丙烯醯胺、環氧丙醯胺及其谷胱甘肽加合物於Sprague-Dawley大鼠之毒物動力學研究

Toxicokinetics of Acrylamide, Glycidamide, and Their GSH Conjugates in Sprague-Dawley Rats

龍泰穎(Tai-Ying Long)

指導教授 ： 吳焜裕

共同指導教授 ： 蔡詩偉

國立臺灣大學/公共衛生學院/環境衛生研究所/碩士(2015年)

https://doi.org/10.6342/NTU.2015.00913

摘要

丙烯醯胺(Acrylamide)為廣泛應用的工業用化合物。近年來發現丙烯醯胺普遍存在於高溫油炸食品中，因而在食品安全議題上受到重視。過去研究指出，丙烯醯胺的潛在致癌性主要來自於中間代謝物環氧丙醯胺(glycidamide)，而丙烯醯胺與環氧丙醯胺可以與體內廣泛分布的谷胱甘肽(glutathione, GSH)結合而降低其毒性。本研究的目的在於建立固相萃取前處理法與電灑離子化搭配液相層析串聯質譜儀分析方法，以此方法去分析Sprague-Dawley大鼠血液樣本中經由Phase-II reactions的三種丙烯醯胺-谷胱甘肽加合物(GSH conjugates)，分別為AAGSH、GA2GSH和GA3GSH，藉由WinNonlin模型軟體分析此三種加合物來了解其毒物動力學資料，其數據可用來提供研究物種間代謝機制差異，以評估丙烯醯胺對於人體的致癌性。本研究經由腹腔注射法，分別給予雄性、雌性大鼠0.1 mg/kg-wt、1.0 mg/kg-wt及5.0 mg/kg-wt的丙烯醯胺。本研究在低劑量0.1 mg/kg-wt下沒有偵測出GA2GSH，除此之外皆有測到AAGSH、GA2GSH和GA3GSH。曲線下面積(AUC, Area under the curve)、代謝速率常數(keli)以及半衰期(T1/2)等毒物動力學參數經由Win-Nonlin模型軟體計算出來。AAGSH、GA2GSH和GA3GSH曲線下面積(AUC)相對於AA給藥劑量的比率分別為0.97-1.78 %、0.03-0.07%和0.17-0.25%。∑ [GAGSH/AAGSH] 比率為0.13-0.18。從GA3GSH曲線下面積大於GA2GSH曲線下面積來看，可以證實谷胱甘肽與環氧丙烯胺上較少取代基的碳容易產生鑑結。本研究的動力學數據除了進一步了解丙烯醯胺的代謝機制外，能夠對於致癌性風險評估上有所幫助。

關鍵字

丙烯醯胺 ； 谷胱甘肽 ； 毒物動力學 ； 液相層析-三段四極柱質譜儀 ； 固相萃取前處理法

並列摘要

Acrylamide (AA) is a widely-used industrial chemical. For the past few years, AA was found in high-temperature fried food, so the food safety has been concerned. According to previous researches, the potential carcinogensis of AA mainly come from its metabolite: glycidamide (GA), which can be reacted with glutathione and further decreased the toxicity. The study is aimed to develop a solid phase extraction (SPE) pretreatment with LC-ESI-MS/MS method. With this analytical method, we can quantify the GSH conjugates (AAGSH, GA2GSH, and GA3GSH) through Phase-II reactions in the blood of Sprague-Dawley rats. The male/female Sprague-Dawley rats were treated with AA of 0.1 mg/kg-wt, 1.0 mg/kg-wt, and 5.0 mg/kg-wt by intraperitoneal injection, respectively (N=5). The toxicokinetic parameters including AUC0-∞, elimination rate constant (keli), and half life (T1/2) were calculated by Win-Nonlin modeling software. Although the concentration of GA2GSH were not detected from the SD rat dosed 0.1 mg/kg-wt AA, Molar percentage for AAGSH, GA2GSH, and GA3GSH of dosing AA accounted for 0.97-1.78 %, 0.03-0.07 %, and 0.17-0.25 %, respectively. Overall, ratio of ∑GAGSH/AAGSH amount excreted were 013-0.18.Based on AUC0-∞, glycidamide were preferred to conjugate with glutathione as GA3GSH than GA2GSH because of attacking least substituted carbon on GA.
This results could be provided for the species differences of AA metabolism and the risk assessments of its carcinogenicity.

並列關鍵字

Acrylamide ； Glutathione ； Toxicokinetics ； LC-MS/MS ； Solid phase extraction

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