目的:建立筛选分析方法评估中药水煎液对铜离子的络合能力,以揭示中药治疗铜离子代谢障碍疾病的物质基础。方法:采用1-(2-吡啶偶氮)-2-萘酚(PAN)为络合剂,乙醇为助溶剂,二氯甲烷为萃取剂,建立分光光度法于600 nm处测定中药水煎液中铜离子的含量。通过外加一定量铜离子来测定回收率。此回收率与铜离子被络合能力负相关,以此评估中药水煎液与铜离子的络合强度。结果:铜离子质量浓度在0.64-6.4 mg·L-1的范围内线性关系良好,重复性较好,检测下限为0.046 mg·L-1。给中药煎液外加铜离子的回收率测定显示出很好的准确性和重复性。以此方法评估了10味中药水煎液对铜离子的络合能力,其中茵陈、黄芩、升麻和鱼腥草4味中药对Cu2+的络合能力较强(56.19%-69.00%)。结论:本文建立的方法能够很好地应用于中药煎液对铜离子络合能力的评估,可用于中药治疗铜离子代谢障碍疾病物质基础的研究。
Objective: To establish an in vitro screening method to evaluate the chelating ability of constituents in decoction of Chinese materia medica ( CMM ) with copper ions. Methods: The copper ions in the decoction ofCMM were chelated with 1- ( 2-pyridylazo ) -2-naphthol ( PAN ), extracted with methylene dichloride-ethanol, and finally determined by spectrophotometry at 600 nm. Then, certain amount of copper ions were added to the decoction, and the recovery of the added amount of copper ions was determined. The recovery was negatively correlated with chelating ability of constituents in decoction of CMM with copper ions. Results: Trace amounts of Cu2+ ( 0. 64-6.4 mg. L-1 ) were determined with good accuracy and reproducibility; the detection limit was about 0. 046 mg. L-1. The recovery of added copper ions in the decoction of CMM was determined with good accuracy and reproducibility. The chelating ability of 10 CMM with copper ions was evaluated. The chelating strength of Artemisia capiUaris Thunb. , Scutellaria baicalensis Georgi. , Cimicifuga foetida L. and Houttuynia cordata Thunb. were much higher ( 56. 19%-69.00% ). Conclusion: The developed method can be used to evaluate the chelating ability of constituents in decoction of CMM with copper ions, and thus can be applied in research of the material basis of the treatment of copper ion metabolism disorders by traditional Chinese medicine.