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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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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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Acknowledgments

The research presented in this manuscript was supported by the National Natural Science Foundation of China (No. 20575047 and 20275029) and the Research Foundation for the Doctoral Program of Higher Education (No. 20030486011).

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Authors and Affiliations

  1. Department of Chemistry, Wuhan University, Wuhan, 430072, China

    Ke-Jing Huang, Min Zhang, Hua-Shan Zhang, Yu-Qi Feng & Hong Wang

  2. Department of Chemistry, Xinyang Normal University, Xinyang, 464000, China

    Ke-Jing Huang & Wan-Zhen Xie

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  1. Ke-Jing Huang
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  2. Min Zhang
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  3. Wan-Zhen Xie
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  4. Hua-Shan Zhang
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  5. Yu-Qi Feng
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  6. Hong Wang
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Correspondence to Hong Wang.

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Huang, KJ., Zhang, M., Xie, WZ. et al. Sensitive determination of nitric oxide in some rat tissues using polymer monolith microextraction coupled to high-performance liquid chromatography with fluorescence detection. Anal Bioanal Chem 388, 939–946 (2007). https://doi.org/10.1007/s00216-007-1283-2

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  • DOI: https://doi.org/10.1007/s00216-007-1283-2

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