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高效液相色谱法测定小鼠肝脏中反式-白藜芦醇含量

姚倩, 侯世祥, 何希辉, 张瑄, 颜军, 苟小军*   

  1. 1. 成都大学 四川省药用微生物资源重点实验室, 四川 成都 610106;
    2. 四川大学华西药学院 药剂学系, 四川 成都 610041;
    3. 北京大学药学院 化学生物学系, 北京 100083
  • 收稿日期:2007-12-10 修回日期:2008-05-10 出版日期:2008-06-15 发布日期:2008-06-15
  • 通讯作者: 苟小军*

Determination of trans-resveratrol in mouse liver by high performance liquid chromatography

Qian Yao, Shi-Xiang Hou, Xi-Hui He, Xuan Zhang, Jun Yan, Xiao-Jun Gou*   

  1. 1. Key Laboratory of Sichuan Province of Medicinal Microorganism Resources, Chengdu University, Chengdu 610106, China;
    2. Department of Pharmaceutics, West China School of Pharmacy, Sichuan University, Chengdu 610041, China;
    3. Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China
  • Received:2007-12-10 Revised:2008-05-10 Online:2008-06-15 Published:2008-06-15
  • Contact: Xiao-Jun Gou*

摘要:

建立一灵敏的HPLC, 用于测定小鼠肝脏中反式-白藜芦醇含量。取小鼠全部肝脏制成匀浆, 用乙酸乙酯提取。分离有机层并挥干, 残渣用0.2 mL流动相溶解, 离心, 取上清液50 μL注入高效液相色谱仪。样品在岛津ODS (150mm × 4.6 mm, 5 μm)上分离, 柱温35 °C, 检测波长305 nm, 流动相为甲醇-0.1 mol/L乙酸溶液(4:6, v/v), 流速为1 mL/min。以信噪比3:1, 白藜芦醇在小鼠肝脏中的检测限为3.0 ng/g; 标准曲线的线性范围为5.0120.0 ng/g; 浓度为61080 ng/g的平均回收率分别为102%96.0%91.2%; 日内、日间精密度的RSD均小于5%。与已报道的其它白藜芦醇生物样品测定方法相比较, 本法更为快速与灵敏, 并具有良好的线性、选择性、准确度与精密度, 可用于小鼠肝脏中白藜芦醇的药物动力学研究。

关键词: 高效液相色谱法, 高效液相色谱法, 高效液相色谱法, 白藜芦醇, 白藜芦醇, 白藜芦醇, 小鼠肝脏, 小鼠肝脏, 小鼠肝脏, , ,

Abstract:

To develop a sensitive high performance liquid chromatography (HPLC) assay for the determination of trans-resveratrol in mouse liver. The whole liver of a mouse was removed from the body, homogenated, and extracted by ethyl acetate. The organic layer was isolated and evaporated to dryness, the residue was reconstituted in 0.2 mL mobile phase for centrifugation, and 50 μL of the supernatant was injected into the HPLC instrument. The sample was separated on a Shimadzu ODS column (150 mm × 4.6 mm, 5 μm) at 35 °C and detected by ultraviolet (UV) detector at the wavelength of 305 nm. The mobile phase consisted of methanol and 0.1 mol/L acetic acid (4:6, v/v) with the flow-rate at 1 mL/min. The limit of detection was 3.0 ng/g in liver homogenate with a signal/noise ratio of 3:1. The linear range of the calibration curve was 5.0-120.0 ng/g. The mean recoveries at the concentrations of 6, 10 and 80 ng/g were 102%, 96.0% and 91.5%, respectively. The RSDs for inter- and intra-day assays were less than 5%. Compared with other reported methods, this method was faster and more sensitive. It was also proved to be of good linearity, selectivity, accuracy and precision, and can be efficiently applied to the pharmacokinetic study of trans-resveratrol in mouse liver.

Key words: High performance liquid chromatography, High performance liquid chromatography, Trans-resveratrol, Trans-resveratrol, Mouse liver, Mouse liver, ,

中图分类号: 

Supporting: The authors thank Postdoctoral Scientific Research Station of Gansu Yasheng Groups for the financial and technical supports.