2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38055Polycyclic aromatic hydrocarbons (PAHs) are considered human carcinogens. Coke oven workers are occupationally exposed to high concentrations of PAHs. Epidemiologic studies demonstrate increased cancer incidence among the workers exposed to PAHs, probably through cumulative oxidative DNA damage. It has been suggested that various reactive oxygen species (ROS) can be generated and can further cause oxidative DNA damage during PAHs metabolism. 8-hydroxy-2’-deoxyguanosine (8-OHdG) is one of the major oxidative DNA damage formed by radical-induced damage at C-8 position of guanine and has been regard as the biomarker of oxidative stress. Previous studies have showed that a significant increase of 8-OHdG in urine among coke oven workers. However, little information was found in the association between the level of 8-OHdG and the major urinary metabolite of PAHs (1-OHP). Several methods have been applied for analysis of 8-OHdG, such as high performance liquid chromatography with electrochemical detection (HPLC-ECD), gas chromatography with mass spectrometry (GC-MS), liquid chromatography with tandem mass (LC-MS/MS), and enzyme-linked immunosorbent assay (ELISA). Among these methods, LC-MS/MS has a high sensitivity and selectivity, and can be applied with the isotope-dilution method, such analytical technique is the most accurate and reliable analytical method. However, it was noted that most previous efforts on the determining of 8-OHdG was made by ELISA rather than LC-MS/MS. In the present study, our aim is to investigate the association between the level of 8-OHdG and exposure biomarker of PAHs (1-OHP). We will conduct a longitudinal study in coke oven workers. Preshift and postshift urine samples were collected consecutively and analyzed for 8-OHdG and 1-OHP. The level of 8-OHdG will be measured by our newly developed isotope dilution LC-MS/MS method. This two-years project is aimed to answer the following questions: 1. The variation of oxidative DNA damage in coke oven workers during working period. 2. The association between urinary 1-OHP and 8-OHdG in coke oven workers, including - the time and order of appearance after exposed to PAHs - the relative amounts 3. The influence of polymorphism on urinary 8-OHdG in coke oven workers (CYP1A1、GSTM1 and GSTT1 genotype) 4. The association between urinary mutagenicity and oxidative DNA damage in coke oven workers多環芳香烴化合物 (PAHs)是已知的人類致癌物。煉焦工人由於長期暴露在 高濃度的PAHs 中，許多流行病學的研究已經發現到煉焦工人罹癌的機率較一般 人高。其一可能致癌的原因是PAHs 暴露造成的基因氧化傷害。PAHs 在體內代 謝的過程中會產生許多活性含氧物質(ROS)有可能導致基因氧化傷害。8-羥基去 氧鳥糞嘌呤核 (8-OHdG)是常見的基因氧化傷害且其定量已被廣泛用來當做體 內基因氧化傷害的生物標記。過去的研究已發現到煉焦工人的尿液中的8-OHdG 明顯高於一般控制組。然而甚少研究探討尿液中8-OHdG 與PAHs 尿液代謝物(1- 羥基芘,1-OHP)兩者間的相關性。 對於生物體內中 8-OHdG 之定量，在過去已發展許多分析方法來準確定量， 包括高效液相層析儀配合電化學偵測(HPLC-ECD)，氣相層析質譜儀(GC-MS)， 液相層析串聯質譜儀(LC-MS/MS) 及酵素連結免疫吸附分析（ enzyme-link immunosorbent assay，簡稱ELISA）。其中以LC-MS/MS 分析方法其特異性及敏 感性高、樣品前處理簡單加上可配合使用同位素稀釋法(isotope dilution)，所以此 方法可說是當前最精準的定量方法。 本研究中將以 90 位煉焦廠員工為測試對象。以縱貫式研究方法（longitudinal study）探討煉焦工人PAHs 暴露之基因氧化傷害，同時試圖瞭解基因氧化傷害與 1-羥基芘 (1-OHP)的相關性。計畫執行時間共需兩年。第一年主要是採用貫時性 的追蹤調查研究方法，針對煉焦爐工廠各暴露族群進行上工前下工後連續尿液採 樣、個人空氣採樣分析(PAH)與問卷調查。並同時開發8-OHdG 自動化連線固相 萃取方法(on-line solid phase extraction)，樣本經自動萃取後連線可直接注入 LC-MS/MS 進行分析。第二年主要是藉由尿液中8-OHdG 與1-OHP 的定量來探 討: 1. PAHs 暴露下，體內DNA 氧化傷害隨工作週期的變化 2. PAHs體內代謝物(1-OHP)與DNA氧化傷害的相關性 (兩者出現在尿液 中的時程與順序) 3. 下一工作週期上工前，尿液中8-OHdG 及1-OHP 的含量是否能降回到 上一工作週期上工前的背景值 4. 各種基因型與尿液中 8-OHdG 間之關連性(CYP1A1、GSTM1 and GSTT1 genotype) 5. 尿液中基因氧化傷害指標 8-OHdG 與尿液致突變性間之關連性8-OHdG尿液煉焦工廠多環芳香烴化合物基因多型性致突變性8-OHdGurinecoke-oven plantPAHspolymorphismmutagenicityProject