Quantitative Analysis of Hispidulin Content in Clerodendrum petasites Roots Distributed in Thailand

Introduction: Clerodendrum petasites (Lour.) S. Moore (locally known as Mai-Thao-Yaai-Mom), belonging to the Verbenaceae family, is widely formulated into multi-herb remedy, Ben-Cha- Lo-Ka-Wi-Chian remedy, possessing antipyretic activity. C. petasites exhibits many biological activities, such as antioxidant, anti-inflammatory, antipyretic, etc. The flavonoid hispidulin is one of the main active compounds present in C. petasites, containing anti-atheromatous, antitumor and antispasmodic effects. Objective: The present study aimed to determine the hispidulin content in the dried roots of C. petasites using HPLC technique. Methods: C. petasites dried roots, collected from twelve different areas, were extracted with ethanol using Soxhlet apparatus, and then subjected to HPLC-PDA to quantify hispidulin content. The quantitative method using HPLC-PDA technique was validated. Results: The optimized HPLC coupling with PDA detector (HPLC-PDA) was validated for the quantitative analysis of hispidulin content in C. petasites roots in terms of linearity (y = 210,200,536.6667x – 448,756.2667; R² = 0.9997), accuracy (88.82-107.69% recovery), precision (0.66% RSD for repeatability precision; 1.17% RSD for intermediate precision), limit of detection (2.30 μg/mL), limit of quantitation (7.00 μg/mL), specificity (peak purity index = 1.0000) and robustness (% RSD < 1). The amount of hispidulin content in the extracts of C. petasites roots conducted from the validated method was found to be 0.0182 ± 0.0109 g/100 g crude drug. Conclusion: The HPLC-PDA analysis was able to effectively determine hispidulin in C. petasites roots. The hispidulin contents in C. petasites dried roots from various areas in Thailand were revealed which could be used for the specification of this crude drug with reference to its chemical marker.