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A new route for simple and rapid determination of hydrogen peroxide in RAW264.7 macrophages by microchip electrophoresis by Xiaocong Gong; Qingling Li; Kehua Xu; Xin Liu; Hongmin Li; Zhenzhen Chen; Lili Tong; Bo Tang; Hongbo Zhong is a Engineering article available to read on EtoBox.

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## Abstract A new method using MCE with LIF detection was developed for the determination of hydrogen peroxide (H~2~O~2~). Bis(p‐methylbenzenesulfonyl)dichlorofluorescein, a new fluorogenic reagent synthesized by our laboratory was employed as a labeling reagent, the derivatization reaction was performed in 0.10 M HEPES buffer (pH 7.4) for 30 min at 37°C. The detection of H~2~O~2~ was accomplished in 55 s, using a 40 mM HEPES buffer, 20% mannitol, pH 7.4, on a glass microchip. The RSDs of migration time and peak area were 1.8 and 3.7%, respectively. Method validation showed the linear response ranging from 0.50 to 50 μM with an LOD (__S__/__N__=3) of 0.20 μM (19.1 amol). The proposed method was applied to determine H~2~O~2~ in phorbol myristate acetate‐stimulated RAW264.7 macrophages, the concentration of H~2~O~2~ was found to be 1.86±0.05 μM; recoveries for macrophage samples were from 96.7 to 97.8%, within‐days and between‐days accuracies were 4.5% (__n__=5) and 6.8% (__n__=5), respectively.

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Author

Xiaocong Gong; Qingling Li; Kehua Xu; Xin Liu; Hongmin Li; Zhenzhen Chen; Lili Tong; Bo Tang; Hongbo Zhong

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley (ISSN 0173-0835)

Published

2009

Language

EN

Field

Engineering (Physical Sciences)