固相萃取和高效液相色谱-荧光法测定普萘洛尔贴剂对家兔给药后血药浓度

 目的建立家兔体内普萘洛尔血药浓度的测定方法。方法用Waters Oasis HLB柱萃取血清中的普萘洛尔,用Xterra C₁₈色谱柱(3.9mm×150mm,5μm),流动相为甲醇-0.05mol·L^-1的碳酸氢铵水溶液(pH9.5),(48:52),流速1.0mL·min^-1。用高效液相色谱-荧光法测定普萘洛尔血药浓度。结果普萘洛尔血药浓度在5～160μm·L^-1内线性良好,相关系数为0.9996,最低检测浓度为2μg·L^-1。低、中、高浓度时普萘洛尔的方法回收率分别为93.25%,94.30%和96.20%,萃取回收率分别为89.31%,92.45%,94.27%。日内和日间精密度RSD分别小于4.0%和7.0%。结论建立了简单、快速和准确的家兔体内普萘洛尔血药浓度的固相萃取和高效液相色谱-荧光测定方法。透皮贴剂中的普萘洛尔可以平稳地透过家兔皮肤,稳定地控释达2d。

Determination of propranolol in rabbit serum by solid phase extraction and HPLC with fluorescence detector

OBJECTIVE To develop a HPLC assay for determining the concentration of propranolol in rabbit serum. METHODS The separation was performed on a Xterra C₁₈ column (3.9mm×150mm,5μm) after the plasma samples were processed by a Waters Oasis HLB cartrdga. The mobile phase consisted of methanol-0.05 mol·L^-1 ammonium hydrogen carbonate solution (48:52), adjusted to pH 9.5 with ammonia solution, with a flow-rate of 1.0 mL·min^-1. RESULTS The calibration curve had good linearity in the range of 5～160μg·L^-1, the limit of detection was 2μg·L^-1. The average recovery was 93.25%, 94.30 % and 96.20 % for 10,40 and 160 μg·L^-1, respectively. RSDs for inter-day and intra-day validation did not exceed 4.0% and 7.0%, respectively. CONCLUSION The solid phase extraction and the HPLC method with fluorescence detection are developed for the determination of propranolol in rabbit serum. It appears to be simple, convenient and precise. The transdermal patch can deliver propranolol to rabbit stably for two days after a single dose.