Saracino Maria Addolorata, Bugamelli Francesca, Conti Matteo, Amore Mario, Raggi Maria Augusta
Laboratory of Pharmaco-Toxicological Analysis, Department of Pharmaceutical Sciences, Faculty of Pharmacy, Alma Mater Studiorum, University of Bologna, Bologna, Italy.
J Sep Sci. 2007 Sep;30(14):2249-55. doi: 10.1002/jssc.200700110.
A liquid chromatographic method with diode array detection (DAD) has been developed for the analysis of the antiepileptic agent lamotrigine (LTG) and its metabolites, lamotrigine 2-N-glucuronide and 2-N-methylated in plasma samples. The analytes were separated on a C8 RP column, using a mobile phase composed of methanol and a 0.45 mM, pH 3.5 phosphate buffer containing 0.17% triethylamine (24:76 v/v). Melatonin was used as the internal standard (IS). The DAD detector was set at 220 nm for the detection of all the analytes. A simple protein precipitation with methanol guaranteed high extraction yield values (>90%) and good purification from matrix interference. Good linearity was obtained in the 0.1-15.0 microg/mL range for LTG and lamotrigine 2-N-glucuronide and in the 0.1-2.0 microg/mL range for lamotrigine 2-N-methylated. The analytical method was validated in terms of precision, extraction yield, and accuracy. These assays gave RSD% values for precision always lower than 4.3% and mean accuracy higher than 80%. The method seems to be suitable for the analysis of plasma samples from patients treated with Lamictal.
已开发出一种采用二极管阵列检测（DAD）的液相色谱法，用于分析抗癫痫药物拉莫三嗪（LTG）及其代谢产物，即血浆样品中的拉莫三嗪2-N-葡萄糖醛酸苷和2-N-甲基化产物。分析物在C8反相柱上进行分离，使用由甲醇和含0.17%三乙胺（24:76 v/v）的0.45 mM、pH 3.5磷酸盐缓冲液组成的流动相。褪黑素用作内标（IS）。DAD检测器设置在220 nm用于检测所有分析物。用甲醇进行简单的蛋白沉淀可保证高提取率值（>90%），并能有效净化基质干扰。拉莫三嗪和拉莫三嗪2-N-葡萄糖醛酸苷在0.1 - 15.0 μg/mL范围内、拉莫三嗪2-N-甲基化产物在0.1 - 2.0 μg/mL范围内获得了良好的线性关系。该分析方法在精密度、提取率和准确度方面进行了验证。这些测定的精密度RSD%值始终低于4.3%，平均准确度高于80%。该方法似乎适用于分析服用拉莫三嗪（Lamictal）患者的血浆样品。
