HPLC determination of hesperidin, diosmin and eriocitrin in Iranian lime juice using polyamide as an adsorbent for solid phase extraction

Solid phase extraction (SPE) and HPLC were used for simultaneous determination of hesperidin, diosmin and eriocitrin in Iranian lime juice samples. The method involved very simple efficient SPE with polyamide cartridge, the use of mixture of water/acetonitrile/acetic acid (78:19:3, v/v) as a mobile phase at a flow rate of 0.8 mL/min and UV detection at 280 nm. Optimum conditions for SPE were achieved using 8 mL water/methanol (85:15, v/v, pH = 3) as the washing solution and 4 mL methanol for elution. SPE parameters, such as maximum loading capacity and breakthrough volume, were also determined for each analyte. Good clean-up and high > 90% were observed for all analytes. Limits of detection, limits of quantification, linear range, recovery, repeatability of retention times, and peak areas for hesperidin, diosmin and eriocitrin were 0.0283–0.0512 μg/mL, 0.0857–0.155 μg/mL, 0.0283–105.0 μg/mL (R2 > 0.99), 93.3–98.1%, 3.2–4.7% and 2.8–3.6%, respectively. The method was applied to analysis of lime juice samples obtained from different locations of Iran.