HPTLC Fingerprinting and Quantitative Estimation of Antioxidant Phytomarkers in Clerodendrum Serratum, a Key Ingredient of Kabasura Kudineer

Clerodendrum serratum (L.) Moon, commonly known as Bharangi, is an important medicinal plant of the family Laminaceae widely used in Ayurveda, Siddha, and traditional systems of medicine for the management of respiratory and inflammatory disorders. Owing to its significant therapeutic relevance and its prominent inclusion in classical formulations such as Kabasura Kudineer, the present study aimed to identify and quantify key antioxidant marker compounds responsible for its pharmacological efficacy using a validated high-performance thin-layer chromatographic (HPTLC) method. Methanolic extracts of authenticated C. serratum raw material were subjected to HPTLC analysis employing a CAMAG HPTLC system. Chromatographic separation was achieved on silica gel 60 F₂₅₄ plates using toluene:ethyl acetate:formic acid:methanol (3:6:1.6:0.4, v/v/v/v) as the optimized mobile phase. Densitometric scanning was performed at 254 nm and 366 nm. The identity of marker compounds was confirmed by comparison of Rf values and densitometric profiles with those of reference standards. Quantitative analysis revealed the presence of major antioxidant phytoconstituents, including rutin (0.17%), mangiferin (0.70%), vitexin (0.02%), kaempferol (4.03%), quercetin (0.02%), gallic acid (0.09%), and luteolin (0.60%). These compounds are well documented for their antioxidant and anti-inflammatory properties mediated through multiple cellular signaling pathways, including NF-κB, MAPK, and Nrf2/HO-1. The findings of this study provide scientific evidence supporting the traditional therapeutic use of C. serratum and establish a reliable HPTLC-based analytical approach for its quality control and standardization. This method may serve as a useful tool for ensuring the consistency, safety, and efficacy of herbal raw materials and formulations containing C. serratum.