Untargeted Metabolomics and Targeted Phytohormone Profiling of Sweet Aloes (Euphorbia neriifolia) from Guyana: An Assessment of Asthma Therapy Potential in Leaf Extracts and Latex.

Background/Objectives:Euphorbia neriifolia is a succulent plant from the therapeutically rich family of Euphorbia comprising 2000 species globally. E. neriifolia is used in Indigenous Guyanese asthma therapy. Methods: To investigate E. neriifolia's therapeutic potential, traditionally heated leaf, simple leaf, and latex extracts were evaluated for phytohormones and therapeutic compounds. Full scan, data-dependent acquisition, and parallel reaction monitoring modes via liquid chromatography Orbitrap mass spectrometry were used for screening. Results: Pathway analysis of putative features from all extracts revealed a bias towards the phenylpropanoid, terpenoid, and flavonoid biosynthetic pathways. A total of 850 compounds were annotated using various bioinformatics tools, ranging from confidence levels 1 to 3. Lipids and lipid-like molecules (34.35%), benzenoids (10.24%), organic acids and derivatives (12%), organoheterocyclic compounds (12%), and phenylpropanoids and polyketides (10.35%) dominated the contribution of compounds among the 13 superclasses. Semi-targeted screening revealed 14 out of 16 literature-relevant therapeutic metabolites detected, with greater upregulation in traditional heated extracts. Targeted screening of 39 phytohormones resulted in 25 being detected and quantified. Simple leaf extract displayed 4.4 and 45 times greater phytohormone levels than traditional heated leaf and latex extracts, respectively. Simple leaf extracts had the greatest nucleotide and riboside cytokinin and acidic phytohormone levels. In contrast, traditional heated extracts exhibited the highest free base and glucoside cytokinin levels and uniquely contained methylthiolated and aromatic cytokinins while lacking acidic phytohormones. Latex samples had trace gibberellic acid levels, the lowest free base, riboside, and nucleotide levels, with absences of aromatic, glucoside, or methylthiolated cytokinin forms. Conclusions: In addition to metabolites with possible therapeutic value for asthma treatment, we present the first look at cytokinin phytohormones in the species and Euphorbia genus alongside metabolite screening to present a comprehensive assessment of heated leaf extract used in Indigenous Guyanese asthma therapy.