Sensitive quantitative analysis of C-peptide in human plasma by 2-dimensional liquid chromatography-mass spectrometry isotope-dilution assay
Sensitive quantitative analysis of C-peptide in human plasma by 2-dimensional liquid chromatography-mass spectrometry isotope-dilution assay
复制标题
二维液相色谱-质谱同位素稀释法灵敏定量分析人体血浆中的 C 肽
DOI：
10.1373/clinchem.2005.063081
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发表时间：
2006-05-01
影响因子：
9.3
通讯作者：
Stein, DT
中科院分区：
文献类型：
--
作者：
Rogatsky, E;Balent, B;Stein, DT
背景：同位素稀释法（IDAs）已被广泛用于生物液体中代谢物或小药物分子的定量分析。由于其增加的特异性，IDAs是一种替代免疫分析法来测量c肽。方法：建立二维液相色谱-质谱（2D LC/MS） IDA方法。样品制备采用离线固相萃取，c肽分离采用Agilent 1100 2D LC系统，采用2个高分辨率反相柱间高压切换的纯化方法。由于c肽的裂解效率较低，因此省略了多反应监测分析，采用选择性离子监测模式进行定量分析。原生和同位素标记（[M+18]和[M+30]）的c -肽在M /z 1007.7、1013.7和1017.7处处于+3状态。结果：该方法线性良好（r(2) = 0.9995），柱上检出限为300 amole （1 pg）。c肽的日内、间CVs分别为
Background: Isotope-dilution assays (IDAs) are well established for quantification of metabolites or small drug molecules in biological fluids. Because of their increased specificity, IDAs are an alternative to immunoassays for measuring C-peptide.Methods: We evaluated a 2-dimensional liquid chromatography-mass spectrometry (2D LC/MS) IDA method. Sample preparation was by off-line solid-phase extraction, and C-peptide separation was performed on an Agilent 1100 2D LC system with a purification method based on high-pressure switching between 2 high-resolution reversed-phase columns. Because of the low fragmentation efficiency of C-peptide, multiple-reaction monitoring analysis was omitted and selective-ion monitoring mode was chosen for quantification. Native and isotope-labeled ([M+18] and [M+30]) C-peptides were monitored in the +3 state at m/z 1007.7, 1013.7, and 1017.7.Results: The assay was linear (r(2) = 0.9995), with a detection limit of 300 amole (1 pg) on column. Inter- and intraday CVs for C-peptide were