臺灣博碩士論文加值系統

環境中存在著一類化學物質，由於其模仿體內荷爾蒙之特性，進入生物體後對其內分泌系統造成干擾，稱之為環境荷爾蒙。其中，烷基酚類（Alkylated phenolic compounds，AP）之雌激素特性更為近年來所重視，被國人使用於塑膠製品之塑化劑與抗氧化劑，及界面活性劑中。本研究欲測定的物質壬基苯酚（4-Nonylphenol，NP）、辛基苯酚（4-tert-Octylphenol，OP）及丁基苯酚（2,4-di-tert-Butylphenol，
BP）即為烷基苯酚聚乙氧基醇類（Alkylphenol polyethoxylates，APnEO）降解所產生的物質。
由於目前此方面人體暴露的研究很少，且可供參考的分析生物樣本方法亦不多，因此無法確切瞭解人體的暴露情形。本研究目的在建立自人體血漿中同時測定壬基苯酚、辛基苯酚及丁基苯酚之研究方法，並應用於一般及特殊暴露族群中。
本研究將血漿稀釋並加入β-Glucuronidase/arylsulfatase酵素後
，使用Varian PH 500mg/3mL之固相萃取管將樣本淨化，所收集之甲醇沖提液則注入HPLC搭配螢光偵測器加以分析。本研究之NP、OP、BP檢量線定量範圍均為0.005~0.385g/L；回收率則分別為114.66±7.84％、107.99±8.31％、108.73±6.04％；偵測極限為0.363mg/L、0.476mg/L、1.050mg/L。
本研究應用於一般族群（學生）、特殊暴露族群（清潔人員）以及捐血者之血袋樣本，發現一般族群之平均NP、OP、BP濃度分別為0.043±0.007、0.013±0.002及0.028±0.005mg/g；特殊暴露族群則分別為0.053±0.050、0.016±0.003、0.026±0.007 mg/g；捐血者之NP與OP平均濃度為0.380±0.148及0.010±0.007mg/g。顯示一般及特殊暴露族群血中之NP、OP及BP濃度相當，而血袋樣本之NP濃度很高，可能來自血袋材質本身之污染或其他原因，值得進一步探討。

Environmental hormones are synthetic chemicals discharged in our environment with the specific characteristics of hormone mimic effect for organisms disturbing the endocrine system. In the past few years, the disclosed estrogenic characteristics of Alkylated phenolic compounds（AP）has attracted attention in Taiwan because it is the chemical widely used as the plastic plasticizer、anti-oxidants or surfactant. This study was to develop method that can simultaneously detect Nonylphenol（NP）、Octylphenol（OP）and Butylphenol（BP）, the products degradated from Alkylphenol polyethoxylates（APnEO）.
There are limited studies on NP、OP and BP exposure in human partly because only limited analytical methods are available for determining the biological samples. Human exposure to those chemicals remains unclear. The aim of our study was to develop a suitable method to simultaneously detect NP、OP and BP in human plasma for general population and specific occupational individuals.
After being diluted, the plasma sample was treated with enzyme β-Glucuronidase/arylsulfatase and cleaned using the Varian PH 500mg/3mL SPE.The collected methanol effluent is injected into HPLC equipped with a fluorescence detector. The calibration curve ranged between 0.005~0.385mg/L, the recoveries for spiked NP、OP and BP were 114.66±7.84％、107.99±8.31％、108.73±6.04％, and the detection limits were 0.363mg/L、0.476mg/L、1.050mg/L, respectively.
The new method in this study has been applied to the general population（students） and the special population（genitors）and some blood donators. The average concentrations of NP、OP and BP for the students are 0.043±0.007、0.013±0.002、0.028±0.005mg/g；in the special population are 0.053±0.050、0.016±0.003、0.026±0.007 mg/g, respectively. The average concentrations of NP and OP the blood donators were 0.380±0.148 and 0.010±0.007mg/g. The results suggests that the differences in concentrations of NP、OP、BP are not significant between students and special populations. Besides, the high NP concentration found in the blood bag sample may suggest that the blood bag is contaminated with NP.

摘要 I
Abstract II
目錄 IV
表目錄 VIII
圖目錄 IX
附錄 IX
第一章 前言 1
第一節 研究緣起 1
第二節 壬基苯酚、辛基苯酚及丁基苯酚對生物體的主要影
響 2
第三節 此方法建立之原因 2
第四節 研究目的 3
第二章 文獻探討 4
第一節 壬基苯酚、辛基苯酚及丁基苯酚之物理化學特性 4
第二節 環境荷爾蒙之定義與特性 4
2-1 定義 4
2-2 常見類型 5
2-3 作用機制 5
2-4 對生物體產生之影響 5
第三節 壬基苯酚、辛基苯酚及丁基苯酚的來源及特性 6
第四節 台灣及其他各國相關規定 6
第五節 環境流佈 7
第六節 對人體的影響及毒性 8
6-1 暴露途徑 8
6-2 代謝與排泄 9
6-3 其他影響 9
第七節 對鼠類的影響及毒性 10
7-1 口腔餵食及腹膜注射 10
7-2 皮下注射 10
7-3 靜脈注射 10
第八節 對魚類的影響及毒性 11
8-1 暴露途徑 11
8-2 代謝與排泄 11
第九節 偵測方式與分析方法之比較 12
第三章 材料與方法 16
第一節 材料 16
1-1 藥品 16
1-2 器材 16
1-3 儀器 17
1-4 試劑配製 17
第二節 分析樣本 18
2-1 回收率與基質效應樣本 18
2-2 一般族群樣本 19
2-3 特殊族群樣本 19
2-4 捐血者樣本 19
2-5 自願者樣本 19
第三節 分析方法 20
3-1 前處理 20
3-2 樣本淨化 20
3-3 分析 20
第四節 分析方法之驗證 21
4-1 線性範圍 21
4-2 血漿樣本分析之基質效應 21
4-3 血漿樣本之添加回收率 22
4-4 血漿樣本分析之再現性 22
4-5 儀器偵測極限 23
4-6 血漿樣本之偵測極限 23
4-7 方法之應用 23
4-8 血袋材質之NP、OP、BP溶出實驗 24
4-9 人體血漿中含NP1EO及NP2EO與否之實驗 24
第四章 結果 25
第一節 壬基苯酚、辛基苯酚與丁基苯酚之HPLC實驗
條件 25
第二節 分析方法之驗證 25
2-1 檢量線 25
2-2 基質效應 25
2-3 血漿樣本添加標準品之回收率 26
2-4 血漿樣本添加標準品之再現性 26
2-5 儀器偵測極限 26
2-6 血漿偵測極限 27
第三節 一般族群與特殊族群血漿中之壬基苯酚、辛基苯酚
及丁基苯酚濃度 27
3-1 一般族群 27
3-2 特殊族群 27
3-3 捐血者 28
3-4 人體血漿中是否含有NP1EO與NP2EO 28
第四節 血袋材質之NP、OP、BP溶出實驗 29
第五章 討論 30
第一節 前處理 30
1-1 樣本稀釋並振碎 30
1-2 加入酵素β-Glucuronidase/arylsulfatase 30
1-3 水浴並震盪 31
第二節 淨化 31
2-1 固相萃取管之選用及淨化改良 31
2-2 樣本酸化 32
2-3 過濾 32
第三節 選用血漿之原因 33
第四節 捐血者結果與血袋溶出實驗之探討 33
第五節 回收率與再現性之探討 34
5-1 回收率 34
5-2 再現性 34
第六節 本研究結果之進階討論與評估 35
6-1 人體中是否偵測到NPnEO及OPnEO之存在 35
6-2 不同族群樣本之比較與清潔人員之暴露量評估 36
第七節 本研究與尿液樣本之回收率與結果比較 37
第八節 此方法建立之重要性及應用 37
參考文獻 38
表目錄
表一、壬基苯酚、辛基苯酚與丁基苯酚的物理化學性質及用途 45
表二、本研究壬基苯酚、辛基苯酚與丁基苯酚檢量線資料 46
表三、血漿樣本添加壬基苯酚、辛基苯酚及丁基苯酚之回收率 47
表四、同日異日回收率之CV值比較 48
表五、一般族群之血漿樣本中壬基苯酚、辛基苯酚及丁基苯酚之
濃度 49
表六、清潔人員血漿中NP、OP、BP濃度描述 50
表七、捐血者血漿之壬基苯酚、辛基苯酚之濃度 51
圖目錄
圖一、APEO好氧與厭氧微生物降解之途徑 52
圖二、NP在鱒魚體內主要之代謝途徑 53
圖三、標準品之HPLC層析圖譜 54
圖四、血漿樣本添加標準品之HPLC層析圖譜 55
圖五、壬基苯酚、辛基苯酚與丁基苯酚之標準溶液檢量線 56
圖六、壬基苯酚、辛基苯酚與丁基苯酚標準溶液之檢量線與添加
標準品於血漿樣本檢量線之比較 57
圖七、血漿樣本之分析圖譜 58
附錄
附錄一、清潔作業人員訪問之問卷 59
附錄二、特殊暴露族群之血漿樣本中NP、OP及BP之濃度 62

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