Determination of hesperidin in Pericarpium Citri Reticulatae by semi-micro HPLC with electrochemical detection

Determination of hesperidin contents in Pericarpium Citri Reticulatae was performed by a simple extraction with methanol and semi-micro high-performance liquid chromatography with electrochemical detection (μHPLC–ECD). Chromatography was performed using a microbore octadecylsilica (ODS) column, methanol–water–phosphoric acid (40:60:0.5, v/v/v), as a mobile phase and applied potential at +0.9 V versus Ag/AgCl. Peak heights were found linearly related to the concentrations of hesperidin injected 9.16 ng/ml to 3.06 μg/ml (r > 0.999). The detection limit (S/N = 3) was 3.06 ng/ml (15.3 pg). Hesperidin of 305 ng/ml was detected with a relative standard deviation (R.S.D.) of 0.79% (n = 5). Hesperidin in Pericarpium Citri Reticulatae was extracted with methanol, diluted with the mobile phase, and injected into the μHPLC–ECD for determination. The hesperidin content of Pericarpium Citri Reticulatae from four different districts in China were determined with R.S.D. of 3.59%, 2.29%, 2.36%, and 2.32% (n = 5), respectively. Recoveries of hesperidin from the four Pericarpium Citri Reticulatae sources were 100.3%, 99.83%, 100.7%, and 100.6%, respectively. This method is useful for the determination of hesperidin in Pericarpium Citri Reticulatae, and especially so for instances when samples are sparse.