Kiljanek Tomasz, Niewiadowska Alicja, Semeniuk Stanisław, Gaweł Marta, Borzęcka Milena, Posyniak Andrzej
Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland.
Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland.
J Chromatogr A. 2016 Feb 26;1435:100-14. doi: 10.1016/j.chroma.2016.01.045. Epub 2016 Jan 22.
A method for the determination of 200 pesticides and pesticide metabolites in honeybee samples has been developed and validated. Almost 98% of compounds included in this method are approved to use within European Union, as active substances of plant protection products or veterinary medicinal products used by beekeepers to control mites Varroa destructor in hives. Many significant metabolites, like metabolites of imidacloprid, thiacloprid, fipronil, methiocarb and amitraz, are also possible to detect. The sample preparation was based on the buffered QuEChERS method. Samples of bees were extracted with acetonitrile containing 1% acetic acid and then subjected to clean-up by dispersive solid phase extraction (dSPE) using a new Z-Sep+ sorbent and PSA. The majority of pesticides, including neonicotionoids and their metabolites, were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) but some of pesticides, especially pyrethroid insecticides, were analyzed by gas chromatography tandem mass spectrometry (GC-MS/MS). The procedure was validated according to the Guidance document SANCO/12571/2013 at four concentration levels: 1, 5, 10 and 100 ng/g bees and verified in the international proficiency test. The analysis of bee samples spiked at the limit of quantification (LOQ) showed about 98% mean recovery value (trueness) and 97% of analytes showed recovery in the required range of 70-120% and RSDr (precision) below 20%. Linearity and matrix effects were also established. The LOQs of pesticides were in the range of 1-100 ng/g. The developed method allows determination of insecticides at concentrations of 10 ng/g or less, except abamectin and tebufenozide. LOQ values are lower than the median lethal doses LD50 for bees. The method was used to investigate more than 70 honeybee poisoning incidents. Data about detected pesticides and their metabolites are included.
已开发并验证了一种用于测定蜜蜂样本中200种农药及其代谢物的方法。该方法涵盖的化合物中，近98%在欧盟被批准用作植物保护产品的活性物质，或养蜂人用于控制蜂箱中狄斯瓦螨的兽药产品。许多重要的代谢物，如吡虫啉、噻虫啉、氟虫腈、甲硫威和双甲脒的代谢物，也能够被检测到。样品制备基于缓冲QuEChERS方法。蜜蜂样本用含1%乙酸的乙腈萃取，然后使用新型Z-Sep+吸附剂和PSA通过分散固相萃取（dSPE）进行净化。大多数农药，包括新烟碱类及其代谢物，通过液相色谱串联质谱（LC-MS/MS）分析，但部分农药，尤其是拟除虫菊酯类杀虫剂，通过气相色谱串联质谱（GC-MS/MS）分析。该方法按照指导文件SANCO/12571/2013在四个浓度水平（1、5、10和100 ng/g蜜蜂）进行了验证，并在国际能力验证测试中得到了验证。对添加了定量限（LOQ）的蜜蜂样本进行分析，结果显示平均回收率（准确性）约为98%，97%的分析物回收率在70 - 120%的要求范围内，相对标准偏差（RSDr，精密度）低于20%。同时还确定了线性和基质效应。农药的LOQ范围为1 - 100 ng/g。所开发的方法能够测定浓度为10 ng/g或更低的杀虫剂，但阿维菌素和虫酰肼除外。LOQ值低于蜜蜂的半数致死剂量LD50。该方法用于调查70多起蜜蜂中毒事件。文中包含了检测到的农药及其代谢物的数据。
