高效液相色谱法测定苯环喹溴铵在Calu-3细胞中的摄取  

Determination of Bencycloquidium Bromide in Calu-3 Cells by HPLC

作　　者：邓万定[1] 金方[1] 张启明[2] 

机构地区：[1]中国医药工业研究总院上海医药工业研究院,上海呼吸系统药物工程技术研究中心,上海200437 [2]中国食品药品检定研究院,北京100050

出　　处：《中国药学杂志》2013年第14期1187-1190,共4页Chinese Pharmaceutical Journal

基　　金：化学新药质量标准研究与评价技术平台资助项目(2011ZX09303-001)

摘　　要：目的建立苯环喹溴铵反相高效液相色谱法,研究其在Calu-3细胞上的摄取特征。方法色谱条件:Discovery C18柱(4.6 mm×250 mm,5μm),流动相为乙腈-水(0.27%磷酸二氢钾,0.3%三乙胺,稀磷酸调节pH至3.0)(35∶65),流速为1.0mL·min-1,检测波长210 nm,柱温25℃。考察了药物浓度、时间对BCQB在Calu-3细胞上摄取的影响。结果 BCQB在0.010 2～1.022 0μg·mL-1内呈良好的线性关系(r=0.999 8),最低定量限为0.010 2μg·mL-1。BCQB在Calu-3细胞中的摄取30 min达峰,后Calu-3细胞中BCQB的含量逐渐下降。浓度处于20～500μmol·L-1之间时,Calu-3细胞对BCQB的摄取量逐渐增加,浓度达到500μmol·L-1后,随着剂量的增加BCQB的摄取量变化不明显。结论该法快速、灵敏、简单。BCQB可被Calu-3细胞迅速摄取,随着剂量的增加,BCQB在Calu-3细胞中的摄取存在饱和现象。ObjectiveTo establish an HPLC assay to determine the uptake characteristics of bencycloquidium bromide （BCQB） in Calu-3 cells. Methods The samples were analyzed with a Discovery C18 column （4.6 mm×250 mm, 5μm） at 25 ℃. The mobile phase was acetonitrile-water containing 0.27% KH2PO4 and 0.3% triethylamine （pH adjusted to 3.0 with H3PO4） （35∶65）. The flow rate was 1.0 mL·min-1. The concentrations of BCQB in Calu-3 cells at various time were measured. Results The linearity of BCQB calibration curve was good in the range of 0.010 2-1.022 0 μg·mL-1 （r=0.999 8）. The minimal quantitation limit was 0.010 2 μg·mL-1. The uptake of BCQB increased rapidly during the first 30 min. When the concentration of BCQB increased from 20 to 500 μmol·L-1, the uptake increased almost linearly. However, the curve showed saturation when the concentration of BCQB reached 500 μmol·L-1. Conclusion This assay is rapid, sensitive and specific, and can be applied to determine BCQB concentrations in Calu-3 cells. BCQB can be uptaken by Calu-3 cells rapidly, and the uptake shows saturation with increase of dosage.

关 键 词：苯环喹溴铵 Calu-3细胞 高效液相色谱法 摄取 

分 类 号：R917[医药卫生—药物分析学]