title: Highly sensitive determination of nitric oxide in biologic samples by a near-infrared BODIPY-based fluorescent probe coupled with high-performance liquid chromatography.
abstract: Nitric oxide (NO) acts as an important regulator and mediator in numerous processes of biological systems. In this work, the analytical potential of a novel near-infrared (NIR, >600 nm) BODIPY-based fluorescent probe for NO, 8-(3,4-diaminophenyl)-4,4-difluoro-4-bora-3a,4a-diaza-di(1,2-dihydro) naphtho[b, g]s-indacene (DANPBO-H) has been evaluated in high performance liquid chromatography (HPLC). In 25 mM pH 6.50 borate buffer, DANPBO-H reacted with NO to give the corresponding triazole, DANPBO-H-T, at 35 °C for 20 min. DANPBO-H-T was eluted using a mobile phase of methanol/tetrahydrofuran/50mM pH 7.00 H3Cit-NaOH buffer (81:7:12, v/v/v) in 4 min on a C8 column and detected with fluorescence detection at excitation and emission wavelengths of 621 and 631 nm, respectively. The limit of detection (LOD) (signal-to-noise=3) reached to 5.50×10(-10) M. Excellent selectivity was observed against other reactive oxygen/nitrogen species. Various representative biological matrixes including the whole blood and organs of mice, the pangen and radical of rice, human vascular endothelial (ECV-304) cells and mouse macrophage (RAW 264.7) cells were used to verify the feasibility and resistance to interfering effects from complex biological sample matrixes of the developed DANPBO-H-based HPLC method. Compared to the existing derivatization-based HPLC methods for NO, the proposed method eliminates interfering effects from complex biological sample matrixes efficiently owing to the fluorescence detection in the NIR region, and is more advantageous and robust for the sensitive and selective determination of NO in complex biological samples.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine, Ministry of Education, Department of Chemistry, Wuhan University, Wuhan 430072, China.
authors.forename: Hui-Xian
authors.initials: HX
authors.lastname: Zhang
authors.forename: Jian-Bo
authors.initials: JB
authors.lastname: Chen
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Guo
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Hua-Shan
authors.initials: HS
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001896:Boron Compounds / Q000737:chemistry*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D042783:Endothelial Cells / Q000737:chemistry / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D008264:Macrophages / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051379:Mice; D009569:Nitric Oxide / Q000032:analysis* / Q000096:biosynthesis; D012275:Oryza / Q000737:chemistry; D012639:Seeds / Q000737:chemistry; D059629:Signal-To-Noise Ratio; D014020:Tissue Extracts / Q000737:chemistry*; D014230:Triazoles / Q000138:chemical synthesis
chemical_list: D001896:Boron Compounds; D005456:Fluorescent Dyes; D014020:Tissue Extracts; D014230:Triazoles; D009569:Nitric Oxide
keywords: Biological samples; DANPBO-H; HPLC; Near-infrared; Nitric oxide