移液枪头式整体柱直接固相微萃取-高效液相色谱法测定尿液中苯二氮卓类药物

Determination of Benzodiazepines in Human Urine by Direct Solid-phase Microextraction Using Monolithic Pipette Coupled with High Performance Liquid Chromatography

本文采用甲基丙烯酸2-羟乙酯(HEMA)与乙二醇二甲基丙烯酸酯在移液器枪头中共聚制备固相微萃取整体柱,用于直接处理尿液中3种苯二氮卓类药物(BZDs)。实验考察了单体用量、聚合时间及固相微萃取条件对BZDs萃取效率的影响,评价了其吸附性能。当HEMA用量为40 mg,聚合温度60℃时,固相微萃取整体柱对目标物吸附效率达98.9%~100%,且对尿液中杂质的吸附率低于10%。取3 mL尿液样品,无需处理,直接上样至该柱,经4 mL纯净水冲洗,1 mL乙腈洗脱,得到的样品结合高效液相色谱仪分析。方法对3种BZDs在2.5~1000 ng/mL浓度范围呈现良好线性关系(r=0.999),检出限(S/N=3)和定量限(S/N=10)为0.8~1.5 ng/mL和2.5~5.0 ng/mL,回收率为86.2%~107%。本方法构筑的萃取柱制备简单,能实现对原始尿液的直接、高效萃取,操作便捷。

A novel solid-phase microextraction(SPME)monolith using 2-hydroxyethyl methacrylate as the monomer and ethylene glycol dimethacrylate as the crosslinker was successfully synthesized in a pipette and applied for direct SPME of three benzodiazepines(bromazepam,lorazepam and midazolam)from crude urine samples prior to high performance liquid chromatography(HPLC).The fabrication condition including the amount of monomer and the reaction temperature was investigated and the adsorption performance of the obtained monoliths was evaluated.Moreover,the effect of extraction parameters on the recovery of benzodiazepines was optimized.The prepared pipette offered the adsorption extraction of 98.9%-100%for benzodiazepines and≤10%for the impurities for urine when 40 mg of monomer was performed at 60℃.The optimized condition for SPME was shown as follows,3 mL of urine for direct loading,4 mL of H_(2)O as washing solution,and 1 mL of acetonitrile as elution solvent.The proposed SPME monolithic pipette coupled with HPLC was developed for the analysis of three benzodiazepines in urine samples.It provided the linear range of 2.5-1000 ng/mL with correlation coefficient(r)of 0.999,the limits of detection(S/N=3)and limits of quantification(S/N=10)of 0.8-1.5 ng/mL and 2.5-5.0 ng/mL,and the recovery of 86.2%-107%.The present monolith not only allowed direct loading of crude urine samples without any filtration or dilution step,but also offered great ability of purification and enrichment for the target with an enhancement factor of 13-16.The developed method proposed an alternative strategy for the accurate and convenient determination of benzodiazepines in urine samples.