ABSTRACT
Objective: To establish a rapid analytical method for volatile components in Tibetan medicine Heracleum millefolium and to determine the volatile components from its roots, stems, leaves, and flower parts, respectively. Methods: Headspace sampling incorporation with gas-chromatography-mass spectrum (HS-GC-MS) determination was introduced to analyze the powder directly. Static headspace equilibration was performed at 100℃ for 40 min, and 1 mL of the headspace gas was injected in split mode of 10:1. The split inlet temperature was 260℃. The carrier gas was He at a constant flow rate of 1.8 mL/min. The column oven temperature was initially set at 50℃ for 2 min, then increased to 100℃ at 2℃/min, held for 6 min, then increased to 300℃ at 10℃/min and held for 2 min. The GC/MS interface temperature was maintained at 280℃. The solvent delay time was 3 min (to bypass the solvent peak). The volatile components were confirmed by NIST11.L database, and volatile organic compounds from roots, stems, leaves, and flower parts were compared. Results: The types of compounds in the roots, stems, leaves, and flowers of H. millefolium are mainly aldehyde, benzene, alcohols, and alkene. Octanal, hexanal, and γ-terpinene are the main components in the roots and stems. While o-isopropyltoluene and terpinolene are the main components in the leaves and flowers. Conclusion: HS-GC-MS method is easy, simple, and feasible, and can be widely used in other Chinese materia medica samples for analysis of volatile components.
ABSTRACT
Objective: To separate and purify coumarins from Heracleum millefolium by high-speed counter-current chromatography (HSCCC). Methods: HPLC was used to analyze and optimize the solvent system and hexane-ethyl acetate-water-methanol (1.5:2.5:2:1.5) was chosen as the two-phase solvent system of HSCCC, in which the upper phase was used as the stationary phase, while the lower phase was used as the mobile phase with flow rate of 3 mL/min, the revolution speed was set at 850 r/min, and detected at 323 nm. Three compounds were obtained as crystals by vacuum concentration at 50℃, and determined through HPLC, then their structures were identified by IR, ESI-MS, 1H-NMR, and 13C-NMR. Results: Columbianetin acetate (6.3 mg), osthole (10.6 mg), and columbianadin (6.4 mg) were obtained from crude sample (300 mg) extracts and their purity were above 96.0%. Conclusion: HSCCC is a powerful technique for the rapid separation and purification of coumarins from crude extract of H. millefolium.