A Simple and Rapid HPLC Technique for Determination of Arecoline in Areca Nut (Areca catechu L.) Extract

Abstract

A simple and rapid high-performance liquid chromatographic (HPLC) method for determination of arecoline extract from areca nut is reported. Arecoline is a major component cholinomimetic alkaloid found in areca nuts. It was extracted from dried seed powder to obtain high purity before analysis. The contents of arecoline in unripe and ripe areca nuts were compared. The analytical conditions for reverse-phase HPLC with UV detection were as follows: column, a 250 × 4.6 mm I.D., particle size 5 mm, Inertsil^® ODS-3 (GL sciences Inc., Tokyo, Japan); mobile phase, a 88:12 (%v/v) mixture of acetonitrile: phosphate buffer (pH 5.9); column temperature, 25 °C; flow rate, 1 mL/min; and detection at 254 nm. The retention time of arecoline was 5.0 min. The calibration curve of the method was linear (r² > 0.99) in the 10 - 200 mg/mL range. Method inter-day precision (RSD) was 0.42 - 1.15 %, and % recovery was 103.23 ± 5.76 % indicating high precision and accuracy of the method. The contents of arecoline in unripe and ripe areca nuts were 0.1434 ± 0.0016 and 0.0944 ± 0.0002 %w/w of dried seed powder, respectively. The method was simple, rapid, precise, accurate and selective to determine arecoline in areca nut extract.