多功能离子液体分散液液微萃取结合高效液相色谱法检测人尿中5种邻苯二甲酸酯代谢物

引用本文: 孙倩, 戴浩强, 陈佩佩, 佘慧, 武佳. 多功能离子液体分散液液微萃取结合高效液相色谱法检测人尿中5种邻苯二甲酸酯代谢物[J]. 色谱, 2020, 38(8): 929-936. doi: 10.3724/SP.J.1123.2019.09026 shu

Citation: SUN Qian, DAI Haoqiang, CHEN Peipei, SHE Hui, WU Jia. Combination of dispersive liquid-liquid microextraction using multifunctional ionic liquids with high performance chromatography for determination of phthalate ester metabolites in human urine sample[J]. Chinese Journal of Chromatography, 2020, 38(8): 929-936. doi: 10.3724/SP.J.1123.2019.09026 shu

多功能离子液体分散液液微萃取结合高效液相色谱法检测人尿中5种邻苯二甲酸酯代谢物

孙倩 ^1,2, ,
戴浩强 ^2, ,
陈佩佩 ^2, ,
佘慧 ^2, ,
武佳 ^{2,
,}

1.
宁波市医疗中心李惠利医院检验科, 浙江宁波 315040;
2.
温州医科大学, 浙江省医学遗传学重点实验室, 浙江温州 325035

通讯作者: 武佳, E-mail:popjia1107@126.com

基金项目:
浙江省自然科学基金（LQ17H260005）.

摘要: 将多功能离子液体与分散液液微萃取（DLLME）技术相结合，建立了测定尿液中5种邻苯二甲酸酯类（PAEs）物质代谢产物的高灵敏度新方法。对影响DLLME效率的各单因素进行了优化，包括萃取剂的种类及体积、分散剂的种类及体积、萃取温度、超声时间、冷却时间、离心时间和盐效应等条件，经过严格的优化，最佳的萃取条件分别为：萃取剂[C₈MIM]PF₆]35 μL，分散剂[BSO₃HMIm]OTf]30 μL和[C₄MIM]BF₆]120 μL，萃取温度为35℃，超声时间5 min，冷却时间5 min，离心时间5 min，盐析剂NH₄PF₆ 0.1 g。在最佳的萃取条件下，5种PAEs代谢物在0.5~1000 μg/L范围内具有良好的线性关系，决定系数（R²）均大于0.9955，方法检出限为0.16~0.19 μg/L，尿液中添加低中高水平（5、20、100 μg/L）的PAEs代谢物，其回收率为92.9%~105.0%，日内精密度及日间精密度的相对标准偏差（RSD）均小于5.96%，方法学验证各指标及稳定性均符合分析要求。对所采集的10份糖尿病患者的尿液进行检测，并对该人群PAEs代谢物的暴露水平进行评价。结果表明，各PAEs代谢物均有检出，其中邻苯二甲酸单（2-乙基己基）酯（MEHP）的检出率为100%。总之，该方法萃取过程中未添加有毒的有机试剂，均使用多功能离子液体作为萃取剂、分散剂和盐析剂，萃取过程绿色环保，简单高效；方法的灵敏度较高，稳定性较好，适用于人体尿液中痕量PAEs代谢物的检测。

关键词:

English

Combination of dispersive liquid-liquid microextraction using multifunctional ionic liquids with high performance chromatography for determination of phthalate ester metabolites in human urine sample

SUN Qian ^1,2, ,
DAI Haoqiang ^2, ,
CHEN Peipei ^2, ,
SHE Hui ^2, ,
WU Jia ^{2,
,}

1.
Clinical Laboratory, Lihuili Hospital, Ningbo Municipal Medical Center, Ningbo 315040, China;
2.
Wenzhou Medical University, Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou 325035, China

Corresponding author: WU Jia, popjia1107@126.com

Received Date: 14 October 2019
Fund Project:
Zhejiang Provincial Natural Science Foundation (No. LQ17H260005).

Abstract: A novel sensitive method for the determination of the five primary metabolites of phthalate esters (PAEs) in urine was developed by combining dispersive liquid-liquid microextraction (DLLME) using ionic liquids with high performance liquid chromatography (HPLC). The factors affecting the efficiency of DLLME were optimized. The types and proportions of extraction solvent and dispersants, as well the ultrasonic extraction time, cooling time, and centrifugal time, were determined. The optimal conditions were as follows:extraction solvent[C₈MIM]PF₆] 35 μL; dispersants[BSO₃HMIm]OTf] 30 μL,[C₄MIM]BF₆] 120 μL; NH₄PF₆ 0.1 g, extraction at 35℃, ultrasonic dispersion for 5 min, cooling in ice water for 5 min, centrifugation at 4000 r/min for 5 min. After optimization, the five primary metabolites of PAEs were determined. The method showed a good linear relationship within the concentration range of 0.5-1000 μg/L. The determination coefficients (R²) were greater than 0.9955. The detection limit was in the range of 0.16-0.19 μg/L. Under the optimized conditions, the extraction recoveries for the PAEs were 92.9%-105.0%, and the relative standard deviations (RSDs) of the intra- and inter-day precisions were < 5.96%. Urine samples collected from 10 diabetic patients were tested, and the exposure level of the population to the PAE metabolites was evaluated. All the PAE metabolites were detected in these samples, and the detection rate of 2-ethylhexyl hydrogen phthalate (MEHP) was 100%. In conclusion, no toxic organic reagents were added during the extraction process in this method, and multifunctional ionic liquids were used as the extraction agent, dispersant, and salting-out agent. In other words, the extraction process was demonstrated to be green, simple, and efficient. The developed method has high sensitivity and stability, and it is suitable for the determination of trace PAE metabolites in human urine.

Key words:

dispersive liquid-liquid microextraction (DLLME)
/ high performance liquid chromatography (HPLC)
/ phthalate metabolites
/ urine
/ multiple functional ionic liquids

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多功能离子液体分散液液微萃取结合高效液相色谱法检测人尿中5种邻苯二甲酸酯代谢物

通讯作者: 武佳, E-mail:popjia1107@126.com

English

Combination of dispersive liquid-liquid microextraction using multifunctional ionic liquids with high performance chromatography for determination of phthalate ester metabolites in human urine sample

Corresponding author: WU Jia, popjia1107@126.com

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

多功能离子液体分散液液微萃取结合高效液相色谱法检测人尿中5种邻苯二甲酸酯代谢物

通讯作者: 武佳, E-mail:popjia1107@126.com

English

Combination of dispersive liquid-liquid microextraction using multifunctional ionic liquids with high performance chromatography for determination of phthalate ester metabolites in human urine sample

Corresponding author: WU Jia, popjia1107@126.com

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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