大鼠尿中木兰碱高效液相色谱测定法的建立

ESTABLISHMENT OF HPLC METHOD FOR DETERMINATION OF MAGNOFLORINE IN RATS URINE

目的建立高效液相色谱测定法测大鼠尿中木兰碱浓度。方法采用Agilent ODSC18柱(4.6mm×250mm,5μm),柱温25℃,流动相为甲醇?KH2PO4溶液(0.05 mol.L-1)=24:76(v:v),流速:0.8 mL.min-1。尿液样品用生理盐水稀释后与甲基叔丁基醚混合,上清液用40℃氮气吹干,残渣用流动相溶解进样。结果标准曲线在0.05～6.00μg.mL-1范围内线性良好,最低检测限为0.01μg.mL-1,木兰碱及内标盐酸小檗碱的保留时间分别为7.95 min和11.02 min,日内和日间精密度RSD均小于11%,提取回收率在92%以上。结论本法简便可靠,适用于大鼠尿液中木兰碱的测定及其药物动力学研究。

Objective To establish a HPLC method for the determination of Magnoflorine in rat urine. Methods Analysis was performed on an Agilent Cls analytical column （4.6mm ×250mm,5μm） with mobile phase consisting of methanol -0.05mol ·min^-1 potassium dihydrogen phosphate （24 ： 76, v ： v） at a flow rate of 0.8mL·min^-1. The urine sample was diluted by physiological saline and extracted with methyl tert - butyl ether. After evaporation of the organic layer, the residue was dissolved with mobile phase and injected into the column. Results The standard curve was linear over the concentration range of 0.05 to 6.00 μg·mL^-1The lowest concentration of detection in urine was 0.01 μg·mL^-1. The retention time for Magnoflorine and Berberine was 7.95 min and 11.02 min respectively. The intra and inter day RSD were less than 11%, the extraction recovery was over 92%. Conclusion The method is simple and accurate and suitable for the determination of Magnoflorine in rat Is urine and pharmacokinetics study.