Analyte and internal standard cross signal contributions and their impact on quantitation in LC–MS based bioanalysis

分析物 生物分析 化学 校准 线性 色谱法 加权 信号(编程语言) 校准曲线 样品制备 航程(航空) 检出限 质谱法 蛋白质沉淀 选择性反应监测 分析化学(期刊) 准确度和精密度 定量分析(化学) 生物系统 重复性 再现性 线性范围 统计 计算机科学 数学 材料科学 放射科 物理 复合材料 医学 量子力学 程序设计语言 生物
作者
Aimin Tan,Isabelle A. Lévesque,François Limoge Et Laure Viel,Nadine Boudreau,Ann Lévesque
出处
期刊:Journal of Chromatography B [Elsevier BV]
卷期号:879 (21): 1954-1960 被引量:42
标识
DOI:10.1016/j.jchromb.2011.05.027
摘要

Cross signal contributions between an analyte and its internal standard (IS) are very common due to impurities in reference standards and/or isotopic interferences. Despite the general awareness of this issue, how exactly they affect quantitation in LC–MS based bioanalysis has not been systematically evaluated. In this research, such evaluations were performed first by simulations and then by experiments using a typical bioanalytical method for tiagabine over the concentration range of 1–1000 ng/mL in human EDTA K3 plasma. The results demonstrate that when an analyte contributes to IS signal, linearity and accuracy can be affected with low IS concentration. Thus, minimum IS concentrations have been obtained for different combinations of concentration range, percentage of cross contribution, and weighting factor. Moreover, while impurity in analyte reference standard is a factor in cross signal contribution, significant systematic errors could exist in the results of unknown samples even though the results of calibration standards and quality controls are acceptable. How these systematic errors would affect stability evaluation, method transfer, and cross validation has also been discussed and measures to reduce their impact are proposed. On the other hand, the signal contribution from an IS to the analyte causes shifting of a calibration curve, i.e. increase of intercept, and theoretically, the accuracy is not affected. The simulation results are well supported by experimental results. For example, good inter-run (between-run) accuracy (bias: −2.70 to 5.35%) and precision (CV: 2.07–10.50%) were obtained when runs were extracted with an IS solution containing 1-fold of the lower limit of quantitation.
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