Simultaneous Determination of Flavanones, Hydroxycinnamic Acids and Alkaloids in Citrus Fruits by HPLC-DAD-ESI/MS

A simple and accurate method has been developed to simultaneously separate and determine 10 bioactive compounds in citrus fruits by high-performance liquid chromatography coupled with diode array detection and electrospray ionisation mass spectrometry (HPLC-DAD-ESI/MS). This HPLC assay was performed on a reversed-phase C18 column with acetonitrile and 0.1% (v/v) aqueous formic acid as mobile phase. DAD has been performed at 273, 283 and 324nm for quantification of the alkaloids, flavanones and hydroxycinnamic acids. MS was also employed to identify the each analyte. Ten analytes (naringin, hesperidin, neohesperidin, ferulic acid, p-coumaric acid, chlorogenic acid, caffeic acid, octopamine, synephrine and tyramine) demonstrated good linearity (r⩾0.9990) in a relatively wide concentration range. The method revealed high average recovery (range, 92.1-97.9%) and good precision with interday and intraday variations with less than 4.71%. The limits of detection (LODs) ranged from 0.02 to 0.11μg/ml, while the limits of quantification (LOQs) were defined in the range of 0.08-0.39μg/ml. The proposed method has been successfully applied to analyse three types of bioactive constituents in eight citrus hybrids pulps and eight citrus hybrids peels, which has been successfully cultivated in China.