三聚氰胺定量檢測方法之比較

Comparison study of methods for quantification analysis for melamine

沈庭羽(Ting-Yu Shen)

指導教授 ： 錢偉鈞

朝陽科技大學/理工學院/應用化學系/碩士(2014年)

摘要

三聚氰胺 (Melamine， 2,4,6-triamino-1,3,5-triazine)，已證實對人體導致腎及肝臟上的疾病。不肖業者藉加入三聚氰胺導致凱氏定氮法（Kjeldahl protein ）中誤以為蛋白質含量增加。然而目前常用於檢測三聚氰胺儀器為液相層析串聯式質譜儀(liquid chromatograph /tandem mass spectrometer，LC-MS/MS)和高效液相層析(high performance liquid chromatography，HPLC)，因此本研究探討以LC-MS/MS及HPLC檢驗三聚氰胺在動物奶粉中殘留量，本研究另外以核磁共振光譜（Nuclear Magnetic Resonance ，NMR）中擴散係數實驗探討三聚氰胺檢測方法，並將三種方法互相比較。 HPLC和LC-MS/MS最佳條件如下：流速為0.4 ml/min；注射量為20 μl；UV波長為240nm；移動相A為5mM甲酸胺溶於水中、B相為5mM甲酸胺溶於乙腈中；比例為5：95。LC-MS/MS中質譜優化測定前驅離子及子離子之質荷比(m/z)分別為m/z 127→85作為定量；m/z 127→68作為定性。 以最佳條件分析出LC-MS/MS與HPLC檢量線範圍各為0.1-0.0005μg/mL和10-0.2μg/mL；判斷線性係數(R2)值各為0.9999和0.9997。進一步進行樣品添加驗證數據結果如下： HPLC和LC-MS/MS標準偏差(Standard Deviation)值為0.746μg/mL和2.429μg/mL；樣品變異係數(coefficient of variance，CV)為1.513 ％和11.258 ％。
核磁共振光譜分析，化學位移值5.947處為三聚氰胺中氫原子共振訊號，將訊號及濃度製成檢量線，其濃度線性範圍為1000μg/mL - 1 μg/mL，判斷線性係數(R2) 為0.99937、偵測極限為0.1 μg/mL。

關鍵字

高效液相層析 ； 核磁共振光譜 ； 液相層析串聯式質譜儀 ； 三聚氰胺 ； 飼料奶粉

並列摘要

Melamine (2,4,6-triamino-1,3,5-triazine) exposure can cause kidney stones and kidney failure. The culprits was added melamine led to Kjeldahl protein contained was raised. Analysis of melamine and its related instrument were containing liquid chromatograph -tandem mass spectrometer(LC-MS/MS) and high performance liquid chromatography(HPLC). In this study , we develop a rapid analysis method by HPLC and LC-MS/MS. We are also developing new NMR methods for the analysis of food additive. The HPLC eluents were 5mM ammonium formate in water / 5mM ammonium formate in acetinrile (5:95) pumped at flow rate of 0.4ml/min with an injection volume of 20 μl. the detection wavelength was set at 240nm. The m/z values of the precursor ions and product ions were m/z 127→85 used as quantitative and m/z 127→68 used as qualitative , respectively. Calibration graphs of melamine standard were constructed for LC-MS/MS and HPLC in the range 0.1 - 0.0005 μg/mL and 10 - 0.2 μg/mL , respectively. and their correlation coefficients calculated for LC-MS/MS and HPLC at 0.9999 and 0.9980, respectively. The relative standard deviations for LC-MS/MS and HPLC analysis were 0.746μg/mL and 2.429μg/mL , respectively. The coefficient of variance for LC-MS/MS and HPLC analysis at 1.513% and 11.258% , respectively. 1H NMR chemical shifts (in ppm) for melamine value 5.947 , and calibration graphs of melamine standard in the range 1000 μg/mL - 1 μg/mL . The correlation coefficients calculated for NMR at 0.99937. The detection limit was 0.1μg/mL.

並列關鍵字

HPLC ； NMR ； LC-MS/MS ； melamine ； Milk powder animal feed

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