HPLC Analysis of Salsola Phytochemicals

To analyze and detect the phenol and flavonoid compounds in the Salsola extract, a specific protocol was followed (30 (link)), with little modification, using the Agilent 1260 Infinity HPLC Series (Agilent, Santa Clara, CA 95051, USA), equipped with a quaternary pump. Kinetex® 5 μm EVO C18 100 × 4.6 mm (Phenomenex, Torrance, CA 90501-1430, USA) was used as the column and was operated at 30°C (31 (link)). The separation method was carried out using a ternary linear elution gradient with (A) 2% HPLC-grade water and H₃PO₄ (v/v; 1:1), (B) methanol, and (C) acetonitrile. Then, a final volume of 20 μL was injected. For the detection of phenols and flavonoids, an AVWD detector set at 284 nm was used.