Comparative Phytochemical Screening and Solvent-dependent Extraction of Hispidulin from Clerodendrum Serratum Leaves
Eswar Tony Darsi *
Faculty of Pharmacy, Parul University, Waghodia, Vadodara, Gujarat, 391760, India.
Komal Patel
Department of Pharmacy, Parul Institute of Pharmacy, Faculty of Pharmacy, Parul University, Waghodia, Vadodara, Gujarat, 391760, India.
Rama Rao Nadendla
Chalapathi Institute of Pharmaceutical Sciences, Guntur, Andhra Pradesh, 522034, India.
*Author to whom correspondence should be addressed.
Abstract
Background: Medicinal plants are important sources of bioactive phytoconstituents with potential therapeutic applications. Clerodendrum serratum (Bharangi) is traditionally used in Ayurveda and contains several pharmacologically important secondary metabolites. Comparative evaluation of solvent-dependent extraction and marker-based quantification is essential for its phytochemical standardisation.
Objective: The present study aimed to investigate the preliminary phytochemical composition of Clerodendrum serratum leaf extracts prepared using different solvents and to compare their hispidulin content through chromatographic techniques. Methodology: Dried leaves of Clerodendrum serratum were extracted using chloroform, ethanol, methanol and aqueous solvents. Preliminary phytochemical screening was performed to identify major secondary metabolites. FTIR and HPTLC were employed for functional group characterisation and chromatographic identification of hispidulin, respectively. Quantitative estimation was performed using reverse-phase HPLC with a C18 column, employing acetonitrile and 0.1% formic acid in water (45:55, v/v) as the mobile phase at a flow rate of 1.0 mL/min. Detection was carried out at 340 nm.
Results: Preliminary phytochemical screening revealed the presence of flavonoids, alkaloids, glycosides, tannins, saponins, and phenolic compounds. HPTLC analysis showed chromatographic peaks corresponding to the hispidulin reference standard at an Rf value of approximately 0.61. HPLC analysis demonstrated a characteristic peak at a retention time of 7.88 ± 0.05 min. The estimated hispidulin content was highest in the methanolic extract (96.12 mg/g), followed by the ethanolic (54.74 mg/g), chloroform (35.22 mg/g), and aqueous extracts (14.67 mg/g).
Conclusion: The findings indicate that solvent selection influences the recovery of hispidulin from Clerodendrum serratum leaves, with methanol yielding the highest estimated marker content among the investigated extracts. These findings provide an analytical basis for further phytochemical characterisation and standardisation. Comprehensive validation of the quantitative HPLC method is necessary to establish the accuracy, precision and reproducibility of the reported estimates.
Keywords: Clerodendrum serratum, phytochemical screening, hispidulin, solvent extraction, HPTLC, HPLC, herbal drug standardisation