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A Novel Solid Substrate Room Temperature Phosphorimetry for the Determination of Trace Cytochrome C and Forecast of Human Diseases

DOI: 10.5402/2012/918093

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Abstract:

The reaction between fullerenol (F-ol) and cytochrome C (Cyt C) could be carried out to form a nonphosphorescence compound using tween-80 as photosensitizer which causes the sharp quenching of room temperature phosphorescence (RTP) of F-ol. Bearing this in mind, a novel solid substrate room temperature phosphorimetry (SSRTP) for the determination of trace Cyt C has been proposed in this study. Under the optimum conditions, the linear range of this method is – ?? , which is directly proportional to of Cyt C-F-ol–tween-80 system, and the detection limit (DL) is ? . It has been applied to the determination of Cyt C in human serum and forecast of diseases, and the result matches with the enzyme-linked fluorescence immunoassay (ELISA). Meanwhile, the reaction mechanism of SSRTP for the determination of Cyt C and the enhancing effect of tween-80 on RTP of F-ol were also discussed. 1. Introduction In recent years, the studies have been found that the incidence of leukemia and the execution of chemotherapy drugs to leukemia cells are closely related to cell apoptosis [1]. Studies on the clinical examination of Cyt C and the researches about its proapoptotic activity regulating molecular mechanism and the release mechanism have become a focus [2]. There are many methods for the determination of Cyt C, such as synchronous fluorescence spectroscopy (linear range: – ?g?mL?1) [3], CdTe/CdS quantum dots resonance rayleigh scattering spectroscopy (DL: ?g?mL?1) [4], electrochemical probe voltammetric method (DL: ?g?mL?1) [5], cyclic voltammetry [6], adsorption method (DL: ?g mL?1) [7], ED/Au/I? modified ultramicroelectrode method (DL: ?g?mL?1) [8], capillary zone electrophoretic method (DL: ?g?mL?1) [9], amperometric method (DL: ?g?mL?1) [10], protein blotting method (DL: ?g?mL?1) [11], and ELISA (DL: ?g?mL?1) [12]. All these methods cannot meet the needs of determining the low content of Cyt C in biological samples due to low sensitivity. Therefore, searching for a high sensitive and accurate method for the determination of Cyt C in biological samples and discussing the relativity between the content of Cyt C and cell apoptosis have become research focus with high academic value and great significance. There have been many reports on the synthesis of multihydroxyl C60 derivatives, F-ol derivatives, aminophenol derivatives, dendritic fullerene derivatives [13–15], the fluorescent property of water-soluble F-ol [16], and the determination of alkaline phosphatase [17, 18], glucose [19], As (V) [20], and Mn2+ [21] based on phosphorescent property of F-ol, showing broad

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