Preparative separation of helvolic acid from the endophytic fungus Pichia guilliermondii Ppf9 by high-speed counter-current chromatography.
High-speed counter-current chromatography (HSCCC) was applied for preparative separation of helvolic acid from the crude extract of the endophytic fungus Pichia guilliermondii Ppf9, associated with the medicinal plant Paris polyphylla var. yunnanensis for the first time. The two-phase solvent system consisted of n-hexane-ethyl acetate-methanol-water (4.5:4.5:5.0:5.0, v/v) appending with phosphoric acid (0.2%, v/v) was employed. The revolution speed of the separation column, flow rate of the mobile phase and separation temperature of the apparatus were 800 rpm, 3 ml min(-1) and 25°C, respectively. About 6.8 mg of helvolic acid was successfully obtained from 450 mg of the crude extract by HSCCC within 4 h separation procedure, and its purity reached to 93.2% according to the HPLC analysis. The product was further characterized by MS, (1)H-NMR and (13)C-NMR spectra.
Department of Plant Pathology, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
Jianglin
J
Zhao
Yan
Y
Mou
Tijiang
T
Shan
Yan
Y
Li
Shiqiong
S
Lu
Ligang
L
Zhou
D002845:Chromatography / Q000379:methods*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D060026:Endophytes / Q000737:chemistry*; D005672:Fusidic Acid / Q000031:analogs & derivatives* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D019684:Magnoliopsida / Q000382:microbiology; D008969:Molecular Sequence Data; D010843:Pichia / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D012997:Solvents / Q000737:chemistry
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D012997:Solvents; D005672:Fusidic Acid; C011267:helvolic acid