高效液相色谱法测定A549细胞内新藤黄酸含量方法的建立

目的:建立高效液相色谱法测定肿瘤细胞内新藤黄酸含量的方法.方法:应用高效液相色谱法测定细胞内新藤黄酸的含量.色谱柱为COSMOSIL C18柱,流动相为甲醇-0.1％磷酸水溶液(90:10 V:V),流速为1.0 mL/min,柱温为30℃,检测波长为360 nm,进样量为20μL.结果:在0.05～30μg/mL内新藤黄酸样品溶液质量浓度与峰面积比线性关系良好(P<0.01).精密度、稳定性的RSD<10％,准确度均在实测值的10％以内,特异性良好,同一浓度的新藤黄酸孵育相同时间样品测定结果重复性好.结论:该方法简单方便,准确度高,适用于定量测定肿瘤细胞内新藤黄酸的含量.

Establishment of HPLC method measuring the gambogenic acid in A459 cells

Objective:To establish an HPLC method for the content determination of gambogenic acid in tumor cells. Methods:The gambogenic acid in cells was measured using HPLC. The chromatographic column was COSMOSIL C18 column,the mobile phase was 90％ methanol:0. 1％ aqueous phosphoric acid(90:10,V:V),the flow rate was 1. 0 mL/min,the column temperature was 30 ℃,the detection wavelength was 360 nm,and the injection volume was 20 μL. Results:The linear relationship between the 0. 05 to 30 μg/mL of gambogenic acid and peak area was good(P<0. 01). The precision and stability of RSD was lower than 10％. The accuracy of the measure value was less than 10％,the specificity was good. The repeatability of sample determination at the same time and concentration of gambogenic acid was good. Conclusions:The method is simple, convenient and accurate, which is suit for the determination of gambogenic acid in tumor cells.