Validation of an off line solid phase extraction liquid chromatography-tandem mass spectrometry method for the determination of systemic insecticide residues in honey and pollen samples collected in apiaries from NW Spain

The use of pesticides to protect crops against plagues and insects is one of the most important ways to assure agricultural quality and productivity. However, bad application practices may cause the contamination of different environmental compartments and animal species, as a consequence of migration or accumulation of those compounds. Fipronil, imidacloprid and thiametoxam are systemic or systemic-like insecticides widely used in maize crops. Their heavy action in the nervous system of target insects also means a high toxicity to nontarget pollinator insects such as honey bees which can get in touch with them through pollen and nectar during foraging activities. These insecticides have even been suspected to cause a significant decrease of honeybee colonies that has been observed in many countries since the past decade. Since September 1 st 2008, the European Commission set new MRLs in food and feed of plant and animal origin. The pesticides included in this study have MRLs in honey and pollen between 10 and 50 ng g(-1). In the present work, an analytical method was developed with the aim of determining residues of fipronil and some of its metabolites (fipronil sulfone, fipronil sulfide, fipronil desulfinyl and fipronil carboxamide), thiamethoxam and imidacloprid in honey and pollen samples. The extraction optimization was performed using a Doehlert experimental design by studying two factors, the mixture and the ratio of solvents used. Prior to the extraction procedure, raw hive samples containing honey, pollen and wax were centrifuged at 4000 rpm. The upper solid material was removed, and 1 g of the lower phase was mixed with 3 mL of the optimized mixture of methanol/water (10/90). The extract was passed through a florisil cartridge and the target compounds were eluted with methanol and analysed by LC-MS/MS in selective reaction monitoring (SRM) mode. The method was validated according to the guidelines included in the SANCO/10684/2009 document and the ISO 11843 standard for the following parameters: decision limit (CC alpha), detection capability (CC beta), recovery, repeatability and reproducibility at 0.5, 1 and 1.5 folds the MRLs. Ion suppression/enhancement effects into the ion source were also assessed. The CC beta values were included between 0.83 and 4.83 ng g(-1), well below the current MRLs. The validated method was applied to the determination of the target pesticides in 91 samples collected in colonies from 73 apiaries of NW Spain (two sampling campaigns during 2008). None of the target insecticides were detected among all the collected samples. (C) 2010 Elsevier BM. All rights reserved.