Analytical Validation of Bacopa monnieri Extract through High-performance Thin-layer Chromatography with Emphasis on Spasmolytic Phytomarkers

Background: Bacopa monnieri (Brahmi) is a traditional Ayurvedic herb widely reported for memory-enhancing, anti-inflammatory, and antispasmodic properties.Fingerprint profiling of extracts by high-performance thin-layer chromatography (HPTLC) ensures quality control and supports correlation with pharmacological activity.Objective: To establish an HPTLC fingerprint of ethanolic extract of Bacopa monnieri and correlate the observed bands with phytochemicals relevant to anti-inflammatory and anti-colicky activity.Methods: Soxhlet extraction of Brahmi powder (80 gm) was performed using ethanol (500 mL) for 24 cycles to yield 400 mL of extract.HPTLC was carried out on silica gel 60 F254 plates using the mobile phase toluene:ethyl acetate:formic acid (4:3:1).Densitometric scanning was performed at 254 nm and 366 nm.Results: Track 1 (Brahmi ethanolic extract) revealed multiple peaks.At 254 nm, the major band was observed at Rf 0.674 (41.8%), with additional peaks at Rf 0.540 (20.3%) and Rf 0.793 (11.2%).At 366 nm, prominent bands appeared at Rf of 0.542 (14.8%),Rf of 0.637 (24.0%), and Rf of 0.804 (20.8%).These Rf values correspond to bacosides, flavonoids, alkaloids, and sterols.Conclusion: The HPTLC fingerprint established for Brahmi ethanolic extract highlights phytochemicals contributing to anti-inflammatory (flavonoids, bacosides, phenolic acids) and anti-colicky (flavonoids, alkaloids, sterols) activities.The band at Rf ~0.54-0.67 is particularly significant, likely representing bacoside-flavonoid fractions responsible for smooth muscle relaxation and intestinal anti-inflammatory action.