高效液相色谱法测定黄芩素葡萄糖醛酸化产物的浓度

Determination on the concentration of metabolites of baicalein in liver microsomes incubation systems by HPLC

目的建立黄芩素葡萄糖醛酸化代谢产物B-7G和B-6G的高效液相色谱方法。方法用Kromasil C18柱（4．6mm×150mm，5μm），柱温40℃，流动相为乙腈（A）和0．2％磷酸水溶液（B），梯度洗脱，检测波长270nm，流速1mL·rain-1，进样量20乩。结果黄芩素葡萄糖醛酸化产物B一7G和B一6G分别在0．1～1000．0，0．1～500．0μmol·L。具有良好的线性关系，r分别为0．9993和0．9995。日间、日内精密度（RSD）均小于10％，回收率和稳定性均符合要求。结论本研究方法简便、快速、准确，适用于大样本连续分析，可以为黄芩素的体外代谢研究提供平台。

Objective To establish a reliable analytical method for the study of baicalein glucuronidation metabolism by a high performance liq- uid chromatography （HPLC） system. Methods Liquid chromatography was performed on a Kromasil C18 column （4. 6 mm × 150 mm,5 μm）. The column temperature was set at 40 ℃. The mobile phase was a mixed system consistingof acetonitrile and 0. 2% phosphoric acid aqueous solu- tion with gradient elution and a flow - rate 1 mL · min-1 The wave- length was at 270 nm and the volume of sample was 20 μL. Results The calibration curves of B -7G and B -6G were linear over a concen- tration of 0. 1 - 1000. 0 μmol · L-1 and 0. 1 - 500. 0 ·mol · L-1 （ r 〉 0. 999）, respectively. The intra- day RSD and inter- day RSD were both less than 10%. The extraction recovery rates were in line with the experimental requirements. Conclusion The method is convenient, sensitive, accurate, and applicable for continous analysis of large sam- ples. It can provide platform for baicalein glucuronidation metabolism studies in vitro.