基于溶剂诱导相变萃取的在线固相萃取-高效液相色谱法测定血清中多环芳烃含量
Determination of polycyclic aromatic hydrocarbon levels in serum by high performance liquid chromatography with online solid phase extraction based on solvent induced phase transition extraction
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摘要:
背景 目前血中多环芳烃(PAHs)的检测方法存在操作烦琐费时、有机溶剂消耗量大、测定多环芳烃种类少等不足，有待改进。
目的 建立简便有效、绿色环保的在线固相萃取-高效液相色谱方法测定血中多种PAHs含量。
方法 比较甲醇-水系统和乙腈-水系统对PAHs的分离效果，并进一步优化液相色谱条件；比较Hypersep Hypercarb柱、Dionex InGuard HRP柱、Hypersep Retain PEP柱以及C18在线SPE柱对PAHs的亲和力以选择SPE柱并确定切阀程序；此外，比较了不同诱导剂(二氯甲烷、三氯甲烷和甲苯)的诱导效果，并采用4因素3水平正交试验考察了乙腈、二氯甲烷以及无机盐氯化钠的用量对诱导相变萃取效率的影响。并进行方法学验证，将所建立的方法应用于8名机械工人和6名普通人群的血样PAHs测定。
结果 采用C18在线SPE柱，用甲醇-水系统进行洗脱时对PAHs分离效果良好。正交试验结果显示乙腈0.50 mL、二氯甲烷0.05 mL及不用氯化钠为最佳前处理条件。结果表明15种多环芳烃在1.00~50.00 μg/L范围内线性关系良好，决定系数均大于0.999；方法检出限(信噪比S/N=3)为0.01~0.20 μg/L，平均加标回收率为90.8%~107.0%，相对标准偏差(n=6)为1.6%~4.4%。采用该法测定了8名机械工人和6名普通人群的血样，低环PAHs的检出率高，高环PAHs的检出率低；机械工人比普通人群多检出3种高环PAHs。
结论 所建方法能同时测定血清中15种多环芳烃的含量，具有简便高效、准确可靠及环境友好等优点，可为开展多环芳烃的暴露风险评估和毒理学研究提供一定的方法学支持。
Abstract:Background Existing methods for the detection of polycyclic aromatic hydrocarbons (PAHs) in blood have some shortcomings, such as complex and time-consuming operation, large consumption of organic solvents, and few detected kinds of PAHs, which need to be improved.
Objective This method ological study aims to establish a simple, effective, and environmentally friendly online solid phase extraction (SPE)-high performance liquid chromatography (HPLC) method for the determination of multiple PAHs in blood.
Methods The separation effects of methanol-water system and acetonitrile-water system on PAHs were compared, and the liquid chromatography conditions were further optimized. The affinities of Hypersep Hypercarb column, Dionex InGuard HRP column, Hypersep Retain PEP column, and C18 online SPE column for PAHs were compared to select SPE column and determine valve switching procedure. In addition, the induction effects of different inducers (dichloromethane, trichloromethane, and and toluene) were compared, and the effects of different dosages of acetonitrile, dichloromethane, and inorganic sodium chloride on induced phase change extraction efficiency were investigated by four-factor and three-level orthogonal test. Method ological validation was performed by applying the established method to the determination of PAHs in blood samples of eight mechanical workers and six general people.
Results The C18 online SPE column showed good separation of PAHs when being eluted with methanol-water system. The results of orthogonal test showed that 0.50 mL acetonitrile, 0.05 mL dichloromethane, and no sodium chloride were the best pretreatment conditions. The linear relationships of 15 PAHs were good in the range of 1.00~50.00 μg/L, the determination coefficients were above 0.999, and the detection limits (signal-to-noise ratio, S/N=3) were 0.01-0.20 μg/L. The average recovery rates were 90.8%-107.0%, and the relative standard deviations (n=6) were 1.6%-4.4%. The blood samples of eight mechanical workers and six general people were determined by this method, detection rates of low-ring PAHs were high, and the detection rates of high-ring PAHs were low; three more high-ring PAHs were detected in the mechanical workers than in the general subjects.
Conclusion The proposed method can simultaneously determine the contents of 15 PAHs in serum, and is simple, efficient, accurate, reliable, and environmentally friendly. It can provide method ological support for exposure risk assessment and toxicology research of PAHs.
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