A high-throughput and eco-friendly method for determination of hesperidin in Citri Reticulatae Pericarpium by solid phase extraction and microplate reader

Abstract

Objective: To develop a rapid, cost-effective, and eco-friendly method that combines solid-phase extraction (SPE) with microplate reader technology for quantifying hesperidin in Citri Reticulatae Pericarpium (CRP) slices, thereby enhancing quality evaluation standards for CRP slices. Methods: Ultrasonic methanol extraction of CRP slices, purification on octadecylsilyl (ODS) SPE, and direct microscale detection by a microplate reader to enable high-throughput quantification without chromatographic separation. Results: The method achieved 96.48 % recovery (relative standard deviation, RSD = 5.88 %) with linearity (correlation coefficient, r > 0.998), and its results are comparable to Pharmacopoeia HPLC standards. Conclusions: The SPE–microplate reader approach is simple, rapid, and sustainable for hesperidin determination in CRP slices, advancing high-throughput quality assessment of traditional Chinese medicines and promoting green analytical practices. It provides a scientific basis for improving CRP slices quality standards and supports broader applications in herbal medicine analysis.