Determination of nitric oxide in hydrophytes using poly(methacrylic acid-ethylene glycol dimethacrylate) monolith microextraction coupled to high-performance liquid chromatography with fluorescence detection.
Nitric oxide (NO) is a bioactive molecule that has recently emerged as a cellular messenger in numerous physiological processes in plants. A novel high-performance liquid chromatography (HPLC) method combined with poly(methacrylic acid-ethylene glycol dimethacrylate) (MAA-EGDMA) monolith microextraction (PMME) is developed for sensitive determination of NO in hydrophytes. NO is derivatized using a fluorescent probe, 1,3,5,7-tetramethyl-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacene (DAMBO), and then the derivatives are extracted with PMME and analyzed by high-performance liquid chromatography (HPLC) with fluorescence detection. The conditions for the derivatization and the subsequent extraction of NO derivatives are optimized in detail. The detection limit (S/N=3) of NO is determined to be 2x10(-12)mol L(-1). Close correlation coefficient and excellent method reproducibility are obtained for the analyte over a linear range of 9x10(-11)-4.5x10(-8)mol L(-1). The inter- and intraday relative standard deviations (R.S.D.s) are less than 5%. The proposed method is successfully applied to the determination of NO levels in hydrophytes samples.
Huang
Ke-Jing
KJ
Department of Chemistry, Wuhan University, Wuhan 430072, PR China.
Zhang
Min
M
Xie
Wan-Zhen
WZ
Zhang
Hua-Shan
HS
Feng
Yu-Qi
YQ
Wang
Hong
H
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005026:Ethylene Glycols / Q000737:chemistry*; D008689:Methacrylates / Q000737:chemistry*; D009569:Nitric Oxide / Q000032:analysis*; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005026:Ethylene Glycols; D008689:Methacrylates; C502776:poly(methacrylic acid-ethylene glycol dimethacrylate); D009569:Nitric Oxide