CN100473986C - Method for measuring low-speed electric interstifial flow in chip capillary cataphoresis - Google Patents
Method for measuring low-speed electric interstifial flow in chip capillary cataphoresis Download PDFInfo
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- CN100473986C CN100473986C CNB200710021018XA CN200710021018A CN100473986C CN 100473986 C CN100473986 C CN 100473986C CN B200710021018X A CNB200710021018X A CN B200710021018XA CN 200710021018 A CN200710021018 A CN 200710021018A CN 100473986 C CN100473986 C CN 100473986C
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Abstract
A method can measure low speed electro-osmotic flow in chip capillary electrophoresis. First of all, measure effective mobility of probe substance by sample zonal method on quick electro-osmotic flow microchip A which being as a reference. The method is simplified as: add a quickly-migrating substance (probe) which can be tested into background electrolyte. Detect electrophoretic pattern when taking hormone background electrolyte with different density from background electrolyte as sample. Based on the peaking time in pattern, effective mobility (Mu eff) of probe substance on reference chip can be calculated. Then, taking same background electrolyte solution without probe as buffer solution and probe substance as sample, detect routine capillary electrophoresis on microchip B with unknown electro-osmotic flow. Apparent mobility (Mu npp) of probe on B can be calculated and curious effective mobility (Mu EOF) can be achieved by the following formula: Mu EOF=Mu npp-Mu eff.
Description
Technical field
The present invention relates to the measurement of low speed electroosmotic flow.Specifically, be based on an electromigratory material and measure the low speed electroosmotic flow indirectly in the constant principle of the ducted effective mobility of microchip.
Background technology
The micro-fluid control chip electrophoretic technology began to develop rapidly [referring to Manz from nineteen nineties, A., Graber, N., Widmer, H.M., Sens.Actuators B 1990, B1 (1-6), 244-248.], in order to satisfy the application of various aspects, often microchannel is carried out chemistry or physical modification, reduce the interaction of solute and tube wall and suppress electroosmotic flow.Therefore the electroosmotic flow property representation to microchannel just seems extremely important.
Commonly used electroosmotic flow assay method has neutral label method [referring to Jorgenson, J.W., Lukacs, K.D., Anal.Chem.1981,53,1298-1302.] and the current monitoring method [referring to: Huang, X., Gordon, M.J., Zare, R.N., Anal.Chem.1988,60,1837-1838.].The most commonly used is neutral label method, utilizes exactly to move under the effect of a kind of neutral compound in electric field and measures electroosmotic flow; The current monitoring method also is the very high method of a kind of utility ratio, the effect that drives by electric osmose in pipeline is with the solution in the alternative kapillary of a same electrolyte solution (comparing with the solution in the kapillary) that slightly dilutes, by electric current time history plot in the observation circuit, calculate electroosmotic flow according to whole times when replaced of pipeline middle and high concentration solution.
When needs are measured low electroosmotic flow or zero electroosmotic flow, difficulty has just appearred, and in neutral label method and current monitoring method, after through very long transit time, signal can become not obvious or not have signal.Up to the present, also not about hanging down the report of electroosmotic flow or zero electroosmotic flow assay method in the microchip capillary electrophoresis.
Summary of the invention
The purpose of this invention is to provide a kind of method of measuring low electroosmotic flow.
Technical scheme of the present invention is as follows:
The measuring method of low speed electroosmotic flow in a kind of chip capillary cataphoresis, it be at first one as the quick electroosmotic flow microchip A of reference on sample area band method [referring to Wang, W., Zhao, L., Jiang, L.P., Zhang, J.R., Zhu, J.J., Chen, H.Y., Electrophoresis 2006,27,5132-5137.] finish the mensuration of the effective mobility of probe substance, be summarized as follows, in back-ground electolyte, add a fast transferring and material can be determined (probe), after the class background electrolyte that is different from background electrolyte concentration is as sample feeding, obtain electrophoretogram, utilized the appearance time among the figure can calculate the effective mobility (μ of probe substance on the reference chip
Eff), then, utilize the identical background electrolyte that does not contain probe as damping fluid, on the microchip B of unknown electroosmotic flow, finish conventional Capillary Electrophoresis with probe substance as sample and detect, can calculate the apparent mobility (μ of probe on microchip B
App), can calculate the electroendosmotic mobility (μ that seeks knowledge according to following formula
EOF), μ
EOF=μ
App-μ
Eff
The effective mobility of a specific probe substance in the damping fluid of determining is invariable, and just the effective mobility of probe substance on reference chip and electroosmotic flow chip to be measured equates,
Therefore, μ
EffA1=μ
EffB1
μ wherein
EffA1And μ
EffB1Be respectively the effective mobility among microchip A and the microchip B.The effective mobility of this probe substance can calculate from its apparent mobility and electroosmotic flow,
μ in the above formula
AppA1, μ
EOFAAnd V
ABe respectively apparent mobility, electroendosmotic mobility and the separation voltage of the probe substance among the microchip A, t
A1And t
A2Be respectively the appearance time of microchip A middle probe material and electroosmotic flow.μ
AppB1, μ
EOFBAnd V
BRepresent the respective amount among the microchip B respectively.L
A, L
Aeff, L
BAnd L
BeffBe respectively length and the effective length of microchip A and microchip B.Therefore, μ
EOFBCan represent with following formula,
If L
A=L
B=L, L
Aeff=L
Beff=L
EffAnd V
A=V
B=V, μ
EOFBCan be calculated as follows and get.
Four, 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 buffer pool, 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.。
The electrophoresis synoptic diagram that Fig. 2 records with indirect method on reference chip A for the present invention, t
A1And t
A2Be respectively the appearance time of microchip A middle probe material and electroosmotic flow.
The electrophoresis synoptic diagram that Fig. 3 records with direct method on electroosmotic flow chip B to be measured for the present invention, t
B1And t
B2Be respectively the transit time of microchip B middle probe material and electroosmotic flow.
Embodiment
The measurement of the electroosmotic flow of poly dimethyl silica silicon (PDMS) microchip of embodiment 1.Brij56 modified
Electroosmotic flow when being full of phosphate buffer (20mM PBS pH7.00) in the PDMS microchip pipeline of measurement Brij56 modified, with 0.1 mM 3,4-dihydroxy benzylamine (DHBA) is as probe substance, 20 mM PBS are electrolyte as a setting, not make the PDMS microchip of modification as reference chip A, wherein: AE=0.3cm; EB=3.6cm; CE=ED=0.5cm.In the Capillary Electrophoresis process, separation voltage is set at 800V, detects current potential+1.2V (vs.Ag/AgCl electrode), according to the transit time of measuring gained, t
B1=59.2s, t
A1=41.2s, t
A2=65.0s, the size that calculates electroosmotic flow is (1.40 ± 0.04) * 10
-4Cm
2/ (Vs).Consistent with the document guess value [referring to: Dou, Y.H., Bao, N., Xu, J.J., Meng, F., Chen, H.Y., Electrophoresis 2004,25,3024-3031.].
The measurement of the electroosmotic flow of the PDMS microchip pipeline of embodiment 2. polyvinyl alcohol (PVA) (PVA) modified
Electroosmotic flow when being full of phosphate buffer (20mM PBS pH7.00) in the PDMS microchip pipeline of measurement PVA modified, with 0.1mM3,4-dihydroxy benzylamine (DHBA) is as probe substance, 20mM PBS is electrolyte as a setting, not make the PDMS microchip of modification as reference chip A, wherein: AE=0.3cm; EB=3.6cm; CE=ED=0.5cm.In the Capillary Electrophoresis process, separation voltage is set at 800V, detects current potential+1.2V (vs.Ag/AgCl electrode), according to the transit time of measuring gained, t
B1=59.2s, t
A1=41.2s, t
A2=65.0s, the size that calculates electroosmotic flow is (0.22 ± 0.02) * 10
-4Cm
2/ (Vs).Consistent with the document guess value [referring to: Wu, D., Luo, Y., Zhou, X., Dai, Z., Lin, B., Electrophoresis 2005,26,211-218.].
Claims (2)
1. the measuring method of low speed electroosmotic flow in the chip capillary cataphoresis is characterized in that: the effective mobility μ that at first records probe substance on a quick electroosmotic flow microchip A as reference
Eff, record the apparent mobility μ of probe substance on the microchip B of electroosmotic flow to be measured then
App, calculate unknown electroendosmotic mobility μ according to following formula
EOF,
μ
EOF=μ
app—μ
eff。
2. the measuring method of low speed electroosmotic flow in the chip capillary cataphoresis according to claim 1, it is characterized in that: with sample area band method on quick electroosmotic flow reference chip once the property transit time of finishing probe substance and electroosmotic flow measure, directly calculate effective mobility.
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CN104122316B (en) * | 2014-06-30 | 2016-05-25 | 中国科学院化学研究所 | Utilize distributed DC electric field to measure the mobility of particle and the method for dielectric mobility simultaneously |
CN108007994B (en) * | 2017-06-27 | 2019-12-03 | 北京理工大学 | A method of measurement Capillary Electrophoresis electroosmotic flow |
CN114660283B (en) * | 2022-05-24 | 2022-09-13 | 佛山微奥云生物技术有限公司 | Immunoassay plate type chip based on electrical acceleration and preparation method thereof |
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2007
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Non-Patent Citations (8)
Title |
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EOF measurement by detection of a sampling zone withend-channel amperometry in microchip CE. Wang W.等.Electrophoresis,Vol.27 No.24. 2006 |
EOF measurement by detection of a sampling zone withend-channel amperometry in microchip CE. Wang W.等.Electrophoresis,Vol.27 No.24. 2006 * |
Fast, Accurate Mobility Determination Method forCapillaryElectrophoresis. Williams B. A.等.Anal. Chem.,Vol.68 No.7. 1996 |
Fast, Accurate Mobility Determination Method forCapillaryElectrophoresis. Williams B. A.等.Anal. Chem.,Vol.68 No.7. 1996 * |
Method for the Accelerated Measurement of ElectroosmosisinChemically Modified Tubes for Capillary Electrophoresis. Sandoval J. E.等.Anal. Chem.,Vol.68 No.17. 1996 |
Method for the Accelerated Measurement of ElectroosmosisinChemically Modified Tubes for Capillary Electrophoresis. Sandoval J. E.等.Anal. Chem.,Vol.68 No.17. 1996 * |
毛细管电泳同时测定酚类系列化合物的表观离解常数. 陈晓峰等.兰州大学学报,第41卷第1期. 2005 |
毛细管电泳同时测定酚类系列化合物的表观离解常数. 陈晓峰等.兰州大学学报,第41卷第1期. 2005 * |
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