LC-MS_MS法测定裸鼠肝脏中6-羟基爵床定A的含量.docVIP

  LC-MS/MS 法测定裸鼠肝脏中 6-羟基爵床 定 A 的含量# 覃禹1,2，仇峰1，罗祖良1,2，毛晓丽2，毕明刚1，杨美华1，覃洁萍2** 5 10 15 20 25 30 35 40 （1. 中国医学科学院药用植物研究所，北京 100193； 2. 广西中医学院，南宁，广西 530001） 摘要：目的：建立测定裸鼠肝组织中 6‘-羟基爵床定 A（HJA）浓度的 LC-MS/MS 法，并 用于其裸鼠组织分布研究。方法：色谱柱为 Agilent Zorbax SB-C18 柱（50 mm × 2.1 mm， 3.5 μm）。流动相为水（含 0.1%甲酸）和乙腈（含 0.1%甲酸），梯度洗脱，流速 0.5 mL · min-1，柱温为室温。内标为丁螺环酮。采用 ESI 源，以多反应监测（MRM）方式分别监 测离子对 m/z 410.8→136.9（6‘-羟基爵床定 A）和 m/z 386.1→122.0（内标丁螺环酮）。肝 组织匀浆后采用蛋白沉淀法进行预处理。结果：6‘-羟基爵床定 A 在裸鼠肝组织中的线性范 围为 1.0-2000 ng/mL，日内精密度（RSD%）小于 8.61%；日间精密度（RSD%）小于 8.15%， 回收率在 89.3~98.3%范围内。结论：该方法简单、灵敏、准确，适用于裸鼠肝组织中 6‘- 羟基爵床定 A 的浓度测定及其组织分布研究。 关键词：6‘-羟基爵床定 A；LC-MS/MS；肝脏；裸鼠 中图分类号：R917 Determination of 6-Hydroxy Justicidin A in Nu/Nu Mice Liver by Liquid Chromatography Tandem Mass Spectrometry QIN Yu1,2, QIU Feng1, LUO Zuliang1,2, MAO Xiaoli2, BI Minggang1, YANG Meihua1, QIN Jieping2 (1. Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Beijing 100193, China; 2. Guangxi Traditional Chinese Medical University, Nanning 530001, China) Abstract: Objective: To develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) for the determination of 6-Hydroxy Justicidin A (HJA) in nu/nu mice liver. Method: An Agilent Zorbax SB-C18 column (50 × 2.1 mm I.D.，3.5 μm) was used with water (0.1% formic acid)-acetonitrile (0.1% formic acid) as the mobile phase at a flow rate of 0.5 mL· min-1. The column temperature was set at room temperature。LC-MS/MS was performed in the multiple reaction monitoring (MRM) mode using target ions at m/z 410.8→136.9 (6-Hydroxy Justicidin A) and m/z 386.1→122.0 (Buspirone, IS). The nu/nu mice liver homogenates were treated using protein precipitation method. Result: The calibration curve of 6-Hydroxy Justicidin A was linear in the range of 1.0-2000 ng/mL in liver homogenate. Inter-day RSDs was less than 8.15% and intra-day RSDs was less than 8.61%. The recovery was calculated to be 89.3~98.3 % in the liver. Conclusion: The method is simple, sensitive and accurate for the