新型胆碱酯酶抑制剂J39136大鼠体内血药浓度测定方法的建立及组织分布研究

Establishment of an HPLC-DAD method to quantify a new cholinesterase inhibitor J39136 in rat plasma and tissues and study on its tissue distribution

目的:建立一种稳定、灵敏的高效液相色谱法,检测新型胆碱酯酶抑制剂异黄酮衍生物J39136的血药浓度,考察其在大鼠体内的药动学特征,为临床药动学研究提供参考。方法:先导化合物J39136大鼠血浆样品及组织样品经乙酸乙酯萃取处理后,采用HPLC-DAD法检测。色谱柱为Agilent SB-C18柱(150 mm×4.6mm,5μm),流动相为乙腈-0.2%磷酸溶液(25∶75),流速为1 mL·min-1,检测波长为250 nm,柱温为30℃。结果:本实验所建立的测定大鼠血浆及组织中J39136含量的HPLC-DAD法,在一定浓度范围内线性良好,r>0.999。日内、日间精密度、回收率、稳定性均符合要求,并成功地应用于检测大鼠体内血浆J39136浓度和组织分布。结论:本实验所建立的检测大鼠体内血浆J39136浓度的方法具有较好的准确度和灵敏度,方法可靠,可用于该化合物的体内药动学研究,对于其类似物的血药浓度检测将提供重要参考信息。

Objective：To establish an HPLC-DAD method to quantify J39136 in rat plasma and different tissues, which is an isoflavone derivative with potent cholinesterase inhibitory activity. Methods：Chromatographic separation was achieved on a reversed phase-HPLC column（Agilent SB-C18,150 mm×4.6 mm,5 μm）. The UV absorbance at 250 nm was recorded. A mixture of acetonitrile-0.2% phosphoric acid aqueous solution（25∶75）was used as the mobile phase with a flow rate at 1 mL？min-1 at 30℃. Results：The calibration curve was linear （r〉0.999）. The intra- and inter-day precisions,and recovery and stability of J39136 in plasma were in accordance with the requirements. This HPLC method was successfully applied to assess the distribution of J39136 in tissues of Sprague-Dawley rats. Conclusion：This HPLC method is accurate,sensitive,and reliable for quantifying the flavonoid derivative J39136 in rat plasma,which is suitable to investigate the pharmacokinetics of J39136 and will provide valuable information for the quantitative analysis of its analogs in plasma. 