食品中防腐劑多重分析方法之開發   全文下載

Development of a Multiple Analytical Method for Preservatives in Foods

現行公告之「食品中防腐劑之檢驗方法（MOHWA0020.03）」係以高效液相層析儀搭配光二極體陣列檢出器（High Performance Liquid Chromatograph/ Photodiode Array Detector, HPLC-DAD）進行12項防腐劑之分析，本研究規劃以超高效液相層析串聯質譜儀（Ultra-high Performance Liquid Chromatograph/tandem Mass Spectrometer, UPLC-MS/MS）開發多重分析方法。本方法之分析物於原檢測之12品項外，新增規定外6項酯類及2項其它類防腐劑，共計20項防腐劑。本研究以含1%甲酸之乙腈溶液及QuEChERS萃取粉劑進行萃取，離心後取上清液以去離子水定容，經濾膜過濾後上機分析，使用ACQUITY UPLC BEH C18層析管柱，移動相採用0.1%甲酸溶液及含0.1%甲酸之甲醇溶液進行梯度沖提，20項防腐劑之質譜參數均進行最佳化設定，檢量線之線性範圍，除苯甲酸、二苯氧乙醇及鄰苯基苯酚等3項防腐劑為100-1,000 ng/mL外，其餘17項防腐劑均為10-100 ng/mL，其相關係數（r）均大於0.990。確效試驗與定量極限評估：於空白基質中添加2種濃度之20項防腐劑，結果顯示，平均回收率介於87.0-104.8%之間，同日內變異係數介於1.8-9.3%之間，異日間變異係數介於1.6-13.8%之間，定量極限介於2-20 mg/kg，均符合衛生福利部食品藥物管理署公布之食品化學檢驗方法之確效規範。另以市售烘焙類食品10件驗證所建立之檢驗方法適用性，結果均與產品標示相符且無基質干擾之情形。

The“Method of Test for Preservatives in Foods (MOHWA0020.03)”announced by the Ministry of Health and Welfare in Taiwan is based on high performance liquid chromatography coupled with a diode-array detector (HPLC-DAD) to analyze 12 preservatives. The aim of this study was to develop a multiple analytical method using ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS) to analyze 20 preservatives. An ACQUITY UPLC BEH C18 column was used, and 0.1% formic acid and methanol containing 0.1% formic acid used as the mobile phase for gradient elution. The mass spectrometric parameters of 20 preservatives were optimized. The linear ranges of the 20 preservatives in this method, except for benzoic acid, 2-phenoxyethanol and o-phenylphenol, which were from 100 to 1,000 ng/mL, were from 10 to 100 ng/mL for the rest of the 17 preservatives. Their linear correlation coefficients (r) were all greater than 0.990. The sample was extracted with acetonitrile containing 1% formic acid and QuEChERS extraction powder, after centrifugation, the supernatant was diluted to a constant volume with deionized water. Twenty preservatives at 2 concentrations were added to the blank samples, and the accuracy, precision and limit of quantification were evaluated. The results showed that the average recoveries ranged from 87.0 to 104.8%. The intraday and interday coefficients of variation ranged from 1.8 to 9.3% and from 1.6 and 13.8%, respectively, and the limits of quantification were between 2 and 20 mg/kg. The above results all complied with the validation guideline of the food chemical testing methods published by TFDA. In addition, ten commercial bakery products were used to verify the applicability of this method. The results were consistent with the product labeling and there were no matrix interference.