Sensitive determination of nitric oxide in some rat tissues using polymer monolith microextraction coupled to high-performance liquid chromatography with fluorescence detection.
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 lambda (ex)/lambda (em) = 498/507 nm. The baseline separation of TMDABODIPY and its NO derivative is performed under simple conditions in which a C(18) 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 x 10(-12) mol L(-1) (S/N = 3). The linearity range of the method was 9 x 10(-11)-4.5 x 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%.
Huang
Ke-Jing
KJ
Department of Chemistry, Wuhan University, Wuhan, China. wanghong@chem.whu.edu.cn
Zhang
Min
M
Xie
Wan-Zhen
WZ
Zhang
Hua-Shan
HS
Feng
Yu-Qi
YQ
Wang
Hong
H
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D019856:Ethanolamine / Q000737:chemistry; D005453:Fluorescence; D006321:Heart / Q000502:physiology; D007668:Kidney / Q000502:physiology; D008099:Liver / Q000502:physiology; D008297:Male; D000432:Methanol / Q000737:chemistry; D009569:Nitric Oxide / Q000032:analysis* / Q000493:pharmacokinetics; D011092:Polyethylene Glycols / Q000737:chemistry*; D011109:Polymethacrylic Acids / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide / Q000737:chemistry*; D052617:Solid Phase Microextraction; D013997:Time Factors; D014018:Tissue Distribution / Q000502:physiology*
D011109:Polymethacrylic Acids; C099679:poly(methacrylic acid-ethylene glycol); D009569:Nitric Oxide; D011092:Polyethylene Glycols; D019856:Ethanolamine; D012822:Silicon Dioxide; D000432:Methanol