ANALYTICAL STANDARDIZATION AND MULTI-STAGE AYURVEDIC SHODHANA (~PURIFICATION) OF VACHA RHIZOMES (ACORUS CALAMUS LINN.): HPTLC FINGERPRINTING, β-ASARONE DETOXIFICATION, AND PHYTOCHEMICAL PROFILING

Background: Vacha (Acorus calamus Linn.) is an important Ayurvedic herb with prominent neurotherapeutic properties. However, the Vacha rhizomes contain β-asarone – a major phytoconstituent which is toxic and carcinogenic, necessitating detoxification before safe use. Ayurvedic multi-stage Shodhana (~purification) is traditionally employed for this purpose. Objective: This study aims to develop, validate, and apply HPTLC fingerprinting for analytical standardization of Vacha rhizomes through classical Shodhana (~purification), demonstrating β-asarone elimination and phytochemical transformations with high reproducibility and pharmaceutical relevance. Methods: Standardized multi-stage Shodhana (~purification) was performed on large batch (~16 kg) of authenticated Vacha rhizomes following classical Ayurvedic protocols. Methanolic extracts from each processing stage were subjected to validated HPTLC analysis. Chromatographic fingerprints were generated at 254, 366, and 540 nm, using β-asarone as the primary reference standard. Quantitative and qualitative data were examined with rigorous replication and statistical validation. Results: The characteristic β-asarone peak (Rf ~0.54) present in raw Vacha extracts showed progressive diminution through successive Shodhana stages and became entirely undetectable following Gorakhamundi processing. Concurrently, new phytochemical bands appeared at Rf ~0.57 and ~0.68,indicating successful integration of bioactive compounds from processing media. Method validation parameters met international standards (ICH Q2(R1)) for linearity, precision, and accuracy. Data evidenced reproducible detoxification and phytochemical enrichment. Conclusion: The developed HPTLC methodology provides a reliable analytical framework for characterizing the multi-stage Ayurvedic Shodhana (~purification) effects on Vacha rhizomes. This analytical approach significantly enhances pharmaceutical quality control and supports regulatory acceptance by ensuring safety and standardization of Vacha rhizomes-based formulations.