HPLC-DAD法测定铁观音中芦丁的含量

The Determination of Rutin Content in Tie Guanyin (铁观音) by HPLC-DAD

目的:建立HPLC-DAD法测定铁观音中芦丁的含量。方法:以芦丁为指标,采用HPLC法测定铁观音中的芦丁含量。色谱条件为:Phenomenex Luna-C_(18)(250 mm×4.6 mm,5μm)色谱柱;以乙腈-0.4%磷酸为流动相进行梯度洗脱;进样量20μL;检测波长为354 nm;柱温为30℃。并选择提取溶剂、料液比、超声功率和超声时间为考察因素,用单因素试验确定因素水平。在单因素试验的基础上,采用正交试验优化铁观音中芦丁的超声提取条件。结果:铁观音中芦丁含量在10.570~105.700μg·m L^(-1)的浓度范围内呈良好的线性关系(r=0.9997)。铁观音样品中芦丁成分的平均含量为0.931 mg·g^(-1)。铁观音中芦丁的最佳超声提取条件:提取溶剂为80%乙醇,料液比1∶35,超声功率350 W,超声提取时间40 min。结论:该优选提取方法提取率高;HPLC-DAD法条件稳定、可控、重复性好。研究结果可为铁观音的质量控制及进一步开发提供参考。

Objective： To determine the content of rutin in Tie Guanyin by HPLC-DAD. Methods： With the content of rutin as index, HPLC was used to determine the content of rutin. Phenomenex Luna - C18（250 mm×4.6 mm,5μm） was used, the mobile phase consisted of acetonitrile and 0.4% phosphoric acid with gradient elution; the injection volume was 20μL： column temperature was 35 ℃： the detective wavelength was 354 nm. Extraction solvent, ratio of liquid-material, ultrasonic power and ultrasonic time were selected as factors; factors and levels were determined by single-factor test; Orthogonal test was used to optimize extraction conditions on the base of single-factor test. Results： The rutin in Tie Guanyin showed good linearity （r=0.9997） in a relatively wide concentration range （10.570-105.700μg·mL-1）. The average content of rutin in Tie Guanyin sample is 0.931 mg·g-1 The optimum extraction conditions were as follows： the extraction solvent was 80 % ethanol; The solid-liquid ratio was 1：35; ultrasonic power was 350 W and ultrasonic time was 40 minutes. Conclusion： The optimized conditions was reliable with high extraction rate; The HPLC-DAD method is stable, reliable and precise. The results can provide reference for quality control and further development of Tie Guanyin.