高效液相色谱-荧光法测定大鼠血浆中罗丹明123浓度

Determination of Rhodamine 123 in Rat Plasma by HPLC-Fluorescent

目的测定罗丹明123(R123)的血药浓度。方法采用高效液相-荧光检测法,色谱柱为BTD4703Packing C18柱,流动相为乙腈-1%三乙胺(28:72),流速1.0 mL.min-1;荧光检测器波长为485 nm(激发波长),546 nm(发射波长)。结果R123检测浓度在15.625～500.000 ng.mL-1范围内线性关系良好(r=0.999 7),最低检测限为5.000 ng.mL-1。血浆样品的提取回收率为(83.05±2.71)%。日内与日间RSD均<8%。样品提取后24 h内及3次冻融稳定性良好。结论该方法操作简便,灵敏度和精确度高,重复性好,适于血浆R123浓度测定和药动学研究。

Objective To establish a HPLC-fluorescent detector method for determining the concentration of rhodamine 123 in plasma. Methods The chromatographic separation was carried out on BTD4703 Packing C18 column with mobile phase consisted of acetonirile-1% triethylamine （28：72） and a flow rate of 1.0 mL · min^-1. The Ex was set at 485 nm and Em was set at 546 nm. Results The linear range of R123 was 15. 625 - 500. 000 ng · mL^-1 （ r = 0. 999 7 ）, with the lowest detectable limit at 5. 000 ng · mL-1. The recovery of extraction was （83.05 - 2.71 ）% . Both the intra-day RSD and interday one were less than 8%. The sample showed a good stability within 24 hours after extraction and three freeze-thaw cycling.Conclusion The method is simple, sensitive, accurate and reproducible, and which is suitable for pharmacokinetic study of R123 in rat plasma.