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1 5 4 Vol. 5 No Journal of Food Safety and Quality Apr 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., ; 2., ; 3. ( ), ; 4., ) 2 摘要 : 目的 - GA 方法 QuEChERS, 1%, C 18 -, C 18, Agilent AJS ESI (MRM), 结果 mg/l, GA3 (S/N=10) mg/kg, mg/kg mg/kg 3, GA % 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 , China; 2. Inspection and Quarantine Technology Center, Xiamen Entry-Exit Inspection and Quarantine Bureau, Xiamen , China; 3. Agilent Tchnologies (China) Limited, Beijing , China; 4. Inspection and Quarantine Technology Center, Ningbo Entry-Exit Inspection and Quarantine Bureau, Ningbo , 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 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 :,,,, [email protected] *Corresponding author: XU Dum-Ming, Professor, Senior Engineer, Technical center of Xiamen Entry-Exit Inspection and Quarantine Bureau, No.2165, Haicang Jiangang Road, Xiamen , China. [email protected]

2 mg/l, with the correlation coefficients higher than for the 3 pesticides. The limit of quantification (LOQ) were mg/kg for GA3, 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 ), , 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 mg ), 1 min r/min 5 min, 0.22 μm,

3 4, : 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 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(%) 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 C 10 H 9 NO * * * 1- C 12 H 10 O * Fig.1 MS/MS spectrum of 3 pesticides

4 Fig.2 TIC of 3 pesticides, - -, 3.4 空白样品筛查 Fig % 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,, 线性范围 基质效应和定量限,,,,, : 1):, = / 100%; 2):,, [20],,, GA3 ; IAA NAA, GA3

5 4, : Fig.4 TIC of background and ultrapure water a. ; b. IAA NAA,, 3,,,, GA3 NAA IAA μg/l (LOQ, S/N=10) 准确度和精密度 10.0 g( 0.01 g), 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% mg/kg TIC MRM 5 4 结论 - 3 QUChERS, 1%, C 18 MgSO 4, HPLC-MS/MS GA3 (S/N=10) mg/kg, IAA NAA mg/kg 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

6 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 Y=13.51X GA3 Y=20.49 X Y=2.43X Y=9.44X Y=183.15X Y=148.68X IAA Y=81.33X Y=55.90X Y=131.43X Y=183.47X Y=182.99X NAA Y=190.40X Y=121.98X Y=186.69X 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) GA IAA NAA

7 4, : 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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