HPLC法测定大鼠血浆中EGCG的血药浓度

目的:建立快速、高效、灵敏的HPLC法测定大鼠血浆中的EGCG血药浓度。方法:以睾丸酮为内标物,血浆经盐酸和乙酸乙酯等去除蛋白。采用Agilent 20RBAX SB-C18色谱柱,乙腈-0.3%乙酸为流动相,流速1.0ml/min,检测波长为280nm,柱温40℃。结果:EGCG和内标的出峰时间分别为12.21min和15.4min。EGCG在0.05μg/ml-100μg/ml范围内线性关系良好(R2=0.9920),高、中、低3种浓度下的日内、日间精密度RSD均低于15%,相对回收率与绝对回收率均在100±15(%)范围内,稳定性好。结论:本方法灵敏、准确、高效,适用于大鼠血浆中EGCG的血药浓度检测。

Objective:A rapid, effective, sensitive HPLC method was develop for detecting EGCG in SD rat plasma. Method: The analytes in plasma were extracted for protein precipitation using hydrochloric and ethyl acetate, with testosterone as the internal standard. The analytes was separated at Agilent 20RBAX SB-C18 column whereas acetonitrile-0.3% acetic acid was the mobile phase. The flow rate was 1.0ml/min. The UV detection wavelength was 280nm. The column temperature was 40℃. Results: the retention time of EGCG and internal standard were 12.2min and 15min, respectively. The calibration curve were linear in the range of 0.05μg/ml-100μg/ml(R2=0.9920). The intra- and inter- day variations were less than 15%. Conclusion: the established method is sensitive, accurate and high performance for detecting the concentration of EGCG in SD rat plasma.