Chemical Markers and Bioactivity of Algerian Royal Jelly: Influence of Bee Subspecies, Geography, and Lyophilization.

Royal jelly (RJ) is a chemically complex bee-derived secretion whose composition and biological properties are influenced by honeybee genetics, geography, and processing. Native Algerian honeybee subspecies of the African lineage (Apis mellifera intermissa and Apis mellifera sahariensis) represent an underexplored source of RJ with potential chemical distinctiveness. In this study, fresh and lyophilized RJ samples from three geographically contrasting Algerian regions were characterized using selected chemical markers to assess biodiversity- and processing-related effects. Chemical analyses revealed marked regional variability in 10‑hydroxy‑2‑decenoic acid, with fresh samples containing 1.9%-3.0% (w/w) and lyophilized samples reaching 4.9%-8.4% (w/w). Estimated total phenolic and flavonoid contents also varied by region. Fresh and lyophilized samples showed comparable patterns for the measured markers, supporting the use of both forms for comparative studies. Biological evaluation revealed weak antiradical and antioxidant activities. In cell-based assays, lyophilized RJ selectively reduced Reh cell viability, while Jurkat cells were largely unaffected, with effects attributable to nonspecific cytotoxicity. RJ showed little to no inhibition of leukotriene biosynthesis in a whole-cell 5‑lipoxygenase assay, indicating limited anti-inflammatory activity via this pathway. By integrating selected chemical characterization with biologically relevant cellular assays, this work provides a balanced and mechanistically informed assessment of RJ bioactivity.