Single cell analysis method |
| CN108333266A (en) * | 2017-12-28 | 2018-07-27 | 西北工业大学 | A kind of high pressure resistant minicore chip liquid chromatogram |
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| CN110987916A (en) * | 2019-12-17 | 2020-04-10 | 中国科学院半导体研究所 | Microfluidic chip and detection method thereof |
| CN111714931A (en) * | 2020-06-28 | 2020-09-29 | 清华大学深圳国际研究生院 | Transmission type solid phase micro extraction micro-fluidic device and manufacturing method thereof |
| CN112858177A (en) * | 2019-11-26 | 2021-05-28 | 武汉理工大学 | Heavy metal ion on-line measuring chip based on micro-fluidic extraction technique |
| CN113447578A (en) * | 2021-05-24 | 2021-09-28 | 华东理工大学 | Magnetic solid-phase extraction micro-fluidic chip-mass spectrometry combined device and application thereof |
| CN113945653A (en) * | 2021-09-27 | 2022-01-18 | 深圳职业技术学院 | Chip type living body solid phase micro-extraction device system and method for nano-drug pharmacokinetics precise analysis by using same |
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2016
- 2016-03-23 CN CN201620228337.2U patent/CN205484232U/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107607608A (en) * | 2017-09-08 | 2018-01-19 | 武汉大学 | A kind of Single cell analysis method |
| CN108333266A (en) * | 2017-12-28 | 2018-07-27 | 西北工业大学 | A kind of high pressure resistant minicore chip liquid chromatogram |
| 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 |
| CN112858177A (en) * | 2019-11-26 | 2021-05-28 | 武汉理工大学 | Heavy metal ion on-line measuring chip based on micro-fluidic extraction technique |
| CN110987916A (en) * | 2019-12-17 | 2020-04-10 | 中国科学院半导体研究所 | Microfluidic chip and detection method thereof |
| CN111714931A (en) * | 2020-06-28 | 2020-09-29 | 清华大学深圳国际研究生院 | Transmission type solid phase micro extraction micro-fluidic device and manufacturing method thereof |
| CN111714931B (en) * | 2020-06-28 | 2024-04-16 | 清华大学深圳国际研究生院 | Permeation type solid phase microextraction micro-fluidic device and manufacturing method thereof |
| CN113447578A (en) * | 2021-05-24 | 2021-09-28 | 华东理工大学 | Magnetic solid-phase extraction micro-fluidic chip-mass spectrometry combined device and application thereof |
| CN113945653A (en) * | 2021-09-27 | 2022-01-18 | 深圳职业技术学院 | Chip type living body solid phase micro-extraction device system and method for nano-drug pharmacokinetics precise analysis by using same |
| CN113945653B (en) * | 2021-09-27 | 2023-08-18 | 深圳职业技术学院 | Chip type living body solid phase microextraction device system and analysis method thereof |
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