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采用电子转移基质辅助激光解吸/电离质谱(ET-MALDI-MS)对细胞、提取物和完整叶绿体进行比较分析。

Comparative profiling of cells, extracts, and intact chloroplasts using electron transfer matrix-assisted laser desorption/ionization mass spectrometry (ET-MALDI-MS).

机构信息

Escuela de Química, Universidad Industrial de Santander, Bucaramanga, Santander, 680002, Colombia.

出版信息

Anal Methods. 2024 Aug 22;16(33):5652-5664. doi: 10.1039/d4ay00846d.

DOI:10.1039/d4ay00846d
PMID:39109659
Abstract

The intricate composition of microalgal pigments plays a crucial role in various biological processes, from photosynthesis to biomarker identification. Traditional pigment analysis methods involve complex extraction techniques, posing challenges in maintaining analyte integrity. In this study, we employ Electron Transfer Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (ET-MALDI-MS) to compare the pigmentary profiles of intact cells, chloroplasts, and solvent extracts. We aim to obtain comprehensive extracts rich in polar and non-polar compounds using ultrasound-assisted and supercritical fluid extraction methods. Additionally, intact chloroplasts are isolated using a lysis buffer and sucrose density gradient centrifugation. Our ET-MALDI-MS analysis reveals distinct compositional differences, highlighting the impact of extraction protocols on microalgal pigment identification. We observe prominent signals corresponding to radical cations of key pigments, including chlorophylls and carotenoids, which are crucial for identification. Furthermore, ET-MALDI-MS facilitates the identification of specific lipids within chloroplast membranes and other organelles. This study underscores the rapid and precise nature of ET-MALDI-MS in microalgal biomarker analysis, providing valuable insights into phytoplankton dynamics, trophic levels, and environmental processes. emerges as a promising model for studying pigment composition and membrane lipid diversity, enhancing our understanding of microalgal ecosystems.

摘要

微藻色素的复杂组成在各种生物过程中起着至关重要的作用,从光合作用到生物标志物鉴定。传统的色素分析方法涉及复杂的提取技术,在保持分析物完整性方面存在挑战。在本研究中,我们采用电子转移基质辅助激光解吸/电离质谱(ET-MALDI-MS)来比较完整细胞、叶绿体和溶剂提取物的色素特征。我们旨在使用超声辅助和超临界流体提取方法获得富含极性和非极性化合物的综合提取物。此外,使用裂解缓冲液和蔗糖密度梯度离心分离完整的叶绿体。我们的 ET-MALDI-MS 分析揭示了明显的组成差异,突出了提取方案对微藻色素鉴定的影响。我们观察到与关键色素,包括叶绿素和类胡萝卜素的自由基阳离子对应的突出信号,这些色素对于鉴定至关重要。此外,ET-MALDI-MS 有助于鉴定叶绿体膜和其他细胞器内的特定脂质。这项研究强调了 ET-MALDI-MS 在微藻生物标志物分析中的快速和精确性质,为浮游植物动态、营养水平和环境过程提供了有价值的见解。该模型为研究色素组成和膜脂多样性提供了一种有前途的方法,增强了我们对微藻生态系统的理解。

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