A dispersive liquid-liquid microextraction followed by reverse-phase high-performance liquid chromatography for QuEChERS determination of chlorogenic acid.

Abstract

The presence of chlorogenic acid (CGA) not only imparts a characteristic flavour to foods but also makes them useful against chronic diseases and metabolic syndromes, especially diabetes mellitus and asthma. The present study was designed to develop a quick, easy, cheap, effective, rugged, and safe (QuEChERS) and pragmatic method to analyse CGA in plant-based products by applying dispersive liquid-liquid microextraction (DLLME) followed by reverse-phase high-performance liquid chromatography under a diode array detector (RP-HPLC-DAD). The DLLME was carried out using Triton X-100 as a dispersant and ethanol as an extraction solvent, while method development and validation activities were performed on a Shimadzu 10-AT HPLC-DAD system equipped with C-18 columns as stationary phases. The well-resolved peak for the standard CGA was observed at 2.92 min for the mobile phase comprising 0.1% aqueous formic acid-ethanol (22:78 v/v) of pH 3.0 programmed under an isocratic flow rate of 1.0 mL/min. The developed method was found to be linear (R2 = 0.9976) over 1-500 μg/mL of CGA concentration at 328 nm. Moreover, the assay was found to be repeatable with RSD ≤ 5.0, and the limit of detection (LOD) and limit of quantification (LOQ) were 0.0281 and 0.0853 μg/mL of CGA, respectively. The DLLME offered an overall recovery rate between 97.60% and 99.54% at an acceptable level of reproducibility [relative standard deviation (RSD) ≤ 5.0]. The developed assay was found to be a QuEChERS and pragmatic choice for the high-throughput analysis of CGA in plant-based products/foods. Finally, the analysis revealed the presence of an ample level of CGA in guava fruit in addition to coffee beans and black tea.