Sensitive determination of nitric oxide in some rat tissues using polymer monolith microextraction coupled to high-performance liquid chromatography with fluorescence detection |
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Authors: | Ke-Jing Huang Min Zhang Wan-Zhen Xie Hua-Shan Zhang Yu-Qi Feng Hong Wang |
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Affiliation: | (1) Department of Chemistry, Wuhan University, Wuhan, 430072, China;(2) Department of Chemistry, Xinyang Normal University, Xinyang, 464000, China |
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Abstract: | A simple, sensitive, selective, and low-cost method is proposed for rapidly determining nitric oxide (NO) in some rat tissues. Polymer monolith microextraction (PMME) using a poly(methacrylic acid–ethylene glycol dimethacrylate) (MAA-EGDMA) monolithic column was combined with derivatization of NO using 1,3,5,7-tetramethyl-8-(3′,4′-diaminophenyl)-difluoroboradiaza-s-indacene (TMDABODIPY), and this was used to analyze the derivatives of NO by high-performance liquid chromatography (HPLC) with fluorescence detection at λ ex/λ em = 498/507 nm. The baseline separation of TMDABODIPY and its NO derivative is performed under simple conditions in which a C18 column is used and eluted with 50 mmol L−1 ethanolamine and methanol. The conditions for the extraction of NO derivatives were optimized. The limit of detection of NO was 2 × 10−12 mol L−1 (S/N = 3). The linearity range of the method was 9 × 10−11−4.5 × 10−8 mol L−1. The interday and intraday relative standard deviations were less than 5%. The proposed method was successfully applied to the determination of NO levels in some rat tissue samples including heart, kidney, and liver with recoveries varying from 87.1 to 95.2%. |
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Keywords: | Poly(methacrylic acid– ethylene glycol dimethacrylate) monolith microextraction Nitric oxide Derivatization 1,3,5,7-Tetramethyl-8-(3′ ,4′ -diaminophenyl)-difluoroboradiaza-s-indacene (TMDABODIPY) High-performance liquid chromatography (HPLC) |
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