Development of an Accelerated Solvent Extraction-Ultra-Performance Liquid Chromatography-Fluorescence Detection Method for Quantitative Analysis of Thiamphenicol, Florfenicol and Florfenicol Amine in Poultry Eggs.

Bo Wang,Zhixiang Diao,Tao Zhang,Kaizhou Xie,Xing Xie,Genxi Zhang,Guojun Dai,Xia Zhao

doi:10.3390/molecules24091830

Bo Wang, Zhixiang Diao + Show 6 more

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https://doi.org/10.3390/molecules24091830

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Abstract

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.

Highlights

Poultry eggs contain high levels of essential amino acids and vitamins as well as various major and trace elements required by the human body, and as a result they have become an increasingly popular consumer product [1]
When detecting CAPs with a fluorescence detector, to enhance the retention of florfenicol amine (FFA) on a C18 column, an ion-pair reagent is usually added to the aqueous phase to react with the FFA to form a weakly bound ion pair, and the pH is adjusted with a buffer to keep the whole system weakly acidic and prevent dissociation of the ion pair [14,32]
Sodium heptane sulfonate was added to the mobile phase, and the target compounds were well separated

Summary

Introduction

Poultry eggs contain high levels of essential amino acids and vitamins as well as various major and trace elements required by the human body, and as a result they have become an increasingly popular consumer product [1]. Thiamphenicol (TAP) and florfenicol (FF) are synthetic chloramphenicols (CAPs) used as broad-spectrum antibiotics, and they have chemical structures and efficacies similar to that of CAP as well as good therapeutic effects on various bacterial strains common in poultry. These compounds are widely used in actual production [3]. TAP can inhibit the formation of red blood cells, white blood cells and platelets; it has a strong immunosuppressive effect; and it has weaker antibacterial effects than CAP, which limit its practical use [4,5,6] Many countries use these compounds as veterinary drugs, but they are banned from use in food animals.

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