Evaluation of Alternate Isotope-Coded Derivatization Assay (AIDA) in the LC-MS/MS analysis of aldehydes in exhaled breath condensate
Evaluation of Alternate Isotope-Coded Derivatization Assay (AIDA) in the LC-MS/MS analysis of aldehydes in exhaled breath condensate
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DOI：
10.1016/j.jchromb.2010.02.010
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发表时间：
2010-10-01
影响因子：
3
通讯作者：
Niessen, Wilfried M. A.
中科院分区：
文献类型：
--
作者：
Manini, Paola;Andreoli, Roberta;Niessen, Wilfried M. A.
我们提出了一种新的同位素稀释方法，称为交替同位素编码衍生法（AIDA），用于呼气冷凝物（EBC）样品中丙二醛（MDA）和4-羟基烯醛（4-HNE）的腙衍生物的液相色谱-串联质谱分析。AIDA是基于分析物与两种纯同位素形式（分别为“轻”和“重”）的试剂的交替衍生化，通过使用轻衍生化的标准来定量重衍生化的分析物，反之亦然。为此，合成了2,4-二硝基-3,5,6-三氘苯基肼（d(3)-DNPH），并与商业“轻”DNPH结合作为“重”试剂使用。使用AIDA方法，可以在不需要校准曲线的情况下计算出基体中任何未知的分析物浓度。为了进行比较，还研究了一种外部校准方法。DNPH和d3-DNPH衍生物的稳定性通过排除任何相互交换的腙来验证。在具有生物学意义的浓度范围内，例如MDA为2-40 nM， 4-HNE为0.5-10 nM， MDA和4-HNE与DNPH和d3-DNPH的衍生化反应表现出重叠动力学和相似的产率。DNPH和d3-DNPH腙的MS响应相似。以%RSD计算，MDA和4-HNE的准确度分别在3.2-8%和4.5-11%之间。通过分析加标的EBC池样本来测试准确性，在空白减去后，通过AIDA获得了可接受的结果（MDA的准确度在98-108%之间，4-HNE的准确度在93-114%之间），这是不可忽略的。用AIDA法对EBC样品中MDA和4-HNE的定量分析结果与用外标法对相同样品的定量分析结果一致。(C) 2010 Elsevier B.V.版权所有
We present the application of a novel isotope dilution method, named Alternate Isotope-Coded Derivatization Assay (AIDA), to the quantitative analysis of hydrazone derivatives of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) in exhaled breath condensate (EBC) samples using liquid chromatography-tandem mass spectrometry. AIDA is based on the alternate derivatization of the analyte(s) with reagents that are available in two pure isotopic forms, respectively "light" and "heavy", by using light-derivatized standards for the quantification of the heavy-derivatized analytes, and vice versa. To this purpose, 2,4-dinitro-3,5,6-trideuterophenylhydrazine (d(3)-DNPH) has been synthesized and used as "heavy" reagent in combination with commercial "light" DNPH. Using the AIDA method, any unknown concentration of the analyte in the matrix can be calculated without the need of a calibration curve. An external calibration method has been also investigated for comparative purpose. The stability of DNPH and d3-DNPH derivatives was verified by excluding any exchange of hydrazones with each other. In the range of concentrations of biological interest, e.g., 2-40 nM for MDA and 0.5-10 nM for 4-HNE, the derivatization reactions of MDA and 4-HNE with DNPH and d3-DNPH showed overlapping kinetics and comparable yields. The MS response of both DNPH and d3-DNPH hydrazones was similar. The precision of AIDA, calculated as %RSD, was within 3.2-8% for MDA and 4.5-11% for 4-HNE. Accuracy was tested by analyzing a spiked EBC pool sample and acceptable results (accuracy within 98-108% for MDA and 93-114% for 4-HNE) were obtained by AIDA after subtraction of the blank, which was not negligible. The results of quantitative analysis of MDA and 4-HNE in EBC samples obtained by AIDA assay with four analyses per sample were in good agreement with those obtained by external calibration method on the same samples. (C) 2010 Elsevier B.V. All rights reserved.