- 學位論文
利用質譜儀建立對羥基苯甲酸酯之快速定量方法及分析其引起之蛋白質表現
Rapid quantitation of parabens and parabens-induced protein expression analysis by mass spectrometry
摘要
對羥基苯甲酸酯(parabens)是對羥基苯甲酸(4-hydroxybenzoic acid)的烷基酯類，為一類防腐劑，經常添加在食品、化妝品及藥品中，常見包括methylparaben、ethylparaben、propylparaben和butylparaben[1]。 本研究利用基質輔助雷射脫附游離飛行時間質譜儀 (matrix-assisted laser desorption ionization time-of-flight mass spectrometry; MALDI-TOF MS)建立一套快速、靈敏的分析方法，可同時檢測methylparaben、ethylparaben、propylparaben及butylparaben的含量，應用於檢測藥物和化妝品等市售產品，此項技術正進行專利申請(PK13059)。 細胞實驗結果顯示，parabens會誘發Huh-7細胞中部分共同細胞及誘發蛋白質表現，例如肝臟代謝、解毒反應及能量生成作用相關的蛋白質。而HaCaT細胞實驗的結果指出，本研究以100 μM之parabens進行培養，屬於low dose的劑量範圍對HaCaT所造成的影響不明顯。然而，經由UVB照射後確實引發與皮膚疾病相關蛋白質表現增加的情形，尤其加入MP及PP再配合光照後所引發與過敏相關蛋白質的表現，證實MP及PP具有觸發過敏反應的特性。 有鑑於parabens的廣泛使用，發展一套快速簡單的方法對市售產品的parabens添加量做快速檢測有其必要性。
並列摘要
Parabens are esters of para-hydroxybenzoic acid, include methylparaben (MP), ethylparaben (EP), propylparaben (PP) and butylparaben (BP). They are widely used as preservatives in food, pharmaceuticals and personal care products. In this study, a rapid and sensitive analytical method was developed for simultaneous quantification of MP, EP, PP, BP in pharmaceuticals and personal care products. Analysis was carried out by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The patent is under process (PK13059). In vitro experiments showed that parabens treatment for 24 hoours may induce cell protein expression, in Huh-7 cells; such as liver metabolism, detoxification reactions and energy production-related proteins. HaCaT keratinocytes treated with low-dose parabens concentration for 24 hours had no effect on protein expression. However, further cultured with UVB exposure for 2 hour significantly induced protein expression associated with dermatitis and allergies. According to the widespread used of parabens as preservatives, it is neccessary to developed a rapid analytical method for determination of parabens in daily use products.