Abstract
Hesperidin (HSP) is a bioactive flavanone glycoside, present abundantly in the variety of citrus fruits. The environmental safety and sustainability of the reported analytical assays of HSP analysis have not been considered in the literature. Hence, a sensitive and sustainable "reversed-phase high-performance thin-layer chromatography (RP-HPTLC)" method has been developed and validated for HSP analysis in traditional (TE) and ultrasound-based (UBE) extracts of four different varieties of citrus fruit peels and its commercial tablet dosage forms. The binary combination of green solvents such as ethanol-water (50:50, v v(-1)) was used as the mobile phase. The detection of HSP was performed at 287 nm. The sustainable RP-HPTLC method was linear in 20-2000 ng band(-1) range. The studied validation parameters, including accuracy, precision, robustness, sensitivity were acceptable for HSP analysis. The content of HSP in TE of four different varieties of citrus fruits including grapefruit peels (Citrus paradisi), mosambi peels (Citrus limetta), lemon peels (Citrus lemon), and orange peels (Citrus sinensis) was detected as 8.26, 6.94, 5.90, and 6.81% w w(-1), respectively. The content of HSP in TE of commercial formulations A and B was detected as 5.31 and 5.55% w w(-1), respectively. However, the content of HSP in UBE of grapefruit peels, mosambi peels, lemon peels, and orange peels was detected as 11.41, 8.86, 7.98, and 8.64% w w(-1), respectively. The content of HSP in UBE of commercial formulations A and B was detected as 6.72 and 6.92% w w(-1), respectively. The greenness score of the sustainable RP-HPTLC method was predicted as 0.83 using analytical GREEnness (AGREE) metric approach, indicated the excellent greenness profile of the RP-HPTLC method. UBE procedure for HSP was superior over its TE procedure. These observations and results suggested that the present RP-HPTLC method can be successfully used for the quantitative estimation of HSP in the variety of citrus fruit peels and its commercial formulations. In addition, this method is simple, rapid, precise, accurate, and economical compared to the reported analytical methods of HSP analysis. It is also safe and sustainable method due to the use of ethanol-water solvents systems, as both the solvents are green solvents compared to the solvents used in reported analytical methods of HSP analysis.