5 4 Vol. 5 No. 4 2014 4 Journal of Food Safety and Quality Apr. 2014 - 黄何何 1,2, 徐敦明 2*, 周昱 2, 罗佳 1, 吕美玲 3, 陈树宾 4, 王连珠 (1., 350002; 2., 361026; 3. ( ), 100102; 4., 315000) 2 摘要 : 目的 - GA3-3- 1-3 方法 QuEChERS, 1%, C 18 -, C 18, Agilent AJS ESI (MRM), 结果 3 0.005 1.000 mg/l, GA3 (S/N=10) 0.010 mg/kg, -3-1- 0.005 mg/kg 0.020 0.050 0.100 mg/kg 3, GA3-3- 1-75.4% 109.8% 79.1% 109.3% 79.6% 112.9%, (n=6) 3.5% 17.4% 6.5% 15.8% 4.8% 13.9% 结论,, 3 关键词 : - ; ; Determination of several natural plant growth regulators in fruits by high performance liquid chromatography-tandem mass spectrometry HUANG He-He 1,2, XU Dun-Ming 2*, ZHOU Yu 2, LUO Jia 1, LV Mei-Ling 3, CHEN Shu-Bin 4, WANG Lian-Zhu 2 (1. Plant Protecet College, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2. Inspection and Quarantine Technology Center, Xiamen Entry-Exit Inspection and Quarantine Bureau, Xiamen 361026, China; 3. Agilent Tchnologies (China) Limited, Beijing 100102, China; 4. Inspection and Quarantine Technology Center, Ningbo Entry-Exit Inspection and Quarantine Bureau, Ningbo 315000, China) ABSTRACT: Objective To develop a method for the simultaneous detection of GA3, IAA and NAA in fruits by QuEChERS-HPLC-ESI-MS/MS. Methods The samples were extracted with acetonitrile with 1% acetic acid, purified using powder of magnesium sulfate and C 18. Three pesticides were separated on a C 18 column with the mobile phases of water and acetonitrile and finally detected with negative electrospray ionization in MRM mode. The matrix-matched external standard calibration curves were used for quantitative analysis. Results The linearities of GA3, IAA and NAA were in the concentration ranges of 0.005 1.000 基金项目 : (2011J01386) (2012C37074) Fund: Supported by Science and Technology Planning Project of Fujian Province (2011J01386) and Public Technology Applied Research Projects of Zhejiang Province (2012C37074) * 通讯作者 :,,,, E-mail: Xudm@xmciq.gov.cn *Corresponding author: XU Dum-Ming, Professor, Senior Engineer, Technical center of Xiamen Entry-Exit Inspection and Quarantine Bureau, No.2165, Haicang Jiangang Road, Xiamen 361026, China. E-mail: Xudm@xmciq.gov.cn
1134 5 mg/l, with the correlation coefficients higher than 0.9990 for the 3 pesticides. The limit of quantification (LOQ) were 0.010 mg/kg for GA3, 0.005 mg/kg for IAA and NAA. For all the samples, the mean spiked recoveries of GA3, IAA and NAA ranged from 75.4% 109.8%, 79.1% 109.3% and 79.6% 112.9%, respectively, and the relative standard deviations (, n=6) were in the range of 3.5% 17.4%, 6.5% 15.8% and 4.8% 13.9%, respectively. Conclusion The method is quick, easy, effective, sensitive and accurate, and it can meet the requirements for the determination of GA3, IAA and NAA pesticide residues in fruits. KEY WORDS: high performance liquid chromatography-tandem mass spectrometry(hplc-ms/ms); natural plant growth regulator; fruits 1 引言 1- (α-naphthaleneacetic acid; NAA) -3- (indole-3-acetic acid; IAA) (gibberellin A3; GA3), [1],, [2] NAA [3] ; IAA, [4] ; GA3,,, [5],,, NAA [6] ; IAA ; GA3, [7] 3,, NAA 0.05 mg/kg, NAA IAA GA3, (ELISA) [8-9] (GC) [10-11] (HPLC) [12-13] - (GC-MS) [14-15] - (LC-MS/MS) [16-19], ; -, ; -, QuEChERS, LC-MS/MS NAA IAA GA3 2 材料与方法 2.1 仪器与试剂 - : Agilent 1290, Agilent 6460 ( Agilent ), AJS ESI ; IKA T18 Basic ( IKA ); ( ); ; ; ; MilliQ (, Merck ); ( ) 1- (α-naphthaleneacetic acid; NAA) -3- (indole-3-acetic acid; IAA) (Gibberellin A3; GA3)( Dr. Ehrensterfer ); Agilent SampliQ (, Agilent ), 1- -3-, 100 mg/l 4,, 2.2 样品前处理 10.0 g( 0.01 g) 50 ml, 10 ml 1%, 2 min, (6 g 1.5 g ), 1 min, 4000 r/min 3 min, 2.0 ml ( 25 mg C 18 150 mg ), 1 min 16000 r/min 5 min, 0.22 μm,
4, : - 1135 2.3 LC-MS/MS 条件 : Agilent ZORBAX SB-C 18 (100 mm 2.1 mm, 1.8 μm) A:, B: ; 1; : 3 min; : 0.2 ml/min; : 40 ; : 2 μl (ESI),, (MRM) : 45 psi; : 500 V; : 325, 8 L min -1 ; : 300, 10 L min -1 ; : 3500 V 2 3 1 50 μg/l 2 3 结果与讨论 3.1 提取溶剂的选择 GA3 IAA NAA 1% [17] [18] [19],, 1%,, 1% 85%, 3,,, 1% -,,, 1% 3 表 1 3 种农药流动相梯度洗脱条件 Table 1 Gradient conditions for the analysis of 3 pesticides T(min) (ml/min) H20() Acetonitrile(%) 0 0.2 98 2 2 0.2 98 2 2.1 0.2 60 40 5 0.2 60 40 5.1 0.2 2 98 8 0.2 2 98 8.1 0.2 98 2 10 0.2 98 2 表 2 3 种农药质谱分析参数 Table 2 MS/MS parameters of 3 pesticides (m/z) (m/z) (ev) (V) C 19 H 22 O 6 345.3 C 10 H 9 NO 2 174.0 142.9* 29 221.3 24 130.1* 7 128.1 20 170 70 * 1- C 12 H 10 O 2 185.0 141.1* 20 50 1 3 Fig.1 MS/MS spectrum of 3 pesticides
1136 5 2 3 Fig.2 TIC of 3 pesticides, - -, 3.4 空白样品筛查 Fig. 3 3 1% Extraction efficiency with acetonitrile and acetonitrile with 1% acetic acid 3.2 净化条件优化 QuEChERS, N- (PSA)+MgSO 4 N- (PSA)+C 18 EC+MgSO 4 MgSO 4 +C 18, MgSO 4 +C 18, 80%,, GA3 NAA IAA, N- (PSA), 3.3 色谱条件优化 MRM, -5 mmol/l -0.1% -,,,,, MRM,, 4 3.5 线性范围 基质效应和定量限,,,,, : 1):, = / 100%; 2):,, [20],,, GA3 ; IAA NAA, GA3
4, : - 1137 4 Fig.4 TIC of background and ultrapure water a. ; b. IAA NAA,, 3,,,, GA3 NAA IAA 5 1000 μg/l (LOQ, S/N=10) 3 3.6 准确度和精密度 10.0 g( 0.01 g), 0.02 0.05 0.1 mg/kg 3 3, 1.2, HPLC-MS/MS 4,, GA3 75.4% 109.8%, () 3.5% 17.4%; IAA 79.1% 109.3%, () 6.5% 15.8%; NAA 79.6% 112.9%, () 4.8% 13.9% 3 0.05 mg/kg TIC MRM 5 4 结论 - 3 QUChERS, 1%, C 18 MgSO 4, HPLC-MS/MS GA3 (S/N=10) 0.010 mg/kg, IAA NAA 0.005 mg/kg 0.020 0.050 0.100 mg/kg 3, GA3 IAA NAA 75.4% 109.8% 79.1% 109.3% 79.6% 112.9%, 3.5% 17.4% 6.5% 15.8% 4.8% 13.9%,, GA3 IAA NAA
1138 5 Table 3 表 3 不同水果中 3 种农药的基质添加标准曲线 相关系数 基质效应和定量限 Calibration curves, correlation coefficients (r 2 ), matrix effects and LOQs of 3 pesticides in fruits r 2 LOQ(mg/kg) Y=15.71X-106.72 0.9991 0.101 0.010 Y=13.51X-97.03 0.9995 0.043 0.010 GA3 Y=20.49 X +108.10 0.9992 0.411 0.010 Y=2.43X-93.28 0.9997 0.032 0.010 Y=9.44X-20.82 0.9993 0.071 0.10 Y=183.15X-1283.45 0.9992 5.750 0.005 Y=148.68X-1084.33 0.9995 4.575 0.005 IAA Y=81.33X+782.75 0.9993 0.739 0.005 Y=55.90X+57.15 0.9993 0.416 0.005 Y=131.43X-38.10 0.9996 0.630 0.005 Y=183.47X+1279.02 0.9990 0.922 0.005 Y=182.99X-378.28 0.9999 1.299 0.005 NAA Y=190.40X +174.13 0.9993 0.669 0.005 Y=121.98X+58.96 0.9999 0.128 0.005 Y=186.69X-358.36 0.9995 0.668 0.005 Table 4 表 4 3 种农药的添加回收率及相对标准偏差 (n=6) Recoveries and precisions of 3 pesticides spiked in fruits (n=6) (mg/kg) 0.02 88.6 14.4 16.3 79.5 7.8 9.8 15.4 3.5 3.5 99.8 8.6 8.6 98.9 8.5 8.6 GA3 0.05 90.1 19.0 10.3 77.2 11.0 14.3 89.0 13.6 15.3 88.8 16.2 17.4 86.0 14.3 12.9 0.10 84.0 10.1 12.0 82.1 8.8 10.7 93.7 10.5 11.2 85.9 12.6 14.7 109.8 9.9 12.4 0.02 109.3 8.6 8.2 88.0 4.9 6.5 98.2 4.4 10.2 84.9 6.8 7.6 79.2 9.2 11.6 IAA 0.05 93.6 10.8 8.8 86.3 5.6 7.5 97.9 8.2 11.6 82.9 5.1 13.3 85.2 5.0 12.7 0.10 92.7 7.9 7.7 79.1 8.3 13.6 98.1 10.8 8.2 75.2 7.7 15.8 85.8 8.0 9.7 0.02 112.9 8.9 7.6 90.8 5.7 5.4 79.6 10.0 5.5 96.7 6.5 7.0 86.9 9.2 10.5 NAA 0.05 91.1 9.0 13.9 82.0 6.5 6.8 85.5 11.4 9.5 94.5 11.0 5.4 91.1 14.1 4.8 0.10 94.6 7.1 8.3 88.5 10.8 9.3 88.1 8.1 12.3 89.9 8.1 9.0 88.5 7.4 9.0
4, : - 1139 5 3 0.05 mg/kg TIC MRM Fig. 5 TIC and MRM chromatograms of 3 pesticides spiked in grape at 0.05 mg/kg
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