以固相微萃取技術與衍生方法搭配氣相層析串聯質譜儀分析個人保健用品中的抗菌防腐成分

Abstract

在日常生活中，人們每天皆會使用各種不同個人保健用品，例如：洗面乳、洗髮精與乳液等；這類產品通常含水量高，且具有醣類、蛋白質或脂質等成份，而若使用或儲存不當，可能會造成微生物孳生。因此，避免污染及延長保存期限，於個人保健用品添加抗菌防腐劑是個常見的方法 (例如：對羥基苯甲酸酯 (parabens)及三氯沙 (triclosan)等)。另一方面，由於塑膠製品常做為個人保健用品之容器，因此使用相關產品時亦可能因為釋出雙酚A (bisphenol A)而造成暴露。 為了保障消費者權益及健康，台灣及歐盟等國家規範了抗菌防腐劑之濃度，而雙酚A亦雖規定不能在奶瓶中添加，但在其他容器並無要求；儘管如此，市面上還是有很多具有對羥基苯甲酸酯或三氯沙等物質。由於臺灣個人保健用品含有抗菌防腐劑以及容器可能釋出的雙酚A的資訊非常有限，因此為了瞭解民眾暴露到這些物質的健康風險，建立一個具敏感及簡易的方法同時分析待測物是必要的。 本研究利用頂空固相微萃取(HS-SPME)前處理步驟搭配氣相層析串聯質譜儀(Triple Quadrupole GC/MS/MS)同步偵測4種對羥基苯甲酸酯、三氯沙及雙酚A。為了增加敏感度，本研究以乙酸酐 (Acetic anhydride)作為衍生試劑進行衍生反應，同時添加氯化鈉 (sodium chloride)產生鹽析反應。樣本分析前，先用超純水稀釋，而萃取步驟使用 65μm PDMS / DVB 萃取纖維搭配頂空萃取模式，以100℃與搖晃 250 rpm萃取 20 分鐘，在達平衡後直接將萃取纖維插入氣相層析儀之進樣口進行 1 分鐘之熱脫附。 6種分析物因物化特性其定量範圍不同，檢量線呈現良好的線性關係 (R > 0.99)，因此此方法足夠敏感可以分析各種個人保健用品的濃度，考量到產品中複雜基質，樣本稀釋以及添加擬標為常見降低其受基質效應影響的方法。 收集155項樣本，共分為12種品項分類進行定量分析，結果顯示對羥基苯甲酸酯較常當作防腐劑出現在產品中，其中對羥基苯甲酸甲酯與對羥基苯甲酸丙酯頻率較高，可能是因為一同添加在產品中增加其抗菌防腐能力。對羥基苯甲酸酯較常檢測於停留性的個人保健用品，例如：身體乳液與臉部乳液等，在研究中，各個物質檢測出最高濃度分別為：對羥基苯甲酸甲酯3704.64 μg/g、對羥基苯甲酸乙酯645.59 μg/g、對羥基苯甲酸丙酯1733.29 μg/g與對羥基苯甲酸丁酯299.91 μg/g，三氯沙只有在牙膏中測得，最高濃度為2905.47 μg/g，產品中雙酚A的檢出率低以及測得濃度也相當低6.02 μg/g，除了一項臉部乳液產品中對羥基苯甲酸丙酯，大多產品中防腐劑濃度皆在規範濃度內，嬰兒及敏感性肌膚專用之產品在本實驗中尚未偵測到濃度，此外，本研究利用所測得的產品中待測物濃度，進行估算皮膚吸收暴露，分為三個族群：成人女性、小孩與嬰兒評估其劑量，對於成人女性暴露的結果顯示0.00001-0.78208 μg/kg-bw/day，另外，對羥基苯甲酸酯於不同類型產品的貢獻量，身體乳液與臉部乳液占較大比例，其中最高暴露量分別為8.54618和3.49533 μg/kg-bw/day。 本研究利用固相微萃取與衍生方法搭配氣相層析串聯質譜儀之檢測方法，提供個人保健用品的抗菌防腐劑與潛在釋出雙酚A濃度，在未來研究中，可以再進一步調查在不同環境介質中分布與濃度，與評估對於民眾暴露到抗菌防腐劑的健康風險。

Preservatives are commonly added to consumer products to ensure the safety during use and storage. Among the preservatives, parabens (methylparaben(MP), ethylparaben(EP), propylparaben(PP), butylparaben(BP)) and triclosan (TCS) are widely used in personal care products (PCPs). In addition, most of the containers for PCPs are made of plastic materials, which might cause the release of bisphenol A (BPA) into the products. Various health concerns related to the exposures of parabens, triclosan, and BPA, including contact allergy, endocrine system disruption, and reproductive system effects, have been reported. Preservatives in the cosmetics have been regulated by the European Union (EU). However, there are still lots of parabens or triclosan-containd products on the market. Hence, there is a need to develop a simple and sensitive analytical method in order to determine the distributions of parabens, triclosan, and BPA in PCPs. In this study, analytes and surrogate standards were prepared in methanol as the stock solution. Further dilutions and mixtures of PCPs were prepared in the same solvent. To enhance sensitivities, acetic anhydride and potassium carbonate were added for the in situ derivatization. Analytes were extracted at 90℃ for 20 minutes with 250 rpm. Headspace solid-phase microextraction (SPME) procedure using 65μm PDMS/DVB was performed. After extraction, the fiber was inserted into the GC injector port coupled with triple quadruple gas chromatography with tandem mass spectrometry (GC/MS/MS). The SPME procedure coupled with GC/MS/MS analysis for the determinations of the above metioned compounds in PCPs was established in this study. Parameters which might have the effects on the SPME extractions were evaluated, including extraction time, extraction temperature, agitator speed, volume of acetic anhydride and concentration of sodium chloride. By adding 100μl of the derivative reagents and 20% (w/v) sodium chloride, the method established was sensitive enough to determine the target compounds. 155 PCPs samples were analyzed in this study. The linear range was 0.02-2 ng/ml for MP and PP, 0.05-1 ng/ml for EP, 0.02-1 ng/ml for BP, 0.02-5 ng/ml for TCS and BPA. The method detection limits (MDLs) of analytes were 0.007-0.017 ng/ml and RSDs showed 1.15-17.63% at low concentration and 0.39-4.94% at high concentration. Good linearity and precision were presented and the recoveries ranged from 65% to 150%. This study provided a survey regarding the concentration of parabens, triclosan and BPA in personal care products in Taiwan, including national or foreign brands. The results revealed methylparaben and propylparaben have the higher frequencies in PCPs. The high concentrations of parabens were mainly found in leave-on products, like body lotion and facial lotion. There is methylparaben for 3704.64 μg/g, ethylparaben for 645.59 μg/g, propylparaben for 1733.29 μg/g and butylparaben for 299.91 μg/g. Triclosan was detected only in toothpaste (2905.47 μg/g) and BPA had the low level (6.02 μg/g) and detection rate in PCPs. In addition, daily dermal exposure was estimated, based on the mean and maximum concentration of analytes in this study. The target group was divided into three types, adult females, toddlers and infants. The results indicated that body lotion and facial lotion were major sources of human exposure. With the findings of this study, the possible health risks due to the exposures can be assessed.