Lobas Anna A, Solovyeva Elizaveta M, Saparbaev Erik, Gorshkov Mikhail V, Boyarkin Oleg V
Laboratoire de Chimie Physique Moléculaire, École Polytechnique Fédérale de Lausanne, CH-1015, Lausanne, Switzerland; V.L. Talrose Institute for Energy Problems of Chemical Physics, Federal Research Center of Chemical Physics, RAS, Moscow, Russia.
Laboratoire de Chimie Physique Moléculaire, École Polytechnique Fédérale de Lausanne, CH-1015, Lausanne, Switzerland; V.L. Talrose Institute for Energy Problems of Chemical Physics, Federal Research Center of Chemical Physics, RAS, Moscow, Russia; Moscow Institute of Physics and Technology (National Research University), Dolgoprudny, Russia.
Talanta. 2021 Sep 1;232:122412. doi: 10.1016/j.talanta.2021.122412. Epub 2021 Apr 24.
Identification of isomeric biomolecules remains a challenging analytical problem. A recently developed spectroscopic method that combines UV photofragmentation and mass spectrometry for fingerprinting of cold ions (2D UV-MS), has already demonstrated its high performance in the library-based identification and quantification of different types of biomolecular isomers. The practical use of the method has been hindered by a slow rate of data acquisition, which makes the fingerprinting incompatible with high-throughput analysis and online liquid chromatography (LC) separation. Herein we demonstrate how the use of a few pre-selected wavelengths can accelerate the method by two orders of magnitude without a significant loss of accuracy. As a proof of principle, 2D UV-MS fingerprinting was coupled to online LC separation and tested for quantification of isomeric peptides containing either Asp or isoAsp residues. The relative concentrations of the peptides mixed in solution have been determined, on average, with better than 4% and 6% accuracy for resolving and non-resolving gradients of LC separation, respectively.
鉴定同分异构生物分子仍然是一个具有挑战性的分析问题。最近开发的一种光谱方法，它结合了紫外光解离和质谱用于冷离子指纹识别（二维紫外-质谱），已经在基于文库的不同类型生物分子异构体的鉴定和定量中展示了其高性能。该方法的实际应用受到数据采集速率缓慢的阻碍，这使得指纹识别与高通量分析及在线液相色谱（LC）分离不兼容。在此我们展示了如何通过使用几个预先选择的波长将该方法加速两个数量级，同时不会显著损失准确性。作为原理证明，二维紫外-质谱指纹识别与在线LC分离相结合，并用于定量含有天冬氨酸（Asp）或异天冬氨酸（isoAsp）残基的同分异构肽。对于LC分离的洗脱和非洗脱梯度，混合在溶液中的肽的相对浓度平均分别以优于4%和6%的准确度得以确定。
