View More View Less
  • 1 Ministry of Agriculture Feed Safety and Bio-availability Evaluation Center, China Agricultural University, Beijing 100094, P.R. China
  • 2 Department of Applied Chemistry, China Agricultural University, Beijing 100094, P.R. China
Restricted access

Summary

A new HPLC method has been established for determination of 3-monoiodotyrosine (MIT), 3,5-diiodotyrosine (DIT), 3,5-diiodothyronine (T2), 3,3′,5-triiodothyronine (T3), 3,3′,5′-triiodothyronine (rT3), and thyroxine (T4) produced by hydrolysis of iodinated casein with barium hydroxide. The hydrolytic stability of each analyte was evaluated. Iodinated casein was hydrolyzed with saturated barium hydroxide solution for 16 h at 110°C and the barium ions were then removed as barium sulfate. Reversed-phase HPLC was performed on a 2.1 mm × 150 mm, 5 μm particle, C18 column with a mixture of acetonitrile and 0.1% (v/v) formic acid as mobile phase at a flow rate of 0.2 mL min–1. Acetonitrile was maintained at 5% (v/v) for 5 min and then increased linearly to 50% (v/v) within 35 min. All analytes were quantified by measuring the absorbance at 280 nm. Validation data indicated the method was linear, with regression coefficients (R2) > 0.998, in the concentration ranges investigated. Sensitivity was adequate—limits of detection (LOD) were 0.04–0.38 μg mL–1 and limits of quantification (LOQ) were 0.05–0.38 μg mL–1. Accuracy and precision were acceptable — for all the analytes recovery was 82.0–93.0% and repeatability, as relative standard deviation, was 1.0–3.0%. Hydrolytic stability tests indicated MIT and DIT are much more stable than the other analytes. rT3 was not released directly from iodinated casein but was formed by deiodination of T4 during hydrolysis. The method could be used to identify iodinated casein, to evaluate its activity and quality, and for supervision and regulation of feed additives.

  • [1]. E.P. Reineke and C.W. Turner, Missouri Agric. Exp. Sta. Res. Bull., 355 (1942).

  • [2]. D.M. Seath C. Branton A.H. Groth 1945 J. Dairy Sci. 28 509.

  • [3]. H.R. Wilson M.A. Boone A.S. Arafa D.M. Janky 1983 Poultry Sci. 62 811.

  • [4]. E.P. Reineke C.W. Turner G.O. Kohler R.D. Hoover M.B. Beezly 1945 J. Biol. Chem. 161 599.

  • [5]. T.W. Mischler E.P. Reineke 1970 J. Dairy Sci. 53 233.

  • [6]. Z.Y. Wang L.Y. Zhang D.F. Li W.J. Yang T. Dong Z.Q. Zhou 2003 Chin. J. Anal. Chem. 31 1187.

  • [7]. M.A. Faircloth A.D. Williams W.H. Florsheim 1965 Anal. Biochem. 12 437.

  • [8]. J.E. Stouffer 1969 J. Chromatogr. Sci. 7 124.

  • [9]. N.N. Nihei M.C. Gershengorn T. Mitsuma L.R. Stringham A. Cordy B. Kuchmy C.S. Hollander 1971 Anal. Biochem. 43 433.

  • [10]. A. de la Vieja M. Calero P. Santisteban L. Lamas 1997 J. Chromatogr. B 688 143.

  • [11]. H. Gika M. Lämmerhofer I. Papadoyannis W. Lindner 2004 J. Chromatogr. B 800 193.

  • [12]. H.G. Gika V.F. Samanidou I.N. Papadoyannis 2005 J. Chromatogr. B 814 163.

  • [13]. G.O. Kohler and R.D. Hoover, US patent no. 3674770 (1972).

  • [14]. E.P. Reineke C.W. Turner 1943 J. Biol. Chem. 149 563.

  • [15]. E.P. Renée C.W. Turner G.O. Kohler R.D. Hoover M.B. Beezly 1945 J. Biol. Chem. 161 599.

  • [16]. Z.Y. Wang B.Y. Chang L.Y. Zhang W.J. Yang T. Dong 2005 Fenxi-Ceshi-Xuebao (Chin. J. Instrum. Anal.) 24 86.

  • [17]. Y. Zheng Y. Sun J. Ren 2006 Talanta 69 107.

  • [18]. T.J. Carne R.E. Huber P. Davitt L.A. Edwards 1986 J. Chromatogr. 367 393.

  • [19]. C.S. Yoon , Ph. D. Dissertation, University of Missouri, Columbia, Missouri, USA, Part II: 4698 (1974).

Monthly Content Usage

Abstract Views Full Text Views PDF Downloads
Jun 2020 0 7 2
Jul 2020 0 0 0
Aug 2020 3 0 0
Sep 2020 6 0 0
Oct 2020 0 0 1
Nov 2020 0 2 1
Dec 2020 0 0 0