Journal of Chromatography A, 1209 (2008) 17–21 Contents lists available at ScienceDirect Journal of Chromatography A journal homepage: www.elsevier.com/locate/chroma New multiresidue method using solid-phase extraction and gas chromatography–micro-electron-capture detection for pesticide residues analysis in royal jelly Emmanouel Karazafiris a,b , Urania Menkissoglu-Spiroudi a,∗ , Andreas Thrasyvoulou b a Laboratory of Pesticide Science, Faculty of Agriculture, Aristotle University of Thessaloniki, GR-54124 Thesaloniki, Greece b Laboratory of Apiculture-Sericulture, Faculty of Agriculture, Aristotle University of Thessaloniki, GR-54124 Thesaloniki, Greece article info Article history: Received 21 July 2008 Received in revised form 31 August 2008 Accepted 4 September 2008 Available online 11 September 2008 Keywords: Solid-phase extraction GC ECD Royal jelly Pesticides Residues abstract Royal jelly, one of the most important bee products, can be contaminated with pesticide and/or antibiotic residues resulting from treatments applied either inside beehives or in the agricultural environment. A new multiresidue method was developed and validated for analysis of nine pesticides in royal jelly. Solid-phase extraction RP-C 18 cartridges were used for sample purification and isolation of analytes. Final solution was analyzed with GC and micro-electron-capture detection. Four synthetic acaricides used by beekeepers (bromopropylate, coumaphos, malathion and -fluvalinate), and moreover one pyrethroid, two organochlorine, and two organophosphate insecticides were tested. Linearity is demonstrated for the range of 0.0025–1 mg kg -1 , with correlation coefficients ranging from 0.99991 to 0.99846, depending on the analyte. Overall recovery rates from royal jelly blank samples spiked at five fortification levels ranged from 80.8% (lindane) to 91.3% (ethion), well above the range defined by the SANCO/10232/2006 and EC/675/2002 documents. The limit of quantification was <0.003–0.005 mg kg -1 depending on the analyte, and the reporting level of the method, defined as the lowest recovery level, was 0.005 mg kg -1 . © 2008 Elsevier B.V. All rights reserved. 1. Introduction Royal jelly is one of the most important beehive products because of its pharmaceutical and nutritive properties for humans. Furthermore, due to its recently increased market demand, which reflected directly to its price, royal jelly has become a product of high economic importance for beekeepers. Since small quantities are produced and consumed worldwide the establishment of qual- ity control criteria for royal jelly is rather insufficient. Although it is considered that royal jelly is not a widely consumed prod- uct, there is a concern, however, that those who do take it as a food supplement are likely to consume it every day and they are therefore high-level consumers. Beehive products such as honey, wax, propolis and royal jelly, could be contaminated with residues resulting from treatments applied either inside the beehives or on the plants where bees collect nectar or pollen and several studies report on pesticide residues in honey and wax [1–3]. A review pub- lished in 2006 [1] has highlighted all the sources of bee products contamination, concluding that the main contamination sources are originated from apicultural practices, while honey and wax ∗ Corresponding author. Tel.: +30 2310998835; fax: +30 2310998835. E-mail address: rmenkis@agro.auth.gr (U. Menkissoglu-Spiroudi). are the most liable to contamination. To our knowledge, regard- ing the royal jelly contamination with residues, only a few studies exist, which indeed report on the presence of antibiotic residues in royal jelly [1,4,5]. One study deals with the presence of the aca- ricides coumaphos and -fluvalinate in royal jelly [6], but none on pesticide residues resulting from the surrounding agricultural envi- ronment. The presence of the acari Varroa destructor Anderson and Trueman in bee colonies, initiates obligatory the use of acaricides by beekeepers, which if not properly applied, cause residues in bee products [1–3,7]. Moreover, the use of a variety of pesticides for agricultural purposes, many of them with systemic action, in the environment of bees, increases the possibility of royal jelly contam- ination with residues. European Union (EU) legislation requires that all active substances used in veterinary medicines for food produc- ing animals must be assessed so that a maximum residue limit can be set. Since MRLs for royal jelly have not been established either at EU or National level, the presence of any (acaricide, pesticide and antibiotic) residues could result in a non-compliant product [8]. Consequently, proper analytical methods developed and validated according to analytical quality control (AQC) performance criteria established by EU [8,9], are necessary for control purposes. A large number of analytical methods report on pesticide residues determination in honey and beeswax [10–15], but there is not any multiresidue method published in the literature espe- 0021-9673/$ – see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.chroma.2008.09.018