胶束萃取-高效液相色谱法同时测定葛根粉中5种异黄酮

Simultaneous Determination of Isoflavonoids in Kudzu Powder by High Performance Liquid Chromatography Coupled with Micelle-mediated Extraction

目的:基于表面活性剂作为萃取剂,应用胶束萃取-反相高效液相色谱法测定葛根粉中葛根素、大豆苷、染料木苷、大豆苷元和染料木素5种异黄酮的含量。方法:以非离子表面活性剂Triton X-114为萃取剂,采用Phenomenex C18色谱柱,甲醇-体积分数0.25%冰醋酸溶液为流动相,梯度洗脱,利用紫外检测器在250nm波长处进行检测。考察色谱测定条件,优化胶束萃取和浊点预富集参数。结果:采用3g/100mL Triton X-114表面活性剂,液固比为100:1(mL/g),40℃超声萃取30min,所得的萃取率最大。在表面活性剂提取液中,加入一定量的氯化钠进行浊点预富集,可以提高萃取率和预富集因子。该方法的定性检测限范围是0.016～0.079μg/mL(RSN=3),平均回收率为93.7%～98.86%,预富集因子平均值可达到12.5。结论:该方法简便快速、灵敏可靠、绿色环保,适用于葛根粉中葛根素等5种异黄酮成分含量的测定和产品质量控制。

Objective：A reverse-phase high performance liquid chromatography（RP-HPLC） coupled with micelle-mediated extraction method was developed for the simultaneous determination of five isoflavonoids such as puerarin,daidzin,daidzein,genistin and genistein from kudzu powder.Methods：The non-ionic surfactant Triton X-114 was used as the extraction solvent.The chromatographic separation of samples was performed on a Phenomenex C18 column using methanol-0.25% glacial acetic acid as the mobile phase by means of gradient elution and a UV detector set at 250 nm was used for detection.The chromatographic conditions were explored to optimize the micelle-mediated extraction and enrichment of cloud points.Results：The highest extraction rate of isoflavonoids was achieved using 3 g/100 mL Triton X-114 as the surfactant for ultrasonic-assisted extraction at 40 ℃ for 30 min with a material/liquid ratio of 1：100（g/mL）.Sodium chloride was added to the extract to improve extraction rate and cloud point enrichment.The limit of detection（LOD） of this developed method was in the range of 0.016-0.079μg/mL（RSN=3） with average recovery rates ranging between 93.7% and 98.86%.The enrichment factor was approximately 12.5.Conclusion：This developed method is characteristics of simple operation,low cost,high extraction efficiency and less pollution to environment,which is suitable for the quality control of five isoflavonoids in kudzu powder.