Development of QuEChERS method for simultaneous detection of cyromazine and melamine residues in eggs

A quick, easy, cheap, effective, rugged and safe (QuEChERS) method was developed for detection of cyromazine (CYR) and melamine (MEL) residues using High Pressure Liquid Chromatography with UV detector (HPLC-UV). Analytes were separated on Chromasol (R) C18 column (4.0 x250 mm, pore size 5 mu m) using mobile phase of 0.05% trifluoroacetic acid (TFA) and acetonitrile (ACN) delivered in gradient mode at a flow rate of 1 mL min - 1. Homogenized egg sample was extracted using solution comprising 99.5% ACN and 0.5% of 0.1 M hydrochloric acid in the presence of MgSO4, calcium silicate and celite 545 as adsorbents. Extract was subjected to liquidliquid partitioning with n -hexane for the removal of fats. Optimization of quantity of adsorbent chemicals was done by Box-Behnken statistical design (BBD). For CYR, the Limit of Detection (LOD) and Limit of Quantitation (LOQ) of the method were found to be 5.3 and 16.0 mu g kg - 1, respectively; and for MEL 11.4 and 34.7 mu g kg - 1, respectively. The average recoveries of the analytes were 87.3% and 102.4% for MEL and CYR, respectively, indicating good accuracy while, Percent Relative Standard Deviation (% RSD) of less than 10% indicated good precision. Further, the R2 values greater than 0.9998 indicated good linearity of the newly developed method. The analysis of real samples reveled that CYR could be detected in eggs only if layer farms had a history of the usage of CYR in the poultry feed. Overall, the performance of the method was optimum and less expensive to serve the purpose of routine monitoring of CYR and MEL residues in eggs.