Razvoj SPE-HPLC-DAD metode za određivanje florfenikola i florfenikol amina u cerebrospinalnoj tekućini svinja

Abstract

A study of florfenicol (FF) and its metabo- lite florfenicol amine (FFA) in pig cerebrospinal fluid was conducted following repeated intramuscular administration of the original (reference) and a generic veterinary medicinal product (VMP) under the same experimental conditions (20 mg FF/kg body weight, 48-hour interval). Both VMPs are solutions for injection containing FF as an active substance in the concentration of 300 mg/mL and have been authorized in Croatia for use in cattle and pigs. In this study, clinically healthy pigs were randomly divided into three groups. The first group was treated with the reference VMP, the second with the generic VMP, while the third served as the control group. Animals were sacrificed at 216, 288 and 384 hours after the first drug administration. Cerebrospinal fluid samples were analysed by the optimized and validated high-performance liquid chromatography-diode array detector method (HPLC-DAD). The solid-phase extraction (SPE) technique was chosen for sample preparation. The HPLC-DAD method provides good linearity over the concentration range of 0.05 to 5.00 μg/mL for FF and FFA. Limits of detection were 0.0023 μg/mL for FF and 0.0100 μg/mL for FFA. Extraction recoveries of FF were from 86.6% to 111.8%, and of FFA from 91.7% to 98.8%. The SPE-HPLC-DAD method has been demonstrated to be a selective, sensitive and suitable analytical method for the determination of FF and FFA in cerebrospinal fluid. The present study was based on a preliminary study that quantified FF in pig plasma at 216 hours after the first application of reference or generic VMP. However, FF and FFA were not detected in any of the cerebrospinal fluid samples during the experimental period. According to the nature of biological fluids, the SPE-HPLC-DAD method can be suitable for further pharmacokinetic studies of FF in pig plasma and serum after intramuscular administration of VMPs.

Development of a QuEChERS–HPLC–FLD Procedure for the Simultaneous Detection of Residues of Florfenicol, Its Metabolite Florfenicol Amine, and Three Fluoroquinolones in Eggs

“…The sensitivity of the LC–MS/MS method may not necessarily be lower than that of the LC method we have established [ 14 , 15 ]. In the detection of FF and fluoroquinolone residues, the majority of studies have primarily focused on a single class of compounds when employing LC methods [ 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 ]. FLD is commonly utilized in LC methods for the detection of FF and FFA [ 19 , 20 , 21 ].…”

A Green HPLC Approach to Florfenicol Analysis in Pig Urine

“…Moreover, it is an amphenicol that should not expected to be found in pig urine samples since it has been banned in food-producing animals for over 25 years. The conditions of the HPLC-DAD method described in our previous experimental study of FF and FFA in pig cerebrospinal fluid [ 24 ] were chosen to achieve good peak symmetry and resolution of FF and CAP in urine analysis.…”

“…Urine samples were taken from untreated pigs that served as a control group and pigs treated with the VMP during the animal experiment described in the pig cerebrospinal fluid experimental FF’s study [ 24 ]. The experimental research on crossbred fattening pigs was approved by the Ethics Committee of the Faculty of Veterinary Medicine, the University of Zagreb, Croatia (reference 640-01/06-17/30, File no.…”

“…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 ].…”

Florfenicol and Florfenicol Amine Quantification in Bull Serum and Seminal Plasma by a Single Validated UHPLC‐MS/MS Method

“…In recent years, several analytical methods based on high-performance liquid chromatography (HPLC) [7,9], high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) [3,[10][11][12][13], gas chromatography (GC) [14], and gas chromatography-mass spectrometry (GC-MS) [15] have been reported for the determination of FF and its metabolites in water, feed, and animal-derived food. On the other hand, the analysis of animal biological fuids has been less frequent, mainly involving serum [16], plasma [13,17], synovial fuid [18], and cerebrospinal fuid [19], but no study has focused on the pharmacokinetics of FF and FFA in seminal plasma. Since no information is available about their distribution in the genital tract, which would allow us to defne the most correct treatment protocols, our research aimed to develop and validate a simple and quick approach to be applied to serum and seminal plasma samples collected during a pharmacokinetic study in bulls.…”