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The preliminary experiments presented in this paper demonstrate that the floating electrode ECL detector can be used successfully for the detection of Ru(bpy) and Ru(phen) following electrophoretic separation

A wireless electrochemiluminescence detector applied to direct and indirect detection for electrophoresis on a microfabricated glass device.

ANALYTICAL CHEMISTRY, no. 14 (2001): 3282-3288

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摘要

A novel electrochemiluminescence (ECL) detector is presented in this article. The detector is applied for micellar electrokinetic chromatographic separation of dichlorotris(2,2'-bipyridyl)ruthenium(II) hydrate [Ru-(bpy)] and dichlorotris(1,10-phenanthroline)ruthenium-(II) hydrate [Ru(phen)] on a microfabricated glass device. It consists o...更多

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简介
  • The detector is applied for micellar electrokinetic chromatographic separation of dichlorotris(2,2′-bipyridyl)ruthenium(II) hydrate [Ru(bpy)] and dichlorotris(1,10-phenanthroline)ruthenium(II) hydrate [Ru(phen)] on a microfabricated glass device
  • It consists of a microfabricated “U”-shape floating platinum electrode placed across the separation channel.
  • Ru(bpy) has been the most studied and exploited inorganic ECL compound to date because of its capability of undergoing ECL at room temperature in aqueous buffered solutions, in the presence of dissolved oxygen and other impurities
  • It has been widely used in ECL flow cells and probe injection analysis systems to detect various chemicals and biochemical assays
重点内容
  • A novel electrochemiluminescence (ECL) detector is presented in this article
  • The detector is applied for micellar electrokinetic chromatographic separation of dichlorotris(2,2′-bipyridyl)ruthenium(II) hydrate [Ru(bpy)] and dichlorotris(1,10-phenanthroline)ruthenium(II) hydrate [Ru(phen)] on a microfabricated glass device. It consists of a microfabricated “U”-shape floating platinum electrode placed across the separation channel
  • The required potential difference for the ECL reaction is generated at the Pt electrode by the electric field available in the separation channel during electrophoretic separation
  • Preliminary results show the indirect detection of three amino acids
  • Chemiluminescence (CL) and ECL have advantages over fluorescence because they do not require a light source, which results in a simple instrumentation and low or zero background signal
  • The preliminary experiments presented in this paper demonstrate that the floating electrode ECL detector can be used successfully for the detection of Ru(bpy) and Ru(phen) following electrophoretic separation
方法
  • A Photomultiplier tube (PMT, model R3896, Hamamatsu, Hamamatsu City, Japan), operating at 1250 V, was placed inside the metal box for data collection.
  • The emitted light from the reaction reaches the PMT window through a 2-mm aperture at the top of the metal box.
  • A personal computer with a data acquisition card and Lab View (V 5.0) was used to control high-voltage power supplies.
  • The output signal from the PMT was recorded using a Pico analog-digital convertor (ADC12, Pico Instrument).
  • A vacuum pump (Charles Austen Pumps Ltd., Surrey, England) was used to fill, empty, or clean the channels with buffer solution and to inject sample into the double-T region
结果
  • A U-shaped Pt wire shows the same behavior (Figure 1C), the leg of the electrode facing the anode behaving as a cathode and the leg facing the cathode behaving as an anode
  • This phenomenon can be explained by considering that the Pt wire short-circuits the solution over its length, creating a potential difference between its ends proportional to the fraction of the total distance short-circuited
结论
  • The preliminary experiments presented in this paper demonstrate that the floating electrode ECL detector can be used successfully for the detection of Ru(bpy) and Ru(phen) following electrophoretic separation.
  • It was shown that nonlabeled analytes following electrophoretic separation in Ru(bpy)TPA buffer can be detected indirectly.
  • Optimization of the distance between the floating electrode and the injection point will decrease the band broadening and will improve the detection limit for individually injected samples
表格
  • Table1: Separation Fields Strength and Floating Electrode Detection Voltage as a Function of Employed Separation Voltage voltage applied separation field strength (V/cm)
Download tables as Excel
基金
  • Authors thank Astra Zeneca and SmithKline Beecham for their financial support, the Worshipful Co. of Scientific Instrument Makers for its WCSIM Burser 1997/98 Award, Dr John Crabtree of the Alberta Microelectronic Corp. for fabricating the microstructures, Dr Darwin Reyes for help with photographs, Dr Martin Arundell for constructing the CE setup, Dr Chao-Xuan Zhang of the Imperial College, London, and Dr Martin Boutelle of the King’s College, London, for the valuable personal discussions, and finally editor Dr Royce W
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