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標題: QuEChERS萃取技術結合液相層析串聯質譜術於穀類產品中真菌毒素之分析
Analysis of Mycotoxins in Grain Products by QuEChERS Combined with Liquid Chromatography-Tandem Mass Spectrometry
作者: 尤品人
Yu, Pin-Jen
關鍵字: Mycotoxin;真菌毒素;Liquid Chromatography-Mass Spectrometry;Grain product;液相層析串聯質譜術;穀類產品
出版社: 化學系所
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由實驗結果顯示,以QuEChERS萃取技術結合液相層析質譜技術,使用正、負離子電灑游離法將分析物離子化後,再以選擇反應偵測(Selected reaction monitoring, SRM)進行偵測,取0.8 g穀類產品粉末加入2.5 mL含1%甲酸之乙腈:水(40:60, v/v)萃取溶劑,用震盪機以750 rpm震盪三分鐘進行萃取,加入300 mg之氯化鈉和500 mg無水硫酸鎂於離心管中,以750 rpm震盪三分鐘後,再以4000 rpm離心五分鐘後取上層液至另一離心管,加入45 mg之C18作為吸附劑和50 mg 無水硫酸鎂去除雜質,將離心管以1400 rpm震盪三分鐘,再以14000 rpm轉速離心五分鐘後,將萃取液通過0.22 μm濾膜,取20 μL注入液相層析質譜儀分析。
由實驗結果顯示,黃麴毒素與赭麴毒素檢測線性範圍在1.5~100 ng/g之間,棒麴毒素於150~800 ng/g,橘黴素則為25~1000 ng/g,線性相關係數均0.992以上,偵測極限定義為訊號三倍雜訊比,分別介於0.4 ng/g(黃麴毒素B1)至28 ng/g(棒麴毒素)之間。同日間與異日間之相對標準偏差分別為1.6%至17.1%與5.8%至19.4%,回收率介於72.5~112.4%之間,使用此方法對於一些真實樣品進行偵測,可測得橘黴素在某傳統市場穀類產品之含量為433.4 ng/g。

QuEChERS combined with LC-MS/MS was used to determine mycotoxins in grain products. The 8 mycotoxins were separated on a Waters XTerraTM MS C18 column in 25 minutes. The optimum parameters of MS/MS for eight mycotoxins and conditions of QuEChERS preparation were also discussed.
0.8g of homogenous grain product was precisely weighted and put into a 15mL centrifuge tube. Subsequently, 2.5 mL of mixed extraction solvent (acetonitrile:H2O:formic acid/40:60:1 v/v/v) was added into the same tube. After vigorously shaking for 3 min, 300 mg of sodium chloride and 500 mg magnesium sulfate were added. The tube was shaken intensely for 3 min then centrifuged at 4000 rpm for 5 min. The supernatant was transferred to 1.5 mL centrifuge tube then 45 mg octadecyl bonded silica gel and 50 mg magnesium sulfate were added. After intensely shaking for 3 min then centrifuging at 14000 rpm for 5 min, the supernatant was passed through a 0.22 μm syringe filter, then the extract was analyzed by LC-MS/MS.
The analytical linearity for aflatoxins, ochratoxins, citrinin and patulin was from 1.5 to 100 μg/kg, 25 to 1000 μg/kg and 150 to 800 μg/kg, respectively. The limits of detection were from 0.4 to 1.0 μg/kg for aflatoxins, 0.4 to 0.6 μg/kg for ochratoxins, 6 μg/kg for citrinin and 28 μg/kg for patulin. The relative standard deviations of intra-day and inter-day analysis were from 1.7% to 17.1% and from 5.8% to 19.4%. The analysis of trace mycotoxin in grain products were also determined, the concentration of citrinin was detected at 433.4 ng/g. The results demonstrate the suitability of QuEChERS combined with LC-MS/MS approach to analyze trace amount of mycotoxins in grain products.
其他識別: U0005-1507201110522100
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