Comprehensive Methodology for the Isolation and Identification of Triterpenes from Rosemary Leaves

This study focused on optimizing the extraction of antioxidants from rosemary (Rosmarinus officinalis) using different solvents and evaluating the efficacy of various purification techniques. Triterpenes, a class of compounds known for their diverse biological activities, are abundant in many plants, including rosemary, where they contribute significantly to its medicinal properties. Ethanol, acetone, and ethyl acetate were used for the initial extraction, with ethanol exhibiting the highest antioxidant activity, as determined by DPPH and ABTS assays. Fractionation of the ethanol extract yielded four fractions, the latter being enriched in triterpenes, as confirmed by thin-layer chromatography (TLC) analysis. Hexane and methanol were identified as optimal solvents for further purification, allowing efficient isolation of nonpolar terpenes while minimizing impurities. Advanced purification techniques, such as liquid-liquid extraction, were employed to improve the yield and purity of triterpenes. Gas chromatography-mass spectrometry (GC-MS) analysis of the final purified fraction revealed the presence of key triterpenes, such as ursolic acid, erythrodiol, and betulinic acid, highlighting the effectiveness of the extraction and purification methodology used. The results demonstrated that the combined use of appropriate solvents and advanced purification techniques allows for an optimized extraction of triterpenes from rosemary, with high purity of the active compounds.