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,5 μm),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1.0 mL·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 0 μg·mL^-1内呈良好的线性关系(r=0.999 8),最低定量限为0.010 2 μg·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20~500 μmol·L^-1之间时,Calu-3细胞对BCQB的摄取量逐渐增加,浓度达到500 μmol·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.