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1 4 4 Vol. 4 No Journal of Food Safety and Quality Aug., 刘华良 *, 阮丽萍, 刘德晔, 周明浩 (, ) 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 , 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 : (BE ) (ZX201109) Fund: Supported by Jiangsu Provincial Science and Technology Support Project (BE ) and Jiangsu Provincial 12th Five-Year Health Development Project with Science and Education (ZX201109) * 通讯作者 :,, LHL-1@163.com *Corresponding author: LIU Hua-Liang, Associate Researcher, Jiangsu Provincial Center for Disease Control and Prevention, No. 172, Jiangsu Road, Nanjing , China. LHL-1@163.com

2 (aconitine),, CAS , C 34 H 47 NO 11, 1, 0.2 mg, 3~5 mg [1], 1 Fig. 1 Molecular structure of aconitine ,, 3 [2] , 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,,,, μm (, ); (1.5 ml, Eppendorf ); (, ); ( ) / (Acquity/Quattro micro, Waters ); ( 1 14, Sigma ) 1.2 实验方法样品前处理 1 g( g) 10 ml, -, 10 min, 30 min, 1 ml Eppendorf, r/min 15 min, 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; : mbar 1 Table 1 (m/z) 表 1 化合物的质谱条件 Mass spectrometric acquisition parameters (, ev) (20) (, ev) 553.7(30) 525.7(30) 367.8(40) (V) 40

3 4, : 测定, 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 m/z m/z ( 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 准确度与精密度, 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

4 Fig. 3 3 MRM ( 5000 μg/kg) MRM chromatogram of aconitine in spiked sesame paste at the spiked level of 5 mg/kg 2.4 实际应用 ,,, 3,, 3 1, 11 2, 3 4, [3],,,,, 3 结论 TLC [4,5] HPLC [6-8], HPLC-MS/MS, HPLC-MS/MS,, 参考文献 [1]. [M]. :, Wen YL. Drugs and chemical toxicity data [M]. Tianjin: Tianjin Science and Technology Publishing House, [2] [DB/OL]. cq.xinhuanet.com/news/ /14/content_ htm Murderer of North Chongqing poisonous wine is aconitine [DB/OL] /14/content _ htm [3] [DB/OL]. longhoo.net/ /04/content_ htm Nanjing Qinhuai poisonous sesame powder incident arised from aconitine[db/ol] htm [4]. [M]. :, Chinese Pharmacopoeia Commission. Chinese Pharmacopoeia [M]. Beijing: Chinese Medical Science and Technology Press, [5],, 贠,. [J]., 2008, 18 (9): Wei ZW, Liu J, Yun KM, et al. Rapid diagnosis of aconites poisoning by TLCs [J]. Chin J Health Lab Technol, 2008, 18(9): [6],,. [J]., 2005, 21(1): Li HH, Yu LS, Jin M. Separation and determination of aconitine in medicine wine by reversed-phase high performance liquid chromatography [J]. J Forensic Med, 2005, 21(1):

5 4, : [7],,,. HPLC [J]., 2009, 44(12): Su G, Liao LC, Yan YY, et al. Simultaneous determination of mesaconitine, aconitine and hypaconitine in blood and liver speciman by HPLC [J]. Chin Pharm J, 2009, 44(12): [8],,,. HPLC [J]., 2012, 27(12): Zhou J, Ye L, Tang L, et al. Simultaneous content determination of aconitine, mesaconitine and hypaconitine in fuzi by HPLC [J]. China J Tradit Chin Med Pharm, 2012, 27(12): [9],,. - [J]., 2012, 33(2): Chen XG, Lai YQ, Cai ZW. Simultaneous analysis of aconitine and its metabolites by liquid chromatography-electrospray ion trap mass spectrometry [J]. J Chin Mass Spectrom Soc, 2012, 33(2): [10],,,. UPLC-MS 3 [J]., 2012, 32(5): Cui P, Yang L, Xiong AZ, et al. Quantification of three aconitine-type diester alkaloids in xiaozhong tablets by UPLC-MS [J]. Chin J Pharm Anal, 2012, 32(5): [11],,. [J]., 2011, 17(2): Li Z, Tan P, Wang CG. The ESI-MS study on the fragmentation phthways of the aconitine, mesaconition, hypaconitine and their hydrolyzates [J]. Mod Instrum, 2011, 17(2): [12],,,. 3 RRLC-MS/MS [J]., 2010, 30(9): Cheng XL, Bai ZL, Xiao XY, et al. RRLC-MS/MS simultaneous determination of 3 diester diterpenoid alkaloids in processed products of Aconitum [J]. Chin J Pharm Anal, 2010, 30(9): [13],,,. [J]., 2006, 21(2): Wang CH, Wen J, Chen YH, et al. Determination of metabolites of aconitine in rat urine by HPLC/MS [J]. Chin J Forensic Med, 2006, 21(2): [14] Gaillard Y, Pepin G. Poisoning by plant material: review of human cases and analytical determination of main toxins by high-performance liquid chromatograpgy-(tandem)mass spectrometry[j]. J Chromatogr B, 1999, 733(1): 作者简介 ( 责任编辑 : 赵静 ) 刘华良, 副研究员, 主要研究方向为食品安全保障技术 LHL-1@163.com

材料与方法仪器 ACQUITY UPLC Xevo TQ-S 三重四级杆液质联用仪 ( 美国 Waters 公司 ); 电喷雾离子源 (ESI), Masslynx 4.1 数据处理系统 (Waters);Thermo 低温离心机 ;KQ

材料与方法仪器 ACQUITY UPLC Xevo TQ-S 三重四级杆液质联用仪 ( 美国 Waters 公司 ); 电喷雾离子源 (ESI), Masslynx 4.1 数据处理系统 (Waters);Thermo 低温离心机 ;KQ [ 应用纪要 ] Rapid identification of 5 illegal additives in slimming health food by Xevo TQ-S 郝刚苏州市食品药品检验所, 苏州 15104 关键词超高效液相色谱 - 质谱, 多反应监测, 减肥类保健食品非法添加, 快速定性定量