Eurasian Journal of Analytical Chemistry, 2018, 13(5), em38 ISSN:1306-3057 OPEN ACCESS Research Paper https://doi.org/10.29333/ejac/92537 © 2018 by the authors; licensee Modestum Ltd., UK. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/). christogeorge84@yahoo.gr sarakaio@yahoo.gr samanidu@chem.auth.gr (*Correspondence) Cyromazine Determination in Poultry based Animal Feedstuffs by HPLC/DAD using QuEChERS Methodology Christos Christogiorgos 1 , Ioannis Sarakatsianos 2 , Victoria Samanidou 1* 1 Laboratory of Analytical Chemistry, University of Thessaloniki, GR 54124 Thessaloniki, GREECE 2 Dept. of Chemical Engineering, Aristotle University of Thessaloniki, 54 124 and C Military Veterinary Hospital, 57 001, Thermi, Thessaloniki, GREECE Received 24 March 2018 ▪ Revised 5 June 2018 ▪ Accepted 13 June 2018 ABSTRACT The extensive and uncontrolled use of the larvicide cyromazine used for the control of fly species in poultry operations may lead to residues in poultry based products as well as in higher melamine levels because of the in vivo metabolism of cyromazine. This has already caused the death of cats and dogs and therefore the tolerable daily intake has been re-evaluated by legislation authorities. The aim of this study was to develop a validated HPLC method for the determination of cyromazine in commercial poultry based animal feedstuffs. Solid phase dispersion applying the QuEChERS approach was used. Chromatographic analysis was performed by an Inertsil ODS -3 analytical column (5 μm, 250 × 4 mm) using a mobile phase of MeOH – H2O (60-40% v/v), delivered isocratically. Cyromazine was detected at 220 nm. Validation was performed in terms of linearity, sensitivity, specificity, accuracy, repeatability and stability. The LOD of the method was found at 0.03 mg / kg, the LOQ was found 0.1 mg / kg, while the linearity extends up to 10 mg / kg. Absolute recovery achieved by the proposed methodology was 78 (±3) %. Several samples from the local market were examined, however no cyromazine residues were identified. Keywords: cyromazine, melamine, animal feedstuff, HPLC, QuEChERS INTRODUCTION The larvicide cyromazine (N-Cyclopropyl-1,3,5-triazine-2,4,6-triamine) is a cyclopropyl derivative of melamine (2,4,6-Triamino-1,3,5-triazine). Their chemical structures can be found in Figure 1. Cyromazine is an insecticide approved for use in the EU and in other countries. It is highly soluble in water and relatively volatile. It may be persistent in soil and water systems depending on local conditions. It has a low mammalian toxicity, hence it may cause adverse effects on reproduction. Its use as an insecticide and insect growth regulator is based to its activity by disturbing the nervous system of the immature larval stages of certain insects, mainly Diptera larvae and flies on livestock and other insect pests in the field and greenhouse. Therefore, it is used to control fly species, which develop in poultry manure and refuse. Moreover cyromazine may yield melamine as metabolism product. The toxicity of melamine is due to the crystal formation with endogenous uric acid or, cyanuric acid, a structural analogue of melamine, in renal tubules and this leads to acute renal failure [1,2]. Failure to follow directions for use of cyromazine (the active ingredient of Larvadex®) and respective precautions found on the label may result not only in poor fly control, but in illegal residues in the meat or eggs as well. Moreover suggested withdrawal times have to be strictly followed prior to slaughter [3,4]. A residual maximum of 50 ng/g for cyromazine in the edible parts of eggs and poultry meat are allowed as stipulated by the US Code of Federal Regulations (1987); however, residual levels and cyromazine inclusion levels in animal diets differ between countries depending on their application as a veterinary drug [5,6].