This study investigates the phytochemical composition and acetylcholinesterase (AChE) inhibitory activity of Anabasis salsa (C.A. Mey.) Benth. ex Volkens (Chenopodiaceae), with a particular focus on secondary metabolites present in its extracts. The plant extracts were profiled using HPLC-ESI-QTOF-MS/MS, revealing a diverse composition including organic acids, phenolic compounds, flavonoids, and alkaloids. The latter were represented by anabasine and lupinine that were identified as significantly present compounds. Depending on the type of extract and plant part, lupinine was present in the extracts at a quantity range from 0.0033 to 6.35%, and its highest concentration was noted in the acidified macerate from the overground parts of the plant. Centrifugal partition chromatography (CPC) operated using the solvent system composed of MTBE:ACN:n-BuOH:H₂O, 2:2:1:5 (v/v/v/v), was applied for the fractionation of the most promising extract, enabling efficient separation using a biphasic solvent system. The obtained fractions were evaluated for AChE inhibitory activity, and the most active fractions were found to contain lupinine, which was successfully isolated in a purified fraction using CPC. These findings highlight lupinine as a significant bioactive compound with neuroprotective potential and demonstrate the applicability of CPC as an efficient and scalable method for the isolation of alkaloids from complex plant matrices.