高效液相色谱法测定离子导入后兔眼房水中哌仑西平浓度

Detemination of pirenzepine in the aqueous humor of rabbits after iontophoresis by HPLC

目的HPLC法测定兔眼房水中哌仑西平的浓度,离子导入法促进哌仑西平的吸收.方法氰基键合硅胶为固定相,流动相为甲醇/0.014mol·L-1三乙胺(pH=5.0)=70/30,检测波长为280nm,丙咪嗪为内标.结果哌仑西平浓度在0.2～10μg·mL-1浓度内线形良好(r=0.999 0),日内RSD为1.20%,日间RSD为1.52%,低、中、高浓度回收率分别为(97.92±2.83)%、(99.19±2.64)%、(101.3±0.94)%,定量限为0.20μg·mL-1,检测限为0.10μg·mL-1.离子导入能明显提高哌仑西平的透过率,pH值对哌仑西平的离子导入有极显著影响.黏度、浓度、电极种类和电流强度对其影响均不是很明显.结论所建立的HPLC法测定兔眼房水中哌仑西平的含量方法准确可靠.

OBJECTIVE To develop an HPLC method to determine the concentrations of pirenzepine in the aqueous humor of rabbits. To investigate the efficiency of pirenzepine permeation enhanced by iontophoresis. METHODS The concentrations of pirenzepine were determined by HPLC using an Inertsil CN-3 column （250mm × 4.6mm,5μm ）. The mobile phase was a mixture of 70% methanol and 30% 0. 014mol · L^-1 triethylamine adjusted to a pH of 5. The detection wavelength was at 280nm. Imipramine was the intemal standard. RESULTS The linear calibration curve was obtained over the range of 0.2μg · mL^-1 to 10μg · mL^-l of pirenzepine in aqueous humor（ r = 0. 999）. The within-day and between-day coefficient of variation were 1.2% and 1.5%, respectively. The recoveries of spiked low, medium and high concentrations of pirenzepine in aqueous humor were 97.9% , 99.2% and 101.3% , respectively. The limit of quatifieation was 0. 20μg · mL^-1, and the limit of detection was 0.10μg · mL^-1. Iontophoresis could improve the penetration of pirenzepine across rabbit eomea obviously, pH of the eye drops had the influence on iontophoretie penetration, while viscosity, strength of current and electrode type had no obvious effects. CONCLUSION This method is simple, rapid, accurate and reproducible, and has been applied to determine iontophoretieally transmitted pirenzepine in aqueous humor of rabbits.