Characterization of bioactive compounds in American elderberry (Sambucus nigra subsp. canadensis) juices
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American elderberry (Sambucus nigra subsp. canadensis) is an emerging specialty crop in North America, valued for its rich composition of bioactive compounds, which have demonstrated nutritional and therapeutic potential. However, comprehensive chemical and functional characterization of American elderberry remain limited. This dissertation integrates advanced metabolomic analysis, bioassay screening, and molecular mechanism studies to elucidate the chemical diversity and antiviral activity of American elderberry. An untargeted ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC–HRMS) metabolomics analysis, combined with high-throughput bioassay screening, was employed to characterize juices from 18 propagated accessions and three established American elderberry cultivars. More than 100 putative bioactive compounds were identified, revealing genotype-dependent differences in metabolite composition. Cultivar Ozark and several wild accessions, such as accession 1911, displayed distinct chemical signatures. Among 32 tested compounds, 14 exhibited strong antioxidant activity, and six demonstrated antiviral effects against HIV-1, highlighting elderberry's potential in functional food and nutraceutical applications. This study further investigated the antiviral properties of American elderberry against the Influenza A Polymerase Acidic (PA) protein using fluorescence resonance energy transfer (FRET)-based endonuclease assay. Four cultivars, Ozark, accession 1892, 1196, and 2084, showed inhibition activity greater than 65% against PA activity, surpassing the European cultivar ‘Haschberg'. Several phenolic compounds, including gallic acid, myricetin, caffeic acid, and luteolin, displayed potent inhibition of PA endonuclease activity (IC₅₀ < 30 μM), suggesting their role as key bioactive constituents underlying elderberry's anti-influenza potential. A targeted ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) method was developed and validated to quantify the bioactive compounds in American elderberry across 21 cultivars. The optimized UHPLC–MS/MS platform achieved excellent detection limits (< 1 ng/mL), minimized matrix effects, and provided accurate quantification of complex juice extracts. This study revealed genotype-dependent variation in metabolite composition, highlighting opportunities for targeted breeding and cultivar selection to enhance desired phytochemical traits, such as antioxidant capacity, antiviral bioactivity, and overall nutritional quality. Notably, wild-propagated accession 1199 exhibited high levels of cyanidin-based anthocyanins, suggesting its utility as a natural food color source. Phenolic acid compounds were found most abundant in accession 1196, whereas the flavonoid isorhamnetin 3-rutinoside was found highest in cultivar Ozark. Collectively, this work establishes a robust, comprehensive, analytical, and bioactivity framework for the characterization of American elderberry. By integrating metabolomic profiling, targeted quantification, and antiviral mechanism studies, these findings advance the understanding of elderberry phytochemistry and support its development as a source of functional and bioactive compounds for nutraceutical applications.
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Ph. D.