Institute of Chemistry, St. Petersburg State University, Universitetskaya nab. 7/9, St. Petersburg, 199034, Russian Federation.
Almazov National Medical Research Centre, Akkuratova 2, St. Petersburg, 197341, Russian Federation.
Anal Bioanal Chem. 2022 Feb;414(4):1723-1737. doi: 10.1007/s00216-021-03805-7. Epub 2022 Jan 3.
A broadband collision-induced dissociation (bbCID) fragmentation mode was proposed for liquid chromatography-mass spectrometric targeted analysis of tryptic peptides obtained from proteins in samples of decoration paint coating. In this approach, a mass spectrometric dataset contains the information on the parent and all fragment ions. This maintains a balance between the quantity of simultaneously acquired data and the sensitivity of the method, which is beneficial under coupling with analytical chromatography. In this study, characteristic peptides were selected for casein, ovalbumin, and collagen, which are the most commonly used binder proteins in the artworks. A simplified sample preparation protocol including only protein extraction and trypsinization was tested and successfully implemented. The combination of analytical chromatography with bbCID MS technique is a lower cost alternative to the use of high-end nano-LC-MS approaches in the investigation of cultural heritage objects of regional or local importance, e.g., prior to and/or during restoration works. It was demonstrated that, for the paint coating samples, the required level of sensitivity could be acquired through the data-independent MS/MS strategy. The proposed approach was tested on a sample obtained during the restoration work at the Gromov cottage in the Lopukhin Garden (middle of the XIX century). As a result, the main protein component, collagen, was identified using 6 characteristic peptides, which may indicate the use of gelatin-based glue. For instance, the identification of the peptide GVQGPPoxGPAGPR of the incoming collagen composition α-1 was undertaken by three parameters: m/z of the precursor ion of 553.2910, m/z of the fragment ion y9 of 821.4238, and retention time of 1.9 min.
提出了一种宽带碰撞诱导解离（bbCID）碎裂模式，用于对来自装饰漆涂层样品中蛋白质的胰蛋白酶肽进行液相色谱-质谱靶向分析。在这种方法中，质谱数据集包含母离子和所有碎片离子的信息。这在与分析色谱耦合时保持了同时采集数据的数量和方法的灵敏度之间的平衡，这对于耦合分析是有益的。在本研究中，选择了用于酪蛋白、卵清蛋白和胶原蛋白的特征肽，它们是艺术品中最常用的结合蛋白。测试并成功实施了仅包括蛋白质提取和胰蛋白酶消化的简化样品制备方案。与使用高端纳升 LC-MS 方法相比，分析色谱与 bbCID MS 技术的结合是调查具有地区或地方重要性的文化遗产对象（例如，在修复工作之前和/或期间）的一种低成本替代方法。结果表明，对于漆涂层样品，可以通过数据非依赖性 MS/MS 策略获得所需的灵敏度水平。该方法在洛普金花园格罗莫夫小屋（19 世纪中叶）修复工作期间获得的样品上进行了测试。结果，使用 6 种特征肽鉴定了主要蛋白质成分胶原蛋白，这可能表明使用了基于明胶的胶。例如，通过三个参数对来自传入胶原蛋白组成 α-1 的肽 GVQGPPoxGPAGPR 进行了鉴定：前体离子 m/z 为 553.2910、碎片离子 y9 的 m/z 为 821.4238 和保留时间为 1.9 分钟。
