Can Statistical Evaluation Tools for Chromatographic Method Development Assist in the Natural Products Workflow? A Case Study on Selected Species of the Plant Family Malpighiaceae

Abstract

Proper chromatographic methods may reduce the challenges inherent in analyzing natural product extracts, especially when utilizing hyphenated detection techniques involving mass spectrometry. As there are many variations one can introduce during chromatographic method development, this can become a daunting and time-consuming task. To reduce the number of runs and time needed, the use of instrumental automatization and commercial software to apply Quality by Design and statistical analysis automatically can be a valuable approach to investigate complex matrices. To evaluate this strategy in the natural products workflow, a mixture of nine species from the family Malpighiaceae was investigated. By this approach, the entire data collection and method development procedure (comprising screening, optimization, and robustness simulation) was accomplished in only 4 days, resulting in very low limits of detection and quantification. The analysis of the individual extracts also proved the efficiency of the use of a mixture of extracts for this workflow. Molecular networking and library searches were used to annotate a total of 61 compounds, including O-glycosylated flavonoids, C-glycosylated flavonoids, quinic/shikimic acid derivatives, sterols, and other phenols, which were efficiently separated by the method developed. These results support the potential of statistical tools for chromatographic method optimization as an efficient approach to reduce time and maximize resources, such as solvents, to get proper chromatographic conditions.

UHPLC-(ESI)-HRMS and NMR-Based Metabolomics Approach to Access the Seasonality of Byrsonima intermedia and Serjania marginata From Brazilian Cerrado Flora Diversity

“…Previous phytochemical studies conducted on the leaf extracts of the plant species mentioned above have shown that B. intermedia has phenolic acids, oligomeric proanthocyanidins, and flavonoids as its main constituents ( Pereira et al, 2015 ; Fraige et al, 2018 ; Mannochio-Russo et al, 2020 ) and S. marginata has flavonoids, oligomeric proanthocyanidins and saponins as its main constituents ( Heredia-Vieira et al, 2015 ).…”

Untargeted Metabolomics Sheds Light on the Diversity of Major Classes of Secondary Metabolites in the Malpighiaceae Botanical Family

“…On the other hand, C -glycosylated flavonoids present a very distinct fragmentation pathway, with many more fragments observed between the precursor ion and the aglycone. The loss of the glycosidic portion is usually reached by the consecutive losses of water molecules and 120 Da, characteristic of C -glycosylated flavonoids ( Mannochio-Russo et al, 2020 ). Therefore, the distinction of these two molecular families in the molecular networking analysis is expected.…”

“…The Malpighiaceae plant family is an excellent example of both the high diversity of secondary metabolites produced by plant species, and a flowering plant family with all its genera sampled in DNA-based phylogenetic studies ( Davis and Anderson, 2010 ; Mannochio-Russo et al, 2020 ). Malpighiaceae is one of the 36 families of flowering plants placed in Malpighiales by several phylogenetic studies based on chloroplast genes ( Angiosperm Phylogeny Group, 1998 , 2003 , 2009 ; Angiosperm Phylogeny Group et al, 2016 ), being also one of the most important and diverse orders of angiosperms in tropical forests ( Xi et al, 2012 ; Cai et al, 2019 ).…”

Insulin-Mimic Components in Acer truncatum Leaves: Bio-Guided Isolation, Annual Variance Profiling and Regulating Pathway Investigated by Omics

“…It has a high sensitivity to detect trace substances but also can determine chemical structures simultaneously. Furthermore, with the continuous improvement of reference databases and data mining methods, LC-MS/MS provides more and more reliable analysis in metabolomics [ 28 , 29 ], which is often used in exploring biological metabolites [ 30 ] and natural products in traditional medicines or foods [ 31 , 32 ].…”