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1 7 10 Vol. 7 No Journal of Food Safety and Quality Oct., 叶雅真 1 1,, 骆和东 2*, 张璨雯 3 1,, 黄三发 2, 蔡伟鹏 (1., ; 2., ; 3., ) 2 摘要 : 目的 - (electrospray ionization-ion mobility spectrometry, ESI-IMS) 5 方法, 90% ESI-IMS,, 结果 5, mg/l, 78.7%~113.8%, (RSD, n=6) 10%, - (liquid chromatography-tandem mass spectrometry, LC-MS/MS) 结论, 5 关键词 : -; ; ; Rapid screening of 5 kinds of illegal drugs in improving sleep health foods and traditional Chinese medicines by electrospray ionization-ion mobility spectrometry YE Ya-Zhen 1, LUO He-Dong 1, 2*, ZHANG Can-Wen 3, HUANG San-Fa 1, 2, CAI Wei-Peng 2 (1. Xiamen Institute for Food and Drug Quality Control, Xiamen , China; 2. Xiamen Center for Disease Control and Prevention, Xiamen , China; 3. College of Biology Engineering, Jimei University, Xiamen , China) ABSTRACT: Objective To establish a method for rapid screening of diazepam, midazolam, barbital, nitrazepam and alprazolam in health foods and traditional Chinese medicines by electrospray ionization-ion mobility spectrometry (ESI-IMS). Methods The samples were extracted by ultrasound with acetonitrile and the supernatant was diluted by 90% acetonitrile, and then detected by IMS with ESI + mode for diazepam and midazolam, with ESI - mode for barbital, nitrazepam and alprazolam. The matrix-matched external standard method was used for quantification. Results Good linearities of 5 kinds of compounds were obtained (r>0.99) and the detection limits of diazepam, midazolam, barbital, nitrazepam and alprazolam were 0.20, 0.50, 2.00, 8.00, 8.00 mg/l, respectively. 基金项目 : (2014D005) Fund: Supported by the Project of Natural Science Foundation of Fujian Province (2014D005) * 通讯作者 :,, luohedong@126.com *Corresponding author: LUO He-Dong, Chief Technician, Xiamen Institute for Food and Drug Quality Control, Xiamen , China. luohedong@126.com

2 10, : The recoveries were 78.7%~113.8% with the relative standard deviations below 10% (n=6). The results were in accordance with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Conclusion The established method is rapid and accurate, and can be applied in detection of illegal drugs for improving sleep products of diazepam, midazolam, barbital, nitrazepam and alprazolam. KEY WORDS: electrospray ionization-ion mobility spectrometry; health foods for improving sleep; illegally added; rapid detection 1 引言,,,,, [1-4] 2012 () [5],,,,,, 5 1,, [6,7],, [8] [9,10] - [11,12],,,, [13-15], 5 - (electrospray ionization-ion mobility spectrometry, ESI-IMS) 5,, Fig Structures of 5 kinds of compounds

3 材料与方法 2.1 仪器与试剂 GA2100, (ESI)( Excellims ); Agilent6460 (); KQ-500E ( ); 3-18K ( Sigma ); WH-2 ( ); HY-2 ( ) 5 : (>99%, )(, Tedia ); 0.22 μm (); Milli-Q 2.2 实验方法 样品前处理 (), 0.30 g( 0.01 g) 25 ml, 10 ml, 20 min, 8000 r/min 10 min, 90%(V:V) 50, 0.22 μm, 标准溶液和基质匹配标准曲线的配制 (1) :, 2 ml,, mg/l mg/l,,, 2, 90%(V:V), : 2, 2.2.1,,, 80 mg/l 20 mg/l,,, (2) :, 2 ml,, mg/l, mg/l, mg/l,,, 3, 90% (V:V), : 3, 2.2.1,,, 30 mg/l,,, 分析条件 (1) IMS : (ESI + ); : 2000 V; : 8300 V; : 180 ; : 200 ; : 0.90 L/min : (ESI - ); : 2000 V; : 8200 V; : 190 ; : 180 ; : 0.90 L/min (2) - (liquid chromatography-tandem mass spectrometry, LC-MS/MS) : Agilent Eclipse XDB-C 18 (4.6 mm 50 mm, 1.8 μm); A: 0.1%, B: 40%, : 0.5 ml/min, : 5 μl (ESI), ; (MRM) ; : 325, : 12 L/min, : 300, : 12 L/min, : 4000 V; 1 表 1 质谱分析参数 Table 1 MS/MS parameters (m/z) (m/z) (V) (V) * * * * * 80 5 : *

4 10, : 结果与分析 3.1 电喷雾离子迁移谱条件优化 正负工作模式的选择 5,,,, 溶剂的选择 (60% 70% 80% 90% 100%) 5, 5, 2.2.3,, 2 3,, 5 ;, 5 90%,, 90% (V:V), 90%(V:V) 电喷雾离子迁移谱检测条件的优化,,, ESI-IMS 5 ( V) ( 4),, 2000 V, 5, 8300 V,, 8200 V (160~200 ) (160~200 ) (0.5~0.9 L/min) ( 5~7), ; ,, 0.9 L/min 5,, 2.2.3(1), 90% (V:V), 2 3 Table 2 表 2 不同浓度甲醇水溶液中各物质的响应值 Response values of 5 kinds of compounds at different concentrations of aqueous methanol solution 60% 70% 80% 90% 100% Table 3 表 3 不同浓度乙腈水溶液下各物质的响应值 Response values of 5 kinds of compounds at different concentrations of aqueous acetonitrile solution 60% 70% 80% 90% 100%

5 Table 4 表 4 不同源电压下各目标物响应值 Response values of 5 kinds of compounds at different source voltages 1600 V 1700 V 1800 V 1900 V 2000 V Table 5 表 5 不同进样口温度下各目标物响应值 Response values of 5 kinds of compounds at different inlet temperatures Table 6 表 6 不同迁移管温度下各目标物响应值 Response values of 5 kinds of compounds at different drift tube temperatures Table 7 表 7 不同迁移气体流量下各目标物响应值 Response values of 5 kinds of compounds at different drift gas flow rates 0.5 L/min 0.6 L/min 0.7 L/min 0.8 L/min 0.9 L/min

6 10, : Fig. 2 2 IMS chromatogram of diazepam and midazolam under the optimal conditions : 1: ; 2: Note: 1: diazepam; 2: midazolam Fig. 3 3 IMS chromatogram of barbital, nitrazepam and alprazolam under the optimal conditions : 1: ; 2: ; 3: Note: 1: barbital; 2: nitrazepam; 3: alprazolam 3.2 样品前处理条件优化, [9,12] [16],,,,, (60% 70% 80% 90% 100%), 5,, 5,,,,, ( min) 5, 20 min, 20 min, 8000 r/min 10 min, 90% 50, 0.22 μm, 3.3 基质效应 ESI-IMS,,,, 50, 5,

7 , 90%(V:V), (ME) (1), 8 空白基质匹配溶液响应值基质效应 ( ME ) 100% (1) 90% 乙腈匹配溶液响应值 85%~105% [17], 85%~105%, 2, %~153.76%, 85.48%~126.92% %~144.86%, ; 50.43%~69.35% 65.34%~96.62%,, 3.4 准曲线 检出限和定量限 2.2.2, IMS, C(mg/L) X, A Y,, r S/N=3 (LOD); S/N=10 (LOQ) 回收率及精密度实验 0.30 g( 0.01 g), 3, 2.2.1, 6, (RSD, n=6), %~113.8%, RSD 7.17% Table 8 表 8 改善睡眠类样品中 5 种目标化合物的基质效应 (n=3) Matrix effect of 5 kinds of compounds in improving sleep samples (n=3) 90% (mg/l) RSD(%) RSD(%) (%) Table 9 表 9 空白基质中 5 种物质的基质添加标准曲线 相关系数 检出限和定量限 Calibration curves, correlation coefficients, limits of detection and limits of quantitation of the matrix spiked with improving sleep drugs (mg/l) (mg/l) (mg/l) Y=0.0466X ~ Y=0.0575X ~ Y=0.0010X ~ Y=0.0021X ~ Y=0.0021X~ ~

8 10, : 表 10 5 种改善睡眠类化学药品的加标平均回收率和精密度 (n=6) Table 10 Recoveries and relative standard deviations (RSDs) of 5 kinds of compounds in improving sleep samples (n=6) (mg/g) (%) RSD(%) 3.6 方法验证 ,, 2.2.1, HPLC-MS/MS IMS, 11 表 11 LC-MS/MS 和 IMS 检测结果比对 Table 11 Comparison of the test results between LC-MS/MS and IMS (mg/g) (mg/g ) LC-MS/MS IMS 实际样品测定, 8, LC-MS/MS 5,, 4 结论, ESI-IMS 5,, 14 ms IMS, HPLC-MS/MS,, 5 参考文献 [1],,. UPLC-MS/MS [J]., 2010, 24(4): Chen AZ, Yang Z, Wang ZW. Detection of illegal mixture in traditional Chinese medicines preparation and health foods of Phenobarbital by UPLC-MS/MS [J]. Chin Pharm Aff, 2010, 24(4): [2],,,. GC-MS [J]., 2011, 1(2): Liu JJ, Xiao LH, Guan XY, et al. Determination of 14 sedative-hypnotic drugs illegal mixed in traditional Chinese medicines and health foods by GC-MS [J]. Chin J Pharm Anal, 2011, 1(2): [3],,,. - 9 [J]., 2013, 22(6): Tian L, Zhang JC, Chen R, et al. Detection of 9 kinds of illegally added chemical medicines in traditional Chinese medicine phenobarbital preparations and health foods by UPLC-MS/MS[J]. China Pharm., 2013, 22(6): [4],,, [J]., 2013, 31(7): Zhu L, Ruan LP, Liu HL, et al. Simultaneous determination of 23 sedative drugs in health foods by high performance liquid chromatography tandem mass spectrometry [J]. Chin J Chromatogr, 2013, 31(7): [5] () [EB/OL] Notice of the list of substances that may be illegally added in health food(the first batch)[eb/ol] html [6],,,. [J], 2012, (2): Jia Y, Zhang DH, Feng CM, et al. Research progress on hazard and detection technology of diazepam residue [J]. Qual SafAgro-Prod, 2012,

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