OPTIMIZA TION AND VALIDATION OF QUINOLONES DETERMINATION IN POULTRY MEAT BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY AND SECONDARY CONFIRMATION WITH FOURIER TRANSFORMINFRAREDSPECTROPHOTOMETERY

Abstract

A multiresidue method for the determination of a range of eight different quinolones including ofloxacin (OFL), norfloxacin (NOR), ciprofloxacin (CIP), enrofloxacin (ENR), sarafloxacin (SAR), difloxacin (DIF), oxolinic acid (OXO), and flumequine (FLU) was optimized and validated. The method used 0.1 % trifuoroacetic acid in methanol followed by solid phase extraction with CI8 cartridges for the maximum recovery and purification of analytes. Chromatographic separation and detection was achieved using C8 HPLC column with a mobile phase consisting of 10 mM citric acid under gradient mode. A good limit of detection (LOD) and quantification (LOQ) were achieved which varied between 3.74 to 9.10 and 12.47 to 30.34 respectively and were below the maximum residue limits (MRLs). The method was linear for all the quinolones under a wide range with correlation coefficients between 0.9989 and 0.9997. Once the method was setup, this was followed by a survey for the detection of quinolones residues in poultry meat samples which were collected from various local poultry shops in Rawalpindi and Islamabad. Overall, out of 60 samples tested, 11.7, 36.7, 48.4, 48.5, 10, 8.4, 6.7 and 15% samples were found to be contaminated with OFL, NOR, CIP, ENR, SAR, DIF, OXO and FLU respectively with 29.1 , 77.3, 84.9, 50, 0, 33.3 and 32.5% of these samples being above MRL set by EU/F AO. For secondary confirmation all the samples tested with HPLC were also analyzed by fourier transform infrared spectrophotometer (FTIR). A good correlation was found between the HPLC and FTIR results, however, 7 of 60 (11.7%) samples detected with HPLC were found negative with FTIR. The presence of a large proportion of quinolones residues in poultry meat is a concern and requires attention by the legislative bodies and calls for raising of awareness among the farmers and other people linked to this industry. This will not only protect consumer health but also will have positive impact on export of poultry meat.