The progressive decline in pollinator populations, particularly bees, has become a pressing concern in recent years. One of the key contributors to this phenomenon is the infestation by parasitic mites such as Varroa destructor, which compromises bee health and compels apiculturists and agricultural producers to intensify the use of acaricides. As a result, residues of the previous mites can be transferred into the food supply via edible bee-derived products and agricultural commodities. Ensuring consumer safety in relation to said toxic products requires the implementation of stricter pesticide regulation and improved apicultural management, which in turn depends on robust analytical monitoring of hive products.
In this context, a gas chromatography mass spectrometry-based method was developed and validated for the simultaneous determination of seven commonly applied pesticides in Spain, atrazine, chlorpyrifos, chlorfenvinphos, α-endosulfan, bromopropylate, coumaphos, and τ-fluvalinate, specifically in royal jelly. The extraction procedure employed a mixture of hexane and isopropanol, followed by a clean-up using primary and secondary amine (PSA) as a sorbent. Chromatographic separation was achieved on a DB-5MS capillary column using a temperature gradient program. The method was validated according to current EU guidelines, evaluating parameters such as selectivity, detection and quantification limits, linearity, matrix effects, accuracy, and precision. Analysis of nine royal jelly samples revealed the presence of chlorfenvinphos in eight samples and α-endosulfan in one, with six samples exceeding the maximum residue limits set by European legislation.
Acknowledgements:
This research was supported by the National Plan for Scientific and Technical Research and Innovation 2013–2016, National Institute for Agricultural and Food Research and Technology-INIA–FEDER (Spain), grant number RTA2017-00004-C02-02. Ana Jano thanks the Spanish Ministry of Education and Vocational Training for her collaboration grant, and Adrián Fuente-Ballesteros to the University of Valladolid for his PhD contract.
