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Highlights • Extraction and isolation of calystegines from leaves, flowers and seeds of Hyoscyamus albus growing in Bejaia (Algeria). • Chromatographic analysis led to the identification of several calystegines in all samples with a new one calystegine B 4 never related in the plant. • Antioxidant tests showed a greater efficiency of seeds extract. • In-vivo anti-inflammatory investigation demonstrated potent potential of all extracts with pic of activity achieved 2 h after injection of carrageenan.