Development and Validation of Simultaneous Analysis Method of Brazilin and 6-Gingerol in the Combined Extracts of Sappan Wood (Caesalpinia sappan) and Ginger Rhizome (Zingiber officinale) Using RP-HPLC

Abstract

Brazilin and 6-gingerol, bioactive compounds found in sappan wood (Caesalpinia sappan L.) and ginger rhizome (Zingiber officinale Rosc.) extracts, offer various potential pharmacological benefits. The combination of these extracts has shown promising antithrombotic and antihyperlipidemic properties, suggesting the potential use of this combination in herbal products. Quantitative analysis is required to ensure the quality control of herbal products. However, the simultaneous quantification of Brazilin and 6-gingerol using an HPLC method is currently unavailable. To address this gap, this study aimed to develop and validate a simultaneous quantification method for Brazilin and 6-gingerol in combined extracts of sappan wood and ginger using RP-HPLC. Chromatographic analysis was performed using a reverse-phase C18 Inertsil ODS3 column (4.5 × 250 mm; particle size 5 μm) at room temperature, with detection at 282 nm using a UV detector. The mobile phase consisted of acetonitrile (A) and water containing 0.1% acetic acid (B), with gradient elution optimized as follows: 0–12 min 15% A: 85% B; 12–16 min 30% A: 70% B; 16–21 min 45% A: 55% B; 21–35 min 60% A: 40% B, at a flow rate of 1 mL/min with an injected volume of 20 μL. The developed method demonstrated acceptable system suitability (peak resolution, tailing factor, theoretical plate number, selectivity) and validated parameters. Both Brazilin and 6-gingerol displayed linear calibration curves (R2?> 0.999), high intraday and interday precision (%RSD < 2%) and accuracy (93–106%). This study successfully developed and validated a rapid RP-HPLC method for simultaneous quantifying Brazilin and 6-gingerol in combined extracts of sappan wood and ginger rhizome. This method provides a reliable means for quality control analysis and could facilitate the development of herbal products incorporating these bioactive compounds.