Hydroxychavicol and Eugenol Profiling of Betel Leaves from Piper betle L. Obtained by Liquid-Liquid Extraction and Supercritical Fluid Extraction

Abstract

The chemical components of liquid-liquid extraction (LLE) and supercritical fl uid extraction (SFE) obtained from dried betel leaves (Piper betle L.) were determined and evaluated for the percentage extraction of hydroxychavicol (HC) and eugenol (EU). SFE was conducted at 60 °C and a working pressure of 8 MPa (SFE1) and 6 MPa (SFE2. The chemical profiles were determined using high performance liquid chromatography (HPLC) with a method developed and validated for quantitative determination of HC and EU in the extracts from betel leaves. The analysis was carried out using HPLC on a Hypersil GOLD column (C18 with 250 × 4.6 mm) using methanol-water (70:30 volume per volume) as the mobile phase under an isocratic system. The fl ow rate was 0.7 mL.min-1 and the detection was evaluated at the wavelength of 280 nm with a UV detector. The validation using hydroxychavicol (HC) and eugenol (EU) as a standard demonstrated a linear relationship (correlation coeffi cient = 0.997 and 0.998, respectively), with precision (relative standard deviation < 4%) in the concentration range of 40–100 μg.mL-1 For HC and EU, the limit of detection was 1.0 and 0.1μg.mL-1, respectively, and the limit of quantifi cation was 2.0 and 0.2 μg.mL-1, respectively. The highest extraction yield (15.6% weight per weight, w/w) and a high content of HC (9.1% w/w) were obtained by using ethyl acetate refl uxed extraction and LLE. A similar chemical profi le, regarding the HPLC fi ngerprint showed quality consistency by SFE1 with lower extraction yields.