醬油中雙十二烷基硫酸硫胺明之檢驗方法開發   全文下載

Development of a Quantification Method for Thiamine Dilaurylsulfate in Soy Sauce

本研究建立以超高效液相層析串聯質譜儀（Ultra-high Performance Liquid Chromatograph/tandem Mass Spectrometer, UPLC-MS/MS）檢測醬油中雙十二烷基硫酸硫胺明之檢驗方法。檢體經乙腈萃取後以儀器分析，使用Hypersil GOLD C8之層析管柱（1.9μm，2.1×150 mm），以0.15%甲酸水溶液與含0.15%甲酸之乙腈溶液作為移動相溶液，流速0.45 mL/min進行梯度流洗，並以醬油空白檢體製備之空白檢液，添加標準品製作基質匹配檢量線進行定量。基質匹配檢量線之線性範圍25-500 ng/mL，相關係數（r）為0.9998。以醬油空白基質添加雙十二烷基硫酸硫胺明0.001及0.01 g/kg（以十二烷基硫酸鹽計）進行準確度、精密度與定量極限之評估，結果顯示，平均回收率分別為91.8及94.9%，同日內變異係數為4.8及1.9%，異日間變異係數為5.2及5.1%，本檢驗方法之定量極限為0.001 g/kg。後續並以市售醬油7件驗證所建立之檢驗方法適用性，結果均未檢出且未有基質干擾之情形。相關檢驗方法已公開於衛生福利部食品藥物管理署網站，提供各界參考引用。

An analytical method for determining thiamine dilaurylsulfate in soy sauce by an ultra-high performance liquid chromatograph/tandem mass spectrometer (UPLC-MS/MS) was developed. Samples were extracted with acetonitrile and fractionated on a Hypersil GOLD C8 column (1.9μm, 2.1×150 mm) with gradient elution of acetonitrile containing 0.15% formic acid and deionized water containing 0.15% formic acid at a flow rate of 0.45 mL/min. Quantification was performed using the matrix-matched calibration, which was found linear in the concentration range of 25 to 500 ng/mL, and the correlation coefficient was 0.9998. The accuracy, precision and limit of quantification were evaluated by spiking thiamine dilauryl sulfate into the blank matrix at 0.001 and 0.01 g/kg, which was estimated as lauryl sulfate. For the soy sauce sample, the average recoveries of lauryl sulfate were 91.8 and 94.9%, the intraday coefficients of variation were 4.8 and 1.9%, and the inter-day coefficients of variation were 5.2% and 5.1%, respectively. The limit of quantification was 0.001 g/kg. This analytical method was further applied to 7 commercial soy sauce samples. The results showed that no dilauryl sulfate was detected and no matrix interference was found in all samples. This method has been published as“Method of Test for Thiamine Dilaurylsulfate in Soy Sauce (TFDAA0096.00)”on the TFDA official website.