Study on Ratiometric Determination of Peroxynitrite and Its Scavenger Based on Synchronous Fluorescence Spectroscopy
DU Jun-ou,GUO Xiang-qun*
Department of Chemistry, College of Chemistry and Chemical Engineering, Key Laboratory of Analytical Sciences of the Ministry of Education, Key Laboratory of Chemical Biology of Fujian Province, Xiamen University, Xiamen 361005, China
Abstract:A ratiometric fluorimetry is proposed for the determination of peroxynitrite (ONOO-) and the evaluation of its scavenger based on synchronous fluorescence spectroscopy.L-tyrosine, an intrinsic fluorescent aminoacid, reacts with peroxynitrite and carbon dioxide, yielding a highly fluorescent dimer of tyrosine in pH 8.5 PBS buffer solution.By synchronous fluorescence scanning, both monomer and dimer emission bands of L-tyrosine appeared at 364 and 406 nm,respectively.The ratio of F406/F364 is quantitatively related to the concentration of peroxynitrite, where F406 represents the intensity of synchronous emission band of dimer and F364 represents that of monomer.The method has been showing merit of being insensitive to the changes in experimental parameters and offers a higher sensitivity and a broader responding linear range of the analyte concentration, compared to fluorimetric method, giving a LOD of 1.84×10-8 mol·L-1 and a linear range of 1.60×10-7 mol·L-1- 6.00×10-6 mol·L-1 for ONOO-, respectively, with a deviation of 2.4% for 1.00×10-6 mol·L-1 ONOO-(n=8).A IC50 of 0.065 μg·mL-1 for mitoxantrone,an antioxidant and anticancer drug,was also obtained.This method has proved to be simple and speedy.It would be easily used in the determination of ONOO- and its scavenger.
杜俊鸥,郭祥群*. 同步荧光光谱比率法检测过氧亚硝酸根及其清除剂的研究[J]. 光谱学与光谱分析, 2008, 28(08): 1875-1878.
DU Jun-ou,GUO Xiang-qun*. Study on Ratiometric Determination of Peroxynitrite and Its Scavenger Based on Synchronous Fluorescence Spectroscopy. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2008, 28(08): 1875-1878.
[1] TIAN Yi-ling, CHEN Guan-hua, CUI Tong(田益玲, 陈冠华, 崔 同).Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2005, 25(4):617. [2] QU Peng, LI Guan-liang, XU Mao-tian(瞿 鹏, 李贯良, 徐茂田).Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2004, 24(11):1407. [3] Ara J, Przedborski S, Naini A B, et al.Proc.Natl.Acad.Sci.U.S.A., 1998, 95:7659. [4] Coddington J W, Hurst J K, Lymar S V.J.Am.Chem.Soc., 1999, 121:2438. [5] Squadrito G L, Pryor W A.Free Radical Biol.Med., 1998, 25:392. [6] Wiseman H, Halliwell B.Biochem.J., 1996, 313:17. [7] Kossenjans W, Eis A, Sahay R, et al.Am.J.Physiology:Heart and Circulatory Physiol., 2000, 278:1311. [8] White C R, Brock T A, Chang L, et al.Proc.Natl.Acad.Sci.U.S.A., 1994, 91:1044. [9] Wang H, Joseph J A.Free Radic.Biol.Med., 1999, 27:612. [10] Daiber A, Oelze M, August M, et al.Free Radical Research., 2004, 38:259. [11] Dikalov S, Grigorev I A, Voinov M, et al.Biochem.Biophys.Res.Commun., 1998, 248:211. [12] Li Bei-bei, Gutierrez P L, Blough N V.Anal.Chem., 1997, 69:4295. [13] Xue J, Ying X Y, Chen J S, et al.Anal.Chem., 2000, 72:5313. [14] Valeur B.Molecular Fluorescence:Principles and Applications;Wiley-VCH:Weinheim, 2002. [15] Singh Y, Gulyani A, Bhattacharya S.FEBS Letters, 2003, 541:132. [16] Stromberg N, Hulth S.Sensors and Actuators B-Chemical., 2003, 90:308. [17] Van Arman S A, Sisk T M, Zawrotny D A.Letters in Organic Chemistry, 2005, 2(1):54. [18] Lymar S V, Jiang Q, Hurst J K.Biochemistry, 1996, 35:7855. [19] ZHANG Hou-wen, Squadrito G L, Pryor W A.Nitric Oxide, 1997, 1:301. [20] Thomson L, Trujillo M, Telleri R, et al.Arch.Biochem.Biophys., 1995, 319:491. [21] Hughes M N, Nicklin H G.J.Chem.Soc.A, 1968, (2):450. [22] CHEN Guo-zhen, HUANG Xian-zhi, ZHENG Zhu-zi, et al(陈国珍, 黄贤智, 郑朱梓, 等).The Method of Fluorescence Analysis(荧光分析法).Beijng:Science Press(北京:科学出版社), 1990. [23] YANG Xiao-feng, GUO Xiang-qun.Analyst, 2001, 126:928. [24] Fernandes E, Gomes A, Costa D, Lima José L F C.Life Sciences, 2005, 77:1983.