Rapid Chromatographic Determination and Structural Confirmation of beta-Hydroxy Acid Form of Lovastatin in the Fermentation Broth of Aspergillus Terreus PM03

摘要

Lovastatin, a fungal secondary metabolite, competitively inhibits 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase, which catalyzes the rate limiting step in the biosynthesis of cholesterol. Preparative thin-layer chromatography (TLC) followed by RP-HPLC was used for determination of lovastatin in the fermentation broth of Aspergillus terreus PM03. The TLC separated bands were cut, recovered, and analyzed by HPLC. The proposed HPLC method used a mobile phase comprising 1 : 1 mixture of acetonitrile and deionized water at a flow rate of 2 mL/min and spectrophotometric detection at a wavelength of 238 nm. The chromatograph revealed a single, sharp and symmetric peak at a retention time of 6.8 min that corresponded to an open beta-hydroxy acid form and was comparable to the peak of standard beta-hydroxy acid form of lovastatin with respect to both retention time and optical absorption at 238 nm. The data of UV-VIS and FTIR spectroscopy confirmed the presence of lovastatin in the purified extract. The structure of the obtained product was also confirmed by LC-MS and NMR analyses.