Isolation and Characterization of Zerumbone Isolated from Zingiber aromaticum

Zerumbone, a monocyclic sesquiterpene compound predominantly found in the rhizomes of lempuyang (Zingiber aromaticum), exhibits diverse biological activities including anticancer, antibacterial, anti-inflammatory, immunomodulatory, hepatoprotective, and gastroprotective effects. This study presents an optimized method for isolation, characterization, and purity analysis of zerumbone from the ethanolic extract of Z. aromaticum. The isolation process employed column chromatography with a gradient eluent system (n-hexane:ethyl acetate in ratios of 19:1 and 9:1) to separate compounds based on polarity differences. The extract and fractions were monitored using thin layer chromatography (TLC) under UV light at 254 nm and 366 nm. Characterization of isolated zerumbone was performed using multiple spectroscopic techniques, including TLC against reference standard, UV-Vis spectrophotometry, infrared (IR) spectroscopy, liquid chromatography-mass spectrometry (LC-MS/MS). The purified isolate demonstrated a yield of 11.2%, representing a significant improvement over previously reported methods (0.87–2.26%). TLC analysis with three different solvent systems consistently showed single spots, confirming high purity. UV-Vis analysis revealed maximum absorption at 252 nm, identical to the zerumbone standard. LC-MS/MS analysis identified a molecular ion peak at m/z 219.26 [M+H]+ with matching retention time and fragmentation pattern to the reference standard. IR spectroscopy identified functional groups characteristic of zerumbone, such as carbonyl and double-bond stretches. This efficient isolation method, yielding a highly purified compound, demonstrate its potential to support zerumbone’s application in pharmaceutical development.