7 10 Vol. 7 No. 10 2016 10 Journal of Food Safety and Quality Oct., 2016-5 叶雅真 1 1,, 骆和东 2*, 张璨雯 3 1,, 黄三发 2, 蔡伟鹏 (1., 361012; 2., 361021; 3., 361021) 2 摘要 : 目的 - (electrospray ionization-ion mobility spectrometry, ESI-IMS) 5 方法, 90% ESI-IMS,, 结果 5, 0.99 0.20 0.50 2.00 8.00 8.00 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 361012, China; 2. Xiamen Center for Disease Control and Prevention, Xiamen 361021, China; 3. College of Biology Engineering, Jimei University, Xiamen 361021, 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 :,, E-mail: luohedong@126.com *Corresponding author: LUO He-Dong, Chief Technician, Xiamen Institute for Food and Drug Quality Control, Xiamen 361012, China. E-mail: luohedong@126.com
10, : - 5 4051 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. 1 1 5 Structures of 5 kinds of compounds
4052 7 2 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 2.2.1 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, 2.2.2 标准溶液和基质匹配标准曲线的配制 (1) :, 2 ml,, 441.2 mg/l 442.0 mg/l,,, 2, 90%(V:V), : 2, 2.2.1,,, 80 mg/l 20 mg/l,,, (2) :, 2 ml,, 517.2 mg/l, 412.8 mg/l, 462.8 mg/l,,, 3, 90% (V:V), : 3, 2.2.1,,, 30 mg/l,,, 2.2.3 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) 285.0 193.0* 135 35 154.0 135 25 326.0 291.0* 145 25 249.0 145 40 282.0 236.0* 125 25 180.0 125 40 316.0 269.9* 140 25 213.9 140 40 183.1 140.0* 80 5 : *
10, : - 5 4053 3 结果与分析 3.1 电喷雾离子迁移谱条件优化 3.1.1 正负工作模式的选择 5,,,, 3.1.2 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) 3.1.3 电喷雾离子迁移谱检测条件的优化,,, ESI-IMS 5 (1600 1700 1800 1900 2000 V) ( 4),, 2000 V, 5, 8300 V,, 8200 V (160~200 ) (160~200 ) (0.5~0.9 L/min) ( 5~7), 180 200 ; 190 180,, 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% 0.180 0.137 0.083 0.220 0.254 0.166 0.073 0.094 0.197 0.300 0.069 0.091 0.090 0.090 0.086 0.101 0.095 0.076 0.082 0.118 0.055 0.066 0.034 0.044 0.065 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% 0.183 0.128 0.112 0.248 0.120 0.162 0.145 0.140 0.346 0.244 0.082 0.070 0.071 0.090 0.022 0.095 0.104 0.099 0.115 0.076 0.070 0.061 0.063 0.065 0.038
4054 7 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 0.179 0.191 0.219 0.214 0.218 0.352 0.336 0.346 0.415 0.424 0.013 0.045 0.060 0.059 0.086 0.022 0.067 0.078 0.085 0.113 0.006 0.037 0.027 0.037 0.058 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 160 170 180 190 200 0.271 0.241 0.257 0.246 0.257 0.467 0.474 0.581 0.566 0.571 0.040 0.059 0.028 0.062 0.033 0.087 0.104 0.079 0.102 0.088 0.052 0.053 0.046 0.063 0.053 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 160 170 180 190 200 0.250 0.196 0.200 0.198 0.251 0.373 0.374 0.391 0.416 0.486 0.063 0.047 0.055 0.042 0.035 0.079 0.083 0.088 0.082 0.088 0.045 0.038 0.044 0.044 0.045 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 0.222 0.238 0.241 0.226 0.251 0.452 0.460 0.444 0.419 0.486 0.055 0.057 0.058 0.036 0.058 0.087 0.095 0.090 0.097 0.102 0.059 0.069 0.055 0.058 0.068
10, : - 5 4055 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,,,,, (5 10 15 20 30 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,
4056 7 2.2.1, 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, 116.84%~153.76%, 85.48%~126.92% 107.55%~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) 9 3.5 回收率及精密度实验 0.30 g( 0.01 g), 3, 2.2.1, 6, (RSD, n=6), 10 78.7%~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(%) (%) 2.0 0.099 8.81 0.116 1.32 116.84 10.0 0.204 0.49 0.314 4.28 153.76 1.0 0.104 8.33 0.132 7.6 126.92 4.0 0.411 1.89 0.351 0.87 85.48 4.0 0.078 6.31 0.039 3.88 50.43 10.0 0.149 3.03 0.103 7.82 69.35 8.0 0.069 5.40 0.067 8.26 96.62 20.0 0.151 5.00 0.099 3.10 65.34 8.0 0.053 5.80 0.057 6.33 107.55 20.0 0.107 8.86 0.155 1.12 144.86 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+0.0393 0.40~10.00 0.9986 0.20 0.40 Y=0.0575X+0.0466 1.00~10.00 0.9975 0.50 1.00 Y=0.0010X+0.0445 5.00~30.00 0.9914 2.00 5.00 Y=0.0021X+0.0006 10.00~30.00 0.9988 8.00 10.00 Y=0.0021X~0.0026 10.00~30.00 0.9980 8.00 10.00
10, : - 5 4057 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 0.027 78.7 7.17 0.333 113.8 4.72 0.667 95.4 1.46 0.067 85.5 5.06 0.333 101.8 2.63 0.667 103.3 1.09 0.333 97.0 1.69 1.333 99.2 1.88 2.000 95.3 1.71 0.667 88.0 5.67 1.333 100.9 3.57 2.000 106.9 1.96 0.667 108.7 6.95 1.333 96.4 2.17 2.000 102.6 2.59,, 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 0.067 0.073 0.060 0.133 0.132 0.126 0.067 0.052 0.045 0.133 0.118 0.122 0.667 0.598 0.580 1.333 1.068 1.022 0.667 0.572 0.602 1.067 1.014 0.960 0.667 0.540 0.524 1.067 0.896 0.872 3.7 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): 383 391. 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): 383 391. [2],,,. GC-MS [J]., 2011, 1(2): 376 379. 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): 376 379. [3],,,. - 9 [J]., 2013, 22(6): 66 68. 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): 66 68. [4],,,. - 23 [J]., 2013, 31(7): 709 713. 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): 709 713. [5] () [EB/OL]. http://www.sda.gov.cn/ws01/cl0847/69914.html. 2012-03-16. Notice of the list of substances that may be illegally added in health food(the first batch)[eb/ol]. http://www.sda.gov.cn/ws01/cl0847/ 69914.html. 2012-03-16. [6],,,. [J], 2012, (2): 47 50 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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