Simplified miniaturized ultrasound-assisted matrix solid phase dispersion extraction and high performance liquid chromatographic determination of seven flavonoids in citrus fruit juice and human fluid samples: Hesperetin and naringenin as biomarkers

• SM-USA-MSPD was applied for the first time to determine seven flavonoids.
• This is an efficient method for analysis of flavonoids in human fluids by HPLC.
• This method is faster and more accurate than the previous literatures.
• The flavonoids were well separated with appropriate retention times.
• Hesperetin and naringenin can be good biomarkers of citrus fruit juice intake.

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), accuracy (recovery = 84.6–101.5%), and precision (repeatability: intra-day precision < 5.9%, and inter-day precision < 7.2%). At the end, SM-USA-MSPD method was successfully applied to estimate the levels of hesperetin and naringenin in plasma and urinary excretion -after ingestion of orange, grapefruit and lime juices- and the obtained results confirmed that these compounds could be used as good biomarkers of citrus fruit juice intake.