Quantification of Bergenin from Mallotus philippinensis by HPTLC-MS and Study on Different Extraction Methods

A simple and accurate high-performance thin-layer chromatographic (HPTLC) method has been established for the determination of bergenin in the roots and stem-bark powder of Mallotus phillippensis. It has been reported to have a wide array of biological activities like antioxidant, anti-HIV, antiarrhythmic, hepatoprotective, anti-inflammatory and anti-microbial. Methanolic extract of the stem-bark and root powder was used for the experimental work. Separation was performed on 20 x 10 silica gel 60 F-254 using ethyl acetate-methanol-acetic acid-formic acid (8:1:0.5:0.5, v/v) as mobile phase and scanned using densitometry at 284 nm. Extracts were prepared by conventional soaking, and accelerated solvent extractor (ASE) was used for the analysis. The results indicate that bergenin was found to be maximum in root ASE extract (6.0% w/w), while the minimum concentration of bergenin was found in stem-bark cold extract (2.9% w/w). The determination was carried out using the densitometric absorbance mode at 284 nm. The method was validated in terms of selectivity, linearity, precision, accuracy, and robustness. Additionally, peak identity was confirmed by mass spectrometry. The electron spray ionization (ESI) mass spectra showed the [M + Na](+) ions for bergenin detected at m/z 351 being acquired directly from the sample bands by an elution-based interface. By considering the validation results, the method was found to be very simple, accurate, precise, fast, and economical and can be used for routine quality control.