臺灣博碩士論文加值系統

合成皮製造工廠作業勞工經由吸入與皮膚接觸而暴露於製程所使用的有機溶劑二甲基甲醯胺（N,N-dimethylformamide, DMF）。對於DMF的暴露可以採集生物檢體進行生物偵測，以有效評估勞工DMF暴露量。DMF在體內代謝過程中會與glutathione結合形成NMG（N-methylcarbamoylglutathione），而NMG與NMF會與血紅素結合，形成蛋白質鍵結物（protein adducts）N-methylcarbamoylated valine-globin （NVH），若能測定NVH在血紅素的量，將更直接評估DMF暴露勞工的暴露量。本研究進行DMF暴露動物實驗模擬勞工DMF暴露，雄性Sprague Dawley大鼠被餵食不同劑量(0.0016X、0.008X、0.04X、0.2X和1X LD50)的DMF溶液，並於連續餵食暴露後第14天進行血液樣本採集，然後使用高效能液相層析法(high-performance liquid chromatograph, HPLC)開發一個新的方法，來分析DMF暴露動物血液中血紅素鍵結物(hemoglobin adducts, 簡稱Hb adducts)的含量。本研究所開發的方法對老鼠血紅素NVH含量分析和DMF餵食劑量呈現統計上顯著相關(r = 0.34)，此方法的偵測極限為73.8 ppb，對血紅素NVH的萃取回收率為94.18%。以本研究所開發的方法分析不同DMF暴露勞工血液樣本，其血紅素NVH含量與DMF空氣濃度值並未呈現統計上顯著相關(r=0.02)，此方法應用到人類血液可能需要進一步的研究。對於血紅素NVH的萃取回收率，為決定此分析方法是否穩定的關鍵因素。對DMF暴露者血液中NVH的測定，將有助於對DMF暴露健康效應危害機轉的瞭解。

Workers in synthetic leather manufacturing plants are exposed to N,N-dimethylformamide (DMF) by inhalation and skin contact. Biological samples are collected to conduct biological monitoring and assess workers’ exposure doses. In the metabolic process of human body, DMF can combine with glutathione to form N-methylcarbamoylglutathione (NMG). NMG and NMF can attach to the hemoglobin of red blood cells to form protein adducts, N-methylcarbamoylated valine-globin (NVH). It has been considered that the quantification of NVH in the hemoglobin of red blood cells can more directly estimate workers’ DMF exposure doses. In this study, animal experiments were conducted to simulate workers’ DMF exposures. Different doses (0.0016X, 0.008X, 0.04X, 0.2X and 1X of LD50) of DMF in aqueous solutions were given by gavage to male Sprague Dawley rats. After 14 days of DMF exposure, blood samples of the rats were collected for analysis. A high-performance liquid chromatograph (HPLC) approach was developed to quantify the amount of hemoglobin adducts (Hb adducts) in the blood samples. The method developed in this study showed that the linear relationship between the amounts of NVH in the blood samples and the exposure doses for the rats was statistically significant (r = 0.34). The limit of detection was 73.8 ppb. The recovery efficiency was about 94.18%. This method was used to analyze the blood samples of workers with different concentrations of DMF exposure. There was no statistically significant linear relationship (r = 0.02) between the amount of NVH in the blood samples and the DMF exposure concentrations. This indicated that more researches are needed for applying this method to quantify human DMF exposure. The recovery efficiency of NVH from blood samples was the critical factor relevant to the stability of this HPLC approach to the quantification of Hb adducts. By means of this fast and accurate approach to measuring the NVH in blood samples, it is possible to obtain more understanding on the mechanism of adverse health effect due to DMF exposure.

目錄 i
圖目錄 iii
表目錄 iv
中文摘要 v
Abstract vi
第一章 緒論 1
1-1 研究背景 1
1-2 研究意義和重要性 2
1-3 研究目的 2
第二章 文獻探討 3
2-1 DMF的物化特性與其應用 3
2-2 DMF的代謝及其健康效應 3
2-2-1 DMF的代謝 3
2-2-2 DMF的健康效應 5
2-3 DMF之生物偵測 5
2-4 HPLC的儀器原理 6
第三章 材料與方法 8
3-1 實驗室之分析方法開發 8
3-1-1 DMF暴露動物實驗 8
3-1-2 MIH標準品之合成 9
3-1-3 實驗藥品 10
3-1-4 實驗器具 11
3-1-5 分析儀器 11
3-2 血液樣本前處理 12
3-2-1 血液樣本前處理實驗步驟 12
3-3 儀器分析之條件 13
3-4 樣本分析品質管制(quality control, QC)和品質保證(quality assurance, QA) 14
3-5 資料分析 15
第四章 結果與討論 16
4-1 血液前處理與方法開發 16
4-2 檢量線的建立 17
4-3 偵測極限 19
4-4 儀器精確度、分析準確度 19
4-5 樣本回收率 20
4-6 實驗動物血液樣本分析結果 22
第五章 結論 33
第六章 研究限制 34
第七章 參考文獻 35

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