正交试验优选黄芩苷血药浓度测定中大鼠血浆的处理条件

Optimization of Processing Conditions of Rat Plasma in Blood Concentration Determination of Baicalin by Orthogonal Test

目的:优选黄芩苷血药浓度测定中大鼠血浆的处理条件。方法:采集大鼠空白血浆,加入黄芩苷,采用高效液相色谱法测定黄芩苷血药浓度。以黄芩苷萃取回收率为指标,以磷酸量、甲醇量、超声萃取时间、两次离心转速、两次离心时间为因素,通过L1(837)正交试验对含黄芩苷大鼠血浆处理条件进行优选并验证。结果:最优提取条件为加入磷酸50μl、甲醇2ml,涡旋混合2min,超声萃取10 min,以离心半径为4 cm、3 000 r/min离心10 min,取上清液氮气流下吹干,取200μl流动相复溶,涡旋混合1 min,以离心半径为3 cm、14 000 r/min离心5 min。验证试验得出平均萃取回收率为91.8%,RSD为2.6%。结论:本试验优化了含黄芩苷血浆的处理条件,阐明了各因素水平对黄芩苷萃取回收率的影响,优选的处理条件稳定可行。

OBJECTIVE ： To optimize the processing conditions of rat plasma in blood concentration determination of baicalin. METHODS： Blank rat plasma were collected and treated with baicalin. The blood concentration of baicalin was determined by HPLC. The processing conditions of baiealin in rat plasma was optimized by L,8（37） orthogonal test using extraction recovery rate of baicalin as index, the amount of phosphate and methanol, ultrasonic extraction time, centrifugal rotational speed （twice） and cen- trifugal time （twice） as factors. RESULTS： The optimal extraction conditions were as follows： adding phosphoric acid 50 μl and methanol 2 ml, vortex mixing 2 min, ultrasonic extracting for 10 min, centrifugal semidiameter of 4 cm, centrifuging for 10 min at speed of 3 000 r/min, drying up supernatant under a stream of nitrogen, dissolving with the mobile phase 200 μl, vortex mixing 1 min, centrifugal semidiameter of 3 cm, centrifuging for 5 min at speed of 14 000 r/min. Validation test showed that average ex- traction recovery rate was 91.8% （RSD=2.6%）. CONCLUSIONS： The trial optimizes the processing conditions of plasma contain- ing baicalin, and introduces the effects of various factors and levels on the extraction recovery rate ofbaicalin. The optimized pro- cessing conditions are stable and feasible.