Simultaneous determination of isoline and its two major metabolites using high-performance liquid chromatography.
Simultaneous determination of isoline and its two major metabolites using high-performance liquid chromatography.
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DOI：
10.1093/jat/28.1.11
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发表时间：
2004
影响因子：
2.5
通讯作者：
Jun Tang;Mian Zhang;Zheng-Tao Wang;T. Akao;N. Nakamura;M. Hattori
中科院分区：
文献类型：
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作者：
Jun Tang;Mian Zhang;Zheng-Tao Wang;T. Akao;N. Nakamura;M. Hattori
建立了一种简便、可靠的高效液相色谱(HPLC)同时测定吡咯里西丁生物碱异丙氨酸及其两种主要代谢物M1(双线)和M2(双线内酯)的方法。后两种代谢产物是在大鼠和小鼠微粒体酶系统对异丙氨酸的体外代谢过程中产生的。通过将经等体积冰冷甲醇处理的微粒子反应混合物的上清液的等量直接进样到常规反相分析柱(150 x 4.6 mm)上进行分析。采用梯度洗脱，流动相A(0.01M二氢磷酸二氢钾，pH 4.8)和B(乙腈)。该方法表现出极好的精密度和准确度，日内和日间变异小于10%，总准确度高于94%。所建立的高效液相色谱方法成功地用于大鼠和小鼠肝微粒体孵育产生的完整等值线及其两种pH和热不稳定代谢物的测定。
A simple and reliable high-performance liquid chromatographic (HPLC) assay was developed for a simultaneous determination of isoline, a potent hepatotoxic pyrrolizidine alkaloid, and its two major metabolites, namely M1 (bisline) and M2 (bisline lactone, a new pyrrolizidine alkaloid). The latter two metabolites were produced during in vitro metabolism of isoline by rat and mouse microsomal enzyme systems. The analysis was conducted by a direct injection of aliquots of supernatant of the microsomal reaction mixture treated with the equal volume of ice-cold methanol onto a conventional reversed-phase analytical column (150 x 4.6 mm). The analytes were separated by a gradient elution with mobile phases A (0.01 M dihydro-potassium phosphate, pH 4.8) and B (acetonitrile). The assay has shown excellent precision and accuracy with less than 10% of overall intra- and interday variations and higher than 94% of overall accuracy. The developed HPLC method was successfully applied for the determination of the intact isoline and its two pH- and thermally labile metabolites produced in rat and mouse liver microsomal incubations.