Quantitative Determination of 14 Major Constituents in the Herbal Preparation Luan-Pao-Prescription Using HPLC Coupled with Photodiode Array Detection

Abstract

High performance liquid chromatography coupled with photodiode array detection (HPLC–DAD) was employed for the determination of 14 major constituents in traditional Chinese herbal formula ‘Luan-Pao-Prescription’. Two different chromatographic determinations for the 14 constituents were carried out to control the quality of this herbal formula. The sample was extracted under ultrasonic extraction with 75% methanol at the frequency of 42 kHz for 45 min at room temperature. The chromatographic separation was performed on a Zorbax SB-C₁₈. The mobile phase was linear gradient elution of water with 0.5% (v/v) acetic acid (A) and acetonitrile (B) column with linear gradient elution procedure. The flow rate was set at 1.0 mL min⁻¹ and the column temperature was kept at 30 °C. The calibration curve for the determination of all the analytes showed good linearity over the investigated ranges (R ² > 0.999). The intra- and inter-day assay precision (RSD) of 14 analytes were in the range of 0.2–3.9% and the recoveries were between 92.4 and 104.4%. The validated method was successfully applied to the determination of 14 major constituents in LPP.