4 4 Vol. 4 No. 4 2013 8 Journal of Food Safety and Quality Aug., 2013 - 刘华良 *, 阮丽萍, 刘德晔, 周明浩 (, 210009) 摘要 : 目的 -(HPLC-MS/MS) 方法 0.1% -2 mmol/l,, -, C 18 结果 1~1000 μg/l, 0.999, (LOQ) 10 μg/kg, 50~5000 μg/kg 82%~94%, (RSD) 4.9%~7.4% 结论 关键词 : - ; ; ; Determination of aconitine in food poisoning samples by high pressure liquid chromatography-tandem mass spectrometry LIU Hua-Liang *, RUAN Li-Ping, LIU De-Ye, ZHOU Ming-Hao (Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China) ABSTRACT: Objective To establish an analytical method for determination of aconitine in poisoning sesame paste samples by high pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods Samples were extracted by formic acid (0.1%)-ammonium formate (2 mmol/l) buffer. The extract was centrifuged and filtered by 0.45 μm nylon membrane filters before being analyzed with HPLC-MS/MS. A Waters Acquity BEH C 18 column (2.1 mm 50 mm, 1.7 μm) was adopted with the mobile phase of methanol and formic acid (0.1%)-ammonium formate (2 mmol/l) buffer (1:1, v/v). Aconitine was detected by high pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) under multiple reactions monitoring (MRM) mode. Results The method showed a good linearity in the range of 1~1000 μg/l with correlation coefficients of 0.999, and the LOQ was 10 μg/kg. The average recoveries were 82%~94% at the spiked level of 50~5000 μg/kg with the relative standard deviation (RSD) of 4.9%~7.4%. Conclusion This method can meet requirements of fast and accurate determination of aconitine in food poisoning samples. KEY WORDS: high pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS); aconitine; food poisoning samples; sesame paste 基金项目 : (BE2010745) (ZX201109) Fund: Supported by Jiangsu Provincial Science and Technology Support Project (BE2010745) and Jiangsu Provincial 12th Five-Year Health Development Project with Science and Education (ZX201109) * 通讯作者 :,, E-mail: LHL-1@163.com *Corresponding author: LIU Hua-Liang, Associate Researcher, Jiangsu Provincial Center for Disease Control and Prevention, No. 172, Jiangsu Road, Nanjing 210009, China. E-mail: LHL-1@163.com
1142 4 (aconitine),, CAS 302-27-2, C 34 H 47 NO 11, 1, 0.2 mg, 3~5 mg [1], 1 Fig. 1 Molecular structure of aconitine 2010 12 11,, 3 [2] 2013 2 28, 1,, [3],,,,, (TLC) [4,5] (HPLC) [6-8] - (HPLC-MS/MS) [9-14] HPLC-MS/MS, [8,10,12,13] [5,7,9] [6], HPLC-MS/MS, 1 材料与方法 1.1 试剂与仪器 (, TEDIA ); ( ) - : 1 mol/l 1.0 ml 0.5 ml 500 ml, HPLC- MS/MS, (1000 mg/l): 10.0 mg ( 0.1 mg) 10 ml,,,, 4 0.45 μm (, ); (1.5 ml, Eppendorf ); (, ); ( ) / (Acquity/Quattro micro, Waters ); ( 1 14, Sigma ) 1.2 实验方法 1.2.1 样品前处理 1 g( 0.001 g) 10 ml, -, 10 min, 30 min, 1 ml Eppendorf, 12000 r/min 15 min, 1.2.2 仪器条件 : Waters Acquity BEH C 18 (2.1 mm 50 mm, 1.7 μm); : 40 ; : A, B -, A B ; : 0.25 ml/min; : 5 μl : ESI + ; : 3.00 kv; : 105 ; : 350 ; : 500 L/h; : 40 L/h; : 3.5 10-3 mbar 1 Table 1 (m/z) 表 1 化合物的质谱条件 Mass spectrometric acquisition parameters (, ev) 645.6 585.7(20) (, ev) 553.7(30) 525.7(30) 367.8(40) (V) 40
4, : - 1143 1.2.3 测定, 1 5 10 50 100 500 1000 μg/l,, -,,, 2 结果与分析 2.1 液相色谱 - 质谱条件的优化 ESI + [9-14] MS1,, m/z 645.6; MS 2, 1 m/z 585.7, 3 m/z 553.7 m/z 525.7 m/z 367.8 ( 2) 4, m/z 367.8, [9,11-13],, 4 ( 2 ) 2.2 提取方法的选择 [4],,,,,,, -, -,,,, 2.3 方法学验证 2.3.1 线性范围和定量限, Y=85.6X 4.638, r=0.999, 1~1000 μg/l 5000 μg/l, S/N =10 1 μg/l, 10, (LOQ) 10 μg /kg 2.3.2 准确度与精密度, 50 500 5000 μg/kg,,, 6 82% 92% 94%, (RSD) 7.4% 6.2% 4.9% 5000 μg/kg MRM 3 Fig. 2 2 Characteristic ions scan mass spectrum of aconitine
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