Utilization of HPTLC in standardization and quality control of Sphaeranthus indicus Linn. Using complementary solvent...

Sphaeranthus indicus Linn. is a well-known medicinal plant in Ayurveda, traditionally used for its anti-inflammatory, hepatoprotective, and neuroactive properties. Its therapeutic potential is attributed to a wide range of phytoconstituents including sesquiterpene lactones, flavonoids, and sterols. Accurate chemical profiling is essential for its authentication and quality control.

To develop a comprehensive and reproducible HPTLC fingerprint of Sphaeranthus indicus using a Complementary Development Solvent (CDS) system across different solvent extracts.

Five extracts - methanol, n-hexane, acetone, petroleum ether, and 50% hydroethanolic - were prepared. Samples were applied on silica gel 60 F254 HPTLC plates and developed using the CAMAG HPTLC PRO system. Three solvent systems were employed: low polarity (LPDS), medium polarity (MPDS), and high polarity (HPDS). Visualization was performed under UV light (254 and 366nm) and white light, before and after derivatization with anisaldehyde sulphuric acid and 10 % methanolic sulphuric acid. Method validation parameters including precision, repeatability, and accuracy were evaluated to ensure the reliability of the fingerprinting process.

All solvent systems provided distinctive chromatographic fingerprints. LPDS resolved non-polar terpenoids and steroids, MPDS effectively separated flavonoids and mid-polar compounds, while HPDS facilitated the detection of polar compounds post-derivatization. Consistent Rf values at 0.15 and 0.43 were observed across multiple extracts. Derivatization enhanced visibility and color differentiation of phytochemical classes. Method validation demonstrated high repeatability of Rf values and spot intensity, with satisfactory precision and accuracy for quantitative comparisons.

The developed HPTLC method using CDS provides a robust and detailed fingerprint profile for Sphaeranthus indicus. It supports effective quality control and can be employed for standardization and identification of marker compounds in herbal formulations.