| In the chapter one, first of all, the development and the present situation of capillary electrophoresis (CE) was introduced briefly. Then the development and application of capillary electrophoresis with electrochemiluninescence (ECL)was reviewed. Finally, the analysis of high-throughput single cell by CE including the single cell analysis and high-throughput single cell detection was summarized.In the chapter two, glutathione (GSH) and ascordic acid (AA) and dopamine (DA) were separated and detected by capillary electrophoresis with electrochemiluninescence in the experiments. The effects of the kinds of running buffer,running buffer pH and concentration , separation voltage , detected potential and concentration of Ru(bpy)32+ on CE-ECL were studied. The optimum conditions of separation and detection for the three materials were 50 mmol·L-1 NaH2PO4- Na2HPO4 (pH 7.8) for the buffer solution, 15 kV for the separation voltage, 2×10-3 mol·L-1 for the concentration of Ru(bpy)32+, 1.3 V (vs.SCE)for the detection potential, 12 kV and 10 s for the injection voltage and the injection time. The concentration limits of detection (signal to noise, S/N =3) of the analytes were 1.0×10-6 mol·L-1 for glutathione, 1.0×10-6 mol·L-1 for ascordic acid and 5.0×10-7 mol·L-1 for dopamine. The response for a series of six injections of 1.0×10-5 mol·L-1 DA ,5.0×10-4 mol·L-1 AA and 1.0×10-5mol·L-1 AA mixed standard solution resulted in a relative standard deviation of 1.1%,0.98% and 1.2% for the migration time ,and 3.3%,2.2%,3.1% for the ECL intensity,respectively.The method was used to detected glutathione, ascordic acid and dopamine in the human red blood cell solution of their injections, The recoveries were GSH 96%, AA 105% and DA 103%. In the chapter three, vanillylmandelic acid (VMA) and homovanillic acid (HVA) were separated and detected by capillary electrophoresis with electrochemiluninescence detection. The effects of the kinds of running buffer,running buffer pH and concentration , separation voltage , detected potential and concentration of Ru(bpy)32+ on CE-ECL were studied. The optimum conditions of separation and detection for the three materials were 50 mmol·L-1 NaH2PO4- Na2HPO4 (pH 8.2) for the buffer solution, 16 kV for the separation voltage, 5 mmol·L-1 for the concentration of Ru(bpy)32+, 1.2 V (vs.SCE)for the detection potential, 10 kV and 10 s for the injection voltage and the injection time. The limits of detection (signal to noise, S/N =3) of the analytes were 5.0×10-7 mol·L-1 for vanillylmandelic acid 6.1×10-7 mol·L-1 for homovanillic acid. The response for a series of six injections of 1.0×10-5 mol·L-1 VMA and 1.0×10-5 mol·L-1 HVA mixed standard solution resulted in a relative standard deviation of 1.1and 0.9% for the migration time 2.7% and 3.1% for the ECL intensity,respectively.The method was used to detected vanillylmandelic acid and homovanillic acid in the human urine solution of their injections, The recoveries were VMA 98%and HVA 99%.In the chapter four, high-throughput single cell counting device was designed.Pressure was adapted control the cells thoughtput. Experiments showed that this detection system was stable and quickly. The optimum conditions of separation and detection are 20 mmol·L-1 NaH2PO4-Na2HPO4 (pH 7.8) for the buffer solution, the concentration of Ru(bpy)32+ is 5.0×10-3 mol·L-1, and 1.3 V(vs.SCE)for the detection potential and the pressure of injection sets 0.6 MPa . Human red blood cells were counted by electrochemiluninescence detection. The single cells were counted at intervals of about 20 s. The throughtput of single cells is accord with the cells circulation observed by microscope, the counting system combined with a cell-dissolve device lay a solid basis for high throughput single-celled analysis. |
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