浊点萃取-高效液相色谱法测定小儿泻速停颗粒中多酚类化合物

Cloud point extraction for the determination of polyphenols in xiaoer xiesuting pellets by high performance liquid chromatography

建立了基于表面活性剂浊点萃取-反相高效液相色谱法同时分离测定小儿泻速停颗粒中没食子酸、儿茶素、表儿茶素的方法。采用phenomenex C18色谱柱,流动相为乙腈-0.2%冰乙酸水溶液,采用梯度洗脱;流速0.8 mL/min;检测波长280 nm。没食子酸、儿茶素和表儿茶素分别在0.0104～223μg/mL,(r=0.9999);0.0152～108μg/mL,(r=0.9999);0.0184～132μg/mL,(r=0.9999)范围内线性关系良好,定性检测限(S/N=3)依次为:0.087 ng/mL,0.43 ng/mL,0.42 ng/mL。加入50 g/L NaCl来进行浊点富集,可提高萃取回收率。平均回收率为91.85%～98.48%,本方法可用于该制剂的质量控制。

A method based on cloud point extraction was developed for the simultaneous separation and determination of gallic acid, catechin, epicatechin from Xiaoer Xiesuting Pellets by reversed-phase high performance liquid chromatography （RP-HPLC）. Three compounds （gallic acid, catechin and epicatechin ） from the Triton X-100 extract have been isolated by using a Phenomenex C18 column with a gradient elution, the mobile phase was acetonitrile-O. 2% acetic acid. The flow rate was 0. 8 mL/min, and UV detection wavelength was set at 280 nm. Chromatographic condition was investigated, and the extraction process was optimized. The calibration curves for gallic acid, catechin and epicatechin were linear between 0. 0104 -223 μg,/mL, 0. 0152 - 108 μg/mL and 0. 0184 - 132 μg,/mL （ r = 0. 9999 ）, respectively. The limits of detection （ LOD ） were 0. 087 ng/mL, 0. 43 ng,/mL, and 0. 64 ng/mL, respectively. Under optimum conditions： 5% of Triton X-100 surfactant （ W/V）, solid/liquid ratio of 1 ： 100 （ g/mL）, ultrasonic-assisted extraction at 30 ℃ for 30 rain, the extraction yield reached the highest value. 5% sodium chloride was added to the solution to facilitate the phase separation and increase the extraction recovery. The average recoveries were 91.85 - 98.48%. Conclusion： The method is simple, rapid, sensitive, low cost and environment-friendly, and suitable for use in the quality control of Xiaoer Xiesuting Pellets.