中药细辛与藜芦配伍化学成分变化的UPLC/Q-TOF-MS研究(3)
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Experimental
Materials and reagents: Chromatographic grade acetonitrile was purchased from Fisher (USA), chromatographic grade formic acid was purchased form Kermel Chemicals Ltd (Tianjin, China). Leucine-enkephalin was from Sigma-Aldrich. The berberine hydrochloride was form National Institute for the Control of Pharmaceutical and Biological Product. Distilled water was purified by MilliQ system (Millipore Corp. Bedford, MA, USA). Millipore filters (0.22m) were purchased from Millipore Corporation. The medicinal substances were purchased from Anguo Medicinal Herbs Co., Ltd (Hebei, China), which had been identified by Professor Zhai Yanjun (Liaoning University of Traditional Chinese Medicine).
Sample preparation: Berberine hydrochloride was used as an internal standard substance during the separations. The concentration of the internal standard was 19.6g/mL dissolved by methanol. Veratrum Nigrum L. sample solution: extract 5.0g crude drug with 100 mL water for 2 h by refluxing. Internal standard solution (3mL) was added to the extraction apparatus before extraction precisely. Concentrate the filtrate to 25 mL, and then dilute the concentrated solution to 50 mL with ethanol precisely. Take the solution as test sample. Herba Asari sample solution: the same to the above mentioned except the weight of the crude drug is 3.0 g. Mixture solutions: extract the 5.0g Veratrum Nigrum L. and 3.0 g Herba Asari with 200 mL water. The other operations were the same to above.
LC/MS: The chromatographic separations were performed on a Waters ACQUITY UPLCTM BEH column (100 mm×2.1 mm, 1.7 μm) using an ultra performance liquid chromatography system (UPLC, Waters, USA) ......
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