Wang Rong, Jia Zheng-Ping, Hu Xiao-Li, Xu Li-Ting, Li Yong-Min, Chen Li-Ren
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, Gansu, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Mar 5;785(2):353-9. doi: 10.1016/s1570-0232(02)00959-5.
A chromatographic method for the separation and determination of D- and L-thyroxine enantiomers (D-, and L-T4) in human serum with a chiral ligand ion-exchange system using a chiral mobile phase additive and a silica column was established. An aqueous eluent containing L-proline (L-pro) sufficiently complexed copper II ions and triethylamine (TEA) was used. It was monitored with a UV detector. The separation was completed in 12 min. The method has acceptable sensitivity, precision and accuracy for analysis. The limit of detection and the limit of quantitation for both D- and L-T4 were 0.1 microg/ml and 0.8 microg/ml, respectively. Calibration curves were linear within 1-100 microg/ml; the mean correlation coefficients were r(D-T4)=0.9986 for D-T4 and r(L-T4)=0.9978 for L-T4. T4 enantiomers were separated on baseline under the optimum condition. L-T4 eluted before D-T4. The concentration of D-T4 and L-T4 in 45 thyroid patients serum (hyperthyroid, hypothyroid, thyroidectomy, goitre or thyroiditis) using HPLC was determined, those results showed that D,L-T4 concentration varied in different thyroid patient. Attention should be paid to this result in treating thyroid disease in the clinic.
建立了一种色谱方法，用于在人血清中分离和测定D-和L-甲状腺素对映体（D-和L-T4），该方法采用手性配体离子交换系统，使用手性流动相添加剂和硅胶柱。使用含有与铜II离子充分络合的L-脯氨酸（L-pro）和三乙胺（TEA）的水性洗脱液。用紫外检测器进行监测。分离在12分钟内完成。该方法对于分析具有可接受的灵敏度、精密度和准确度。D-和L-T4的检测限和定量限分别为0.1μg/ml和0.8μg/ml。校准曲线在1-100μg/ml范围内呈线性；D-T4的平均相关系数r（D-T4）=0.9986，L-T4的平均相关系数r（L-T4）=0.9978。在最佳条件下，T4对映体在基线处分离。L-T4在D-T4之前洗脱。使用高效液相色谱法测定了45例甲状腺疾病患者（甲亢、甲减、甲状腺切除、甲状腺肿或甲状腺炎）血清中D-T4和L-T4的浓度，结果表明不同甲状腺疾病患者血清中D、L-T4浓度存在差异。临床治疗甲状腺疾病时应注意这一结果。
