Analysis of florfenicol in pig plasma using a validated PPT-HPLC-DAD method

Abstract

High performance liquid chromatography (HPLC) has proven to be an effective tool for examining the disposition kinetics of florfenicol (FF), a synthetic broad-spectrum antibiotic used to treat infectious diseases in veterinary medicine. Modification and optimisation of the protein precipitation (PPT) sample preparation procedure and HPLC with diode array detector (DAD) instrumental method were carried out to ensure conditions suitable for FF analyses in pig plasma samples. Stable supernatants with good plasma mean recoveries of FF (99.8% ± 0.7%RSD) were achieved using 1% v/v phosphoric acid solution in methanol and 10% w/v sodium chloride in aqueous solution. The PPT-HPLCDAD method’s detection limit of 0.004 μg/mL and quantification limit of 0.013 μg/mL provides high sensitivity for analyses of FF in plasma samples. In addition, the optimisation of method conditions resulted in shorter extraction and analysis time and less solvent consumption, which stresses the sustainability of this method in analytical chemistry. The optimised and validated PPT-HPLC-DAD method was applied in a comparative study of FF in pig plasma after administration of veterinary medicinal products. The study was conducted on fattening pigs following repeated intramuscular administration of two similar solutions for injection at a dose of 20 mg FF/kg bodyweight (test groups 1 and 2). The solutions for injection contained the same FF concentration, i.e., 300 mg/mL, but differed in excipients. The aim was to examine the influence of administrated solutions for injection on the extent of exposure to FF in pig plasma. The dynamics of kinetic profiles of FF in pig plasma from both treatments correspond to the FF kinetic profiles published in similar studies. However, differences were observed in the concentrations of FF, which were constant throughout the study, and statistical differences between the test groups were confirmed (P<0.05). Though these findings suggest the possible influence of excipients, a full comprehensive conclusion on the influence of administrated solutions for injection on FF exposure in pig plasma requires additional research.

A Green HPLC Approach to Florfenicol Analysis in Pig Urine

“…Analytical methods for determination of the FF and/or its metabolite florfenicol amine (FFA) in blood samples, edible tissues or drug formulations are primarily described in the literature. For example, the following methods were used: voltammetric method [ 13 ], enzyme-linked immunosorbent assay (ELISA) method [ 14 ], ultraviolet-visible (UV-Vis) spectrophotometric method [ 15 ], near-infrared spectroscopy [ 16 ], capillary electrophoresis [ 17 ], and chromatographic methods such as thin-layer chromatography (TLC) [ 18 ], gas chromatography (GC) [ 19 ], (ultra)high-performance liquid chromatography ((U)HPLC) with ultraviolet (UV), diode array detector (DAD) or fluorescence (FL) detector [ 20 , 21 , 22 , 23 , 24 , 25 ], and liquid chromatography-tandem mass spectrometry (LC-MS/MS) [ 26 , 27 , 28 , 29 ]. Only several reported papers included and described the analytical methods for determining the FF in urine samples of animal origin [ 1 , 11 , 12 , 30 , 31 , 32 , 33 ].…”

“…It can also be an alternative sample to edible tissues, for example, for mon antibiotic administration [11] or detecting an appropriate time of animal slaughter (a) (b) Analytical methods for determination of the FF and/or its metabolite flor amine (FFA) in blood samples, edible tissues or drug formulations are primarily des in the literature. For example, the following methods were used: voltammetric m [13], enzyme-linked immunosorbent assay (ELISA) method [14], ultraviolet-visibl Vis) spectrophotometric method [15], near-infrared spectroscopy [16], capillary e phoresis [17], and chromatographic methods such as thin-layer chromatography [18], gas chromatography (GC) [19], (ultra)high-performance liquid chromatog ((U)HPLC) with ultraviolet (UV), diode array detector (DAD) or fluorescence (FL) d [20][21][22][23][24][25], and liquid chromatography-tandem mass spectrometry (LC-MS/MS) [ Only several reported papers included and described the analytical methods for mining the FF in urine samples of animal origin [1,11,12,[30][31][32][33]. However, employ verse and often complex analytical techniques in FF analyses indicated that their on the environment should also be considered.…”