涉案乌头生物碱与1-甲基海因的LC-MS/MS快速检测
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摘要
建立生物检材中1-甲基海因和乌头生物碱的快速分析方法,并提示检材新鲜度与1-甲基海因含量的相对关系。取全血样品经乙腈提取,使用Agilent Poroshell 120 SB-C_(18)(2. 1 mm×100 mm,2. 7μm)色谱柱,以0. 1%甲酸溶液-乙腈(30:70,V/V)为流动相等度洗脱,在多反应监测模式下测定全血样品中1-甲基海因和乌头生物碱成分。用于定量分析的离子对分别为1-甲基海因:m/z 115. 1 [M+H]~+→42. 2,乌头碱:m/z 646. 4 [M+H]~+→586. 3,新乌头碱:m/z 632. 3 [M+H]~+→572. 4和次乌头碱:m/z 616. 3 [M+H]~+→556. 3。4种目标物在各自的浓度范围内线性关系良好(R2≥0. 99),3个添加浓度(5,50,250 ng/mL)下,回收率在85. 7%～95. 2%之间,检出限(S/N=3)在0. 05～0. 5 ng/mL范围内,定量限(S/N=10)在0. 2～1 ng/mL范围内。利用建立的检测方法,发现与检材新鲜度有关的1-甲基海因和乌头生物碱的检出量有密切关系,为法医学鉴定结论的科学性服务。
The goal of this study was to jointly employ liquid chromatography-tandem mass spectrometry( LC-MS/MS) to detect 1-methylhydantoin and aconitine alkaloids in specimen,and to investigate the relative relationship between sample freshness and 1-methylhydantoin content. The aconitum alkaloids and 1-methylhydantoin were extracted from the whole blood by using acetonitrile and then analyzed by using HPLC-MS in multiple reaction monitoring( MRM) mode with positive ionization. The analytical column was Agilent Poroshell 120 SB-C_(18)( 2. 1 mm × 100 mm,2. 7 μm)and the mobile phase were water containing 0. 1% formic acid : acetonitrile( 30∶ 70,V/V) in isocratic elution. The precursor product ion combinations of 1-methylhydantoin: m/z 115. 1 [M+H]~+→42. 2,aconitine: m/z 646. 4 [M+H]~+→ 586. 3,mesaconitine : m/z 632. 3 [M+H]~+→572. 4 and hypaconitine: m/z 616. 3 [M+H]~+→556. 3 were used for quantitative analysis,respectively. The calibration curves were linear in their concentration ranges with the correlation coefficients( R2) more than 0. 99. The average recoveries of the method were 85. 7 % to 95. 2 %. The limits of detection( LODs) were in the range of 0. 05-0. 5 ng/m L( S/N = 3),and the limits of quantification( LOQ)were in the range of 0. 2-1 ng/m L( S/N = 10). It was found that 1-methylhydantoin related to sample freshness was closely related to the detection amount of aconitum alkaloids. This method holdsthe potontials in serving as a scientific service for forensic expert conclusions.
The goal of this study was to jointly employ liquid chromatography-tandem mass spectrometry( LC-MS/MS) to detect 1-methylhydantoin and aconitine alkaloids in specimen,and to investigate the relative relationship between sample freshness and 1-methylhydantoin content. The aconitum alkaloids and 1-methylhydantoin were extracted from the whole blood by using acetonitrile and then analyzed by using HPLC-MS in multiple reaction monitoring( MRM) mode with positive ionization. The analytical column was Agilent Poroshell 120 SB-C_(18)( 2. 1 mm × 100 mm,2. 7 μm)and the mobile phase were water containing 0. 1% formic acid : acetonitrile( 30∶ 70,V/V) in isocratic elution. The precursor product ion combinations of 1-methylhydantoin: m/z 115. 1 [M+H]~+→42. 2,aconitine: m/z 646. 4 [M+H]~+→ 586. 3,mesaconitine : m/z 632. 3 [M+H]~+→572. 4 and hypaconitine: m/z 616. 3 [M+H]~+→556. 3 were used for quantitative analysis,respectively. The calibration curves were linear in their concentration ranges with the correlation coefficients( R2) more than 0. 99. The average recoveries of the method were 85. 7 % to 95. 2 %. The limits of detection( LODs) were in the range of 0. 05-0. 5 ng/m L( S/N = 3),and the limits of quantification( LOQ)were in the range of 0. 2-1 ng/m L( S/N = 10). It was found that 1-methylhydantoin related to sample freshness was closely related to the detection amount of aconitum alkaloids. This method holdsthe potontials in serving as a scientific service for forensic expert conclusions.
引文
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