【作者】 李丽；

【导师】 莫卫民； 吴俐勤；

【作者基本信息】 浙江工业大学 ， 农药学， 2014， 硕士

【摘要】 本文主要研究唑螨酯及其异构体在蜂蜜和蜂王浆中残留检测方法,建立了超高效液相色谱检测蜂蜜中唑螨酯残留量的检测方法、高效液相色谱-串联质谱法测定蜂蜜中唑螨酯残留量的检测方法、高效液相色谱-串联质谱法检测蜂王浆中唑螨酯残留量的检测方法。目前国内研究较多的是蔬菜水果中唑螨酯的残留,对蜂蜜和蜂王浆中唑螨酯残留及检测方法的文献匮乏,所以对唑螨酯在蜂蜜和蜂王浆中的残留检测方法进行开发和制定相应的检测标准显得很有必要和意义,对我国的农产品检测标准作出相应的补充,也为蜂蜜和蜂王浆的出口提供检测依据。1.蜂蜜中唑螨酯残留量超高效液相色谱检测方法的建立建立了超高效液相色谱法测定蜂蜜中唑螨酯残留量的检测方法。样品经提取净化、反相超高效液相色谱分离、二极管阵列检测器检测。检测波长为258nm,色谱柱Waters BEH C 18(1.7μm,2.1 X 50mm),流动相:乙腈+水(70+30,v+v),柱温40℃,流速0.2mL/min,进样量2 μL。通过优化分离条件,(E)-唑螨酯和(Z)-唑螨酯色谱行为良好,二者保留时间分别为2.4min和3.2min。在10～500 μg/L浓度范围内,线性关系良好,相关系数大于0.999。在50、100、200 μg/kg三个添加水平下(E)-唑螨酯和(Z)-唑螨酯的平均回收率分别在98.6～103.9%和101.8～103.4%之间,相对标准偏差在0.51～2.0%和1.8～3.5%之间,检测限分别为4.2μg/kg和4.3 μg/kg。适用性实验测得(E)-唑螨酯和(Z)-挫螨酯在50、100、200μg/kg添加浓度下的平均回收率分别在99.8%～103.0%和97.4%～101.4%之间,RSD分别在0.31%～3.9%和0.74%～4.7%之间。2.蜂蜜中唑螨酯残留量高效液相色谱-串联质谱检测方法的建立建立了蜂蜜中(E)-唑螨酯和(Z)-唑螨酯残留的高效液相色谱-串联质谱(LC-MS/MS)检测方法,样品经过提取净化,液相色谱分离,三重四级杆串联质谱分析,在多反应监测(MRM)模式下进行特征母离子和子离子的信号采集,根据保留时间、母离子和子离子进行定性分析,以离子丰度比最大的m/z366离子作为定量离子。在0.1～100μg/L线性范围内,峰面积与添加浓度的线性关系良好,在5、10、20μg/kg添加水平下,唑螨酯的平均回收率在85.8%～101.8%之间,相对标准偏差在2.4%～5.6%之间,(E)-唑螨酯和(Z)-唑螨酯最低检测限均为0.01μg/kg。在实际样品的测定中,20个蜂蜜样品有2个样品检出唑螨酷残留。3.蜂王浆中唑螨酯残留量高效液相色谱-串联质谱检测方法的建立建立了蜂王浆中(E)-唑螨酯和(Z)-唑螨酯残留的高效液相色谱-串联质谱(LC-MS/MS)检测方法,样品经过正交实验选出最优的提取液种类、固相萃取柱种类、淋洗液种类、洗脱剂种类等前处理条件。(E)-唑螨酯和(Z)-唑螨酷在5～100μg/L浓度内线性良好,相关系数大于0.999,通过讨论优化基质最佳稀释比,确定最佳的基质稀释倍数。在5、10、50μg/kg3个添加水平下回收率范围为86.6%～101.1%,相对标准偏差范围为3.5～7.1%,精密度实验结果测定日内5次实验数值变异系数小于10.8%,日间测定数值变异系数小于10.3%,最低检测限达到0.1μg/kg。在实际样品检测中,检测60份蜂王浆样品,每个样品测定两次,2份蜂王浆样品有(E)-唑螨酯残留出现,最高残留量达到20μg/kg。对蜂王浆中唑螨酯进行膳食暴露评估,结果表明,唑螨酷急性风险值%ARfD均小于100,在安全范围内,其中男性%ARfD值在0.0072～0.0356之间,女性%ARfD值在0.0058～0.0395之间;蜂王浆中唑螨酯,男性的慢性风险值%ADI在0.0030～0.0150之间,女性的慢性风险值%ADI在0.0024～0.0167之间,慢性风险值%ADI均小于100。无论男性还是女性,随着年龄的增长,其急性风险值%ARfD和慢性风险值%ADI越来越大。更多还原

【Abstract】 The objective of this paper was researching a new method for detection,confirmation and quantification of fenpyroximate and its isomer residues in honey and royal jelly.Methods were developed and validated for the determination of fenpyroximate in honey by Ultra performance liquid chromatography(UPLC)and in honey and royal jelly by liquid chromatography-tandem mass spectrometry(LC-MS/MS),respectively.Methods for the determination of fenpyroximate in fruits and vegetables have been in successfully reported.However,research papers concerning the determination of fenpyroximate in honey and royal jelly are very limited.The results of this work fill the blank of the the standards for the agro-products in our country and provide a scientific basis for the validation of the export honey.1.A simple and rapid method of determination of fenpyroximate residue in honey by UPLC was developed.The analytes was extracted,and cleaned-up by solid-phase extraction step.The chromatographic separation of analytes was performed on a BEH C1 8(2.1×50mm,1.7μm Waters)reversed-phase column using an acetone-water(70:30,v/v)as flow rate 0.2mL/min,injection volume is 2μL,the column temperature was set on 40℃,and set ultraviolet detection at 258nm wavelength.The(E)-fenpyroximate and(Z)-fenpyroximate chromatographic behavior is good through optimizing the separation conditions,the retention time were 2.4 min and 3.2 min.The linear ranges was from0.01～0.5 mg/L and the correlation coefficients(r~2)were all greater than 0.999.The recoveries for(E)-fenpyroximate and(Z)-fenpyroximate at three spiked levels(0.05、0.1、0.2mg/kg)were in the range of 98.6～103.9%and97.4～103.4%with relative standard deviations(RSDs,n=5)of 0.51～2.0%and 1.5～3.7%,the respective limits of detection was4.2μg/kg and 4.3μg/kg.Employment and suitability test measured(E)-fenpyroximate and(Z)-fenpyroximate add concentrations at 0.05,0.1,0.2 mg/kg respectively the average recovery between 99.8%～103.0%and 97.4%～101.4%,RSD between 0.31%～3.9%and 0.74%～4.7%respectively.2.An analytical method for the determination of fenpyroximate residue by LC-MS/MS in honey was developed.In the procedure the samples were extracted and purificated,then separated by liquid chromatographic and tandem mass spectrometry analysis.Mass spectral acquisition was applied with multiple reaction monitoring of two diagnostic transition reactions.The qualitative analysis was based on the retention time,the precursor ion and two productions,and the quantitation was carried out with intension of the characteristic ion m/z366.The linear ranges were from0.1～100μg/L for fenpyroximate and the correlation coefficients(r~2)were all greater than 0.999.The average recoveries for fenpyroximate ranged was 85.8%～113.2%with relative standard deviations(RSDs,n=5)of 1.2-4.3%,the detection limits was O.01μg/kg,the fenpyroximate residues were detected in two honey samples among 20 samples.3.A high performance liquid chromatography tandem mass spectrometric(HPLC-MS/MS)method was developed for the determination of fenpyroximate in royal jelly.Select the optimal extracting solution,solid phase extraction column and the elution by orthogonal experiment.The linear range was from5～100μg/L for fenpyroximate and the correlation coefficients(r~2)were all greater than 0.999.In replicate sets of royal jelly samples spiked with the drug concentration of 5,10,and50μg/kg,the average recoveries for fenpyroximate ranged were 88.5%～96.6%with relative standard deviations(RSDs,n=5)of 4.5-8.1%,and the relative standard deviations(RSDs)were less than 10.8%for intra-day and 10.3%for inter-day determinations.The detection limits was 0.1μg/kg.The dietary exposure assessment result of fenpyroximate in royal jelly show that the risk of acute dietary exposure value(%ARfD)and chronic dietary risk value(%ADI)were less than 100.The man and woman acute dietary exposure value(%ARfD)were 0.0072～0.0356 and 0.0058～0.0395,respectively.The man and woman chronic dietary risk value(%ADI)were 0.0030～0.0150 and 0.00240.0167,respectively.As the growth of the age,the risk of%ARfD and%ADI are higher and higher.更多还原