Development of the direct modification method of the ruthenium complex on conductive diamond surfaces and the selective detection of bio-related materials

Abstract

The tris-2,2′-(bipyridine) ruthenium(II) complex derivative, which shows the electrogenerated chemiluminescence (ECL) reaction, was directly immobilized on the boron-doped diamond (BDD) surface with three types of modification to form C–N, C–O and C–C bond between them. The objective of this study is to find the most appropriate modification method for the highest amount of immobilization on diamond and the highest stability for electrochemical reaction in the high potential ranges. By making use of this photofunctional BDD, the establishment of an analytical system for the selective and high-sensitive detection of oxalic acid in human urine using the ECL reaction was attempted. Comparing the stability for the ECL reaction among three modified BDD (MBDD) electrodes using the long-term potential cycling, in the initial of cycling, the ECL peak intensity for modified BDD linked with C–C bond (MBDD(C–C)), with C–N bond (MBDD(C–N)) and with C–O bond after the exposure to O 2 plasma (MBDD(C–O pl)) were in the same range. At MBDD(C–C), the ECL intensity was very stable and initial value was maintained even after 200 min of the potential cycling, but the ECL intensities for other modified BDD were rapidly decreased, indicating that the modification method through C–C bond can provide the high density and stable modification. Using MBDD(C–C), the analysis for oxalic acid in human urine was carried out by the high-performance liquid chromatography (HPLC) and flow injection analysis (FIA) systems in that the electrochemical amperometric detector (ECD) and ECL detector with modified BDD were installed. In the HPLC-ECD chromatogram, many peaks including uric acid and ascorbic acid were observed. On the other hand, only one peak was detected in the HPLC-ECL chromatogram, indicating that oxalic acid could be selectively detected in human urine using the ECL reaction at modified BDD. The linear dynamic range and the lowest detection limit (S/N = 3) for the oxalic acid detection at MBDD(C–C) were achieved to be 2 orders of magnitude from 5 μM to 100 μM and 5 μM, respectively. MBDD(C–C) was proven to have sufficient sensitivity for oxalic acid in human urine (average concentration 500 μM).