建立离子液体[C4MIM]PF6萃取剂,结合超声辅助萃取,利用高效液相色谱法同时测定人工蛹虫草中尿苷、肌苷、鸟苷、腺苷及虫草素含量.采用Inert Sustain C18色谱柱(4.6 mm×150mm,5 μm),流动相为甲醇、0.02 mol/L KH2PO4溶液梯度洗脱,洗脱流速为0.6 mL/min,柱温30℃,检测波长254 nm.5种成分的最佳萃取条件为0.7 mol/L [C4MIM]PF6-20％甲醇溶液、过50目筛、固液比1∶50(g/L)、超声时间50 rain、离心转速3 000 r/min.线性范围分别为0.568～3.408、0.284～1.704、0.264～1.584、0.232～1.392、1.672～10.032 mg/mL,相关系数均在0.999 8以上,加样回收率为97.6％～101.5％,相对标准偏差为1.43％～1.97％.所建方法快速简便、准确可靠、重复性良好,可用于人工蛹虫草中核苷类成分的同时快速分析.%This study aimed to develop a high performance liquid chromatography (HPLC) method for the simultaneous determination of the contents of five nucleoside (uridine,inosine,guanosine,adenosine and cordycepin) in cultured Cordyceps militaris.An ultrasonic-assisted ionic liquid microextraction procedure was proposed using [C4MIM]PF6-20％ methanol as extraction solvent for sample pretreatment.The method was performed on an Inert Sustain C18 column (4.6 mm× 150 mm,5 μm) with methanol-0.02 mol/L KH2PO4 solution as mobile phase at a flow rate of 0.6 mL/min by gradient elution.The column temperature was 30 ℃ and the detection wavelength was set at 254 nm.The highest extraction yields of five nucleosides were obtained by using 0.7 mol/L [C4MIM]PF6-20％ methanol solution as extraction solvent at a solid-to-liquid ratio of 1∶50 (g/L),pulverizing samples to pass through a 50-mesh sieve,ultrasonic irradiation for 50 min,and centrifugation at 3 000 r/min.A good linearity was observed in the range of 0.568-3.408,0.284-1.704,0.264-1.584,0.232-1.392 and 1.672 10.032 mg/mL for uridine,inosine,guanosine,adenosine and cordycepin,respectively,with correlation coefficients above 0.999 8.The average recoveries were in the range of 97.6％ 101.5％,with relative standard deviation (RSDs) ranging between 1.43％ and 1.97％.The method was rapid,simple and sensitive,and it could be applied for rapid analysis of five nucleosides in cultured C.militaris.
展开▼
