HPLC柱切换法血浆直接进样测定糖肾平

采用HPLC柱切换技术,以Merck Lichroprep RP2(25～40μm)为预处理柱填料,以水为预处理流动相,以0.2mol/L乙酸-乙酸铵为净化清洗液,血浆直接进样,在线富集净化样品。血浆样品以0.2ml/min慢速转移通过预处理柱,净化回收率达80%。以ShimpackCLC-ODS为分析柱(15cm×6mm,ID),甲醇-异丙醇-0.2mol/L 乙酸铵(61:5:34)为分析流动相,310nm波长检测,外标法定量测定人体血浆中糖肾平的浓度。血浆样品仅需稀释后即可进样,净化富集样品操作简便,预处理柱可长期反复进样使用。血浆中最低检测浓度为30ng/ml,日内变异系数为2.0%～5.4%,日间变异系数为2.4%～2.7%,在浓度为60～2020ng/ml血浆范围内呈线性关系。

DIRECT INJECTION OF PLASMA TO DETERMINE GLIQUIDONE IN PLASMA USING HPLC COLUMN SWITCHING TECHNIQUE

An automated clean up and concentration method by column switching is described for the assay of gliquidone, a hypoglycemic agent, in plasma. The system uses Merck Lichroprep RP2 (25-40 microns) as short precolumn packing material for on--line clean up and concentration. A Shimpack CLC--ODS (5 microns) is used as analytical column. Water is used as the pretreatment mobile phase and a mixture of methanol--isopropanol--0.2 mol/L ammonium acetate solution (61:5:34) is used as the analytical mobile phase. The plasma is diluted with a solution of 0.2 mol/L acetic acid and injected directly on the precolumn. The low speed of 0.2 ml/min of pretreatment mobile phase during transfer of plasma sample on the precolumn can get recovery of 80.7%. The detection is at 310 nm and the detection limit is 30 ng/ml plasma. The procedure is simple and automatic.