A Holistic Approach to Ayurvedic Gel Stability: Qualitative and quantitative estimation of Bio Markers using HPTLC, HPLC and GC

This research investigates the Ayurvedic Gel Formulation (AGF), an Ayurvedic product, utilizing various analytical techniques to assess its quality, safety, and stability. The AGF underwent stringent packaging in containers, following ICH guidelines Q1A (R2), to undergo extended shelf-life studies at 30°C ± 2°C and 60% RH ± 5% RH. Continuous evaluations, including physico-chemical and marker-based assessments, were conducted, with a control sample. The shelf-life dynamics were scrutinized through withdrawals at zero days and at 3-month intervals. This research article presents a thorough investigation into the HPTLC fingerprint profiling of Mimosa pudica plant and AGF, employing a solvent system of n-hexane: Ethyl acetate: Formic acid (6:4:0.1 v/v) with Linoleic acid as the reference standard. The study utilized UV observation at 254 nm and 366 nm, iodine vapors derivatization, and anisaldehyde-sulphuric acid reagent for detection, yielding comprehensive HPTLC fingerprint profiles. Safety parameters, encompassing microbial load, specific pathogens, and aflatoxin analysis, along with Total Phenolic Content (TPC) (0.3087%w/w) and Total Flavonoid Content (TFC) (0.1828%w/w), were systematically examined. The quantified results disclosed linoleic acid content in AGF (0.0182%) and Mimosa pudica (0.04119%). Palmitic acid analysis in AGF (0.0351%) and Coconut Oil (0.0119%) further validated the formulation. Safety evaluations demonstrated compliance with permissible limits for heavy metals, while microbial analysis highlighted a moderate aerobic count and low fungal count, ensuring high hygiene standards. Aflatoxin levels below quantifiable limits and pesticide residues within detectable limits emphasized adherence to safety standards. Thermal analysis confirmed the stability of AGF, providing a comprehensive understanding of its safety, quality, and compositional attributes in Ayurvedic formulations.