Integrating metabolomics and indigenous uses of Helichrysum kraussii from neighbouring communities

Abstract

Helichrysum kraussii, a plant from the Asteraceae family is traditionally used to treat various ailments such as coughs, colds, tuberculosis and other infections. Despite its widespread use in rural communities as traditional medicine, there is limited scientific information on its bioactive compounds and pharmacological validation. This study explores the ethnobotanical knowledge and chemical diversity of H. kraussii across six villages in the Vhembe district in Limpopo, namely, Gundani, Vuvha, Sambandou, Tshirovha, Phiphidi and Tshilapfene. By integrating qualitative survey data with advanced metabolomic analysis through the ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-qTOF-MS), the research investigates how local names and traditional uses of H. kraussii reflect cultural identity, geographic variation, and chemical diversity of the plant. Based on the survey findings, H. kraussii was known by different names including Tshifulathulo, Tshifhefhewana and Mulalamuvhuda, depending on the village, with Tshifulathulo being the most recognized across all the studied villages, particularly in Vuvha (100%). This study highlights the impact of cultural practices and lineage diversity in shaping how this medicinal plant is perceived and utilized, pointing to the rich diversity of indigenous knowledge that can be found in different villages that are within the same region. Traditional uses of H. kraussii varied across villages and included treatment for flu, fever, skincare, wound healing, digestive issues, dental care, renal ailments, and sexually transmitted infections. Other cultural uses such as spiritual practices, fire-making, broom-making, and toiletry were also documented. Metabolic profiling of methanolic extracts of H. kraussii from the six villages exhibited a generally similar secondary metabolite composition. However, multivariate statistical tools such as principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and hierarchical cluster analysis (HCA) revealed notable metabolomic distinctions, particularly among samples from Gundani and Tshilapfene. These differences are hypothesized to be influenced by local environmental conditions. Volcano plots and PCA biplots identified several unique metabolites, including caffeic acid-O-hexoside, quinic acid derivatives and cinnamic acid derivatives, to be specific to certain villages. Molecular networking approach further uncovered village-specific chemical markers, such as rosmarinic acid-glucoside in Gundani and acetyl hydroxy rosmanol in Tshirovha, which have not been previously reported in H. kraussii. Protein target prediction of these metabolites suggested that each village-specific compound interacts with distinct biological targets, supporting the hypothesis that chemical variation contributes to the differences observed in traditional medicinal uses. This study emphasizes the importance of integrating ethnobotanical data with metabolomic profiling to preserve indigenous knowledge and provide a scientific foundation for the discovery of novel bioactive compounds. The findings also emphasize the need to expand natural product spectral libraries to better characterize the chemical diversity of under-researched indigenous plants like H. kraussii.