以毛細管電泳分離尿液中多環芳香碳氫化合物之代謝物

Separation of polycyclic aromatic hydroxycarbon metabolites in urine by capillary electrophoresis

李建德(Chien-De Lee)

指導教授 ： 蔡錦蓮

高雄醫學大學/健康科學院/職業安全衛生研究所/碩士(2004年)

摘要

本研究以毛細管電泳儀搭配紫外光偵測器，以毛細管區帶電泳之分離模式分析六種多環芳香碳氫化合物(polycylclic aromatic hydrocarbons；PAHs)代謝物3-hydroxybenz(a)anthracene、1-hydroxychrysene、3-hydroxyphenanthrene、3-hydroxybenzo(a)pyrene、1-hydroxypyrene、3-hydroxyfluoranthene，並應用此分析方法檢測職業廚師尿液中PAHs代謝物之含量。
本研究以不同濃度的緩衝液(50mM, 80mM, 100mM CAPS)及不同pH值(pH9.7~11.1)作分析條件的探討，並比較不同種類的有機修飾劑(methanol, acetonitril )及不同的有機修飾劑含量(10﹪~50﹪)來達到最佳化的分析條件，有效的分離出六種PAHs的代謝物，偵測極限為0.05~0.09μm/mL，而滯留時間之變異係數為2.3~2.9 %，積分面積之變異係數為5.5~9.7 %，均小於10%。廚師尿液經過前處理後，以C18 column 搭配自動進樣固相萃取裝置能萃取出五種PAHs的代謝物，其中以3-hydroxybenzo(a)pyrene的含量最高。
PAHs代謝物的分析在先前的文獻中都以膠束電動層析法來分析，本研究以最簡單快速的毛細管區帶電泳法，避免使用膠束及介面活性劑而降低再現性，並加入有機修飾劑來提高分析的效率，結果發現能有效的改善解析度，成功的分離六種PAHs的代謝物。

關鍵字

尿液 ； 代謝物 ； 多環芳香碳氫化合物 ； 毛細管電泳

並列摘要

The aim of study is used capillary electrophoresis and UV detector to separation six polycylclic aromatic hydrocarbons (PAHs) metabolite incluuding 3-hydroxybenzo(a)pyrene、3-hydroxyphenanthrene、1-hydroxychrysene、3-hydroxybenz(a)anthracene、1-hydroxypyrene、3-hydroxyfluoranthene by capillary zone electrophoresis (CZE), and use same conditions to assay concentration of cook urine.
The study is tested the assay conditions of diverse concentration of buffer (50mM, 80mM, 100mM CAPS) and pH value (pH9.7~pH11.1), and compare two organic modification (methanol, acetonitril) and diverse content (10%~50%) to obtain optimization analysis, excellent separation is achieced fo the six metabolites. The detection limits were in the 0.05~0.09μm/mL range, reproducibility of migration time range between 2.3~2.9 % CV, peak areas between 5.5~9.7 %.
The extraction method for determining urine from the cook was consisted of enzymatic hydrolysis, auto-sampling solid-phase extraction by C18 column. In all the cook’s urine sample we investigatede we could determine five PAHs metabolites, the highest level of urine was 3-hydroxybenzo(a)pyrene.
In previous published, the PAHs metabolites was separation by micellar electrokinetic capillary electrophoresis (MEKC). In this study, we use simple and rapid CZE method, the method avoid the poor reproducibility by use of micelles and cyclodextrins. In addition, highly efficient separation of metabolites is achieved with organic modification, the result obtain show great improvement in overall resolution, and six PAHs metabolites could be separate.

並列關鍵字

metabolites ； polycyclic aromatic hydroxycarbon ； capillary electrophoresis ； urine

參考文獻

六、參考文獻

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