Development of a high-performance liquid chromatography method for simultaneously analysis of saponins and flavonoid in materials and dietary supplements containing Hedera helix extracts

Abstract

English Ivy or Hedera Helix is a multi-medicinal functioned plant in nature. Most coughmedicines in Vietnam were extracted from Ivy leaves because of its 5 main active components, in which 4 Saponins included Hederacoside C (predominance), α-Hederin, Hederacoside D, Hederasaponin B are responsible for eliminating congestion (breaking up the phlegm and mucus) and Flavonoid: Kaempferol 3-rutinoside plays the role of reducing inflammations [1]. This study aimed to develop a HPLC-PDA method to simultaneously and fast analyze these 5 compounds in materials and dietary supplements containing hedera helix extract in Vietnam market. After the simple preparation procedure, the analytes were separated by using a C18 column (150 mm × 4.6mm, 5 µm) as stationary phase, and a mixture of 0.1% phosphoric acid and acetonitrile as mobile phase. The detection and quantification were in PDA detector at 205 nm. The method validation followed AOAC criteria. The calibration curves in the range of 0.5 - 200 mg/L for 4 saponins and 0.1 - 100 mg/L for the flavonoid with high correlation coefficient (R² > 0.9999). The MDL (0.03 - 0.15 mg/kg) and MQL (0.15 - 0.50 mg/kg); RSD_r (%) for repeatability (1.01 - 3.90%) and RSD_R reproducibility (1.25 - 6.89%); recoveries (91.3 - 106%) for 5 compounds satisfied the AOAC requirements. The method was applied successfully for determining the content of the analytes in 10 real samples including dried ivy extract powder, dried leaves, and some cough relief products purchased from markets in Hanoi. The levels of the 5 analytes were different in each sample in which Hederacoside C and α-Hederin account for the main proportions.