QuEChERS-LC-MS/MS法测定樱桃中63种农药残留量
于晓慧1*，李俏2，魏爱云2
1山东金特安全科技有限公司，山东 济南 250000；2方圆标志检验检测（山东）有限公司，山东 济南250100
摘要:建立QuEChERS结合了液相色谱-串联质谱法同时测定樱桃中63种农药残留的分析方法。样品经过乙腈提取，采用 SpeedTest-QuEChERS 净化包进行净化，用LC-MS/MS MRM正负离子模式测定，外标法定量。63种农药在1.0~200 μg/L范围内线性关系良好，相关系数(r)均大于0.996，方法定量限为0.12～16.0 μg/kg，63种农药在20、50和100 μg/kg 添加水平的回收率76.9%~106.7%之间，相对标准偏差为0.76%~4.52%（n=6）。该方法前处理过程简单，检测结果准确稳定，适用于大批量的樱桃中农药残留的检测。
关键词：高效液相色谱-串联质谱；QuEChERS；樱桃；农药残留
Determination of 63 kinds of pesticide residues in cherry using QuEChERS-LC-MS/MS
Yu Xiaohui1* ,LI Qiao2,Wei Aiyun2
(1Shandong Giant Safety Technology Co.,Ltd,Jinan 250000, Shandong
2 China Quality Mark Certification(Shan dong)Co.,Ltd, Jinan 250100 ,Shandong)
Abstract:To establish a method for the simultaneous determination of 63 pesticide residues in chrrry by QuEChERS combined with liquid chromatography-tandem mass spectrometry. The samples were extracted with acetonitrile and purified by SpeedTest-QuEChERS, then analyzed by HPLC-MS/MS with MRM under positive ion mode of electrospray ionization .The linear relationship of 63 pesticides was good in the range of 1.0～200 μg/L, the correlation coefficients were more than 0.996, and the limit of quantification were 0.12～16.0 μg/kg. The recoveries of 63 pesticides at 20, 50 and 100 μg/kg levels ranged from 76.9% to 106.7%, and the relative standard deviation was 0.76%～4.52% (n=6). This method has positive signifificance for market access and monitoring of cherry.
Key words: liquid chromatography-tandem mass spectormetry;QuEChERS;cherry; pesticide residues
引 言
近几年来，随着休闲农业发展，种植樱桃的农户数量增加及种植面积单位扩大，随之，当前市面流通买卖的樱桃质量也随之下降，况且樱桃特别容易发生病害，农药的不合理使用，给农药残留带来超标风险。现在对于农业质量的要求越来越严格，对水果中农药残留的检测工作也越来越重视。目前现有的农药残留分析包括样品前处理和仪器数据检测两部分构成，农药的残留分析是一项对复杂混合物中痕量组分进行分析的手段。目前常见的样品前处理新技术有加速溶剂萃取法[1-3]、微波萃取法[4-5]、固相微萃取[6-7]、分子印迹萃取技术[8-9]、QuEChERS法[11-12]等。农药残留检测常用的分析技术有HPLC法[13]、GC[14]、LC-MS/MS[15-16]、GC-MS法[15]。本研究采用QuEChERS的前处理技术，前处理参照QuEChERS前处理技术的分散固相萃取法，使用山东美正生物科技有限公司的SpeedTest-QuEChERS净化包，对样品进行净化后上机操作，该前处理操作简单便捷，省时间，经济廉价，准确度高，结合HPLC-MS/M的高选择性、高灵敏度和强抗干扰能力等特点，建立QuEChERS-HPLC-MS/MS快速测定樱桃中63种农药残留的分析方法。该方法操作简便、灵敏度高、重现性好、分析速度快，适用于樱桃中农药多残留的同时检测，为樱桃农药残留的风险监测提供了技术保证。
QuEChERS-LC-MS/MS法测定樱桃中63种农药残留量
于晓慧1*，李俏2，魏爱云2
1山东金特安全科技有限公司，山东 济南 250000；2方圆标志检验检测（山东）有限公司，山东 济南250100
摘要:建立QuEChERS结合了液相色谱-串联质谱法同时测定樱桃中63种农药残留的分析方法。样品经过乙腈提取，采用 SpeedTest-QuEChERS 净化包进行净化，用LC-MS/MS MRM正负离子模式测定，外标法定量。63种农药在1.0~200 μg/L范围内线性关系良好，相关系数(r)均大于0.996，方法定量限为0.12～16.0 μg/kg，63种农药在20、50和100 μg/kg 添加水平的回收率76.9%~106.7%之间，相对标准偏差为0.76%~4.52%（n=6）。该方法前处理过程简单，检测结果准确稳定，适用于大批量的樱桃中农药残留的检测。
关键词：高效液相色谱-串联质谱；QuEChERS；樱桃；农药残留
Determination of 63 kinds of pesticide residues in cherry using QuEChERS-LC-MS/MS
Yu Xiaohui1* ,LI Qiao2,Wei Aiyun2
(1Shandong Giant Safety Technology Co.,Ltd,Jinan 250000, Shandong
2 China Quality Mark Certification(Shan dong)Co.,Ltd, Jinan 250100 ,Shandong)
Abstract:To establish a method for the simultaneous determination of 63 pesticide residues in chrrry by QuEChERS combined with liquid chromatography-tandem mass spectrometry. The samples were extracted with acetonitrile and purified by SpeedTest-QuEChERS, then analyzed by HPLC-MS/MS with MRM under positive ion mode of electrospray ionization .The linear relationship of 63 pesticides was good in the range of 1.0～200 μg/L, the correlation coefficients were more than 0.996, and the limit of quantification were 0.12～16.0 μg/kg. The recoveries of 63 pesticides at 20, 50 and 100 μg/kg levels ranged from 76.9% to 106.7%, and the relative standard deviation was 0.76%～4.52% (n=6). This method has positive signifificance for market access and monitoring of cherry.
Key words: liquid chromatography-tandem mass spectormetry;QuEChERS;cherry; pesticide residues
引 言
近几年来，随着休闲农业发展，种植樱桃的农户数量增加及种植面积单位扩大，随之，当前市面流通买卖的樱桃质量也随之下降，况且樱桃特别容易发生病害，农药的不合理使用，给农药残留带来超标风险。现在对于农业质量的要求越来越严格，对水果中农药残留的检测工作也越来越重视。目前现有的农药残留分析包括样品前处理和仪器数据检测两部分构成，农药的残留分析是一项对复杂混合物中痕量组分进行分析的手段。目前常见的样品前处理新技术有加速溶剂萃取法[1-3]、微波萃取法[4-5]、固相微萃取[6-7]、分子印迹萃取技术[8-9]、QuEChERS法[11-12]等。农药残留检测常用的分析技术有HPLC法[13]、GC[14]、LC-MS/MS[15-16]、GC-MS法[15]。本研究采用QuEChERS的前处理技术，前处理参照QuEChERS前处理技术的分散固相萃取法，使用山东美正生物科技有限公司的SpeedTest-QuEChERS净化包，对样品进行净化后上机操作，该前处理操作简单便捷，省时间，经济廉价，准确度高，结合HPLC-MS/M的高选择性、高灵敏度和强抗干扰能力等特点，建立QuEChERS-HPLC-MS/MS快速测定樱桃中63种农药残留的分析方法。该方法操作简便、灵敏度高、重现性好、分析速度快，适用于樱桃中农药多残留的同时检测，为樱桃农药残留的风险监测提供了技术保证。
1实验部分
1.1仪器与试剂
仪器设备：SHIMADZU LC-MS/MS 8045三重四级杆液质联用仪，上海舜宇恒平科学仪器有限公司 JA31002型电子天平，博科BK-180J型超声波清洗器，北京优晟联合科技有限公司UMV-1型多管旋涡混合器，湖南凯达科学仪器有限公司KL04A型离心机。
试剂和耗材：山东美正生物科技有限公司SpeedTest-QuEChERS净化包，甲醇、乙腈（色谱纯、fisher），一级水（屈臣氏），甲酸（色谱级、fisher)，樱桃（超市购买），63种农药残留标准品100μg/mL（坛墨质检标准物质中心），50mL聚乙烯离心管。
1.2标准溶液配制
混合标准中间液配制：分别精确移取63种农药的标准物质，用乙腈稀释成浓度为 2 μg/mL 的混合标准中间液，于-18 ℃冰箱保存，待用(有效期为3个月)。
基质混合标准工作溶液：用空白基质溶液将混合标准中间液逐级稀释为 1.0、5、10、20、30、50、100、200 μg/L的基质混合标准工作溶液。
1.3样品的制备及提取
检测样品制备：取樱桃样品食用部分，捣碎，最后捣成匀浆，保存条件为零下20℃到零下16℃，待用。在进行前处理称取样品时，需要对已经冷冻好的样品进行完全解冻，待樱桃样品放置常温后，使樱桃样品混匀后取样。
检测样品提取：准确称取樱桃样品20g（精确至0.01）于聚乙烯的50mL带盖离心管中，精确加入20mL乙腈试剂，拧好离心管盖，在旋涡机上，涡旋3min，在离心管中加入5g氯化钠，拧好离心管盖，涡旋混匀，放置超声机中，超声提取，提取时间为30min，超声完成后，以8000r/min的转速，离心5min，用一次性塑料吸管吸取上层提取液5-10mL，至于SpeedTest-QuEChERS管中，混匀，4000r/min离心3min，取上层清液，过0.22μm有机滤膜至进样小瓶中，待测。
1.4检测方法
1.4.1色谱条件
色谱柱：Shim-pack GIST C1-AQ HP (100mm×2.1 mm，1.9μm)；
仪器柱温：40℃；
流动相流速：0.25mL/min；
仪器进样量：2μL；
流动相：A=2mmol/L甲酸铵0.01%甲酸水；B=2mmol/L甲酸铵甲醇；
梯度洗脱条件：0.01-0.4 min：6%(2mmol/L甲酸铵甲醇)；0.41-1.3 min：6~45%(甲酸铵甲醇)；1.31-3.0min：45~85%(甲酸铵甲醇)； 3.01-6.0min：86~96%(甲酸铵甲醇)；6.01-8.0min：96%(甲酸铵甲醇)；8.01-9.00min：6%(甲酸铵甲醇)。
1.4.2质谱条件
离子源：电喷雾离子源；
扫描方式：正负离子同时扫描；
采集方式：MRM；
离子源接口电压：+4.5 kV；-3.5 kV；
碰撞气：氩气；离子传输管管温度：200℃；接口温度：250℃；
加热模块温度：350℃。
2结果与分析
2.1标准曲线绘制、回归方程、相关系数与方法检出限
用1.4所述的检测方法测定基质混合标准工作溶液（现用现配），进行线性回归处理，测定的63种农药在1.0~200 μg/L线性范围内展现出很好的相关关系，综合63种物质相关系数均在0.9961~0.9999之间，仪器所测得到63种物质方法定量限，定量限低出国家标准的限量值，63种农药的质谱参数见表 2-1、2-2。
表2-1 63种农药的色谱及质谱条件
Table 2-1 The chromatographic and mass spectrometry conditions of 63 pesticides
NO. | Compound | Retention time（min） | Qualitative ion | Qusntitative ion | Serial number | Compound | Retention time（min） | Qualitative ion | Qusntitative ion | |
1 | acetamiprid | 4.176 | 223.10>56.10 | 223.10>126.15 | 33 | myclobutanil | 5.073 | 289.10>125.05 | 289.10>70.05 | |
2 | dimethomoeph | 4.906 | 388.20>165.00 | 388.20>301.00 | 34 | propiconazole | 5.388 | 342.05>205.10 | 342.05>159.10 | |
3 | emancctin benzoate | 4.413 | 886.60>126.30 | 886.60>158.15 | 35 | forchlorfenuron | 4.640 | 248.10>93.10 | 248.10>129.10 | |
4 | phridaben | 6.758 | 365.20>309.10 | 365.20>147.10 | 36 | fenbuconazole | 5.192 | 248.10>93.10 | 336.90>70.00 | |
5 | buprofezin | 3.941 | 292.00>181.10 | 292.00>211.10 | 37 | carbaryl | 4.680 | 202.00>127.20 | 202.00>145.20 | |
6 | carbofuran | 4.611 | 222.20>123.20 | 222.20>165.25 | 38 | fenarimol | 5.074 | 331.00>259.10 | 331.00>268.10 | |
7 | imidacloprid | 4.139 | 255.80>175.10 | 255.80>209.00 | 39 | propamocarb | 0.780 | 189.20>144.05 | 189.20>102.05 | |
8 | carbendazim | 0.790 | 192.10>132.20 | 192.10>160.20 | 40 | tolfenpyrad | 5.961 | 384.10>154.05 | 384.10>197.10 | |
9 | phoxim | 5.637 | 299.00>129.10 | 299.00>77.10 | 41 | buprofezin | 5.902 | 306.10>116.10 | 306.10>201.10 | |
10 | aldicab | 4.396 | 207.80>89.20 | 207.80>116.25 | 42 | indoxacarb | 5.666 | 528.10>292.95 | 528.10>249.10 | |
11 | omethoate | 0.890 | 213.80>108.95 | 213.80>125.00 | 43 | pymetrozin | 0.781 | 218.10>78.10 | 218.10>105.05 | |
12 | methamidophos | 0.850 | 141.80>125.00 | 141.80>93.95 | 44 | carbosulfan | 7.558 | 381.20>160.10 | 381.20>118.10 | |
13 | 3-hydroxycabofuran | 4.101 | 237.50>135.05 | 237.50>181.10 | 45 | chlordimeform | 0.880 | 197.20>125.10 | 197.20>117.10 | |
14 | aldicarb sulfone | 3.609 | 239.70>86.15 | 239.70>223.20 | 46 | chlorantraniliprole | 4.880 | 484.00>285.85 | 484.00>452.90 | |
15 | aldicarb sulfoxide | 0.910 | 206.65>132.15 | 206.65>89.20 | 47 | isoprocarb | 4.821 | 194.10>137.10 | 194.10>95.00 | |
16 | demeton | 4.851 | 259.10>61.00 | 259.10>89.00 | 48 | penconzaole | 5.314 | 284.10>159.00 | 284.10>70.00 | |
17 | trichlorphon | 4.024 | 256.90>220.85 | 256.90>108.95 | 49 | propoxur | 4.605 | 210.10>168.10 | 210.10>111.10 | |
18 | thiacloprid | 4.319 | 253.00>99.00 | 253.00>126.05 | 50 | metolcarb | 4.507 | 166.10>107.10 | 166.10>109.10 | |
19 | clothianidin | 4.083 | 250.00>132.00 | 250.00>169.10 | 51 | triadimenol | 4.940 | 296.10>99.15 | 296.10>70.05 | |
20 | trifloxystrobin | 5.738 | 409.10>145.00 | 409.10>186.05 | 52 | clofentezine | 5.650 | 303.00>102.05 | 303.00>138.05 | |
21 | cadusafos | 5.680 | 271.10>97.00 | 271.10>159.00 | 53 | imazalil | 4.015 | 297.00>201.00 | 297.00>159.00 | |
22 | diethofencarb | 5.030 | 268.10>180.10 | 268.10>226.10 | 54 | epoxiconazole | 5.099 | 330.10>141.10 | 330.10>121.20 | |
23 | mctalaxyl | 4.735 | 280.10>192.20 | 280.10>220.20 | 55 | tebuconazole | 5.375 | 326.10>159.00 | 326.10>70.00 | |
24 | mefenoxam | 4.735 | 280.10>248.10 | 280.10>220.15 | 56 | flusilazole | 5.178 | 316.10>165.10 | 316.10>247.10 | |
25 | isazofos | 5.405 | 314.10>162.00 | 314.10>120.00 | 57 | Thiophate-methyl | 4.509 | 343.05>311.05 | 343.05>151.15 | |
26 | pyraclostrobin | 5.563 | 388.10>163.05 | 388.10>194.05 | 58 | thiabendazole | 0.865 | 202.00>131.10 | 202.00>175.05 | |
27 | dimethoate | 4.165 | 230.00>125.00 | 230.00>198.95 | 59 | chlorobenzuron | 5.298 | 307.00>126.15 | 307.00>154.15 | |
28 | tebuconzole | 5.189 | 308.10>125.00 | 308.10>70.10 | 60 | diflubenzuron | 5.199 | 309.10>156.15 | 309.10>289.05 | |
29 | tebufenozide | 5.347 | 353.20>297.10 | 353.20>133.10 | 61 | novaluron | 5.653 | 491.00>305.10 | 491.00>471.00 | |
30 | fenpyroximate | 6.352 | 422.20>138.10 | 422.20>366.10 | 62 | flonicamid | 3.998 | 228.00>145.90 | 228.00>80.85 | |
31 | kresoxim-methyl | 5.033 | 404.10>329.00 | 404.10>372.05 | 63 | flufenoxuron | 6.087 | 487.10>467.10 | 487.10>156.10 | |
32 | cyromazine | 0.699 | 167.10>60.10 | 167.10>85.10 |
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表2-2 63种农药的线性关系和定量限
Table 2-2 Linear relationship and quantitative limits of 63 pesticide
NO. | Compound | Linear range( μg/L) | Linear equation | r2 | LOQ(μg/kg) |
1 | acetamiprid | 1.0~200μg/L | Y=52469x+12456.23 | 0.9991 | 0.8 |
2 | dimethomoeph | 1.0~200μg/L | Y=78951.14x+9841.98 | 0.9981 | 0.6 |
3 | emancctin benzoate | 1.0~200μg/L | Y=4897.23x-756.1 | 0.9962 | 1.6 |
4 | phridaben | 1.0~200μg/L | Y=6431.462x+2141.3 | 0.9993 | 1.5 |
5 | buprofezin | 1.0~200μg/L | Y=12658.32x-469.2 | 0.9999 | 2.7 |
6 | carbofuran | 1.0~200μg/L | Y=32459.90x-3479.6 | 0.9968 | 0.8 |
7 | imidacloprid | 1.0~200μg/L | Y=15698.12x-5612.21 | 0.9986 | 2.8 |
8 | carbendazim | 1.0~200μg/L | Y=35694.02x+2341.66 | 0.9992 | 0.19 |
9 | phoxim | 1.0~200μg/L | Y=66512.92x+8941.15 | 0.9993 | 16.0 |
10 | aldicab | 1.0~200μg/L | Y=65127.5x-5641.01 | 0.9987 | 9.4 |
11 | omethoate | 1.0~200μg/L | Y=456.02x-81.2 | 0.9969 | 0.6 |
12 | methamidophos | 1.0~200μg/L | Y=32015.01x-2458 | 0.9963 | 1.2 |
13 | 3-hydroxycabofuran | 1.0~200μg/L | Y=13478.4x+890.1 | 0.9986 | 8.3 |
14 | aldicarb sulfone | 1.0~200μg/L | Y=4405.56x-213.09 | 0.9996 | 7.8 |
15 | aldicarb sulfoxide | 1.0~200μg/L | Y=5698.14x-512.21 | 0.9990 | 1.8 |
16 | demeton | 1.0~200μg/L | Y=328.02x+341.39 | 0.9997 | 1.2 |
17 | trichlorphon | 1.0~200μg/L | Y=16912.9x+5612.15 | 09998 | 14.3 |
18 | thiacloprid | 1.0~200μg/L | Y=3204.5x-563.45 | 0.9982 | 2.1 |
19 | clothianidin | 1.0~200μg/L | Y=26401.1x-8108.9 | 0.9986 | 2.4 |
20 | trifloxystrobin | 1.0~200μg/L | Y=61232x+3612.45 | 0.9976 | 0.23 |
21 | cadusafos | 1.0~200μg/L | Y=2341.61x+1012.01 | 0.9969 | 0.21 |
22 | diethofencarb | 1.0~200μg/L | Y=12347.56x-2612.99 | 0.9963 | 1.3 |
23 | mctalaxyl | 1.0~200μg/L | Y=11341.52x-2341.15 | 0.9982 | 0.3 |
24 | mefenoxam | 1.0~200μg/L | Y=21256.4x+341.39 | 0.9993 | 0.3 |
25 | isazofos | 1.0~200μg/L | Y=13569.45x+8912.07 | 0.9978 | 0.7 |
26 | pyraclostrobin | 1.0~200μg/L | Y=4103.65x+234.78 | 0.9969 | 0.12 |
27 | dimethoate | 1.0~200μg/L | Y=23451.21x-712.01 | 0.9961 | 4.0 |
28 | tebuconzole | 1.0~200μg/L | Y=2516.83x+21.22 | 0.9992 | 0.60 |
29 | tebufenozide | 1.0~200μg/L | Y=23451.17x-25.69 | 0.9996 | 0.20 |
30 | fenpyroximate | 1.0~200μg/L | Y=5621.41x-123.56 | 0.9992 | 0.36 |
31 | kresoxim-methyl | 1.0~200μg/L | Y=68901.23x-456.12 | 0.9976 | 0.6 |
32 | cyromazine | 1.0~200μg/L | Y=784.12x+51.09 | 0.9978 | 1.3 |
33 | myclobutanil | 1.0~200μg/L | Y=6512.02x+841.19 | 0.9986 | 2.4 |
34 | propiconazole | 1.0~200μg/L | Y=512.5x-61.09 | 0.9998 | 1.5 |
35 | forchlorfenuron | 1.0~200μg/L | Y=41256.02x-8021.2 | 0.9990 | 2.1 |
37 | fenbuconazole | 1.0~200μg/L | Y=2915.01x-438.22 | 0.9981 | 2.7 |
38 | carbaryl | 1.0~200μg/L | Y=2388.4x+1560.71 | 0.9978 | 2.8 |
39 | fenarimol | 1.0~200μg/L | Y=14805.56x-1213.29 | 0.9992 | 2.4 |
40 | propamocarb | 1.0~200μg/L | Y=6908.14x-1293.21 | 0.9995 | 3.3 |
41 | buprofezin | 1.0~200μg/L | Y=71262.0x | 0.9963 | 0.14 |
表2-2 续
Table 2-2 Continued
NO. | Compound | Linear range( μg/L) | Linear equation | r2 | LOQ(μg/kg) |
42 | indoxacarb | 1.0~200μg/L | Y=1241.61x+212.88 | 0.9966 | 8.9 |
43 | pymetrozin | 1.0~200μg/L | Y=4789.01x-1369.99 | 0.9980 | 1.2 |
44 | carbosulfan | 1.0~200μg/L | Y=1141.72x+7341.15 | 0.9968 | 1.4 |
45 | chlordimeform | 1.0~200μg/L | Y=6156.4x-481.49 | 0.9969 | 1.3 |
46 | chlorantraniliprole | 1.0~200μg/L | Y=7066.65x+9012.37 | 0.9972 | 2.0 |
47 | isoprocarb | 1.0~200μg/L | Y=2103.65x-56.39 | 0.9983 | 2.2 |
48 | penconzaole | 1.0~200μg/L | Y=24051.21x+632.01 | 09977 | 5.4 |
49 | propoxur | 1.0~200μg/L | Y=5613.09x+800.36 | 0.9996 | 3.1 |
50 | metolcarb | 1.0~200μg/L | Y=40051.17x-2135.37 | 0.9981 | 6.2 |
51 | triadimenol | 1.0~200μg/L | Y=6021.41x+1623.76 | 0.9976 | 2.3 |
52 | clofentezine | 1.0~200μg/L | Y=90101.23x-5956.32 | 0.9969 | 0.42 |
53 | imazalil | 1.0~200μg/L | Y=1425.50x-1203.39 | 0.9963 | 1.4 |
54 | epoxiconazole | 1.0~200μg/L | Y=6877.84x-412.21 | 0.9972 | 3.1 |
55 | tebuconazole | 1.0~200μg/L | Y=1348.02x+1341.79 | 0.9983 | 2.3 |
56 | flusilazole | 1.0~200μg/L | Y=2913.9x+682.63 | 09977 | 1.6 |
57 | Thiophate-methyl | 1.0~200μg/L | Y=2903.8x-163.15 | 0.9996 | 0.25 |
58 | thiabendazole | 1.0~200μg/L | Y=35408.1x-5108.22 | 0.9981 | 0.54 |
59 | chlorobenzuron | 1.0~200μg/L | Y=14222x+6012.36 | 0.9976 | 14.3 |
60 | diflubenzuron | 1.0~200μg/L | Y=4041.61x+1012.01 | 0.9964 | 10.3 |
61 | novaluron | 1.0~200μg/L | Y=3407.23x-262.09 | 0.9976 | 12.0 |
62 | flonicamid | 1.0~200μg/L | Y=3910.25x-2041.23 | 0.9979 | 3.2 |
63 | flufenoxuron | 1.0~200μg/L | Y=4613.33x+612.31 | 0.9973 | 5.1 |
2.2回收率和精密度
本试验采用在空白樱桃样品（不含有63种目标物质）中加标的方法进行此方法的精密度和回收率的验证。准确称取18个空白樱桃样品进行加标回收试验，以3水平浓度进行添加试验，测试样品含量为 20、50、100μg/kg的，每个水平做6组加标试验，按照上述的1.3和1.4进行前处理处理和仪器分析测定，试验结果见表2-3。
结果表明，在 20、50、100μg/kg的3水平浓度下，樱桃中63种化合物的回收率分别76.9%~106.7%之间，63种物质的相对标准偏差在0.76%~4.52%之间，回收率达到较高且稳定的效果。
表2-3 63种农药的平均回收率和相对标准偏差（n=6）
Table 2-3 Average recovery and relative standard deviation of 63 pesticides (n=6)
NO. | Compound | Spiked 20μg/kg | Spiked 50μg/kg | Spiked 100μg/kg | |||
Recovery/% | RSD/% | Recovery/% | RSD/% | Recovery/% | RSD/% | ||
1 | acetamiprid | 95.1 | 2.31 | 97.8 | 1.71 | 92.1 | 3.62 |
2 | dimethomoeph | 87.2 | 1.69 | 85.9 | 2.03 | 90.3 | 2.83 |
3 | Emancctin benzoate | 86.6 | 3.22 | 90.6 | 1.06 | 92.6 | 4.30 |
4 | phridaben | 87.7 | 3.46 | 85.7 | 2.71 | 90.8 | 2.59 |
5 | buprofezin | 92.0 | 2.21 | 89.2 | 1.36 | 93.0 | 3.04 |
表2-3 续
Table 2-3 Continued
NO. | Compound | Spiked 20μg/kg | Spiked 50μg/kg | Spiked 100μg/kg | |||
Recovery/% | RSD/% | Recovery/% | RSD/% | Recovery/% | RSD/% | ||
6 | carbofuran | 90.6 | 1.79 | 90.9 | 3.54 | 88.5 | 3.53 |
7 | imidacloprid | 87.0 | 1.34 | 94.5 | 1.66 | 92.1 | 1.78 |
8 | carbendazim | 78.6 | 2.65 | 80.6 | 2.15 | 80.1 | 3.06 |
9 | phoxim | 77.9 | 2.72 | 79.1 | 2.63 | 82.5 | 2.78 |
10 | aldicab | 86.9 | 3.01 | 88.0 | 3.13 | 88.9 | 3.56 |
11 | omethoate | 90.0 | 1.62 | 91.9 | 1.77 | 89.1 | 2.06 |
12 | methamidophos | 82.1 | 4.06 | 84.6 | 3.46 | 86.3 | 2.89 |
13 | 3-hydroxycabofuran | 88.9 | 3.29 | 94.6 | 3.06 | 95.7 | 3.08 |
14 | aldicarb sulfone | 87.3 | 2.13 | 85.6 | 2.76 | 89.4 | 1.96 |
15 | aldicarb sulfoxide | 101.6 | 0.76 | 99.0 | 1.51 | 103.2 | 2.03 |
16 | demeton | 102.3 | 1.52 | 98.7 | 1.79 | 104.6 | 2.81 |
17 | trichlorphon | 89.6 | 1.36 | 85.4 | 2.06 | 87.6 | 3.06 |
18 | thiacloprid | 85.7 | 2.06 | 89.0 | 2.78 | 90.6 | 4.52 |
19 | clothianidin | 88.0 | 2.41 | 92.3 | 2.64 | 93.7 | 4.31 |
20 | trifloxystrobin | 96.3 | 2.36 | 94.5 | 2.64 | 98.1 | 4.01 |
21 | cadusafos | 101.2 | 1.89 | 104.6 | 1.90 | 97.3 | 0.99 |
22 | diethofencarb | 104.3 | 1.54 | 102.9 | 1.64 | 105.6 | 1.34 |
23 | mctalaxyl | 82.0 | 1.63 | 85.3 | 1.77 | 84.9 | 2.06 |
24 | mefenoxam | 89.9 | 3.09 | 87.4 | 3.52 | 91.2 | 2.31 |
25 | isazofos | 85.0 | 4.47 | 83.9 | 4.07 | 86.5 | 2.46 |
26 | pyraclostrobin | 82.6 | 2.06 | 84.7 | 2.16 | 86.6 | 3.52 |
27 | dimethoate | 90.3 | 4.03 | 88.7 | 3.77 | 90.5 | 3.64 |
28 | tebuconzole | 100.7 | 3.21 | 96.4 | 2.62 | 102.6 | 3.41 |
29 | tebufenozide | 103.9 | 0.99 | 99.9 | 1.21 | 104.0 | 2.33 |
30 | fenpyroximate | 106.7 | 1.42 | 104.5 | 1.33 | 103.2 | 2.06 |
31 | kresoxim-methyl | 105.1 | 1.87 | 106.4 | 1.99 | 103.9 | 2.52 |
32 | cyromazine | 95.3 | 2.05 | 96.4 | 2.33 | 96.8 | 1.67 |
33 | myclobutanil | 94.0 | 2.31 | 93.7 | 2.44 | 95.6 | 4.52 |
34 | propiconazole | 93.5 | 2.71 | 94.1 | 2.06 | 93.7 | 3.06 |
35 | forchlorfenuron | 76.9 | 1.60 | 78.6 | 1.74 | 79.3 | 2.45 |
36 | fenbuconazole | 79.0 | 1.77 | 80.1 | 1.89 | 80.5 | 3.14 |
37 | carbaryl | 87.9 | 0.86 | 88.6 | 1.52 | 88.3 | 2.98 |
38 | fenarimol | 88.2 | 2.54 | 87.5 | 2.06 | 88.6 | 2.81 |
39 | propamocarb | 86.2 | 3.36 | 86.7 | 1.68 | 87.2 | 3.06 |
40 | fenpyroximate | 105.4 | 2.40 | 106.1 | 2.34 | 104.6 | 2.41 |
41 | buprofezin | 77.0 | 3.02 | 79.3 | 4.02 | 78.6 | 3.37 |
42 | indoxacarb | 88.6 | 2.65 | 88.9 | 2.15 | 90.1 | 3.06 |
43 | pymetrozin | 87.9 | 1.72 | 89.1 | 2.63 | 90.5 | 2.03 |
表2-3 续
Table 2-3 Continued
NO. | Compound | Spiked 20μg/kg | Spiked 50μg/kg | Spiked 100μg/kg | |||
Recovery/% | RSD/% | Recovery/% | RSD/% | Recovery/% | RSD/% | ||
44 | carbosulfan | 76.9 | 2.01 | 78.0 | 2.13 | 78.9 | 2.56 |
45 | chlordimeform | 90.2 | 1.72 | 89.0 | 3.02 | 92.3 | 3.06 |
46 | chlorantraniliprole | 84.4 | 3.62 | 85.7 | 3.77 | 86.9 | 1.89 |
47 | isoprocarb | 85.2 | 4.50 | 84.9 | 3.16 | 86.3 | 2.08 |
48 | penconzaole | 77.3 | 3.13 | 78.6 | 3.76 | 79.6 | 1.96 |
49 | propoxur | 92.0 | 1.76 | 93.5 | 1.53 | 94.6 | 3.08 |
50 | metolcarb | 99.6 | 1.69 | 97.7 | 1.79 | 96.3 | 2.81 |
51 | triadimenol | 83.6 | 1.32 | 84.4 | 2.05 | 85.6 | 3.06 |
52 | clofentezine | 84.7 | 2.06 | 86.0 | 2.78 | 86.9 | 4.06 |
53 | imazalil | 88.0 | 2.41 | 92.3 | 2.64 | 93.7 | 4.31 |
54 | epoxiconazole | 106.2 | 3.56 | 96.0 | 1.90 | 106.0 | 2.99 |
55 | tebuconazole | 103.6 | 3.69 | 95.0 | 3.37 | 96.3 | 4.03 |
56 | flusilazole | 82.0 | 1.63 | 85.3 | 1.77 | 84.9 | 2.06 |
57 | Thiophate-methyl | 89.6 | 3.09 | 87.6 | 4.52 | 88.9 | 4.31 |
58 | thiabendazole | 85.7 | 3.47 | 84.9 | 4.07 | 83.5 | 2.46 |
59 | chlorobenzuron | 80.6 | 3.06 | 84.7 | 2.16 | 83.6 | 2.52 |
60 | diflubenzuron | 80.3 | 3.03 | 80.7 | 3.17 | 80.5 | 3.64 |
61 | novaluron | 77.9 | 0.86 | 78.6 | 1.52 | 78.3 | 2.98 |
62 | flonicamid | 78.6 | 3.54 | 77.1 | 3.06 | 78.9 | 3.81 |
63 | flufenoxuron | 84.2 | 4.36 | 83.7 | 4.40 | 84.2 | 4.06 |
2.3实际样品测定
随机购买附近市场或者超市的樱桃样品10份，按照该试验的方法进行63种农药物质残留的测定，检测得到的结果均符合农药的残留限量规定。本次检测樱桃样品的63个农残参数中有32个农药品种残留限量不作判定。31种物质有残留限量判定，其中腈菌唑、克百威的有值检出率最高，有两批樱桃样品接近限量值规定；多菌灵在10份樱桃样品中检出率为40%；乐果、甲胺磷和甲氨基阿维菌素苯甲酸盐这三种物质检出率为30%；甲霜灵、氧乐果两种化合物检出率为20%；吡唑嘧菌酯在樱桃样品中的检出率为10%；10份樱桃样品均为合格样品，其他农药参数均未测出数值。结果详见表2-4。
表2-4 10批樱桃样品的检验结果及63种农药的最大残留限量MRL
Table 2-4 Test results of 10 batches of cherry samples and MRL of 63 pesticides maximum residue limit
Analyte | Detection rate% | MRL/mg/kg | Above rate% | Analyte | Detection rate% | MRL/mg/kg | Above rate% |
acetamiprid | 0 | 2.0 | - | myclobutanil | 50 | 3.0 | 0 |
dimethomoeph | 20 | - | - | propiconazole | 0 | - | - |
Emancctin benzoate | 30 | - | - | forchlorfenuron | 0 | - | - |
phridaben | 0 | - | - | fenbuconazole | 0 | 1.0 | 0 |
buprofezin | 0 | 1.0 | 0 | carbaryl | 0 | - | - |
carbofuran | 50 | 0.02 | 0 | fenarimol | 0 | - | - |
imidacloprid | propamocarb | 0 | - | - |
表2-4续
Table 2-4 Continued
Analyte | Detection rate% | MRL/mg/kg | Above rate% | Analyte | Detection rate% | MRL/mg/kg | Above rate% | ||||
carbendazim | 40 | 0.5 | 0 | fenpyroximate | 0 | - | - | ||||
phoxim | 0 | - | - | buprofezin | 0 | 2.0 | 0 | ||||
aldicab | 10 | 0.5 | 0 | indoxacarb | 0 | 1.0 | 0 | ||||
omethoate | 20 | 0.02 | - | pymetrozin | 0 | - | - | ||||
methamidophos | 30 | 0.05 | 0 | carbosulfan | 0 | - | - | ||||
3-hydroxycabofuran | 0 | 0.02 | 0 | chlordimeform | 0 | 0.01 | 0 | ||||
aldicarb sulfone | chlorantraniliprole | 0 | 1.0 | 0 | |||||||
aldicarb sulfoxide | isoprocarb | 0 | - | - | |||||||
demeton | 0 | 0.02 | 0 | penconzaole | 0 | - | - | ||||
trichlorphon | 0 | 0.2 | 0 | propoxur | 0 | - | - | ||||
thiacloprid | 0 | 0.5 | 0 | metolcarb | 0 | - | - | ||||
clothianidin | 0 | 0.2 | 0 | triadimenol | 0 | - | - | ||||
trifloxystrobin | 0 | 3.0 | 0 | clofentezine | 0 | 0.5 | 0 | ||||
cadusafos | 0 | 0.02 | 0 | imazalil | 0 | - | - | ||||
diethofencarb | 0 | - | - | epoxiconazole | 0 | - | - | ||||
mctalaxyl | 20 | - | - | tebuconazole | 0 | - | - | ||||
mefenoxam | flusilazole | 0 | - | - | |||||||
isazofos | 0 | 0.01 | 0 | Thiophate-methyl | 0 | - | - | ||||
pyraclostrobin | 10 | 3.0 | 0 | thiabendazole | 0 | - | - | ||||
dimethoate | 30 | 2.0 | 0 | chlorobenzuron | 0 | - | - | ||||
tebuconzole | 0 | 4.0 | 0 | diflubenzuron | 0 | - | - | ||||
tebufenozide | 0 | - | - | novaluron | 0 | 7.0 | - | ||||
fenpyroximate | 0 | 2.0 | 0 | flonicamid | 0 | - | - | ||||
kresoxim-methyl | 20 | 2.0 | 0 | flufenoxuron | 0 | - | - | ||||
cyromazine | 0 | - | - |
|
|
|
| ||||
3结论
本研究采用 QuEChERS 前处理结合液相色谱串联质谱法同时检测樱桃中63种农药残留，前处理参照QuEChERS前处理技术的分散固相萃取法，使用山东美正生物科技有限公司的SpeedTest-QuEChERS净化包，63种农药在 1.0~200 μg/L范围内线性关系良好，相关系数(r)均大于0.996，方法的定量限为 0.12~16.0 μg/kg。20、50 和 100 μg/kg三个添加水平的回收率在76.9%~106.7%之间，相对标准偏差为0.76%~4.52%（n=6），本研究建立的方法具有前处理快速、目标物提取效率高、实验过程中使用到有机溶剂用量少、检测结果准确、重复性好、实验耗材少、成本低的优点，可以很好地应用于大批量樱桃样品中多种农药残留分析检测。
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