Integrated Omics Center, Life/Health Division, Korea Institute of Science and Technology, Seoul, Republic of Korea.
J Sep Sci. 2010 Dec;33(23-24):3701-9. doi: 10.1002/jssc.201000020.
An analytical method of CE-MS and CE with an online preconcentration technique induced by a dynamic pH junction, addition of organic solvent and large volume injection was developed for sensitive determination of peptides in biological samples. Leucine enkephalin, methionine enkephalin, dynorphin A, β-endorphin and angiotensin II were used as model peptides. The optimal online preconcentration conditions were obtained at a sample matrix consisting of 100 mM borate buffer (pH 10.0) with 50% v/v acetonitrile and a BGE containing 1 M formic acid at pH 2.0, along with a 25-cm injection length. Under the optimized conditions, a 4.0×10(3)-1.1×10(4)-fold increase in peak intensity was achieved without degrading the peak shape. This online preconcentration method was applied to analyze the intracellular angiotensin II within the peptides extracted from HL1 cells and approximately increase of 1×10(4)-fold sensitivity was achieved compared to normal condition. Thus, the developed method could be applied to the analysis of various peptides for peptidomics study in biological samples.
建立了一种采用 CE-MS 和 CE 联用技术，通过动态 pH 结、有机溶剂添加和大体积进样在线浓缩技术，用于生物样品中肽的灵敏检测的分析方法。亮氨酸脑啡肽、甲硫氨酸脑啡肽、强啡肽 A、β-内啡肽和血管紧张素 II 被用作模型肽。在由 100 mM 硼酸缓冲液（pH 10.0）和 50% v/v 乙腈组成的样品基质中，并在 pH 2.0 时含有 1 M 甲酸的 BGE 中，获得了最佳的在线浓缩条件，与 25-cm 的进样长度。在最佳条件下，在不破坏峰形的情况下，峰强度提高了 4.0×10(3)-1.1×10(4)倍。该在线浓缩方法用于分析从 HL1 细胞中提取的肽内的细胞内血管紧张素 II，与正常条件相比，灵敏度提高了约 1×10(4)倍。因此，该方法可用于生物样品中各种肽的分析，用于肽组学研究。
