CN105466905A - Solid phase extraction column integrated surface enhanced Raman detection chip - Google Patents
Solid phase extraction column integrated surface enhanced Raman detection chip Download PDFInfo
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- CN105466905A CN105466905A CN201510706113.8A CN201510706113A CN105466905A CN 105466905 A CN105466905 A CN 105466905A CN 201510706113 A CN201510706113 A CN 201510706113A CN 105466905 A CN105466905 A CN 105466905A
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Abstract
The invention discloses a solid phase extraction column integrated surface enhanced Raman detection chip. The chip comprises an upper substrate, a middle substrate and a lower substrate, the middle substrate is provided with a microfluidic channel (7), a sample detection cell (6) and sieve plate grooves (3), one end of the microfluidic channel (7) is provided with a sample inlet (5), the other end of the microfluidic channel (7) gets close to the sample detection cell (6), and a dam (4) and two sieve plate grooves (3) are arranged between the sample inlet and the sample detection cell; the upper substrate is provided with a sample introduction port (1), and the bottom of the upper substrate is provided with a surface enhanced Raman detection substrate (2), wherein the surface enhanced Raman detection substrate (2) aligns at the sample detection cell (6) of the middle substrate, and the sample introduction port (1) aligns at the sample inlet (5) of the microfluidic channel of the middle substrate; and the lower substrate is provided with a detection cell outlet (10), and the detection cell outlet (10) is positioned right below the sample detection ell and is connected with the sample outlet (8) of the middle substrate. The chip realizes online Raman detection and SPE operation integration of a sample to be detected.
Description
Technical field
The present invention relates to micro-fluidic chip and surface-enhanced Raman detection chip, specifically a kind of chip being detected on one for integrated solid-phase extraction column and surface-enhanced Raman.
Background technology
Solid-Phase Extraction (SPE) technology is widely used in separation/purification, and concentrated or enriched with trace determinand, removing chaff interference, improve concentration and the detection sensitivity of determinand, is a kind of conventional and important sample pretreating method.The microchannel structure utilizing chip micro-processing technology to be formed completes SPE and operates the advantage had less than many conventional SPE technology on microfluidic platform, and such as rate of extraction is fast, and sample/reagent consumes low, can be integrated etc. in other operating units.
Raman spectroscopy is the most important spectral technique analyzed various molecular structure and differentiate substance classes, it has fast, simply, can repeat, undamaged feature and can detecting under all kinds of complex environment He in water solution system, has outstanding advantage compared with other spectral techniques.The Surface enhanced raman spectroscopy technology (SERS) found for 1974 utilizes the enhancing in the surface electrical magnetic field of the surface phasmon induction with the metallic substrates of nanostructured, greatly can promote the detection sensitivity of determinand, thus overcome Raman spectroscopy cannot be used for trace materials detect weakness.
At present, there is the multiple micro-fluidic chip based on SPE technology, can be applicable to extraction and the enrichment of test substance.Also there is the multiple detection chip based on surface-enhanced Raman principle.After eluent after the existing micro-fluidic chip based on SPE technology all needs collection SPE pre-treatment, row detects and can not realize the integration of pre-treatment and detection again, micro-microfluidic channel is micron order (being less than 1mm), exist and make processed complex, passage holds susceptible to plugging shortcoming, and all not integrated surface enhanced Raman detection substrates of aforementioned micro-fluidic chip, this result also in the application of such chip and Detection results is restricted.And based on the detection chip of surface-enhanced Raman principle due to not integrated SPE pre-treatment operating unit, its application also have received restriction.
Summary of the invention
The object of the present invention is to provide a kind of surface-enhanced Raman detection chip of integrated solid-phase extraction column and preparation thereof and practical approach, to being realized the integration of SPE technology and surface-enhanced Raman detection by the design of this kind of novel chip, thus realize determinand more accurate, sensitiveer and detect more easily.
For achieving the above object, the technical solution used in the present invention is:
A surface-enhanced Raman detection chip for integrated solid-phase extraction column, is characterized in that, this chip comprises upper, middle and lower-ranking substrate,
Middle level substrate is provided with microchannel (7), sample detection pond (6) and sieve plate groove (3), microchannel (7) one end is provided with sample inlet (5), the other end is near sample detection pond (6) and establish outlet (8), a dam (4) between outlet (8) and sample detection pond (6), establishing two sieve plate grooves (3) between sample inlet (5) and outlet (8), in sieve plate groove, putting into the sieve plate for preventing filler from being washed away by fluid; Microchannel wherein in the middle of two sieve plate grooves forms filler filling channel;
Upper substrate is provided with injection port (1), bottom surface is provided with surface-enhanced Raman and detects substrate (2), wherein surface-enhanced Raman detects the sample detection pond (6) that middle level substrate is aimed in substrate (2), and injection port (1) is facing to the sample inlet (5) of middle level substrate microchannel;
Underlying basal is provided with detection cell outlet (10), and detection cell outlet (10) to be positioned at immediately below sample detection pond and to be communicated with the outlet (8) in middle level.
In the preferred embodiment, microchannel (7) width is 1-5mm.
In the preferred embodiment, underlying basal is also provided with the filler charging aperture (9) for filling solid phase extraction filler.
In the preferred embodiment, filler charging aperture (9) bottom is provided with cover plate (11).
In the preferred embodiment, the bottom surface of dam (4) and middle level and underlying basal interface contour.
Surface-enhanced Raman detection chip for integrated solid-phase extraction column provided by the invention can realize Solid-Phase Extraction by turning-over of chip and wash-out integrates, and surface-enhanced Raman is detected combined by micro-fluidic design with the step such as Solid-Phase Extraction, simple and quick, and achieve the online Raman detection of testing sample and the integration of SPE operation.
Tool of the present invention has the following advantages:
1, when this chip injection port upward time, sample solution and cleansing solution flow into microchannel by sample inlet, flow through after solid-phase extraction column enrichment by dam structure barrier, then chip is flowed out by the outlet of below, do not enter sample detection pond, Solid-Phase Extraction and the washing of determinand can be realized by this step.
2, when this chip injection port down time, eluent injects microchannel by sample inlet, flowing into sample detection pond, can be realized the wash-out of determinand by this step by covering dam structure after determinand wash-out in solid-phase extraction column.
3, after eluent flows into sample detection pond, determinand is combined with surface-enhanced Raman detection chip, then can carry out Raman detection
4, microchannel structure of the present invention is grade, is different from common micro scale channel, it is simple that grade passage has processing, and cost is low, without the need to the micro-fluidic sample introduction equipment of complex proprietary, only need to utilize the sampling instrument such as liquid-transfering gun or syringe, can fast sample be realized.And millimetre-sized passage has sample introduction speed soon, not easily blocked, chip is prepared into the high advantage of power.
5, simplicity of design of the present invention, is skillfully constructed, and realizes integrating of Solid-Phase Extraction and wash-out by this simple operations of turning-over of chip, and mating surface strengthens Raman detection chip realizes the integration that SPE pre-treatment and surface-enhanced Raman detect.
Accompanying drawing explanation
Fig. 1 is chip structure schematic diagram of the present invention, is wherein divided into upper, middle and lower three layers, and in upper strata, 1 is injection port, and 2 is surface-enhanced Raman detection substrate (being bonded at below).Middle level 3 is sieve plate groove, and 4 is dam structure, and 5 is injection port, and 6 is detection cell, and 7 is filler filling channel, and 8 is outlet.Lower floor 9 is filler charging aperture, and 10 is detection cell outlet, and 11 is filler charging aperture cover plate.
Fig. 2 is one-piece construction schematic diagram of the present invention;
Fig. 3 is testing result figure of the present invention.Wherein solid line is the detection spectrogram of the beverage containing lemon yellow, and dotted line is the detection spectrogram not containing lemon yellow beverage
Embodiment
Embodiment 1
See Fig. 1 and Fig. 2, the present invention is a kind of detects the chip with one for integrated solid-phase extraction column and surface-enhanced Raman, and chip is formed by upper, middle and lower-ranking substrate and cover plate sealing-in, and its upper strata is provided with injection port 1 and surface-enhanced Raman detection chip 2; Middle level is provided with microchannel 7, sample detection pond 6 and sieve plate groove 3, and microchannel 7 one end is provided with sample inlet 5, and the other end establishes outlet 8.Be provided with a dam 4 between outlet 8 and sample detection pond 6, establish two sieve plate grooves 3 between sample inlet 5 and outlet 8, sieve plate groove 3, for putting into polymkeric substance sieve plate, is washed away by fluid for preventing filler; Wherein two middle microchannels of sieve plate groove 3 form filler filling channel.The bottom surface of dam structure and the interface (middle level and underlying basal interface) of substrate contour, dam structure is used for preventing sample clean liquid from entering in detection cell, can strengthen Raman detection substrate 2 by receiving surface in sample detection pond; Underlying basal is provided with detection cell outlet 10 and filler charging aperture 9, and detection cell outlet 10 is in the same size with sample detection pond immediately below sample detection pond, and is communicated with outlet 8.Filler charging aperture 9, immediately below packed column bed passage, for filling solid phase extraction filler use, having been filled rear cover plate 11 and having been sealed filler injection port.
Chip material can be PMMA, PDMS or can be template with them various polymkeric substance and their complex, is filled with the solid phase extraction filler of forward absorption in filling channel.Microchannel 7 width is 1-5mm.
Surface-enhanced Raman detects substrate and following steps can be adopted to make:
1) first golden nanometer particle is synthesized, by 200ml massfraction be 0.01% HAuCl4 aqueous solution be heated to boiling, adding rapidly 1.4ml massfraction is 1% sodium citrate aqueous solution, after about 1min, solution colour faint yellowly becomes black from previous, brownish red is become again after 2 ~ 3min, continue to keep micro-40min that boils, stop reaction, water-bath cools.Namely obtain comprehensive red particle diameter and be about 55nmAu nano particle.
2) get centrifugal 8min under 1000 μ L above-mentioned Au nano particle 4500rpm/min, remove supernatant, namely obtain the concentrating particulate that volume is about 10 μ L.Draw above-mentioned sample 5 μ L to drip on clean silicon wafer, natural drying in air; Surface-enhanced Raman can be obtained and detect substrate.
Chip manufacturing step is as follows:
1) the chip structure figure first designed, as shown in Figure 1, in introducing carbon dioxide laser stepper software, makes the chip part of required PMMA material by lithography according to design drawing,
2) in the sieve plate groove of middle level substrate, put into the polymkeric substance sieve plate of fixed measure, for prevent filler fluid through time be flushed away;
3) according to design drawing order, upper, middle and lower upper substrate is fixed, utilizes hot press, carry out bonding according to setting heating schedule, after the complete wait cooling of bonding, take out chip;
4) forward solid phase extraction filler (Silon) is loaded in 5ml syringe, then utilize syringe to inject in chip channel by filler by the filler charging aperture of chip lower floor;
5) around filler charging aperture, finally drip the methenyl choloride of thin layer, cover plate is placed compacting ten second of getting on, then place five minutes.
6) surface-enhanced Raman made being detected substrate utilizes glue bottom detection cell, can complete the making of whole detection chip.
Embodiment 2
Detecting step:
1) by chip injection port upward, the orange beverage utilizing syringe to get 1ml to contain lemon yellow pigment, slowly injected in chip by injection port, waste liquid by outlet because Action of Gravity Field down drips, simultaneously because dam structure barrier can not flow in detection cell, prevent surface-enhanced Raman from detecting substrate and polluted by sample waste.
2) for washing solid-phase extraction column, impurity on solid-phase extraction column is washed utilizing syringe slowly to inject cleansing solution (1ml aqueous solution and 1ml ethanolic solution) successively;
3) overturn by chip, down, utilize syringe to inject eluent (0.5ml, 1% ammonia ethanol solution) and got off by sample elution, eluent covers dam structure to injection port, flows in detection cell and contacts with surface enhanced Raman substrate.
4) place after 10 seconds, chip is put into below laser Raman spectrometer camera lens, directly detect defocused, sample spectrogram (see Fig. 3) can be obtained.
Claims (6)
1. a surface-enhanced Raman detection chip for integrated solid-phase extraction column, is characterized in that, this chip comprises upper, middle and lower-ranking substrate,
Middle level substrate is provided with microchannel (7), sample detection pond (6) and sieve plate groove (3), microchannel (7) one end is provided with sample inlet (5), the other end is near sample detection pond (6) and establish outlet (8), a dam (4) between outlet (8) and sample detection pond (6), establishing two sieve plate grooves (3) between sample inlet (5) and outlet (8), in sieve plate groove, putting into the sieve plate for preventing filler from being washed away by fluid; Microchannel wherein in the middle of two sieve plate grooves forms filler filling channel;
Upper substrate is provided with injection port (1), bottom surface is provided with surface-enhanced Raman and detects substrate (2), wherein surface-enhanced Raman detects the sample detection pond (6) that middle level substrate is aimed in substrate (2), and injection port (1) is facing to the sample inlet (5) of middle level substrate microchannel;
Underlying basal is provided with detection cell outlet (10), and detection cell outlet (10) to be positioned at immediately below sample detection pond and to be communicated with the outlet (8) in middle level.
2. the surface-enhanced Raman detection chip of a kind of integrated solid-phase extraction column as claimed in claim 1, is characterized in that: microchannel (7) width is 1-5mm.
3. the surface-enhanced Raman detection chip of a kind of integrated solid-phase extraction column as claimed in claim 1, is characterized in that: underlying basal is also provided with the filler charging aperture (9) for filling solid phase extraction filler.
4. the surface-enhanced Raman detection chip of a kind of integrated solid-phase extraction column as claimed in claim 3, is characterized in that: filler charging aperture (9) bottom is provided with cover plate (11).
5. the surface-enhanced Raman detection chip of a kind of integrated solid-phase extraction column as claimed in claim 1, is characterized in that: the bottom surface of dam (4) and middle level and underlying basal interface contour.
6. a surface enhanced Raman detection method, comprises the steps:
1) the surface-enhanced Raman detection chip of the integrated solid-phase extraction column described in any one of claim 1 to 5 is provided;
2), utilize syringe to get testing liquid by chip injection port (1) upward, slowly injected in chip by injection port (1), waste liquid down drips because Action of Gravity Field passes through outlet (8);
3) utilize syringe slowly to inject cleansing solution successively for washing solid-phase extraction column, impurity on solid-phase extraction column is washed;
4) overturn by chip, down, utilize syringe to inject eluent and got off by sample elution, eluent covers dam (4) to injection port (1), detects substrate (2) contact in inflow detection cell with surface-enhanced Raman;
5), after placing a period of time, chip is put into below laser Raman spectrometer camera lens, directly detect defocused, obtain sample spectrogram.
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| CN108152429A (en) * | 2016-12-04 | 2018-06-12 | 中国科学院大连化学物理研究所 | A kind of Solid Phase Extraction microtrabeculae and application |
| CN108496072A (en) * | 2016-04-22 | 2018-09-04 | 惠普发展公司，有限责任合伙企业 | SERS sensor devices with passivating film |
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| CN107036874A (en) * | 2017-05-25 | 2017-08-11 | 华东理工大学 | The encapsulation type enrichment and separation device detected for online solid phase spectra |
| CN107036874B (en) * | 2017-05-25 | 2021-03-16 | 华东理工大学 | A encapsulation type enrichment separator for online solid-phase spectral detection |
| CN114308153A (en) * | 2021-12-17 | 2022-04-12 | 华南农业大学 | Solid-phase extraction micro-fluidic chip for detecting nafil substances and system for detecting nafil substances |
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