高效液相色谱法同时测定人血清中美托洛尔、地尔硫卓及单硝酸异山梨酯

Study of HPLC method for simultaneous separation and determination of metoprolol,diltiazem and isosorbide-mononitrate in human serum

目的建立高效液相色谱法同时分离测定人血清中美托洛尔、地尔硫卓及单硝酸异山梨酯的方法,为临床药学研究提供简单方便的分析技术。方法固定相为Kromasil ODS-C_(18)(250 mm×4.6 mm,5μm)色谱柱;流动相为磷酸盐缓冲液(pH=6.0)和乙腈,梯度洗脱,流速为1.0 mL/min;检测波长210 nm;进样量10μL。结果线性范围分别为:美托洛尔6.8～400μg/mL,r=0.9997(n=6);地尔硫卓6.8～400μg/mL,r=0.9999(n=6);单硝酸异山梨酯5.1～300μg/mL,r=0.9998(n=6)。方法回收率分别为:美托洛尔103.3%,地尔硫卓98.2%,单硝酸异山梨酯91.4%。结论本方法能有效分离美托洛尔、地尔硫卓及单硝酸异山梨酯,适于同时检测人血清中3种物质的含量。

Objective To establish a HPLC method that can be used to separate and determine metoprolol,diltiazem and isosorbide-mononitrate in human serum simultaneously.The method proposed by us could provide a simple analytical tool in the assay of these analytes in clinical pharmacy studies.Methods A HPLC method with stationary phase Kromasil-C_（18）（250 mm×4.6 mm, 5μm） column was used;the mobile phase was composed of phosphoric buffer（pH=6.0） and acetonitrile at different percentages in gradient mode.The flow rate was 1.0 mL/min and the detection wavelength was located at 210 nm and injection volume of sample was 10μL.Results the linear ranges and correlation coefficients（n=6） were 6.8～400,6.8～400,5.1～300μg/mL and 0.999 7, 0.9999,0.9998 for metoprolol,diltiazem,and isosorbide-mononitrate respectively.The recoveries were 103.3%,98.2%,and 91.4%for metoprolol,diltiazem and isosorbide-mononitrate respectively. Conclusion The method can effectively separate metoprolol,diltiazem and isosorbide-mononitrate and it can be used to determine the analytes in human serum.