title: Simultaneous determination of thiamphenicol, florfenicol and florfenicol amine in eggs by reversed-phase high-performance liquid chromatography with fluorescence detection.
abstract: A specific, sensitive and widely applicable reversed-phase high-performance liquid chromatography with fluorescence detection (RP-HPLC-FLD) method was developed for the simultaneous determination of thiamphenicol (TAP), florfenicol (FF) and florfenicol amine (FFA) in eggs. Samples were extracted with ethyl acetate-acetonitrile-ammonium hydroxide (49:49:2, v/v), defatted with hexane, followed by RP-HPLC-FLD determination. Liquid chromatography was performed on a 5 μm LiChrospher C(18) column using a mobile phase composed of acetonitrile (A), 0.01 M sodium dihydrogen phosphate containing 0.005 M sodium dodecyl sulfate and 0.1% triethylamine, adjusted to pH 4.8 by 85% phosphoric acid (B) (A:B, 35:65 v/v), at a flow rate of 1.0 mL/min. The fluorescence detector of HPLC was set at 224 nm for excitation wavelength and 290 nm for emission wavelength. Limits of detection (LODs) were 1.5 μg/kg for TAP and FF, 0.5 μg/kg for FFA in eggs; limits of quantitation (LOQs) were 5 μg/kg for TAP and FF, 2 μg/kg for FFA in eggs. Linear calibration curves were obtained over concentration ranges of 0.025-5.0 μg/mL for TAP with determination coefficients of 0.9997, 0.01-10.0 μg/mL for FF with determination coefficients of 0.9997 and 0.0025-2.50 μg/mL for FFA with determination coefficients of 0.9998, respectively. The recovery values ranged from 86.4% to 93.8% for TAP, 87.4% to 92.3% for FF and from 89.0% to 95.2% for FFA. The corresponding intra-day and inter-day variation (relative standard deviation, R.S.D.) found to be less than 6.7% and 10.8%, respectively.
authors: College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, People's Republic of China.
authors: Kaizhou
authors: K
authors: Xie
authors: Longfei
authors: L
authors: Jia
authors: Yilin
authors: Y
authors: Yao
authors: Dong
authors: D
authors: Xu
authors: Shuqing
authors: S
authors: Chen
authors: Xing
authors: X
authors: Xie
authors: Yan
authors: Y
authors: Pei
authors: Wenbin
authors: W
authors: Bao
authors: Guojun
authors: G
authors: Dai
authors: Jinyu
authors: J
authors: Wang
authors: Zongping
authors: Z
authors: Liu
mesh_terms: D000818:Animals; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation / Q000379:methods; D004531:Eggs / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis; D013839:Thiamphenicol / Q000031:analogs & derivatives* / Q000032:analysis*
chemical_list: C083312:florfenicol amine; C035534:florfenicol; D013839:Thiamphenicol