Simultaneous Measurement of Fluoroquinolones in Eggs by a Combination of Supercritical Fluid Extraction and High Pressure Liquid Chromatography

Abstract

Simultaneous detection of the fluoroquinolone antibiotics ciprofloxacin, enrofloxacin, ofloxacin, and norfloxacin in eggs by a combination of supercritical fluid extraction (SFE) and high pressure liquid chromatography (HPLC) was studied. Lipid matrices that have been considered to result in poor extraction and isolation of fluoroquinolones in eggs were removed first by SFE with supercritical CO(2) alone, and then the fluoroquinolones were extracted by SFE with supercritical CO(2) containing 20% (v/v) methanol for HPLC analysis. A time-course study of the extraction of lipid matrices of eggs suggested that the SFE method successfully removed the matrices within 20 min. When the fluoroquinolones added to control eggs were extracted by SFE, the extraction efficiency was similar to that by the solvent extraction method, giving the recovery percentages from 83 to 96% in a 40 min-extraction time. The fluoroquinolones extracted from eggs by SFE were analyzed simultaneously by HPLC equipped with a fluorescence detector with detection sensitivity at about 10 ppb for the detection limit. The standard calibration profiles of fluoroquinolones showed linear responses to HPLC, showing more than 0.995 for the mean r(2) value. This is the first report of the simultaneous measurement of fluoroquinolones in eggs by a combination of SFE and HPLC. Using the SFE method allowed us to avoid extensive sample preparation such as solvent extraction and chromatographic cleanup that are basically required in extraction of fluoroquinolones.

Monitoring of fluoroquinolone residual levels in chicken eggs by microbiological assay and confirmation by liquid chromatography

“…The analysis of antibiotics in eggs is quite complex, as they may bind to lipoproteins, and the extraction solvents may form emulsions and foams with the egg matrix. The result of this is poor isolation and purification of the antibiotics and, therefore, lipids and proteins should be removed prior to analysis (Shim et al, 2003). Different sample preparation methods have been described for the analysis of FQs in eggs, including either liquid-liquid extraction (LLE; Chu et al, 2002;Schneider and Donoghue, 2003;Garcia et al, 2005;Zeng et al, 2005) or solid-phase (micro) extraction (SPE, SPME; Gigosos et al, 2000;Liu et al, 2004;Heller et al, 2006;Huang et al, 2006).…”

“…Some of these studies used special equipment for sample preparation, e.g. microdialysis (Schneider and Donoghue, 2000;Lolo et al, 2005), supercritical fluid extraction (SFE; Shim et al, 2003), accelerated solvent extraction (ASE; Herranz et al, 2007), or analysis using liquid chromatography-mass spectrometry (LC-MS; Schneider and Donoghue, 2003;Lolo et al, 2005) or liquid chromatography-tandem mass spectrometry (LC/MS/MS; Heller et al, 2006). The advantages of these analytical instruments, in addition to increased analytical speed, include lower cost per sample through reduced solvent consumption, but they require sophisticated equipment that is not always available to laboratories with limited funding.…”

Comparison of different extraction methods for the simultaneous determination of pesticide residues in kiwi fruit using gas chromatography–mass spectrometry

“…The vessel was pressurized with CO 2 to 300 atm and at 80°C for 5 min (static extraction setting on instrument), and the extracts were then flushed from the vessel using a 30 min dynamic extraction process at a flow rate of 3.0 mL/ min. All of the experimental variables were selected on the basis of our previous research findings (Shim et al, 2003;Choi et al, 2006;Kang et al, 2006;Kim et al, 2006;Jeon et al, 2007). The total extract volume (30 mL) was then transferred into 100 mL pear-shaped flasks, and concentrated to approximately 3 mL using a Büchi Rotavapor R-114 at 40°C.…”

“…Additionally, it may have competed with polar analytes for active sites in the matrix and displaced them into the fluid. Finally, it may have acted to swell the matrix and exposed small internal cavities, allowing for better access of the supercritical fluid into the absorbed analytes (Shim et al, 2003;Choi et al, 2006;Kang et al, 2006).…”

Optimization of a phase separation based magnetic-stirring salt-induced liquid–liquid microextraction method for determination of fluoroquinolones in food

“…Several pre-treatment methods, including solid-phase extraction (SPE) (Hermo, Nemutlu, Kir, Barron, & Barbosa, 2008), liquid-liquid extraction (LLE) (Chu et al, 2002), stir bar sorption extraction (SBSE) (Huang, Yuan, & Lin, 2011), microwave-assisted extraction (MAE) (Hermo, Barron, & Barbosa, 2005), cloud point extraction (CPE) (Wu, Zhao, & Du, 2010) and supercritical fluid extraction (SFE) (Shim, Lee, Kim, Lee, & Kim, 2003) have been developed. Major limitations of these methods include timeconsuming extraction procedures, low enrichment factor, tedious operation and creation of a large amount of hazardous organic solvent waste.…”