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The objectives of this study were to compare the serum and seminal plasma pharmacokinetic profiles of florfenicol (FLO) and florfenicol amine (FLA) after the administration of FLO either by IM or SC routes in beef bulls. Four clinically healthy Hereford bulls underwent a comprehensive physical exam, including breeding soundness examination, CBC, and chemistry profile panel. Bulls were healthy and classified satisfactory potential breeders. In one group (n = 2), a single dose of FLO was administered SC in the middle of the neck at a dose of 40 mg/kg of body weight. In the second group (n = 2), a single dose was administered IM in the muscles of the neck at a dose of 20 mg/kg. Concentrations of FLO and FLA in serum and seminal plasma were determined by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Blood and semen samples were collected before the administration of FLO and at 12, 24, 36, 48, 72, 96, 120, 144, and 168 h after injection. The blood was collected from the coccygeal vessels, and semen was collected by electroejaculation. All samples were immediately refrigerated, processed within the first hour after collection, and finally stored at −80 °C. The mean level of total FLO in serum was higher when administered by the SC route (1,415.5 ng/mL) than by the IM route (752.4 ng/mL; P = 0.001). Differences were observed between the percentage of FLA in serum (1.8%; ranging from 1.3 to 2.9) and in seminal plasma (27.5%; ranging from 15.9 to 34.2; P = 0.0001). The mean level (±SD) of FLA was higher in seminal plasma compared to serum (467 ± 466 ng/mL and 18 ± 16 ng/mL, respectively; P = 0.001). The mean level of total FLO in seminal plasma was 1,454.8 ng/mL for the SC route and 1,872.9 ng/mL for the IM route without differences between the two routes (P = 0.51). Differences in the mean level of total FLO between serum and seminal plasma were detected (1,187 ± 2,069 ng/mL and 1,748 ± 1,906 ng/mL, respectively; P = 0.04). From the present investigation, it was concluded that FLO is a suitable antibiotic based on its pharmacokinetic attributes and may be employed for the treatment of bull genital infections when its use is indicated. To study the pharmacokinetics of FLO in seminal plasma, the analysis of FLA should be incorporated. [ABSTRACT FROM AUTHOR]

Florfenicol (FF) is a commonly used antibacterial agent in animals. We investigated the pharmacokinetics of FF and its metabolite florfenicol amine (FFA) in donkeys. Donkeys were administered FF (30 mg/kg bodyweight, p.o.). Pharmacokinetic parameters were calculated using a non-compartmental model. The FF (FFA) pharmacokinetics parameters were characterized by along elimination half-life (t1/2 kz) of 5.92 h (15.95 h), plasma peak concentration (Cmax) of 0.13 μg/mL (0.08 μg/mL), and the time taken to reach Cmax (Tmax) of 0.68 h (0.72 h). The area under plasma concentration-time curve and mean residence time of FF (FFA) in plasma were 1.31 μg·mL-1·h (0.47 μg·mL-1·h) and 10.37 h (18.40 h), respectively. The t1/2 kz of FF and FFA in urine was 21.93 and 40.26 h, and the maximum excretion rate was 10.56 and 4.03 μg/h reached at 25.60 and 32.20 h, respectively. The respective values in feces were 0.02 and 0.01 μg·h-1 reached at 33.40 h. The amount of FF and FFA recovered in feces was 0.52 and 0.22 μg, respectively. In conclusion, FF (FFA) is rapidly absorbed and slowly eliminated after a single oral administration to donkeys. Compared to FF, FFA was more slowly eliminated. FF (FFA) is mostly excreted through urine. [ABSTRACT FROM AUTHOR]

ObjectiveTo establish a method for the determination of chloramphenicol drugs in hen eggs by ultra-performance liquid chromatography-tandem mass spectrometry with Captiva EMR-Lipid pretreatment.MethodsSamples were extracted with ammoniated acetonitrile，then purified with Captiva EMR-Lipid，and finally detected by UPLC-MS/MS with electrospray ionization in multiple reaction monitoring mode via positive and negative ions scanning. Compounds were quantified by internal standard method.ResultsThere were good linear relationships for 3 compounds in the range of 0.03-6.0 μg/L with correlation coefficients （r） not less than 0.995. The limits of detection of chloramphenicol （CAP）， florfenicol （FF） and florfenicol amine （FFA） were 0.02， 0.01， 0.04 μg/kg， respectively. And the limits of quantitation were 0.06， 0.03， 0.12 μg/kg， respectively. The method recoveries were between 92.3%-109.6%，with relative standard deviations（n=6） between 1.4%-8.2%.ConclusionWith simple operation and high accuracy， the method was suitable for the determination of CAP， FF and FFA in hen eggs.

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In two experimental trials; florfenicol pharmacokinetics following a single dose oral administration at 15 mg kg-1 fish body weight and biosafety through extended medicated feeding were studied in the rainbow trout, Oncorhynchus mykiss. The pharmacokinetic trial was conducted for 5 days, whereas the biosafety experiment lasted for a 30-day safety margin followed by a 20-day residual period analysis at 3, 5 and 10 times greater than the therapeutic dose 10 mg kg-1 biomass day-1. Cmax µg kg-1 calculated for florfenicol were found to be 5,360 in intestine, 2,890 in gill, 2,250 in kidney, 973 in liver and 273 in plasma, obtained at Tmax of 16 h. Intestine had utmost area under the concentration-time curve (tissue/plasma) of 13.83 h μg kg-1 and a prolonged half life (t1/2ß) of 28.62 h. The highest apparent metabolic rate value in the kidney (0.327) showed a high level of biotransformation of florfenicol to its metabolite florfenicol amine. The apparent distribution rate of florfenicol amine in muscle, in comparison to the parent drug florfenicol, indicated elimination of the medication mostly in the form of florfenicol amine with t1/2 of 16.75 h. The biosafety of florfenicol orally administered to rainbow trout recorded effective feed consumption, physiological responses, drug tolerance and significantly low drug concentrations in muscle of rainbow trout, thus its usage at 10 mg kg-1 fish body weight is recommended. In the study, the rapid absorption, greater bioavailability, enhanced dispersion, slower elimination and biosafety of the drug form a significant basis for the florfenicol and its metabolite florfenicol amine as a useful antibacterial agent in aquaculture. [ABSTRACT FROM AUTHOR]

Florfenicol is a broad-spectrum and bacteriostatic antibiotic with a time-dependent killing action. It is commonly used to treat respiratory diseases in goats in an extra-label manner. This study aimed to determine the plasma pharmacokinetics and milk residue depletion profiles and calculate the milk withdrawal interval (WDI) of florfenicol and its main metabolite florfenicol amine in lactating goats. Five healthy lactating goats were administered with 40 mg/kg florfenicol by subcutaneous injection, twice, 96 h apart. Plasma and milk samples were collected up to 864 h post the first injection. Non-compartmental analysis was used to estimate the plasma pharmacokinetic parameters. Milk WDIs were calculated using the U.S. Food and Drug Administration (FDA) method and European Medicines Agency (EMA) method. A Monte Carlo simulation was performed to generate simulated data for five virtual animals to meet the data requirement of the FDA method. The calculated milk WDIs based on florfenicol, florfenicol amine, and the combined (the sum of florfenicol and florfenicol amine) were 720.28, 690.45, and 872.69 h after the last injection using the FDA method. In conclusion, this study improves our understanding on the plasma pharmacokinetics and milk residue depletion kinetics of florfenicol and florfenicol amine in lactating ruminants after subcutaneous injections. • Pharmacokinetics of florfenicol in plasma and milk were studied in goats. • Goats received 40 mg/kg florfenicol via subcutaneous injections twice 96 h apart. • Milk withdrawal interval (WDI) based on florfenicol was 720.28 h using FDA method. • Milk WDI based on sum of florfenicol and florfenicol amine was 872.69 h. • The results help derive extralabel milk WDIs of florfenicol in lactating goats. [ABSTRACT FROM AUTHOR]

In two experimental trials; florfenicol pharmacokinetics following a single dose oral administration at 15 mg kg−1 fish body weight and biosafety through extended medicated feeding were studied in the rainbow trout, Oncorhynchus mykiss. The pharmacokinetic trial was conducted for 5 days, whereas the biosafety experiment lasted for a 30-day safety margin followed by a 20-day residual period analysis at 3, 5 and 10 times greater than the therapeutic dose 10 mg kg−1 biomass day−1. Cmax µg kg−1 calculated for florfenicol were found to be 5,360 in intestine, 2,890 in gill, 2,250 in kidney, 973 in liver and 273 in plasma, obtained at Tmax of 16 h. Intestine had utmost area under the concentration–time curve (tissue/plasma) of 13.83 h μg kg−1 and a prolonged half life (t1/2ß) of 28.62 h. The highest apparent metabolic rate value in the kidney (0.327) showed a high level of biotransformation of florfenicol to its metabolite florfenicol amine. The apparent distribution rate of florfenicol amine in muscle, in comparison to the parent drug florfenicol, indicated elimination of the medication mostly in the form of florfenicol amine with t1/2 of 16.75 h. The biosafety of florfenicol orally administered to rainbow trout recorded effective feed consumption, physiological responses, drug tolerance and significantly low drug concentrations in muscle of rainbow trout, thus its usage at 10 mg kg−1 fish body weight is recommended. In the study, the rapid absorption, greater bioavailability, enhanced dispersion, slower elimination and biosafety of the drug form a significant basis for the florfenicol and its metabolite florfenicol amine as a useful antibacterial agent in aquaculture.

Some florfenicol (FF) metabolites have a strong binding affinity towards biomolecules in the edible tissues of some food animals. These bound FF residues cannot be extracted directly from edible tissues with organic solvents and are present in higher concentrations even than solvent extractable residues. In this study, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established to detect the total residues of FF in eggs, by quantifying the metabolite florfenicol amine (FFA). The sample was hydrolyzed at 95–100 °C for 4 hours to release sample-matrix bound residues and convert them all into FFA. The hydrolyzed sample was washed with ethyl acetate to remove interfering substances, extracted with ethyl acetate under alkaline conditions, purified by solid phase extraction and quantified by UPLC-MS/MS. The recoveries of FFA in eggs ranged from 91.2 to 102.4%, with an RSD ≤ 10.9%. The LOD and LOQ were 0.5 and 1.0 μg/kg, respectively. This method can be applied to the quantification of total FF residues in eggs. [ABSTRACT FROM AUTHOR]

Abstract The pharmacokinetic properties and residue elimination of florfenicol (FLO) and its amine were investigated in European seabass Dicentrarchus labrax at 24°C. The trial mainly included analysis of FLO in plasma after a single dose dietary administration of 10 mg/kg and in muscle plus skin following a multiple dosing (10 mg kg−1 day−1 for 7 days) to estimate pharmacokinetics and residue depletion, respectively. The maximum plasma concentration of FLO was measured to be 1.64 μg/ml, 4 hr post administration. The elimination half‐life (t1/2b) and the area under the concentration‐time curve extrapolated to infinity (AUC0‐∞) were calculated to be 13.0 hr and 34.7 μg h−1 ml−1, respectively. Withdrawal times of FLO and its amine were calculated to be 46.3 degree‐days, indicating a fast removal from the edible tissues of treated European seabass. Overall, FLO can be considered as a potentially efficient antibacterial agent for farmed European seabass provided that additional efforts will be devoted towards its in vitro and clinical efficacy.

A monoclonal antibody against florfenicol amine (FFA) was produced, and a competitive enzyme-linked immunosorbent assay (ELISA) was developed for detecting florfenicol (FF) and FFA in eggs. This proposed method exhibited the 50% inhibiting concentrations (IC50) of 0.120 ng/mL for FF and 0.118 ng/mL for FFA under the optimization of experimental parameters. At the fortified levels of 0.4, 0.8, 2.4, and 30.0 μg/kg, the inter-assay recoveries of the target compounds ranged from 78.3 to 103.2%, and the intra-assay recoveries ranged from 81.5 to 104.1%, respectively. The coefficients of variation were all below 12%. Results for the determination of spiked samples by this ELISA showed a good relationship with the high-performance liquid chromatography (HPLC) technique. This newly developed method confirmed to be rapid, user-friendly, and sensitivity for simultaneous detecting FF and FFA in eggs.

The accumulation of antimicrobial residues in edible animal products and aquaculture products could pose health concerns to unsuspecting consumers. Hence, this study aimed to develop a validated method for simultaneous quantification of chloramphenicol (CAP), thiamphenicol (TAP), florfenicol (FF), and florfenicol amine (FFA) in beef, pork, chicken, shrimp, eel, and flatfish using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction method coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Primary-secondary amine (PSA) and MgSO4 were used for sample purification. The analytes were separated on a reversed-phase analytical column. The coefficients of determination for the linear matrix-matched calibration curves were ≥0.9941. Recovery rates ranged between 64.26 and 116.51% for the four analytes with relative standard deviations (RSDs) ≤ 18.05%. The calculated limits of detection (LODs) and limits of quantification (LOQs) were 0.005–3.1 and 0.02–10.4 μg/kg, respectively. The developed method was successfully applied for monitoring samples obtained from local markets in Seoul, Republic of Korea. The target residues were not detected in any tested matrix. The designed method was versatile, sensitive, and proved suitable for quantifying residues in animal-derived products.

When two drugs are combined, drug-drug interactions (DDI) often occur. Metabolic DDI usually occur due to inhibition of the metabolism of one drug by the other. This leads to an increase in the plasma concentration of the drug whose metabolism is inhibited. The objective of this research study was to verify the DDI risk of two antibacterial, florfenicol (FF) and doxycycline (DOX) due to metabolism. Because food containing residues of any pharmacologically active substance could potentially constitute a public health hazard, we selected a food producing animal, goat, goat liver microsomes and recombinant metabolic enzymes, for in vivo and in vitro metabolism studies. In vitro experiments showed that CYP3A was the key enzyme subfamily in FF metabolism, DOX slowed down FF metabolism and R440 was possibly the key amino acid in the metabolic interaction between FF and DOX. In vivo studies in the goats showed that DOX inhibited up-regulation of CYP3A24 gene expression produced by FF; in liver and kidney, DOX slightly slowed down FF metabolism. Quantitative prediction of DDI risk suggest that when DOX is used in combination with FF in veterinary medicine, may result in a clinical significant increase of FF plasma and tissue concentrations, resulting a prevalence of harmful tissue residues of medicinal products in the food chain. Through our experimentation, when DOX is used in combination with FF, the withdrawal period of FF in the kidney was extended by 1 day. Otherwise, an appropriate withdrawal period (20 days) of FF was established for FF and DOX combined use to ensure that the animal can be safely slaughtered for food.

The demand for healthier foods with high nutritional value has resulted in intensive fish farming. In this production system, high-frequency infections occur, and antibiotics are administrated for control. Only two antibiotics are allowed for use in Brazilian aquaculture, one of which is florfenicol. In this work, a bioconcentration assay was performed to assess the accumulation of florfenicol in the muscle of Nile tilapia (Oreochromis niloticus). Tilapia was evaluated as it is the most produced fish species in Brazil. The fish were exposed to florfenicol at a nominal concentration of 10 mg/L, through the water. Muscle and water were collected at 0, 1.5, 3, 6, 24, and 48 h during the exposure phase and at 1.5, 3, 6, 24, 48, and 120 h during the depuration phase. Quantification was performed using an LC-MS/MS. The results showed rapid absorption and elimination of the antibiotic (half-life, t1/2 = 5 h), with low potential for accumulation of florfenicol in tilapia muscles. The study was performed to determine the bioconcentration factor (BCF) and withdrawal period of florfenicol, being 0.05 mL/μg and 1.8 h, respectively. The results contribute to set protocols for the safe use of florfenicol in tilapia transport, avoiding residues in fish that may pose risks to human health. [ABSTRACT FROM AUTHOR]

Florfenicol is a chloramphenicol antibiotic that plays an essential role on bacteriostasis. Long-term use of florfenicol in livestock and poultry can cause immunetoxicity and reproductive toxicity and the consumption of animal-derived products with excessive residues of florfenicol will pose a certain threat to human health. To study residue depletion of oral florfenicol in chickens, 48 healthy 30 -day -old AA broilers received continuous administration of 150 mg/kg/d florfenicol for five days. Muscle, liver, kidney and sebum were collected at 0.16, 1, 3, 5, 7 and 9 days after discontinuation of the drug and detected by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Most previous studies have shown that acid hydrolysis is necessary for tissue sample extraction. However, the steps of acid hydrolysis are tedious and time-consuming. In this experiment, we tried to improve the solvent extraction method to simplify the pretreatment process and the effect of acid hydrolysis was further analyzed and compared. Using the WT1.4 software the withdrawal time in muscle, liver, kidney and sebum obtained by F-QuEChERS-AOAC 3202 method was 7.82, 0.06, 4.38, 13.71 days respectively while acid hydrolysis method were 8.89, 5.84, 5.39 and 9.65 days, respectively. To reduce the residues of florfenicol and ensure the safety of chicken products, it is recommended that chickens taking oral florfenicol should be subjected to a 14 -days withdrawal time. [ABSTRACT FROM AUTHOR]

In the study on Oreochromis niloticus , singular oral gavage of florfenicol (FFC) at 15 mg/kg biomass/day was conducted, mimicking approved aquaculture dosing. Samples of plasma, bile, muscle, intestine, skin, liver, kidney, gill, and brain tissues were collected at 0, 2, 3, 4, 6, 8, 12, 16, 24, 32, 48, 64, 96, and 128 hours (h) after oral gavage. LC-MS/MS analysis revealed FFC concentrations peaked at 12.15 μg/mL in plasma and 77.92 μg/mL in bile, both at 24 hours. Elimination half-lives were 28.17 h (plasma) and 26.88 h (bile). The residues of FFC ranked muscle>intestine>skin>liver>kidney>gill. In contrast, the residues of florfenicol amine (FFA) ranked kidney>skin>liver>muscle>gill>intestine>brain, particularly notable in tropical summer conditions. The minimum inhibitory concentration of FFC was elucidated against several bacterial pathogens revealing its superior efficacy. Results highlight bile's crucial role in FFC elimination. Further investigation, especially during winter when fish susceptibility to infections rises, is warranted. [Display omitted] • Low FFC MIC and MBC against tested strains elucidated superior therapeutic efficacy. • Low absorption half-life in plasma compared to bile. • Biliary excretion pathway of FFC is a significant viable pathway in O. niloticus. [ABSTRACT FROM AUTHOR]

The pharmacokinetic properties and residue elimination of florfenicol (FLO) and its amine were investigated in European seabass Dicentrarchus labrax at 24°C. The trial mainly included analysis of FLO in plasma after a single dose dietary administration of 10 mg/kg and in muscle plus skin following a multiple dosing (10 mg kg−1 day−1 for 7 days) to estimate pharmacokinetics and residue depletion, respectively. The maximum plasma concentration of FLO was measured to be 1.64 μg/ml, 4 hr post administration. The elimination half‐life (t1/2b) and the area under the concentration‐time curve extrapolated to infinity (AUC0‐∞) were calculated to be 13.0 hr and 34.7 μg h−1 ml−1, respectively. Withdrawal times of FLO and its amine were calculated to be 46.3 degree‐days, indicating a fast removal from the edible tissues of treated European seabass. Overall, FLO can be considered as a potentially efficient antibacterial agent for farmed European seabass provided that additional efforts will be devoted towards its in vitro and clinical efficacy. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To assess the effects of the acute inflammatory response (AIR) induced by Escherichia coli lipopolysaccharide (LPS) on plasma and tissue disposition of florfenicol (FFC) and its metabolite florfenicol amine (FFC-a), after its intramuscular (IM) administration, twenty-two New Zealand rabbits were randomly distributed in two experimental groups: Group 1 (LPS) was treated with three intravenous doses of 2 μg LPS/kg bw, before an intramuscular dose of 20 mg/kg FFC twenty-four h after the first LPS or SS injection; Group 2 (Control) was treated with saline solution (SS) in equivalent volumes as LPS-treated group. Blood samples were collected before (T0) and at different times after FFC administration. Acute inflammatory response was assessed in a parallel study where significant increases in body temperature, C-reactive protein concentrations and leukopenia were observed in the group treated with LPS. In another two groups of rabbits, 4 h after FFC treatment, rabbits were euthanized and tissue samples were collected for analysis of FFC and FFC-a concentrations. Pharmacokinetic parameters of FFC that showed significantly higher values in LPS-treated rabbits compared with control rabbits were absorption half-life, area under the curve, mean residence time and clearance /F (Cl/F). Elimination half-life and mean residence time of FFC-a were significantly higher in LPS-treated rabbits, whereas the metabolite ratio of FFC-a decreased significantly. Significant differences in tissue distribution of FFC and FFC-a were observed in rabbits treated with LPS. Modifications in plasma and tissue disposition of FFC and FFC-a were attributed mainly to haemodynamic modifications induced by the AIR through LPS administration., Competing Interests: Declaration of competing interest None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A monoclonal antibody against florfenicol amine (FFA) was produced, and a competitive enzyme-linked immunosorbent assay (ELISA) was developed for detecting florfenicol (FF) and FFA in eggs. This proposed method exhibited the 50% inhibiting concentrations (IC50) of 0.120 ng/mL for FF and 0.118 ng/mL for FFA under the optimization of experimental parameters. At the fortified levels of 0.4, 0.8, 2.4, and 30.0 μg/kg, the inter-assay recoveries of the target compounds ranged from 78.3 to 103.2%, and the intra-assay recoveries ranged from 81.5 to 104.1%, respectively. The coefficients of variation were all below 12%. Results for the determination of spiked samples by this ELISA showed a good relationship with the high-performance liquid chromatography (HPLC) technique. This newly developed method confirmed to be rapid, user-friendly, and sensitivity for simultaneous detecting FF and FFA in eggs. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The previously adopted marker residue for florfenicol (FF) in China was only florfenicol amine (FFA); however, the marker residue has been changed to FF plus FFA since the end of 2017. The previous official withdrawal period determined based on the only concentration of FFA may no longer be suitable. Therefore, the present study aimed to determine the depletion profiles of FF and FFA and further calculate the withdrawal period in the crucian carp (Carassius auratus) based on the new marker residues. Florfenicol was intramuscularly administered at 10 mg/kg bodyweight daily for five consecutive days to crucian carps reared in freshwater at 10°C. After the last dose, plasma and tissue samples were randomly collected from 10 fish at different time points. The FF and FFA concentrations were simultaneously determined by high‐performance liquid chromatography (HPLC) with a fluorescence detector and further subjected to noncompartmental analysis. The elimination half‐life (h) of FF in different tissues decreased as follows: liver (39.1) > kidney (36.3) > skin plus muscle (34.6) > plasma (31.7), whereas that of FFA decreased as follows: kidney (41.4) > skin plus muscle (39.4) > liver (39.3) > plasma (35.7). Considering a maximum residue limit of 1 μg/g for the total concentration of FF and FFA in the skin plus muscle, a withdrawal period of 6 days was calculated based on the upper limit of the one‐sided 95% confidence interval. [ABSTRACT FROM AUTHOR]

Florfenicol (FF) is a commonly used antibacterial agent in animals. We investigated the pharmacokinetics of FF and its metabolite florfenicol amine (FFA) in donkeys. Donkeys were administered FF (30 mg/kg bodyweight, p.o.). Pharmacokinetic parameters were calculated using a non-compartmental model. The FF (FFA) pharmacokinetics parameters were characterized by along elimination half-life ( t 1/2 kz ) of 5.92 h (15.95 h), plasma peak concentration ( C max ) of 0.13 μg/mL (0.08 μg/mL), and the time taken to reach C max ( T max ) of 0.68 h (0.72 h). The area under plasma concentration-time curve and mean residence time of FF (FFA) in plasma were 1.31 μg·mL -1 ·h (0.47 μg·mL -1 ·h) and 10.37 h (18.40 h), respectively. The t 1/2 kz of FF and FFA in urine was 21.93 and 40.26 h, and the maximum excretion rate was 10.56 and 4.03 μg/h reached at 25.60 and 32.20 h, respectively. The respective values in feces were 0.02 and 0.01 μg·h -1 reached at 33.40 h. The amount of FF and FFA recovered in feces was 0.52 and 0.22 μg, respectively. In conclusion, FF (FFA) is rapidly absorbed and slowly eliminated after a single oral administration to donkeys. Compared to FF, FFA was more slowly eliminated. FF (FFA) is mostly excreted through urine., Competing Interests: HQ and JC were employed by Dong-E-E-Jiao Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Liu, Guo, Qu, Dong, Zhao, Fu, Kang, Cheng, Huang, Zhao and Ma.)

• DFC-M is a key oxazoline-containing intermediate in commercial process for the synthesis of Florfenicol. • The unique acidic solution degradation mechanism of DFC-M was elucidated for the first time using both online LC-MSMS and in-situ NMR. • DFC-M was found to mainly generate kinetically preferred product DFA in on-column degradation. • The solvent stability study indicated that aprotic solvents are good choices for preparing DFC-M fresh stock solution for immediate analysis. 2-(dichloromethyl)-5[4-(methylsulfonyl)-phenyl]-4-(fluoromethyl)-oxazoline (DFC-M, 1) is a key oxazoline-containing intermediate in commercial process for the synthesis of Florfenicol (3), a marketed broad spectrum veterinary antibiotic. DFC-M was not stable in solution due to the presence of oxazoline moiety, which provided further hindrance for analytical sample preparation and HPLC analysis. Hence, the mechanistic study on the in-solution degradation of DFC-M was carried out via online and offline UPLC-HR-ESI-MS as well as in-situ NMR, and the degradation pathways were proposed. This mechanistic information, together with the follow-up solution stability study, provided crucial information regarding the solution handling and mobile phase selection for DFC-M analysis during commercial processing. [ABSTRACT FROM AUTHOR]

The pharmacokinetics and residue elimination of florfenicol (FFC) and its metabolite florfenicol amine (FFA) were studied in healthy blunt‐snout bream (Megalobrama amblycephala, 50 ± 10 g). The study was conducted with a single‐dose (25 mg/kg) oral administration at a water temperature of 18 or 28°C, while in the residue elimination study, fish were administered at 25 mg/kg daily for three consecutive days by oral gavage to determine the withdrawal period (WDT) at 28°C. The FFC and FFA levels in plasma and tissues (liver, kidneys and muscle) were analysed using high‐performance liquid chromatography (HPLC). A no‐compartment model was used to analyse the concentration versus time data of M. amblycephala. In the two groups at 18 and 28°C, the maximum plasma concentration (Cmax) of FFC was 5.89 and 6.21 μg/ml, while the time to reach Cmax (Tmax) was 5.97 and 2.84 hr, respectively. These suggested that higher temperature absorbed more drug and more quickly at M. amblycephala. And the elimination half‐life (T1/2kβ) of FFC was calculated as 26.75 and 16.14 hr, while the total body clearance (CL) was 0.09 and 0.15 L kg−1 hr−1, and the areas under the concentration–time curves (AUCs) were 265.87 and 163.31 μg hr/ml, respectively. The difference demonstrated that the elimination rate of FFC in M. amblycephala at 28°C was more quickly than that at 18°C. The results of FFA showed the same trend in tissues of M. amblycephala. After multiple oral doses (25 mg/kg daily for 3 days), the k (eliminate rate constant) of FFA in M. amblycephala muscle was 0.017, the C0 (initial concentration) was 3.07 mg/kg, and the WDT was 10 days (water temperature 28°C). [ABSTRACT FROM AUTHOR]

Violative residues of florfenicol (FF) in porcine edible tissues pose a potential risk for human health. In this study, urine was selected as target matrix for routine residue monitoring of FF in pig, and a thin layer chromatography (TLC)-high-performance liquid chromatography (HPLC) method was developed for simultaneously determining FF and florfenicol amine (FFA) in porcine urine. The urine samples were extracted with ethyl acetate under alkaline environment. The extracts were enriched through evaporation, purified by TLC and analysed by HPLC at 225 nm. A Waters Symmetry C18 column was used for the separation of the two analytes. The mobile phase was acetonitrile-phosphate buffer mixtures (33.3: 66.7, v/v), and was pumped at 0.6 mL/min. The TLC-HPLC method was well validated and successfully applied to residue depletion study. Good analytical specificity was confirmed by the lack of interfering peaks at the retention times of FF and FFA. The standard curves showed good linearity (FF: y = 143064x – 1045.3, r= 0.9999; FFA: y = 275826x + 1888.8, r= 0.9999) over the range of 0.0625–8 μg/mL. The precision ranged from 0.83% to 11.66% and 2.19% to 8.75% for intraday and interday determination, respectively. The corresponding accuracy ranged from −13.38% to 10.78% and −12.15% to 7.14%, respectively. The limits of quantification (LOQs) for FF and FFA were 0.125 μg/mL. The residue depletion study showed that the concentrations of FF and FFA in urine were higher than those in edible tissues at three time points. This method was reliable, simple and cost efficient, and could be used to monitor FF residues in porcine edible tissue without slaughtering animals. TLC showed excellent purification efficiency and is expected to solve matrix interferences in veterinary drug residue analysis. [ABSTRACT FROM AUTHOR]

This manuscript describes a simple, reliable, and sensitive approach that utilizes high-performance liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (HPLC-ESI/MS/MS) for the selective determination of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in poultry eggs (hen eggs, duck eggs, and goose eggs). Samples were extracted with methanol-ammonium hydroxide-ultrapure water (97:2:1, v/v) via accelerated solvent extraction (ASE) at 80 °C and 1500 psi and defatted with hexane saturated with acetonitrile. The experimental method uses ESI (+) and ESI (−) combination mode, full-scan qualitative analysis, and multiple reaction monitoring (MRM) combined with an internal standard for quantitative analysis. This optimized method was validated according to the requirements defined by the European Union and the Food and Drug Administration. The poultry eggs were spiked with the target compounds at four levels, and the average recoveries were all higher than 88.3%, and the relative standard deviations did not exceed 3.9%. The limits of detection and limits of quantification were 0.04–0.5 μg/kg and 0.1–1.5 μg/kg, respectively. The decision limits were 0.37–102 μg/kg, and the detection capabilities were 0.44–103 μg/kg. Finally, the new approach was successfully applied to the quantitative determination of these analytes in 120 commercial poultry eggs from local supermarkets. [ABSTRACT FROM AUTHOR]

Accelerated solvent extraction was investigated as a novel alternative technology for the separation and quantitative analysis of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine from poultry eggs, and the results were compared with the results of liquid–liquid extraction. Rapid quantification of the target compounds was carried out by ultra‐performance liquid chromatography‐electrospray ionization tandem triple quadrupole mass spectrometry. This optimized method was validated according to the requirements defined by the European Union and the United States Food and Drug Administration. Finally, the new approach was successfully applied to the quantitative determination of these analytes in 90 commercial poultry eggs from local supermarkets. [ABSTRACT FROM AUTHOR]

Abstract Florfenicol (FF) is used in cattle to treat respiratory diseases but could result in tissue residues. This study aimed to develop a population physiologically based pharmacokinetic (PBPK) model to predict the concentrations of FF and its metabolite, florfenicol amine (FFA), in cattle after four different routes of administration, and to calculate and compare the withdrawal intervals (WDIs) with approved withdrawal times based on different marker residues and their MRLs or tolerances. A flow-limited PBPK model including both FF and FFA sub-models were developed with published data using acslXtreme. This model predicted FF and FFA concentrations in tissues and plasma/serum after intramuscular or subcutaneous administration. Based on the model, the WDIs of 46 and 58 days were calculated to ensure that total residue concentrations (FF + FFA) in 95th percentile of the population after intramuscular and subcutaneous administration were below the MRL, respectively. WDIs were calculated as 44 and 47 days to ensure that FFA concentrations after intramuscular and subcutaneous administration fell below tolerances in 99th percentile of the population, respectively. WDIs were longer than the corresponding label in China, US, and EU. This model provides a useful tool to predict tissue residues of FF and FFA in cattle to improve food safety. Highlights • A population PBPK model was developed to predict florfenicol residues after different routes of administration in cattle. • Estimated withdrawal intervals (WDIs) were compared with approved withdrawal times in several jurisdictions. • Estimated WDIs were longer than the corresponding approved label withdrawal times in several jurisdictions. [ABSTRACT FROM AUTHOR]

Abstract The pharmacokinetic profiles of florfenicol (FF) and florfenicol amine (FFA) following a single oral dose of FF at 10 mg·kg−1 body weight were compared in crucian carp reared at three different temperatures of fresh water. The concentrations of FF and FFA were determined simultaneously by high-performance liquid chromatography coupled with fluorescence detection, followed by compartmental analysis using a one-compartment open model. At water temperatures of 10 ± 0.5, 20 ± 0.5, and 25 ± 0.5 °C, the maximum concentration (C max) of FF was 2.435, 2.795, and 3.555 μg·mL−1, respectively, while that of FFA was 0.810, 0.945, and 1.063 μg·mL−1, respectively. The absorption half-life (t 1/2ka) of FF was 0.72, 0.73, and 0.70 h, and the elimination half-life (t 1/2kel) was 22.86 12.29, and 9.56 h, respectively. The volume of distribution per fraction absorbed (V d /F) for FF at water temperatures of 10, 20, and 25 °C was 3.67, 2.99, and 2.29 L·kg−1, respectively. For FFA, the formation half-life (t 1/2kf) was 2.13, 3.47, and 5.11 h, respectively, whereas the t 1/2kel was 27.79, 20.33, and 15.44 h at water temperatures of 10, 20, and 25 °C, respectively. At higher water temperatures, FF was absorbed and eliminated more quickly and was more extensively distributed than at lower water temperatures; in addition, the formation and elimination of FFA were faster at higher water temperatures. From the values of time > minimum inhibitory concentration (T > MIC) calculated in the current study at 10 °C, an oral dose of FF at 10 mg·kg−1 body weight administered at 48-h intervals would be sufficient for the treatment of bacterial infection with an MIC of ≤0.5 μg·mL−1, whereas a 24-h interval would be necessary at 20 and 25 °C to treat bacterial infections with the same MIC. Highlights • The plasma concentrations of florfenicol amine were reported for the first time in crucian carp. • The pharmacokinetic profiles of florfenicol and florfenicol amine in crucian carp were compared at three water temperatures. • The dosing regimens were obtained for florfenicol in crucian carp reared at three different water temperatures. [ABSTRACT FROM AUTHOR]

Abstract A reliable liquid chromatography-tandem mass spectrometry method was developed to determine total florfenicol residues in bovine tissues and eel. Florfenicol and its metabolites (florfenicol amine, monochloroflorfenicol, florfenicol oxamic acid, and florfenicol alcohol) were analyzed as the marker residue, florfenicol amine, as defined by several regulatory agencies. After hydrolysis with hydrochloric acid, samples were defatted and subjected to solid-supported liquid extraction and Oasis MCX-cartridge cleanup before analysis. The method was validated for florfenicol and its metabolites at two levels in eel and bovine muscle, fat, and liver. Excellent recoveries were obtained (93–104%), with relative standard deviations of <6% for all compounds. Negligible matrix effects and minimal analyte loss during sample preparation enabled accurate quantification by external calibration using solvent standards. No interfering peaks were observed around the retention time of florfenicol amine, indicating the high selectivity of the method. Retention times in the spiked samples corresponding to that of the calibration standard in solvent did not exceed ±0.1 min. Ion ratios from the spiked sample were within ±10% (relative) of the calibration standards. Calibration curves were linear in the range of 0.5 to 100 ng/mL, with coefficients of determination higher than 0.998. The limits of quantification and limits of detection of the proposed method were estimated to be 0.01 mg/kg and 0.0005 mg/kg, respectively, in all food samples. Thus, the developed method is considered reliable and suitable for regulatory use. Highlights • An LC-MS/MS method was developed for total florfenicol residues in foods. • Florfenicol and its metabolites were analyzed as the marker, florfenicol amine. • Sample hydrolysis, cleanup, and LC-MS/MS conditions were optimized. • The proposed method demonstrated excellent recoveries and precision. [ABSTRACT FROM AUTHOR]

A widely applicable method involving liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry (LC-ESI/MS/MS) was developed for the simultaneous determination of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in eggs. Samples were extracted with ethyl acetate-acetonitrile-ammonium hydroxide (49:49:2, v/v) and defatted with hexane saturated with acetonitrile. The analysis was carried out on a mass spectrometer via an electrospray interface operated in the positive and negative ionization modes using deuterated chloramphenicol-d5 as the internal standard. The limits of detection and limits of quantification were 0.04–0.5 μg/kg and 0.1–1.5 μg/kg in eggs, respectively. The average recoveries of the four analytes from egg samples were 90.84–108.23%, with relative standard deviations of less than 9.61%. The corresponding intra-day and inter-day variations were found to be less than 8.11% and 11.30%, respectively. Finally, the new approach was successfully applied to the quantitative determination of these analytes in 50 commercial eggs from local supermarkets. [ABSTRACT FROM AUTHOR]

Abstract: The pharmacokinetic profiles of florfenicol (FF) or florfenicol amine (FFA) in crucian carp were compared at different water temperatures after single intramuscular administration of FF at 10 mg/kg bodyweight. The concentrations of FF and FFA were determined by a high‐performance liquid chromatography method, and then, the concentration versus time data were subjected to compartmental analysis using a one‐compartment open model. At the water temperatures of 10, 20, and 25°C, the peak concentrations (Cmaxs) of FF were 2.28, 2.29, and 2.34 μg/ml, respectively, while those of FFA were 0.42, 0.71, and 0.82 μg/ml, respectively. And the absorption half‐life (t1/2ka) of FF was 0.21, 0.19, and 0.21 hr, while the elimination half‐life (t1/2kel) was 31.66, 24.77, and 21.48 hr, respectively. For FFA, the formation half‐life (t1/2kf) was 3.85, 8.97, and 12.43 hr, while the t1/2kel was 58.34, 30.27, and 21.22 hr, respectively. The results presented here demonstrated that the water temperature had effects on the elimination of both FF and FFA and the formation of FFA. Based on the T > MIC values calculated here, to treat the infections of bacterial with MIC value ≤ 0.5 μg/ml, FF intramuscularly given at 10 mg/kg bodyweight with a 72‐hr interval is sufficient at the water temperature of 10°C, while the intervals of 60 and 48 hr were needed at 20 and 25°C, respectively. But to treat bacterial with higher MIC values, more FF or FF at 10 mg/kg BW but with shorter intervals should be intramuscularly given to the infected fish. [ABSTRACT FROM AUTHOR]

Antimicrobial residues might persist in products and by-products destined for human or animal consumption. Studies exploring the depletion behavior of florfenicol residues in broiler chicken claws are scarce, even though claws can enter the food chain directly or indirectly. Hence, this study intended to assess the concentrations of florfenicol (FF) and florfenicol amine (FFA)--its active metabolite--in chicken claws from birds that were treated with a therapeutic dose of florfenicol. Furthermore, concentrations of these analytes in this matrix were compared with their concentrations in edible tissues at each sampling point. A group of 70 broiler chickens were raised under controlled conditions and used to assess residue depletion. Sampling points were on days 5, 10, 20, 25, 30, 35, and 40 after ceasing treatment, thus extending beyond the withdrawal period established for muscle tissue (30 days). Analytes were extracted using HPLC-grade water and acetone, and dichloromethane was used for the clean-up stage. Liquid chromatography coupled to mass spectroscopy detection (LC-MS/MS) was used to detect and quantify the analytes. The analytical methodology developed in this study was validated in-house and based on the recommendations described in the Commission Decision 2002/657/EC from the European Union. Analyte concentrations were calculated by linear regression analysis of calibration curves that were fortified using an internal standard of chloramphenicol-d5 (CAF-d5). The depletion time of FF and FFA was set at 74 days in claws, based on a 95% confidence level and using the limit of detection (LOD) as the cut-off point. Our findings show that FF and FFA can be found in chicken claws at higher concentrations than in muscle and liver samples at each sampling point. [ABSTRACT FROM AUTHOR]

A simple, reliable and sensitive liquid chromatography-tandem mass spectrometry-based confirmatory method was redeveloped and validated for the simultaneous determination of chloramphenicol, thiamphenicol, florfenicol and florfenicol amine in chicken muscles. The analytes were extracted from minced chicken muscle with acetonitrile and ammoniated water mixture. A second extraction with ethyl acetate was followed by evaporation and dissolution of the residue in ammoniated methanol before defatting with n-hexane. Finally, the extract was further cleaned up by dispersive solid phase extraction using C-18 end-capped dispersive material. The validation protocol was adapted from the European Commission Decision 2002/657/EC and all the performance characteristics were successfully satisfied. The recoveries of all the analytes were found to be in the range of 86.4-108.1% and the precision values, within day and between days, ranged from 2.7% to 11% and 4.4% to 16.3%, respectively. The method was tested in various incurred samples and applied to analyse a wide range of random poultry market samples (n = 120) collected from three cities of the Punjab, Pakistan. Chloramphenicol and florfenicol residues were detected at low levels in less than 11% of the samples. Chloramphenicol was detected only in 4 samples with the concentration range of 0.17-0.477 µg kg-1, whereas the levels of florfenicol/florfenicol amine residues detected in 9 samples ranged from 8.7 to 32.8 µg kg-1. Moreover, most of the florfenicol residues were identified as tissue bound, extractable only after strong acid hydrolysis. [ABSTRACT FROM AUTHOR]

This study was aimed to determine the bioequivalence of three different preparations of florfenicol using non-drugged broiler chickens. A total of 28 broiler chickens aging 30-day were divided into four equal groups; these were Group I, II, III, and IV. The birds of Group I (for effective substance) were given intravenous (i.v.) administration of florfenicol dosed at 40 mg/kg body weight (b.wt.). The birds of Group II (for reference drug), Group III (for test-1 drug), and Group IV (for test-2 drug) received florfenicol preparations with water (dosed at 40 mg/kg b.wt.) through intracrop administration. Blood samples were collected periodically from the birds of all four groups, and blood plasma was separated. Levels of florfenicol and its metabolite (florfenicol amine) in the plasma were measured by High Performance Liquid Chromatography (HPLC). In this study, the limit of detection (LOD) for florfenicol and florfenicol amine were recorded as 0.017 and 0.78 µg/mL, respectively. On the other hand, the recovery of florfenicol and florfenicol amine were 83.4-84.6 and 82.2-83.8%, respectively. Based on the values of area under the curve (AUC), maximum concentration (Cmax), and time to maximum concentration (Tmax), test-1 drug was found to be acceptable, whereas test-2 drug was remained below the acceptable limits (80-125%) of AUC and Cmax. Thus, it was concluded that test-1 drug was bioequivalent as compared to the reference drug.

A simple, rapid and novel method for the detection of residues of thiamphenicol (TAP), florfenicol (FF) and its metabolite, florfenicol amine (FFA), in poultry eggs by ultra-performance liquid chromatography-fluorescence detection (UPLC-FLD) was developed. The samples were extracted with acetonitrile-ammonia (98:2, v/v) using accelerated solvent extraction (ASE) and purified by manual degreasing with acetonitrile-saturated n-hexane. The target compounds were separated on an ACQUITY UPLC® BEH C18 (2.1 mm × 100 mm, 1.7 μm) chromatographic column using a mobile phase composed of 0.005 mol/L NaH2PO4, 0.003 mol/L sodium lauryl sulfate and 0.05% trimethylamine, adjusted to pH 5.3 ± 0.1 by phosphoric acid and acetonitrile (64:36, v/v). The limits of detection (LODs) and limits of quantification (LOQs) of the three target compounds in poultry eggs were 1.8–4.9 µg/kg and 4.3–11.7 µg/kg, respectively. The recoveries of the three target compounds in poultry eggs were above 80.1% when the spiked concentrations of three phenicols were the LOQ, 0.5 maximum residue limit (MRL), 1.0 MRL and 2.0 MRL. The intraday relative standard deviations (RSDs) were less than 5.5%, and the interday RSDs were less than 6.6%. Finally, this new detection method was successfully applied to the quantitative analysis of TAP, FF and FFA in 150 commercial poultry eggs.

Florfenicol, a broad spectrum bacteriostatic antibiotic belonging to amphenicol class, is widely used in poultry and livestock for the treatment of various infections. The major metabolite of florfenicol in different animal species is florfenicol amine which is exploited as the marker residue for the determination of florfenicol. Analysis of florfenicol merely by solvent extraction cannot determine the accurate amount of the drug present in incurred tissues (muscle, liver and kidney) of treated birds, as indicated by this study. Thus the methods solely based on solvent extraction may lead to false negative results. A reliable LC–MS/MS based confirmatory method for the analysis of florfenicol and its metabolites in chicken muscle was developed and validated according to the European Union Commission Decision 2002/657/EC. The method was based on acid hydrolysis to liberate non-extractable residues having presumably been covalently bound to tissues, and to convert all the florfenicol residues as well as its metabolites into florfenicol amine. The amine was subsequently recovered with ethyl acetate at pH 10.5, defatted and further cleaned up with dispersive solid phase extraction (dSPE). The LC separation was achieved on reverse phase C-18 column with isocratic elution using acetonitrile/water mobile phase and the analysis was performed on linear ion trap mass spectrometer. Calibration curve was obtained over a concentration range of 25–600 μg/kg for chicken muscles. The accuracy values ranged from 84 to 101.4% and the precision values for within day and between days ranged from 1.2-11.7%, respectively. Limit of detection (LOD), limit of quantification (LOD), CCα and CCβ values were 0.98, 3.2, 113 and 126 μg/kg, respectively. The developed method was highly robust and was further applied to estimate the relative distribution of solvent-extractable against solvent-non-extractable florfenicol drug residues in muscle, liver and kidney samples of broiler chicken after 5 days of oral dosing. [ABSTRACT FROM AUTHOR]

simple method of sample preparation, identification, and determination of amphenicols in foodstuffs (chloramphenicol, chloramphenicol succinate, thiamphenicol, thiamphenicol glycinate acetylcysteinate, florfenicol, florfenicol amine) via high-performance liquid chromatography/high resolution quadrupole time-of-flight mass spectrometry by the exact masses of the ion that form in electrospray is proposed. The objects of analysis are meat (beef, pork, poultry), meat by-products, milk and dairy products, honey, and aquaculture products. Sample preparation consisted of extraction of the target components by acetonitrile and dilution of the extract twice with deionized water. Specific features in the determination of amphenicols and their derivatives associated with the emergence of various adducts in the electrospray device in the detection of both positive and negative ions were considered. A method of amphenicol determination by the addition method is proposed; it uses the summation of area signals of all registered adducts. The features and benefits of utilizing the method of standard additions are presented. The ranges of the defined contents are 0.1-10 ng/g for chloramphenicol and 1-100 ng/g for thiamphenicol and florfenicol. The relative standard deviation of the test results did not exceed 0.1; the duration of the analysis was 30-40 min. [ABSTRACT FROM AUTHOR]

A rapid and reliable method using liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS) to identify and quantify chloramphenicol (CAP), thiamphenicol (TAP), florfenicol (FF) and its marker residue florfenicol amine (FFA) has been developed and validated. FF and TAP are both allowed substances and different MRLs have been established for different matrices and species. However, CAP is a well-known forbidden substance with a MRPL of 0.3 μg kg for all matrices. It was relevant to develop this confirmatory method to quantify simultaneously the four amphenicols in kidney and muscle of all species, to make more effective the official control programme for this group of substances. The variety of levels of interest to be studied for the allowed compounds and the different chemical behaviour of FFA have been the main difficulties in the development and validation of the method. The method has been validated in swine, bovine, caprine, equine, porcine, aquaculture species, rabbit and poultry muscle and kidney (except aquaculture species, rabbit and poultry). Performance criteria have been calculated in accordance with Commission Decision 2002/657/EC. The proposed procedure is being applied to the official control in the Valencian Region after being successfully evaluated by ENAC (Spanish national body for the accreditation). [ABSTRACT FROM AUTHOR]

Antimicrobial residues might persist in products and by-products destined for human or animal consumption. Studies exploring the depletion behavior of florfenicol residues in broiler chicken claws are scarce, even though claws can enter the food chain directly or indirectly. Hence, this study intended to assess the concentrations of florfenicol (FF) and florfenicol amine (FFA)—its active metabolite—in chicken claws from birds that were treated with a therapeutic dose of florfenicol. Furthermore, concentrations of these analytes in this matrix were compared with their concentrations in edible tissues at each sampling point. A group of 70 broiler chickens were raised under controlled conditions and used to assess residue depletion. Sampling points were on days 5, 10, 20, 25, 30, 35, and 40 after ceasing treatment, thus extending beyond the withdrawal period established for muscle tissue (30 days). Analytes were extracted using HPLC-grade water and acetone, and dichloromethane was used for the clean-up stage. Liquid chromatography coupled to mass spectroscopy detection (LC–MS/MS) was used to detect and quantify the analytes. The analytical methodology developed in this study was validated in-house and based on the recommendations described in the Commission Decision 2002/657/EC from the European Union. Analyte concentrations were calculated by linear regression analysis of calibration curves that were fortified using an internal standard of chloramphenicol-d5 (CAF-d5). The depletion time of FF and FFA was set at 74 days in claws, based on a 95% confidence level and using the limit of detection (LOD) as the cut-off point. Our findings show that FF and FFA can be found in chicken claws at higher concentrations than in muscle and liver samples at each sampling point.

Despite the benefits to fish farmers, the use of antimicrobials in aquaculture has concerned consumers and competent authorities. The indiscriminate use of such substances promotes the emergence of resistant microorganisms, decreases the effectiveness of treatments, and causes possible toxic effects in humans. In Brazil, florfenicol is the only antimicrobial registered for use in aquaculture and is often used in tilapia in cage creation. Thus, this study aimed to develop a method for determination of florfenicol residues and its metabolite florfenicol amine in tilapia fillet by UPLC–MS/MS. Analytes were extracted with ethyl acetate, followed by liquid-liquid partition clean-up with hexane and SPE. The sorbents C18, phenyl and HLB-Oasis were evaluated by SPE. Phenyl sorbent showed the best results, and the extraction conditions were optimized in the sample matrix with fractional factorial design 2 4−1 . The analytes were separated on a C18 chromatographic column (50 × 2.1 mm × 1.7 μm) using water (A) and acetonitrile (B) as mobile phase at a flow rate of 0.3 mL min −1 with a linear gradient (in% B): 0–2.0 min: 20%; 2.0–2.5 min: increase to 90%; 2.5–3.5 min: 90%; 3.0–3.5 min: decrease to 20%; 4.0–5.0 min: 20%. The analytes were monitored in a MS/MS triple quadrupole system by MRM mode with transitions at m / z 356.1 > 336.1 (florfenicol) and m / z 248.1 > 130.1 (florfenicol amine). The optimized method was validated obtaining LOQ values of 3 and 25 ng g −1 for florfenicol and florfenicol amine, respectively, precision between 20 and 36%, absolute extraction efficiency between 38 and 80%, and adequate linearity. The method was applied to samples intended for human consumption, and within the 15 evaluated samples, only one showed florfenicol residue at 30 ng g −1 , which is below the maximum residue limit established in Brazil. [ABSTRACT FROM AUTHOR]

Pharmacokinetics and elimination of florfenicol and florfenicol amine in grouper held in sea water at 23.3 ± 0.8 °C were studied using HPLC method after they were given a single peroral dose of florfenicol at 24 mg kg−1 body weight. Florfenicol was rapidly absorbed from intestine and distributed extensively to all the tissues examined. The maximum concentrations ( Cmax, μg g−1 or μg mL−1) in plasma and tissues were observed at 2-6 h (the time to reach maximum concentration, Tmax) except for bile ( Tmax = 24 h) and were in the order of intestine (52.02 ± 25.07) > bile (49.41 ± 28.16) > gill (45.12 ± 11.10) > plasma (28.28 ± 5.43) > liver (21.97 ± 12.08) > muscle (21.63 ± 6.12) > kidney (20.88 ± 11.28) > skin (19.10 ± 5.88). The drug distribution level was higher in plasma than in extravascular tissues except for bile, based on the ratios of the area under concentration-time curve between tissue and plasma (AUCtissue/plasma). The elimination of florfenicol was rapid in fish, and the corresponding half-lives ( T1/2β) in the order of magnitude were bile (13.92 h) > muscle or liver (12.31 h) > skin (11.77 h) > plasma (11.57) > gill (11.04 h) > intestine (10.55 h) > kidney (10.05 h). The delayed Tmax, lower Cmax and longer T1/2β for florfenicol amine compared with florfenicol were measured in grouper. [ABSTRACT FROM AUTHOR]

The parent compound florfenicol (FF) is a broad-spectrum antibacterial compound licensed in the UK for use in cattle, pigs and the aquaculture industry. The analysis of porcine tissues in this study demonstrates that significant amounts of solvent non-extractable FF-related residues are present in incurred tissues (kidney and muscle) from treated animals. The results indicate that methods based on solvent extraction alone may carry a significant risk of reporting false-negative results. The use of a strong acid hydrolysis step prior to solvent extraction of tissue samples is necessary for an accurate estimate of the total tissue FF content. A robust and sensitive method for the determination of total FF residue content in kidney samples by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been developed and validated. This method covers the synthetic amphenicol drug FF and its metabolites, measured as the marker residue florfenicol amine (FFA) as per Commission Regulation (EU) No. 37/2010. Non-extractable and intermediate metabolites are converted to the hydrolysis product FFA, and then partitioned into ethyl acetate. Extracts are solvent exchanged prior to a dispersive solid-phase extraction step, then analysed using an alkaline reverse-phase gradient separation by UPLC-MS/MS. The method was validated around the maximum residue levels (MRLs) set out in Regulation (EU) No. 37/2010 for bovine kidney in accordance with Commission Decision No. 2002/657/EC. The following method performance characteristics were assessed during a single laboratory validation study: selectivity, specificity, sensitivity, linearity, matrix effects, accuracy and precision (decision limit (CCα) and detection capability (CCβ) were determined). [ABSTRACT FROM AUTHOR]