Application of screening experimental designs to assess chromatographic isotope effect upon isotope-coded derivatization for quantitative liquid chromatography-mass spectrometry.
Application of screening experimental designs to assess chromatographic isotope effect upon isotope-coded derivatization for quantitative liquid chromatography-mass spectrometry.
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DOI：
10.1021/ac501309s
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发表时间：
2014-07-15
影响因子：
7.4
通讯作者：
Prokai, Laszlo
中科院分区：
文献类型：
--
作者：
Szarka, Szabolcs;Prokai-Tatrai, Katalin;Prokai, Laszlo
同位素效应可能会导致标记（重）和未标记（轻）同位素对的部分色谱分离。与同时发生的基质效应一起，这可能会导致定量液相色谱-质谱分析的准确性不可接受，特别是在使用电喷雾电离时。四种生物学相关的活性醛（丙烯醛、丙二醛、4-羟基-2-壬烯醛和 4-氧代-2-壬烯醛）用轻或重（d3-、13C6-、15N2-或 15N4-标记）2,4-二硝基苯肼衍生化，并用作模型化合物来评估色谱同位素效应。为了全面评估不同梯度反相液相色谱条件下轻/重对之间的保留时间差异，使用包括不对称 (Addelman) 和 Plackett-Burman 方案的实验设计，通过多因素筛选来解决主要色谱参数（固定相、流动相 pH、温度、有机溶剂和梯度斜率）和不同同位素标记，然后进行统计评估。结果证实，避免色谱同位素效应的最有效方法是使用 15N 或 13C 标记代替氘标记，而色谱参数一般无影响。在对生物基质中的一些模型醛进行定量时，使用氘与 15N 标记的替代同位素编码衍生化测定 (AIDA) 的比较给出了不可接受的差异 (>15%)。根据我们的结果，我们建议修改 AIDA 方案，用 15N 或 13C 标记的衍生剂代替 d3-2,4-二硝基苯肼，以避免色谱同位素效应可能产生的不利后果。
Isotope effect may cause partial chromatographic separation of labeled (heavy) and unlabeled (light) isotopologue pairs. Together with a simultaneous matrix effect, this could lead to unacceptable accuracy in quantitative liquid chromatography–mass spectrometry assays, especially when electrospray ionization is used. Four biologically relevant reactive aldehydes (acrolein, malondialdehyde, 4-hydroxy-2-nonenal, and 4-oxo-2-nonenal) were derivatized with light or heavy (d3-, 13C6-, 15N2-, or 15N4-labeled) 2,4-dinitrophenylhydrazine and used as model compounds to evaluate chromatographic isotope effects. For comprehensive assessment of retention time differences between light/heavy pairs under various gradient reversed-phase liquid chromatography conditions, major chromatographic parameters (stationary phase, mobile phase pH, temperature, organic solvent, and gradient slope) and different isotope labelings were addressed by multiple-factor screening using experimental designs that included both asymmetrical (Addelman) and Plackett–Burman schemes followed by statistical evaluations. Results confirmed that the most effective approach to avoid chromatographic isotope effect is the use of 15N or 13C labeling instead of deuterium labeling, while chromatographic parameters had no general influence. Comparison of the alternate isotope-coded derivatization assay (AIDA) using deuterium versus 15N labeling gave unacceptable differences (>15%) upon quantifying some of the model aldehydes from biological matrixes. On the basis of our results, we recommend the modification of the AIDA protocol by replacing d3-2,4-dinitrophenylhydrazine with 15N- or 13C-labeled derivatizing reagent to avoid possible unfavorable consequences of chromatographic isotope effects.
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影响因子：
2
作者：
Jemal, M;Schuster, A;Whigan, DB
通讯作者：
Whigan, DB
影响因子：
2.2
作者：
Hanneman, Sandra K.
通讯作者：
Hanneman, Sandra K.
影响因子：
4.1
作者：
Berg, Thomas;Strand, Dag Helge
通讯作者：
Strand, Dag Helge
DOI：
10.2307/2987937
发表时间：
1983-01-01
期刊：
JOURNAL OF THE ROYAL STATISTICAL SOCIETY SERIES D-THE STATISTICIAN
影响因子：
--
作者：
ALTMAN, DG;BLAND, JM
通讯作者：
BLAND, JM
DOI：
10.1016/j.jchromb.2010.02.010
发表时间：
2010-10-01
影响因子：
3
作者：
Manini, Paola;Andreoli, Roberta;Niessen, Wilfried M. A.
通讯作者：
Niessen, Wilfried M. A.