Isolation, identification, and structure elucidation of Beta-sitosterol from Iraqi Plantago major using GC-MS, HPTLC, NMR, and FTIR

Abstract

Background: Plantago major, a plant well recognized for its therapeutic features, has been widely adopted in several cultural instances and conventionally used due to its anti-inflammatory, antibacterial, and wound-healing characteristics. Recent research efforts have been focused on the identification and validation of the phytochemical elements of P. major in order to establish their association with their corresponding pharmacological effects. The main aim of this study is to precisely define, isolate, purify, and completely analyse the chemical composition of beta-sitosterol, a phytosterol obtained from the Plantago major plant native to Iraq. Methods: In November 2021, P. major samples were procured from the Al-Salihiya Neighbourhood in Baghdad. These samples were afterward verified for authenticity by the Iraqi local Herbarium, located at the Al-Razi institute for alternative medicine. The desiccated botanical matter was subjected to a hexane-based defatting process inside a Soxhlet device, followed by gas chromatography/mass spectrometry (GC/MS) analysis in order to ascertain the presence of bioactive compounds. The evaluation of beta-sitosterol’s presence was afterwards conducted via the use of conventional and preparative thin layer chromatography (TLC) methodologies and high-performance thin layer chromatography (HPTLC). The identification of the molecule was further validated by the use of Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy techniques. Results: The findings revealed that Compound No. 5 had notable attributes that were consistent with those of Beta-Sitosterol. The provided data pertains to mass spectrometry (MS), especially the observation of a molecular ion peak at 414. This finding is consistent with the previously documented characteristics of beta-sitosterol. This finding provides further evidence supporting its classification as a prominent organic component originating from P. major. Conclusions: We effectively determined beta-sitosterol as the primary bioactive component in the P. major samples collected. The compound’s existence highlights the plant’s longstanding therapeutic standing, ascribing it several advantageous pharmacological benefits.