van 't Hoff Institute for Molecular Sciences, Analytical Chemistry Group, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, the Netherlands; Centre for Analytical Sciences Amsterdam (CASA), the Netherlands.
Department of Chemistry, Gustavus Adolphus College, Saint Peter, MN 56082, United States.
J Chromatogr A. 2021 Feb 22;1639:461922. doi: 10.1016/j.chroma.2021.461922. Epub 2021 Jan 21.
A peak-tracking algorithm was developed for use in comprehensive two-dimensional liquid chromatography coupled to mass spectrometry. Chromatographic peaks were tracked across two different chromatograms, utilizing the available spectral information, the statistical moments of the peaks and the relative retention times in both dimensions. The algorithm consists of three branches. In the pre-processing branch, system peaks are removed based on mass spectra compared to low intensity regions and search windows are applied, relative to the retention times in each dimension, to reduce the required computational power by elimination unlikely pairs. In the comparison branch, similarity between the spectral information and statistical moments of peaks within the search windows is calculated. Lastly, in the evaluation branch extracted-ion-current chromatograms are utilized to assess the validity of the pairing results. The algorithm was applied to peptide retention data recorded under varying chromatographic conditions for use in retention modelling as part of method optimization tools. Moreover, the algorithm was applied to complex peptide mixtures obtained from enzymatic digestion of monoclonal antibodies. The algorithm yielded no false positives. However, due to limitations in the peak-detection algorithm, cross-pairing within the same peaks occurred and six trace compounds remained falsely unpaired.
开发了一种用于与质谱联用的二维液相色谱的峰跟踪算法。利用可用的光谱信息、峰的统计矩和两个维度中的相对保留时间，在两个不同的色谱图中跟踪色谱峰。该算法由三个分支组成。在预处理分支中，根据与低强度区域的质谱比较去除系统峰，并应用相对于每个维度的保留时间的搜索窗口，以通过消除不太可能的对来减少所需的计算能力。在比较分支中，计算搜索窗口内峰的光谱信息和统计矩之间的相似性。最后，在评估分支中，使用提取离子电流色谱图来评估配对结果的有效性。该算法应用于在不同色谱条件下记录的肽保留数据，用于保留建模作为方法优化工具的一部分。此外，该算法还应用于从单克隆抗体酶解获得的复杂肽混合物。该算法没有产生假阳性。然而，由于峰检测算法的限制，同一峰内发生了交叉配对，并且有六种痕量化合物仍然错误地未配对。
