Separation of the Enantiomers of Thyroxine by HPLC with Chiral Mobile Phases |
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Authors: | Rong Wang Zheng-Ping Jia Li-Ren Chen Xin Ge Jun Ma Qiang Zhang Hua Xie Yan Ao Juan Wang |
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Institution: | 1. Department of Pharmacy, Lanzhou General Hospital, Lanzhou Command, Gansu, 730050, China 2. Lanzhou Institute of Chemical Physics, The Chinese Academy of Sciences, Gansu, 730000, China
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Abstract: | A high-performance liquid chromatographic (HPLC) method has been developed for separation of the enantiomers of thyroxine (T4). A silica gel column was used in conjunction with a chiral mobile phase (CMP) comprising 35:65 (v/v) acetonitrile–water containing 0.1 mM copper(II) acetate, 0.2 mM L-proline, and 0.5 mM triethylamine (TEA), pH 5.42. The flow rate was 1.0 mL min?1 and the analysis temperature 40 °C. L-T4 was eluted before D-T4 by mobile phase containing L-proline copper complex. Inversion of the chirality of the mobile phase resulted in reversal of this order of elution. A racemic mobile phase containing DL-proline copper complex resulted in no separation. The enantioseparation phenomenon is discussed. When the method was used to determine the concentration of T4 enantiomers in the serum of a patients with thyroid disease, different concentrations of T4 enantiomers were found in different patents. |
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