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In the present study, for the first time, a simplified miniaturized ultrasound-assisted matrix solid-phase dispersion (SM-USA-MSPD) method with a different application for liquid matrices was developed to extract different flavonoids (hesperidin, diosmin, eriocitrin, narirutin, naringin, hesperetin and naringenin) from citrus fruit juice and human fluid samples prior to their determination using high performance liquid chromatography (HPLC). Different effective parameters were studied and under the optimum conditions (including sample volume: 150 μL; solid phase: silica-based C18, 200 mg; eluting solvent: methanol, 500 μL; pH: 4; and sonication: 6 min; at room temperature), limits of detection and limits of quantification were ranged from 23.3 to 46.8 ng mL⁻¹ and 74.8 to 141.5 ng mL⁻¹, respectively. Once optimized, analytical performance of the method was studied in terms of linearity (0.074–198.5 μg mL⁻¹, r² > 0.991 …