Determination of Flavanones in Citrus Byproducts and Nutraceutical Products by a Validated RP-HPLC Method

Abstract

The aim of this work was to develop and validate a specific reversed phase high performance liquid chromatographic method (HPLC) using diode array detection (DAD) for the determination of flavanones, which is the dominant flavonoid class in the genus citrus. The procedures and criteria used for validation of the method including sensitivity, linearity, repeatability, recovery, and robustness were carried out following the EURACHEM guidelines.

The validated RP-HPLC-DAD method was applied for the determination of the mainly flavanone glycosides contained in dried blood orange peel and dried lemon peel, and for the quantitation of flavanone glycosides and hesperetin in nutraceutical products.

Keywords:

• Citrus byproduct
• Flavanones
• Flavonoids
• HPLC-DAD
• Method validation
• Nutraceutical tablets

ACKNOWLEDGMENTS

The Project was funded by the Italian Ministry for the University and Research (MUR) with a PNR 2005–2007 Project n. RBIP06SXMR “Sviluppo di metodologie innovative per l'analisi di prodotti agroalimentari.” The authors gratefully thank Ortogel SpA of Caltagirone (Italy), which provided the samples of dried blood orange and lemon peels.

Notes

Eluent A = 2-propanol-acetonitrile-water (6:10:84, v/v/v) containing 0.5% formic acid.

Eluent B = 2-propanol-acetonitrile (70:30, v/v).

Dbop = dried blood orange peel; Dlp = dried lemon peel.

nq = not quantificable.