pH依赖型悬浮固化液-液微萃取-HPLC测定尿液中2种蒽醌类物质

doi:10.3969/j.issn.1000-1565.2018.06.007

摘要/Abstract

摘要： 建立了一种pH依赖型悬浮固化液-液微萃取-HPLC测定尿液中大黄素和大黄酚的方法.该方法选择辛酸钠为萃取剂前体,在加入酸性样品溶液的过程中原位形成萃取剂辛酸.将100 μL辛酸钠溶液注入到事先酸化的10 mL样品水溶液中,经离心和冰浴后,萃取液固化并浮于溶液上层,取出固化的萃取液并溶解于50 μL甲醇供HPLC分析使用.对作为萃取剂的酸种类、辛酸钠体积、离子强度、离心时间等重要因素进行了优化.在最优条件下,大黄素和大黄酚在5~400 ng/mL内与峰面积间存在良好的线性关系(R² > 0.999),检出限为0.75~0.8 ng/mL,相对标准偏差为2.0%~3.2%(n=5).将该方法应用于添加后的尿液样品分析,回收率为82%~110%.该法萃取过程中避免了有机分散溶剂的使用,萃取时间短,操作简单,成本低,绿色环保,非常适于与液相色谱分析联合使用.

Abstract: A method for the determination of anthraquinones in urine was developed by the combination of pH - dependent solidified floating liquid-liquid microextraction with HPLC. Octanoic acid chosen as extraction solvent was dissolved in alkaline solution, and 100 μL of it was taken and injected into 10 mL aqueous solution which had been acidified. After centrifugation for 4 min and ice bath for 2 min, the extract was floated on the surface of the aqueous sample. The extract was taken out and dissolved with 50 μL methanol for HPLC analysis. The main factors, such as acid type, sodium octanate volume, ion strength and centrifugal time, were systematically investigated. In the optimal conditions, there was a good linear relationship of peak areas for emodin and chrysophanol with their concentrations in the range of 5—400 ng/mL(R² > 0.999),with detection limit 0.75—0.8 ng/mL, and the relative standard deviation- DOI:10.3969/j.issn.1000-1565.2018.06.007pH依赖型悬浮固化液-液微萃取-HPLC测定尿液中2种蒽醌类物质杜雪红, 辛晓莹,王计彭, 张红医,杨冰(河北大学化学与环境科学学院,河北保定 071002 )摘要:建立了一种pH依赖型悬浮固化液-液微萃取-HPLC测定尿液中大黄素和大黄酚的方法.该方法选择辛酸钠为萃取剂前体,在加入酸性样品溶液的过程中原位形成萃取剂辛酸.将100 μL辛酸钠溶液注入到事先酸化的10 mL样品水溶液中,经离心和冰浴后,萃取液固化并浮于溶液上层,取出固化的萃取液并溶解于50 μL甲醇供HPLC分析使用.对作为萃取剂的酸种类、辛酸钠体积、离子强度、离心时间等重要因素进行了优化.在最优条件下,大黄素和大黄酚在5~400 ng/mL内与峰面积间存在良好的线性关系(R² > 0.999),检出限为0.75~0.8 ng/mL,相对标准偏差为2.0%~3.2%(n=5).将该方法应用于添加后的尿液样品分析,回收率为82%~110%.该法萃取过程中避免了有机分散溶剂的使用,萃取时间短,操作简单,成本低,绿色环保,非常适于与液相色谱分析联合使用.关键词:分散液-液微萃取;辛酸;蒽醌类;高效液相色谱法中图分类号:O652.1 文献标志码:A 文章编号:1000-1565(2018)06-0596-07Determination of anthraquinones in urine samples by pH-dependent solidified floating liquid-liquid microextraction-HPLCDU Xuehong,XIN Xiaoying,WANG Jipeng,ZHANG Hongyi,YANG Bing(College of Chemistry and Environmental Science, Hebei University, Baoding 071002,China)Abstract: A method for the determination of anthraquinones in urine was developed by the combination of pH - dependent solidified floating liquid-liquid microextraction with HPLC. Octanoic acid chosen as extraction solvent was dissolved in alkaline solution, and 100 μL of it was taken and injected into 10 mL aqueous solution which had been acidified. After centrifugation for 4 min and ice bath for 2 min, the extract was floated on the surface of the aqueous sample. The extract was taken out and dissolved with 50 μL methanol for HPLC analysis. The main factors, such as acid type, sodium octanate volume, ion strength and centrifugal time, were systematically investigated. In the optimal conditions, there was a good linear relationship of peak areas for emodin and chrysophanol with their concentrations in the range of 5—400 ng/mL(R² > 0.999),with detection limit 0.75—0.8 ng/mL, and the relative standard deviation- 收稿日期:2018-02-18 基金项目:教育部留学回国人员科研启动基金(47批);河北大学校内科研项目(799207022;799207217014);河北大学2018年校级大学生创新创业计划项目及河北大学创新资助项目(hbu2018ss21) 第一作者:杜雪红(1989—),女,河北邯郸人,河北大学在读硕士研究生.E-mail:1043267494@qq.com 通信作者:张红医(1967—),男,河北保定人,河北大学教授,主要从事色谱、样品前处理和识别分析.E-mail:freetime20100401@163.com第6期杜雪红等:pH依赖型悬浮固化液-液微萃取-HPLC测定尿液中2种蒽醌类物质2.0%~3.2%(n=5). The method was applied to the analysis of urine samples, and the recovery after spiking was in the range of 82% and 110%. The extraction process avoids the use of organic dispersive solvents. The advantages of the proposed method are short extraction time, simple operation, low consumption and friendly environment, and very suitable for the combination with liquid chromatography analysis.

Key words: dispersive liquid-liquid microextraction, octanoic acid, anthraquinones, HPLC

中图分类号:

O652.1

杜雪红, 辛晓莹,王计彭, 张红医,杨冰. pH依赖型悬浮固化液-液微萃取-HPLC测定尿液中2种蒽醌类物质[J]. 河北大学学报(自然科学版), 2018, 38(6): 596-602.

DU Xuehong,XIN Xiaoying,WANG Jipeng,ZHANG Hongyi,YANG Bing. Determination of anthraquinones in urine samples by pH-dependent solidified floating liquid-liquid microextraction-HPLC[J]. Journal of Hebei University (Natural Science Edition), 2018, 38(6): 596-602.

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