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运用高效液相色谱手性流动相法(HPLC-CMP)对影响甲状腺素对映体(D-,L-T4)分离方法的因素:三乙胺(TEA)浓度,流动相pH值,铜离子(Cu2+)浓度,L-脯氨酸(L-pro)浓度,柱温以及流动相的流速进行了系统的研究。同时,考察了色谱方法分离T4对映体的线性关系,精密度和准确度。线性响应范围为0.6~3.2 nmol (D-,L-T4),线性相关系数为rD-T4=0.9980,rL-T4=0.9990,日内和日间的精密度分别为RSD<2.3%(n=6),RSD<3.15%(n=5)。结果表明本实验所得的色谱条件较文献报道的优越,分离条件简单,重现性好。HPLC-CMP法测定甲状腺素对映体其意义在于该方法可为定量测定药品及人体血液中D-,L-T4两种异构体,为治疗药物监测(TDM)和药物不良反应监测(ADRs)提供了依据。
The factors influencing the separation of thyroxine enantiomers (D-, L-T4) by high performance liquid chromatography (HPLC): triethylamine (TEA) concentration, mobile phase pH Cu2 + concentration, L-pro concentration, column temperature and mobile phase flow rate were systematically studied. At the same time, the linear relationship, precision and accuracy of chromatographic separation of T4 enantiomers were investigated. The linear response range was 0.6-3.2 nmol (D-, L-T4). The linear correlation coefficients were rD-T4 = 0.9980 and rL-T4 = 0.9990. And daytime precision were RSD <2.3% (n = 6) and RSD <3.15% (n = 5), respectively. The results showed that the chromatographic conditions obtained in this experiment superior to the literature reported, the separation conditions are simple, reproducible. The significance of HPLC-CMP for the determination of thyroxine enantiomers is that the method can be used for the quantitative determination of D-, L-T4 two isomers in the blood of patients and the human body for the monitoring of therapeutic drugs (TDM) and adverse drug reactions (ADRs) provided the basis.