應用氣相層析串聯質譜儀（GC-MS/MS）建立禽畜水產物多氯聯苯的快速分析方法

Application of Gas Chromatography Tandem mass Spectrometry (GC-MS/MS) to Establish a Rapid Analysis Method for Polychlorinated Biphenyls in Aquatic Products of Poultry and Livestock

陳思縈(Sz-Ying Chen) ； 黃鎮華(Chen-Hua Huang) ； 徐慈鴻(Tsyr-Horng Shyu) ； 曾昭銘(Chao-Ming Tsen)

《臺灣農藥科學》 11期 (2021/12) Pp. 105-126

https://doi.org/10.6671/TPS.202112_(11).06

摘要

近年來發生的食品飼料檢出戴奧辛呋喃（PCDD/Fs）超量事件，由於檢驗期程過長或監測廣度不足，而造成不小的經濟損失及社會不安。而與戴奧辛結構相似的多氯聯苯（PCBs），有12種共平面型的同源物同樣具有戴奧辛毒性，稱為戴奧辛類多氯聯苯（DL-PCBs），另外有6種非共平面型同源物為環境中所有多氯聯苯總量的一半以上，因此我們選擇這6種同源物作為監測的依據。為能與戴奧辛分析流程同步的多氯聯苯化合物檢測，更具效率、環保、易操作並保有準確性，本研究探討氣相層析串聯質譜儀（GC-MS/MS）作為偵檢儀器，分析魚粉及奶蛋基質中微量多氯聯苯的可行性，並評估前處理流程自動化的確效。以此建立一種高效準確的快速檢測模式，樣品經由高壓溶劑萃取在30分鐘內完成油脂萃取，相較於傳統索氏萃取的8至24小時縮減90%期程，再以自動淨化系統於3小時內可完成檢液淨化，相較於現行人力操作方式需時2－3天，效率提高15倍以上，以魚粉、雞蛋及乳品等三種基質進行添加回收試驗，以氣相層析串聯質譜儀（GC-MS/MS）分析，所有分析的待測同源物之離子對強度比值＜15%，達到歐盟分析規範，檢量線最低點之濃度與公告方法所用的HRGC/HRMS相同，樣品中所有多氯聯苯同源物的回收率介於95－139%，相對標準偏差（RSD）小於12%，說明本研究導入的自動化前處理在方法確效具有良好的準確性及穩定性。進一步取20件真實樣品進行方法驗證，樣品以本研究的快速方法及GCMS/MS分析的測值，比對本實驗室依認證方法及HRGC/HRMS所得之測值，由配對t檢定結果顯示兩方法間並無顯著差異，且在國際實驗室間分析能力比試中，鮭魚樣品的12種DL-PCBs總毒性當量及魚油ICES-6個別濃度的檢測結果，其標準化分數（Z score）均小於2，驗證本方法應用於真實樣品中多氯聯苯分析的可行性。本流程由單人操作可在1天內完成單件樣品的檢測分析，或在3天內得到一批次6件樣品的檢測數值，相較於現行檢驗期程需時一周左右，本方法除了大幅提升檢測效率，亦維持高度的準確性及穩定度，更具有安全、省力、減碳、易建置、易操作及易維護等優點。

關鍵字

高壓溶劑萃取裝置 ； 氣相層析串聯質譜儀 ； 配對t檢定

並列摘要

In recent years, excessive amounts of dioxinfuran (PCDD/Fs) have been detected in food and feed. Furthermore, long inspection periods or insufficient monitoring, have led to economic losses and social unrest. Polychlorinated biphenyls (PCBs), which share a similar structure with dioxin, have (1) twelve coplanar homologues that possess dioxin toxicity, (called dioxin-like polychlorinated biphenyls DL-PCBs), and (2) six non-coplanar polychlorinated biphenyls (NDL-PCBs). This study sought to develop a more efficient, accurate, user-friendly,and environmentally-sustainable method of detecting environmental PCBs. As polychlorinated biphenyls account for more than half of all PCBs in the environment, we selected these six homologs as the basis for monitoring in synchronization with the analysis process of dioxin, more efficient, environmentally friendly, easy to operate and maintain accuracy, Specifically, we investigated (1) the efficacy and fessibility of using gas chromatography tandem mass spectrometry (GC-MS/MS) to analyze fish meal and trace PCBs in milk and egg matrices, and (2) the effectiveness of automation in the pretreatment process. In so doing, we developed an efficient, accurate, and rapid PCB detection method. Under our method the sample is extracted using high-pressure extraction in less than 30 minutes, which is 90% faster than traditional Soxhlet extraction (8 to 24 hours). Following sample extraction, the test solution is purified using an automatic purification system three hours. The purification of the test solution can be completed. Compared with the current manual operation method, it takes 2-3 days and the efficiency is increased by more than 15 times GC-MS/MS analysis. In addition, the ion-pair intensity ratio of all analyzed homologues was less than 15%, which meets EU analysis standards. The concentration at the lowest point of the calibration curve was the same as that of HRGC/HRMS used in the announced method. In validating our method, the recovery values of chlorobiphenyl homologues were found to range from 95% to 139%, and the relative standard deviation (RSD) was less than 12%. These results indicate that the automated pre-processing process introduced in this study has good accuracy and stability. We further verified our proposed method using 20 samples. The samples were compared with the measured values obtained by the laboratory according to the certified method and HRGC/HRMS using the rapid method of this research and the measured values of GC-MS/MS analysis, and results of paired t-tests were calculated. Values obtained using both methods were not significantly different. Moreover, in an international inter-laboratory analysis ability comparison, the total toxicity equivalent of 12 kinds of DL-PCBs in salmon samples, the detection results of individual concentrations of fish oil ICES-6, and standardized scores (Z score) were all less than 2. Using our proposed detection and analysis method, a single sample can be analyzed by a single person in less than one day, and a batch of 6 samples can be analyzed within 3 days. Current methods require approximately one week for detection and analysis. Thus, our proposed method greatly improves detection efficiency while maintaining a high degree of accuracy and stability. Moreover, our method is safe, reduces labor costs, reduces carbon emission, and is easy to set up, operate and maintain.

並列關鍵字

high-pressure extraction device ； Gas chromatography tandem mass spectrometry (GC-MS/MS) ； paired t test