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采用含十二烷基硫酸钠的聚合物溶液通过毛细管电泳分析乳腺癌细胞中的氨基酸和生物胺。

Analysis of amino acids and biogenic amines in breast cancer cells by capillary electrophoresis using polymer solutions containing sodium dodecyl sulfate.

机构信息

Department of Chemistry, National Taiwan University, 1 Section 4, Roosevelt Road, Taipei 106, Taiwan.

出版信息

J Chromatogr A. 2010 Jan 22;1217(4):582-7. doi: 10.1016/j.chroma.2009.11.069. Epub 2009 Dec 3.

DOI:10.1016/j.chroma.2009.11.069
PMID:20004398
Abstract

We describe simultaneous analysis of naphthalene-2,3-dicarboxaldehyde (NDA)-amino acid and NDA-biogenic amine derivatives by CE in conjunction with light-emitting diode-induced fluorescence detection using poly(ethylene oxide) (PEO) solutions containing sodium dodecyl sulfate (SDS). After sample injection, via EOF 0.1% PEO prepared in 100mM TB solution (pH 9.0) containing 30 mM SDS entered a capillary filled with 0.5M TB solution (pH 10.2) containing 40 mM SDS. Under this condition, 14 NDA-amino acid and NDA-amine derivatives were separated within 16 min, with high efficiency ((1.0-3.2)x10(5) theoretical plates) and sensitivity (LODs at S/N=3 ranging from 2.06 to 19.17 nM). In the presence of SDS and PEO, analytes adsorption on the capillary wall was suppressed, leading to high efficiency and reproducibility. The intraday analysis RSD values (n=3) of the mobilities for the analytes are less than 0.52%. We have validated the practicality of this approach by quantitative determination of 9 amino acids in breast cancer cells (MCF-7) and 10 amino acids in normal epithelial cells (H184B5F5/M10). The concentrations of Tau and Gln in the MCF-7 cells were different than those in the H184B5F5/M10 cells, respectively. Our results show the potential of this approach for cancer study.

摘要

我们描述了通过 CE 与使用含有十二烷基硫酸钠 (SDS) 的聚环氧乙烷 (PEO) 溶液结合的发光二极管诱导荧光检测同时分析萘-2,3-二羧酸醛 (NDA)-氨基酸和 NDA-生物胺衍生物。在样品注入后,通过EOF 0.1% PEO 在含有 30 mM SDS 的 100mM TB 溶液 (pH 9.0) 中进入充满 0.5M TB 溶液 (pH 10.2) 的毛细管,其中含有 40 mM SDS。在这种条件下,14 种 NDA-氨基酸和 NDA-胺衍生物在 16 分钟内得到分离,具有高效率 ((1.0-3.2)x10(5)理论塔板) 和灵敏度 (S/N=3 时的 LODs 范围从 2.06 到 19.17 nM)。在 SDS 和 PEO 的存在下,分析物在毛细管壁上的吸附被抑制,导致高效率和重现性。分析物迁移率的日内分析 RSD 值 (n=3) 小于 0.52%。我们通过定量测定乳腺癌细胞 (MCF-7) 中的 9 种氨基酸和正常上皮细胞 (H184B5F5/M10) 中的 10 种氨基酸验证了这种方法的实用性。MCF-7 细胞中的 Tau 和 Gln 浓度与 H184B5F5/M10 细胞中的浓度不同。我们的结果表明该方法在癌症研究中的潜力。

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