Huang Ke-Jing, Wang Hong, Ma Ming, Sha Mei-Lan, Zhang Hua-Shan
Department of Chemistry, Wuhan University, Wuhan 430072, China.
J Chromatogr A. 2006 Jan 27;1103(2):193-201. doi: 10.1016/j.chroma.2005.11.015. Epub 2005 Nov 23.
To reveal the function of nitric oxide (NO) that has been reported as an intracellular second messenger and a diffusible intercellular messenger, a novel ultrasound-assisted liquid-phase microextraction (ULPME) and high-performance liquid chromatographic method for the determination of NO produced in PC12 cells has been developed. 1,3,5,7-Tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s-indacene (DAMBO-CO(2)Et) has been used for NO trapping to form corresponding triazole (DAMBO-CO(2)Et-T). The enrichment factor of DAMBO-CO(2)Et-T reached 150 in 2.5 min with the proposed ULPME method. NO produced in (200+/-5) PC12 cells has been determined directly with the detection limit (S/N=3) of 2.5 x 10(-13)mol/L (5a mol), which was the lowest in HPLC determination of NO reported yet. The proposed method was sensitive, selective, convenient and rapid, which has great prospect in the qualitative and quantitative determination of NO in minimal biological samples.
为揭示一氧化氮（NO）作为细胞内第二信使和可扩散细胞间信使的功能，开发了一种新型超声辅助液相微萃取（ULPME）结合高效液相色谱法来测定PC12细胞中产生的NO。1,3,5,7-四甲基-2,6-二乙氧羰基-8-（3',4'-二氨基苯基）-二氟硼二氮杂-s-茚满（DAMBO-CO(2)Et）用于捕获NO以形成相应的三唑（DAMBO-CO(2)Et-T）。采用所提出的ULPME方法，DAMBO-CO(2)Et-T在2.5分钟内的富集因子达到150。已直接测定了（200±5）个PC12细胞中产生的NO，检测限（S/N = 3）为2.5×10(-13)mol/L（5a mol），这是迄今报道的HPLC法测定NO中最低的。所提出的方法灵敏、选择性好、方便且快速，在微量生物样品中NO的定性和定量测定方面具有广阔前景。
