Determination of chlorantraniliprole, cyantraniliprole and its metabolites residues in litchi using QuEChERS and high performance liquid chromatography-tandem mass spectrometry
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摘要: 采用高效液相色谱-串联质谱(HPLC-MS/MS)技术，结合改良的QuEChERS法，建立了同时测定荔枝中氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38残留的分析方法。样品用乙腈提取，以N-丙基乙二胺(PSA)和十八烷基键合硅胶(C18)组合吸附剂净化，采用C18反相色谱柱分离，以V(甲酸水溶液) : V(乙腈)=15 : 85为流动相，采用电喷雾正离子扫描(ESI+)，多反应监测(MRM)模式，外标法定量。结果表明：3种目标化合物在0.005~1 mg/L范围内线性关系良好(r>0.99)；方法的定量限(LOQs)为0.0005~0.01 mg/kg；在0.0005~0.5 mg/kg添加水平下，氯虫苯甲酰胺、溴氰虫酰胺及代谢物在荔枝全果和果肉中的平均回收率为76%~98%，相对标准偏差(RSD)为2.5%~9.6%。应用该方法对广州市场中的20批次荔枝样品进行检测，结果显示，氯虫苯甲酰胺检出量为0.001~0.01 mg/kg，溴氰虫酰胺及其代谢物J9Z38未检出。该方法简便、高效、快速，可用于荔枝样品中双酰胺类农药的同时检测。Abstract: An analytical method was established for simultaneous determination of chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 in litchi by high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) combined with a modified QuEChERS procedure. The target compounds were extracted from the samples with acetonitrile and then cleaned up with primary secondary amine (PSA) and octadecylsilane (C18). The analytes were separated on a C18 column using V (formic acid aqueous solution) : V (acetonitrile)=15 : 85 as mobile phase and analyzed in multiple reaction monitoring (MRM) mode with positive electrospray ionization (ESI+). Quantitative analysis was performed by external standard method using matrix-matched calibration curves. The results showed that the linearity of the target compounds was good (r >0.99) in the range of 0.005-1 mg/L. The limits of quantification (LOQs) were 0.0005-0.01 mg/kg. The average recoveries of the three pesticides in litchi (whole fruit and pulp) ranged from 76% to 98% at the spiked levels of 0.0005-0.5 mg/kg, with the relative standard deviations of 2.5% to 9.6%. The method was applied for the detection of twenty commercial litchi samples in Guangzhou City. The results showed that the residues of chlorantraniliprole were 0.001-0.01 mg/kg, and neither cyantraniliprole nor its metabolite J9Z38 were found. The proposed method is convenient, effective and rapid, and is suitable for the simultaneous determination of chlorantraniliprole, cyantraniliprole and its metabolite in litchi.
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图 1 3种提取溶剂对氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38的提取效率 (n=3)
Figure 1. Extraction rate of three extraction solvents for chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 (n=3)
图 2 5种吸附剂组合对氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38的吸附效率
Figure 2. Absorption efficiency of five adsorbent combinations for chlorantraniliprole, cyantraniliprole and its metabolite J9Z38
图 3 不同色谱柱对氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38的分离效果 (0.05 mg/L)
A: Agilent Infinity Lab Poroshell-120 SB-C18 (75 mm × 2.1 mm, 2.7 μm); B: Agilent Zorbax Extend-C18 (150 mm × 4.6 mm, 5.0 μm); C: Agilent Zorbax SB-Aq-C18 (150 mm × 4.6 mm, 5.0 μm); D: 岛津 Shim-pack GIST-HP C18 (100 mm × 2.1 mm, 3.0 μm); E: 岛津 Shim-pack GIST-HP C18 (150 mm × 2.1 mm, 3.0 μm).
Figure 3. Separation efficiency of different chromatographic columns for chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 (0.05 mg/L)
图 4 不同流动相对氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38的洗脱效果 (0.1 mg/L)
Figure 4. Elution effect of different mobile phase composition for chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 (0.1 mg/L)
图 5 氯虫苯甲酰胺(A)、溴氰虫酰胺 (B) 及代谢物J9Z38 (C) 的二级离子质谱图 (0.1 mg/L)
Figure 5. Mass spectrometry of product ions of chlorantraniliprole (A), cyantraniliprole (B) and its metabolite J9Z38 (C) (0.1 mg/L)
表 1 氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38的质谱参数
Table 1. Mass spectra parameters of chlorantraniliprole, cyantraniliprole and its metabolite J9Z38
化合物
Compound母离子
Precursor ion,
m/z子离子
Product ion,
m/z碰撞能量
Collision energy/
V保留时间
Retention time/
min氯虫苯甲酰胺
chlorantraniliprole484 285.9* −12 5.5 453.35 −14 溴氰虫酰胺
cyantraniliprole475.3 285.95* −10 5.1 444.1 −19 代谢物J9Z38
Metabolite J9Z38457.1 188* −27 6.3 299 −35 注(Note)：*定量离子(Quantitative ion)。 
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表 2 吸附剂用量对氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38回收率的影响（n = 3）
Table 2. Effect of adsorbent dosage on the recovery of chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 (n = 3)
吸附剂组合
Adsorbent combination回收率 Recovery/% 代谢物J9Z38
Metabolite J9Z38溴氰虫酰胺
cyantraniliprole氯虫苯甲酰胺
chlorantraniliproleMWCNT 5 mg 4±1.1 10±0.8 14±0.9 MWCNT 10 mg 2±0.8 3±0.6 5±0.4 PSA 50 mg + 5 mg GCB 16±2.4 32±1.0 43±1.6 PSA 50 mg + 10 mg GCB 8±1.4 19±2.7 25±2.1 PSA 50 mg + 20 mg GCB 5±2.9 12±5.0 18±7.1 PSA 50 mg + C18 50 mg + 5 mg GCB 17±1.1 36±0.6 46±0.5 PSA 50 mg + C18 50 mg + 10 mg GCB 9±1.6 19±1.7 29±2.6 PSA 50 mg + C18 50 mg + 20 mg GCB 4±0.6 10±0.4 16±0.4 PSA 25 mg + C18 25 mg + 5 mg GCB 28±2.6 31±3.8 45±1.4 PSA 25 mg + C18 25 mg + 10 mg GCB 7±3.5 17±2.1 24±2.7 C18 25 mg + 5 mg GCB 23±3.9 33±1.2 44±1.8 C18 25 mg + 10 mg GCB 19±4.3 20±2.1 27±3.5 PSA 25 mg + 5 mg GCB 15±2.0 34±1.4 44±4.2 PSA 25 mg + 10 mg GCB 7±2.5 17±1.7 24±2.4 PSA 100 mg + C18 50 mg 95±2.5 81±1.9 81±3.2 PSA 50 mg + C18 100 mg 81±1.4 80±2.1 82±3.5 PSA 50 mg + C18 50 mg 87±1.6 84±1.1 91±2.7 注：表中数据为平均值 ± 标准差。Note: Data of the table are mean ± SD.
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表 3 不同色谱柱下氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38线性方程及基质效应
Table 3. Linear equation and matrix effects of chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 using different chromatographic columns
色谱柱类型
Chromatographic column化合物
Compound基质
Matrix线性方程
Linear equation相关系数
r基质效应
Matrix effect/%A 氯虫苯甲酰胺 chlorantraniliprole 溶剂 Solvent y=1.56609 × 107x + 113324 0.9997 — 果肉 Flesh y=1.36692 × 106x + 3949.26 0.9999 −91.3 全果 Whole fruit y=1.16033 × 106x + 7783.01 0.9997 −92.6 溴氰虫酰胺 cyantraniliprole 溶剂 Solvent y=5.25996 × 106x + 43758.2 0.9995 — 果肉 Flesh y=151406x + 1780.56 0.9997 −97.1 全果 Whole fruit y=148975x−1068.72 0.9979 −97.2 代谢物 J9Z38 Metabolite J9Z38 溶剂 Solvent y=1.03342 × 106x−6121.65 0.9978 — 果肉 Flesh y=577299x + 7626.68 0.9996 −44.1 全果 Whole fruit y=392552x−3513.27 0.9971 −62.1 B 氯虫苯甲酰胺 chlorantraniliprole 溶剂 Solvent y=1.15242 × 107x + 59844.7 0.9987 — 果肉 Flesh y=1.22073 × 107x + 160325 0.9984 5.9 全果 Whole fruit y=8.29595 × 106x + 144793 0.9992 −28.0 溴氰虫酰胺 cyantraniliprole 溶剂 Solvent y=2.93376 × 106x + 15813.9 0.9988 — 果肉 Flesh y=2.59718 × 106x + 35846.4 0.9981 −11.5 全果 Whole fruit y=1.57836 × 106x + 18578.6 0.9988 −46.2 代谢物 J9Z38 Metabolite J9Z38 溶剂 Solvent y=519147x + 2091.2 0.9994 — 果肉 Flesh y=539501x + 4769.44 0.9994 3.9 全果 Whole fruit y=176071x + 3453.94 0.9979 −66.1 C 氯虫苯甲酰胺 chlorantraniliprole 溶剂 Solvent y=1.22326 × 107x + 66452.3 0.9999 — 果肉 Flesh y=1.25802 × 107x + 270196 0.9963 2.8 全果 Whole fruit y=8.05065 × 106x + 121095 0.9999 −34.1 溴氰虫酰胺 cyantraniliprole 溶剂 Solvent y=4.30801 × 106x + 42111.2 0.9999 — 果肉 Flesh y=3.33573 × 106x + 74203.7 0.9959 −22.6 全果 Whole fruit y=2.32339 × 106x + 16241 0.9999 −46.1 代谢物 J9Z38 Metabolite J9Z38 溶剂 Solvent y=645920x + 916.731 0.9999 — 果肉 Flesh y=714497x + 16419.3 0.9959 10.6 全果 Whole fruit y=424131x + 3202.63 0.9999 −34.3 注 (Note): A: Agilent Infinity Lab Poroshell-120 SB-C18 (75 mm × 2.1 mm, 2.7 μm); B: Agilent Zorbax Extend-C18 (150 mm × 4.6 mm, 5.0 μm); C: Agilent Zorbax SB-Aq-C18 (150 mm × 4.6 mm, 5.0 μm).
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表 4 氯虫苯甲酰胺、溴氰虫酰胺及代谢物J9Z38在荔枝中的添加回收率、相对标准偏差及定量限 (n=5)
Table 4. Recoveries, RSDs, and LOQ of chlorantraniliprole, cyantraniliprole and its metabolite J9Z38 in litchi pulp and whole fruits (n=5)
化合物
Compound基质
Matrix添加水平
Spiked
level/
(mg/kg)回收率
Recoveries/
%相对
标准偏差
RSD/
%定量限
LOQ/
(mg/kg)氯虫苯甲酰胺
chlorantraniliprole果肉
Flesh0.0005 80 4.7 0.0005 0.01 87 8.6 0.1 96 5.8 0.5 97 5.4 全果
Whole fruit0.0005 78 8.1 0.0005 0.01 96 6.7 0.1 93 5.3 0.5 84 5.0 溴氰虫酰胺
cyantraniliprole果肉
Flesh0.005 78 5.1 0.005 0.01 88 5.6 0.1 98 3.9 0.5 91 3.5 全果
Whole fruit0.005 76 7.3 0.005 0.01 92 8.5 0.1 95 5.1 0.5 81 2.5 代谢物J9Z38
Metabolite J9Z38果肉
Flesh0.01 88 6.4 0.01 0.1 95 5.6 0.5 92 6.0 全果
Whole fruit0.01 95 9.6 0.01 0.1 92 8.1 0.5 86 6.8
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