CN101034080B - Simple sampling method suitable for portable microchip capillary electrophoresis - Google Patents
Simple sampling method suitable for portable microchip capillary electrophoresis Download PDFInfo
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- CN101034080B CN101034080B CN200710021383A CN200710021383A CN101034080B CN 101034080 B CN101034080 B CN 101034080B CN 200710021383 A CN200710021383 A CN 200710021383A CN 200710021383 A CN200710021383 A CN 200710021383A CN 101034080 B CN101034080 B CN 101034080B
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- infuse
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- 238000000034 method Methods 0.000 title claims abstract description 15
- 238000005251 capillar electrophoresis Methods 0.000 title claims abstract description 8
- 238000005070 sampling Methods 0.000 title claims description 7
- 239000000523 sample Substances 0.000 claims abstract description 52
- 239000007788 liquid Substances 0.000 claims abstract description 26
- 239000002699 waste material Substances 0.000 claims abstract description 13
- 238000000926 separation method Methods 0.000 claims abstract description 10
- 229910019142 PO4 Inorganic materials 0.000 claims abstract description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims abstract description 5
- 239000010452 phosphate Substances 0.000 claims abstract description 5
- 239000012488 sample solution Substances 0.000 claims abstract description 4
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical compound OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 claims description 8
- 239000012530 fluid Substances 0.000 claims description 5
- 239000000872 buffer Substances 0.000 claims description 4
- 238000013016 damping Methods 0.000 claims description 4
- 230000003139 buffering effect Effects 0.000 claims description 3
- 229910021607 Silver chloride Inorganic materials 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims description 2
- 239000008366 buffered solution Substances 0.000 claims 2
- 238000002347 injection Methods 0.000 abstract description 11
- 239000007924 injection Substances 0.000 abstract description 11
- 239000007853 buffer solution Substances 0.000 abstract description 5
- 239000000243 solution Substances 0.000 abstract 4
- 230000000694 effects Effects 0.000 description 7
- 238000001962 electrophoresis Methods 0.000 description 7
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 4
- 238000005370 electroosmosis Methods 0.000 description 3
- 150000005206 1,2-dihydroxybenzenes Chemical class 0.000 description 2
- 206010019133 Hangover Diseases 0.000 description 2
- 229960003638 dopamine Drugs 0.000 description 2
- 101100379079 Emericella variicolor andA gene Proteins 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000005357 flat glass Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000037230 mobility Effects 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000000930 thermomechanical effect Effects 0.000 description 1
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- Sampling And Sample Adjustment (AREA)
Abstract
This invention relates to a Sample injection method of portable type microchip capillary electrophoresis equipment. The invention keeps microchip flat, infuse sample solution into sample cell, and infuse phosphate ( PBS) buffer solution into sample waste liquor pool and balanced solution pool. it also infuse balanced solution into balanced solution waste liquor pool, and keep substantial same liquid level with balanced solution pool, then incliningly infuse microchip: Inclining angle is 10 deg to 25 deg, oblique heading is to make sample cell high and sample waste liquor pool low. 40 second after Sample injection, set level microchip, at the same time applies separating tension to carry out electrophoretic separation. The Sample injection method is simplicity and easy, basically has no need of Sample injection equipments.
Description
Technical field
The present invention relates to be fit to the simple sampling method of portable chip capillary cataphoresis equipment.Specifically, by the inclination microchip, the liquid level in the sample cell is higher than the liquid level of sample waste liquid pool, so produce hydraulic pressure difference, has realized the hydraulic-driven sample introduction.
Background technology
Since designs such as Manz have used the flat glass chip [referring to (a) D.J.Harrison, A.Manz, Z.Fan, H.Luedi and H.M.Widmer, Anal.Chem., 1992,64,1926-1932. (b) A.Manz, D.J.Harrison, E.M.J.Verpoorte, J.C.Fettinger, A.Paulus, H.Ludi and H.M.Widmer, J.Chromatogr.A, 1992,593,253-258.], the microchip capillary electrophoresis is just studied widely to be used [referring to: (a) D.Mijatovic, J.C.T.Eijkel and A.van den Berg, Lab Chip, 2005,5,492-500. (b) T.Vilkner, D.Janasek andA.Manz, Anal.Chem., 2004,76,3373-3386. (c) P.S.Dittrich, K.Tachikawa, and A.Manz, Anal.Chem., 2006,78,3887-3907.].The condition precedent of successfully analyzing is accurately to realize the micro-example sample introduction.Research work has dropped into a large amount of energy and time research and has improved sample injection method, in the input mode that all have been reported, based on the main method of the electrokinetic injection of electroosmotic flow driving, but this input mode has two shortcomings, the influence that the sample introduction situation is subjected to pipeline surface nature component sample introductions very big and for different mobilities exist deviation (bias) [referring to (a) J.P.Alarie, S.C.Jacobson and J.M.Ramsey, Electrophoresis, 2001,22,312-317. (b) B.E.Slentz, N.A.Penner and F.Regnier, Anal.Chem., 2002,74,4835-4840.].For fear of this deviation sample introduction, the pressure-driven sample introduction is used, and this input mode is not subjected to the influence of the character of tube wall surface and sample.Bai etc. have proposed to use the pressure clamp of many logical sampling valves and three syringe pump compositions to flow to sample loading mode, sample size can reach receives upgrading [referring to X.Bai, H.J.Lee, J.S.Rossier, F.Reymond, H.Schafer, M.Wossner and H.H.Girault, Lab Chip, 2002,2,45-49.].Solignac and Gijs have set up a kind of thermo-mechanical drive that utilizes, apply a pressure pulse on the barrier film of liquid storage tank, have realized sample fluid hydrodynamic injection [referring to D.Solignac and M.A.M.Gijs, Anal.Chem., 2003,75,1652-1657.].Zhang etc. set up a kind of method be used for the microchip capillary electrophoresis quantitatively accurately, do not have a sample feeding of the skin upgrading of deviation, the driving of sample is to utilize different liquid level differences produce in negative pressure, electroosmotic flow and the chip liquid storage tank that syringe pump produces hydraulic pressure acting in conjunction [referring to L.Zhang, X.Yinand Z.Fang, Lab Chip, 2006,6,258-264.].Lin seminar has reported on micro-fluidic chip, the input mode that combines with electric power by hydraulic pressure [referring to: H.Gai, L.Yu, Z.Dai, Y.Ma and B.C.Lin, Electrophoresis, 2004,25,1888-1894.], the main shortcoming of this input mode is to exist peak shape hangover, and the influence of hangover is fatal sometimes even to separation, makes that sample can not be separated or quantitative.
Also there is not bibliographical information to use a kind of additional equipment and sample introduction simple sample injection method satisfactory for result of not needing.
Summary of the invention
The simple sampling method that the purpose of this invention is to provide a kind of suitable portable microchip capillary electrophoresis.
Technical scheme of the present invention is as follows:
A kind of simple sampling method of suitable portable microchip capillary electrophoresis, it is made up of the following step:
Step 1. keeps flat microchip, and sample solution is injected sample cell, and phosphate (PBS) buffer solution is injected sample waste liquid pool and buffering liquid pool, also injects buffer solution in the damping fluid waste liquid pool, and with the liquid level basically identical of buffer pool,
With the microchip inclination sample introduction of step 1, the angle of inclination is 10 °-25 ° to step 2. then, and the direction of inclination is to make the sample cell height, and the sample waste liquid pool is low, 40 seconds of sample introduction,
The microchip of step 3. after with step 2 sample introduction set level, and applies separation voltage simultaneously and carries out electrophoretic separation.
Above-mentioned sample injection method, described step 2 can tilt microchip with the wedge of a fixed angle.
Sample injection method of the present invention is simple, and sample introduction is effective, does not need extra sample introduction equipment basically.
Description of drawings
Fig. 1 is an applied microchip synoptic diagram when measuring electroosmotic flow among the present invention, and wherein: A is a buffer pool, and B is the separating pipe outlet, C is a sample cell, and D is the sample waste liquid pool, and E is sample channel and separating pipe crossing, F is the damping fluid waste liquid pool, and WE is an electrochemical working electrode.
Fig. 2 is the separation electrophoresis figure that the mixed solution of 500 μ M dopamines and 500 μ M catechols in the embodiments of the invention is implemented chip electrophoresis behind the hydraulic pressure sample introduction, 5 intensity of receiving peace scale explanation peak height among the figure, and A is a dopamine, B is a catechol.
Embodiment
Embodiment 1. utilizes input mode of the present invention, implements the microchip capillary electrophoretic separation with dopamine and catechol biased sample and measures.
Utilize the standard solution of 500 μ M dopamines and 500 μ M catechols to check the effect of sample injection method of the present invention.
Microchip is kept flat, 500 μ M dopamines and the 500 μ M catechol biased sample solution of 13.5 μ L are put into sample cell (C) (liquid level is approximately 0.27cm), the phosphate of 15 μ L20mM (PBS) buffer solution is put into sample waste liquid pool (D) and buffering liquid pool (A) (liquid level is approximately 0.30cm), also put into the PBS buffer solution of 20mM in the damping fluid waste liquid pool (F), and with the liquid level basically identical of buffer pool (A).The sample introduction that then microchip tilted tilts 17 ° microchip with the wedge of 17 ° of fixed angles.Behind the sample introduction 40 seconds, microchip is set level, applied separation voltage 800V simultaneously and carry out electrophoretic separation, detect at detection current potential+1.4V (vs.Ag/AgCl electrode).Obtain electrophoretogram, see Fig. 2.According to peak shape and separating effect, the sample introduction effect satisfies the requirement of chip capillary cataphoresis fully.
Embodiment 2.
Except the angle with wedge makes 10 ° into, microchip is tilted beyond 10 °, other are all with embodiment 1, and its separating effect is identical with the separating effect of embodiment 1.
Embodiment 3.
Except the angle with wedge makes 25 ° into, microchip is tilted beyond 25 °, other are all with embodiment 1, and its separating effect is identical with the separating effect of embodiment 1.
Claims (1)
1. the simple sampling method of a suitable portable microchip capillary electrophoresis is characterized in that it is made up of the following step:
Microchip is kept flat, 500 μ M dopamines and the 500 μ M catechol biased sample solution of 13.5 μ L are put into sample cell (C), liquid level is 0.27cm, the phosphate buffered solution of 15 μ L 20mM is put into sample waste liquid pool (D) and buffering liquid pool (A), liquid level is 0.30cm, also put into the phosphate buffered solution of 20mM in the damping fluid waste liquid pool (F), and liquid level basically identical with buffer pool (A), sample introduction then tilts microchip, wedge with 17 ° of fixed angles tilts 17 ° microchip, behind the sample introduction 40 seconds, microchip is set level, applying separation voltage 800V simultaneously and carry out electrophoretic separation, is contrast electrode with the Ag/AgCl electrode, and current potential+1.4V detects in detection.
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| CN200710021383A CN101034080B (en) | 2007-04-10 | 2007-04-10 | Simple sampling method suitable for portable microchip capillary electrophoresis |
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| CN200710021383A CN101034080B (en) | 2007-04-10 | 2007-04-10 | Simple sampling method suitable for portable microchip capillary electrophoresis |
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| CN101590389B (en) * | 2009-06-24 | 2011-07-20 | 中国科学院大连化学物理研究所 | Method for forming and capturing liquid drops based on three-layer sandwich disc-type chip and application thereof |
| CN106959332B (en) * | 2017-02-28 | 2019-03-29 | 中国科学院合肥物质科学研究院 | The automatic fine sampling device and its control method of electrophoresis detection instrument |
| CN110252438A (en) * | 2019-07-19 | 2019-09-20 | 南京信息工程大学 | A kind of solution is automatically introduced into type electrophoretic microchip and its working method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2169849Y (en) * | 1993-05-01 | 1994-06-22 | 浙江大学 | Gravity sample feeder for capillary electrophoretic instrument |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2169849Y (en) * | 1993-05-01 | 1994-06-22 | 浙江大学 | Gravity sample feeder for capillary electrophoretic instrument |
Non-Patent Citations (1)
| Title |
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| 凌云扬等.在线衍生微流控芯片电泳和激光诱导荧光法测定单细胞中谷胱甘肽.高等学校化学学报25.2004,25167-168. * |
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