超声辅助离子液体微萃取-反相液相色谱法测定人工蛹虫草活性成分  

Determination of the Active Ingredients in Cultured Cordyceps militaris by Ultrasound-Assisted Ionic Liquid Microextraction Coupled with Reversed Phase Liquid Chromatography

作　　者：张伟[1] 张勇[1] 尹震花[1] 康文艺[1] ZHANG Wei;ZHANG Yong;YIN Zhenhua;KANG Wenyi(Zhengzhou Key Laboratory of Medicinal Resources Research, Huanghe Science and Technology College, Zhengzhou 450063, China)

机构地区：[1]黄河科技学院郑州市药用资源研究重点实验室,河南郑州450063

出　　处：《食品科学》2017年第8期179-183,共5页Food Science

基　　金：河南省产学研合作项目(162107000038);河南省教育厅科学技术研究重点项目(17B360005);2016年度河南省高校科技创新团队项目(16IRTSTHN019)

摘　　要：建立离子液体[C_4MIM]PF_6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 C_(18)色谱柱(4.6 mm×150 mm,5μm),流动相为甲醇、0.02 mol/L KH_2PO_4溶液梯度洗脱,洗脱流速为0.6 m L/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[C_4MIM]PF_6-20%甲醇溶液、过50目筛、固液比1∶50(g/L)、超声时间50 min、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/m L,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 C18column(4.6mm×150mm,5μm)with methanol-0.02mol/L KH2PO4solution as mobile phase at a flow rate of0.6mL/min by gradient elution.The column temperature was30℃and the detection wavelength was set at254nm.The highest extraction yields of five nucleosides were obtained by using0.7mol/L[C4MIM]PF6-20%methanol solution as extraction solvent at a solid-to-liquid ratio of1:50(g/L),pulverizing samples to pass through a50-mesh sieve,ultrasonic irradiation for50min,and centrifugation at3000r/min.A good linearity was observed in the range of0.568?3.408,0.284?1.704,0.264?1.584,0.232?1.392and1.672?10.032mg/mL for uridine,inosine,guanosine,adenosine and cordycepin,respectively,with correlation coefficients above0.9998.The average recoveries were in the range of97.6%?101.5%,with relative standard deviation(RSDs)ranging between1.43%and1.97%.The method was rapid,simple and sensitive,and it could be applied for rapid analysis of five nucleosides in cultured C.militaris.

关 键 词：人工蛹虫草 离子液体 高效液相色谱法 核苷类 

分 类 号：R931.7[医药卫生—生药学]