HPLC法测定甲氨蝶呤片含量的方法学研究

Content Determination of Active Ingredients in Methotrexate Tablets by HPLC

建立一种液相色谱分析方法,测定甲氨蝶呤片的含量。采用色谱柱Inertsil ODS-2 (25 cm×4.6 mm,5μm);以乙腈-磷酸盐缓冲液(5︰95)为流动相;检测波长为302 nm;流速为1.0 mL/min;采集时间为20 min;进样体积为20μL。结果显示甲氨蝶呤线性范围为36.07~108.22μg·mL^(-1)(r=1.000);甲氨蝶呤平均回收率为99.1%,RSD为0.4%。该方法简单准确、重复性好,可用于甲氨蝶呤片的含量测定。

The high-performance liquid chromatography(HPLC) method was established to determine the content of methotrexate tablets. Methotrexate was determined by HPLC as follows: chromatographic column, Inertsil ODS-2(25 cm×4.6 mm, 5 μm);mobile phase, acetonitrile-phosphate buffer(5:95);detection wavelength, 302 nm;flow rate, 1.0 mL/min;acquisition time, 20 min;injection volume, 20 μL. The liner range of methotrexate was 36.07~108.22 μg·mL^(-1)(r=1.000). The average recovery rate of methotrexate(n=9) was 99.1 % with its RSD value of 0.4 %. The method was simple, accurate and repeatable, which was suitable for the content determination of active ingredients in methotrexate tablets.