在线固相萃取-高效液相色谱法同时测定人尿液中的马钱子碱和士的宁  

Sensitive determination of brucine and strychnine in human urine by online solid phase extraction coupled with high performance liquid chromatography

作　　者：刘菲[1] 檀笑昕 韩旭[3] 王玮[2] 康维钧[2] 

机构地区：[1]河北中医学院基础医学院,石家庄050200 [2]河北医科大学公共卫生学院,石家庄050017 [3]河北省疾病预防控制中心,石家庄050000

出　　处：《分析试验室》2017年第8期923-926,共4页Chinese Journal of Analysis Laboratory

基　　金：河北省高等学校科学技术研究项目(Z2014030);河北省食品药品监督管理局科技项目(PT2014054)资助

摘　　要：建立了全自动、快速的在线固相萃取-液相色谱技术用于测定人尿液中的马钱子碱和士的宁的方法。样品离心后直接进样,其中被测组分富集于Hyper Sep Retain MCX柱上,尿液中的大分子基质物质不被保留而排出,在线完成萃取和净化富集过程;再通过阀切换使待测物转移至分析流路,在Acclaim C_(18)色谱柱上分离检测。分析时间约13 min。在线固相萃取流路以0.1%H_3PO_4和乙腈为流动相,1 m L/min流速萃取和洗脱;分析流路以乙腈和0.02 mol/L KH_2PO_4为流动相,1 m L/min流速等度洗脱,检测波长为260 nm。马钱子碱在1.25×10^(-6)~2.5×10^(-4)mol/L浓度范围内相关系数r为0.9930,高、中、低浓度的平均回收率分别为85.7%,81.4%和80.9%;士的宁在2.9×10^(-6)~2.9×10^(-4)mol/L浓度范围内相关系数r为0.9993,高、中、低浓度的平均回收率分别为87.1%,82.4%和78.9%。本法可用于测定人尿液中的马钱子碱和士的宁含量。A method was developed for simultaneous determination of brucine and strychnine in human urine by fully automated online solid phase extraction（ SPE）-high performance liquid chromatography.The target compounds were online concentrated on the Hyper Sep Retain MCX solid phase extraction column after centrifugation,and then transferred by the six-way valve to the Acclaim C（18） column for separation. A single sample analysis cycle took only 13 min. In the experiment,ACN-0. 1% H3PO4 was used as the SPE mobile phase with a flow rate of 1 m L/min and ACN-0. 02 mol/L KH2PO4 was used as the analytical mobile phase with a flow rate of 1 m L/min at 260 nm. The calibration curve of brucine with good linearity（ r = 0. 9930） was obtained in the range of 1. 25 × 10^-6- 2. 5 ×10^-4 mol/L in human urine,and the recoveries at high,medium and low concentration levels were85. 7%,81. 4% and 80. 9%,respectively. The calibration curve of strychnine with good linearity（ r = 0. 9993） was obtained in the range of 2. 9 × 10^-6- 2. 9 × 10^-4 mol/L in human urine,and the recoveries at high, medium and low concentration levels were 87. 1%,82. 4% and 78. 9%,respectively. In conclusion,the established automated online SPE-HPLC method demonstrated good performance in terms of specificity and limits of quantification,and was successfully utilized to quantify brucine and strychnine in human urine.

关 键 词：马钱子碱 士的宁 人尿液 在线固相萃取 高效液相色谱法 

分 类 号：R991[医药卫生—毒理学]