In a continuation of studies on the chemical constituents of Scrophulariaceae family, Scrophularia saharae Batt. & Trab. was investigated. The plant material was collected in Biskra, Algeria, with a voucher specimen deposited. Powdered roots were extracted with cyclohexane and CHCl3, and constituents were isolated via silica gel column chromatography and semi-preparative HPLC. A new iridoid glycoside (scrophuloside B5, C42H48O18) together with four known iridoids (harpagoside, 8-O-acetylharpagide, scropolioside B 1, scropolioside D) were isolated. Their structures were identified using spectroscopic methods (UV, IR, ESI-MS, 1D/2D NMR). Chemotaxonomically, these iridoid glycosides (derived from 6-O-α-L-rhamnopyranosyl catalpol or harpagide derivatives) are markers for Scrophulariaceae and Scrophularia genus, consistent with previous studies on Scrophularia species.