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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 establish a HPLC-DAD method for the determination of rutin in Tieguanyin. Methods: Rutin was used as an index to determine the content of rutin in Tieguanyin by HPLC. The chromatographic conditions were as follows: Phenomenex Luna-C 18 column (250 mm × 4.6 mm, 5 μm); gradient elution with acetonitrile-0.4% phosphoric acid as mobile phase; injection volume 20 μL; detection wavelength at 354 nm; Is 30 ° C. Solvent extraction, solid-to-liquid ratio, ultrasonic power and ultrasonic time were selected as the investigation factors, and single factor test was used to determine the level of the factors. On the basis of single factor experiment, the orthogonal experiment was used to optimize the ultrasonic extraction condition of rutin in Tieguanyin. Results: The rutin content in Tieguanyin showed a good linearity (r = 0.9997) within the range of 10.570 ~ 105.700μg · m L ~ (-1). The average content of rutin in Tieguanyin sample was 0.931 mg · g -1. Tieguanyin rutin best ultrasonic extraction conditions: 80% ethanol extraction solvent, solid-liquid ratio 1:35, ultrasonic power 350 W, ultrasonic extraction time 40 min. Conclusion: The extraction rate of this extraction method is high. The HPLC-DAD method is stable, controllable and reproducible. The results can provide reference for the quality control and further development of Tieguanyin.