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Untargeted and Targeted LC-MS and Data Processing Workflow for the Comprehensive Analysis of Oligopeptides from Cyanobacteria

CHEMOSPHERE(2023)

Univ Napoli Federico II | Laboratory of Photo-Catalytic Processes and Environmental Chemistry | Natl Res Council Canada | Univ Gdansk

Cited 2|Views51
Abstract
Cyanobacteria produce a plethora of structurally diverse bioactive secondary metabolites, including cyanotoxins which pose a serious threat to humans and other living organisms worldwide. Currently, a wide variety of mass spectrometry-based methods for determination of microcystins (MCs), the most commonly occurring and studied class of cyanotoxins, have been developed and employed for research and monitoring purposes. The scarcity of commercially available reference materials, together with the ever-growing range of mass spectrometers and analytical approaches, make the accuracy of quantitative analyses a critical point to be carefully investigated in view of a reliable risk evaluation. This study reports, a comparative investigation of the qualitative and quantitative MCs profile obtained using targeted and untargeted liquid chromatography-mass spectrometry approaches for the analyses of cyanobacterial biomass from Lake Kastoria, Greece. Comparison of the total MCs content measured by the two approaches showed good correlation, with variations in the range of 3.8-13.2%. In addition, the implementation of an analytical workflow on a hybrid linear ion trap/orbitrap mass spectrometer is described, based on combining data-dependent acquisition and a powerful database of cyanobacterial metabolites (CyanoMetDB) for the annotation of known and discovery of new cyanopeptides. This untargeted strategy proved highly effective for the identification of MCs, microginins, anabaenopeptins, and micropeptins. The systematic interpretation of the acquired fragmentation patterns allowed the elucidation of two new MC structural variants, MC-PrhcysR and MC-Prhcys(O)R, and proposal of structures for two new microginins, isomeric cyanostatin B and MG 821A, and three isomeric micropeptins at m/z 846.4715, 846.4711 and 846.4723.
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Key words
Microcystins,Cyanotoxins,Cyanopeptides,Untargeted LC-HRMS,Targeted LC-MS,MS,Data-dependent acquisition
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要点】:比较研究了有针对性和无针对性的液相色谱-质谱方法,用于分析希腊卡斯托利亚湖蓝藻生物质中的微囊藻毒素,发现无针对性策略对于鉴定微囊藻毒素等新型氨基酸肽类化合物非常有效,揭示了一些新的结构变体。

方法】:结合数据依赖性获取和蓝藻代谢物数据库(CyanoMetDB)的强大数据库,在混合线性离子阱/轨道质谱仪上实施分析工作流程。

实验】:比较了通过有针对性和无针对性方法测得的微囊藻毒素总含量,发现两种方法的相关性良好,变动范围为3.8-13.2%。采用无针对性策略有效鉴定了微囊藻毒素、微菌素、黏球藻多肽和微肽,揭示了两种新的微囊藻毒素结构变体,MC-PrhcysR 和 MC-Prhcys(O)R,以及两种新的微菌素结构,同分异构体 cyanostatin B 和 MG 821A,和三种同分异构体的微肽。