RP-HPLC法测定去羟肌苷肠溶胶囊有关物质次黄嘌呤及含量

目的:建立反相HPLC法测定去羟肌苷肠溶胶囊有关物质次黄嘌呤及含量。方法:采用InertSurstain C₁₈(250 mm×4.6 mm,5 μm)色谱柱;以0.386%醋酸铵溶液加氨水调节pH至8.0为流动相A,以乙腈-甲醇(50:50)为流动相B,梯度洗脱,测定去羟肌苷肠溶胶囊有关物质次黄嘌呤;以流动相A-流动相B(90:10)为流动相测定去羟肌苷肠溶胶囊中去羟肌苷含量;检测波长254 nm;流速1.0mL·min^-1;柱温25℃;进样体积为20 μL。结果:在选定的色谱条件下,主成分及其有关物质能完全分离,杂质次黄嘌呤在0.0996~9.960 μg·mL^-1范围内呈良好的线性关系(r=1.000),平均回收率(n=9)为100.7%(RSD=1.9%),次黄嘌呤与去羟肌苷的相对较正因子为0.62;去羟肌苷在0.2423~48.45 μg·mL^-1范围内呈良好的线性关系(r=0.9996),平均回收率(n=9)为99.9%(RSD=1.4%)。结论:本法经方法学验证,可用于去羟肌苷肠溶胶囊的有关物质检查及含量测定。

Objective:To establish a RP-HPLC method for the determination of related substance (hypoxanthine)and assay in the didanosine enteric-coated capsules.Method:An InertSurstain C₁₈ column (250 mm×4.6 mm,5 μm)was adopted with gradient elution.Mobile phase A consisted of 0.386% ammonium acetate(adjusted pH to 8.0 by ammonia water)and mobile phase B consisted of methanol-acetonitrile(50:50); the assay was carried out with the mobile phase consisting of mobile phase A-mobile phase B(90:10);the flow rate was 1.0 mL·min^-1;the detector wavelength was set at 254 nm;the column temperature was maintained at 25℃,and the injection volume was 20 μL.Results:The impurities and didanosine were well separated. The liner range of hypoxanthine was 0.0996-9.960 μg·mL^-1(r=1.000)and the average recovery was 100.7% with RSD of 1.9%(n=9).The relative calibration factor of hypoxanthine with respect to didanosine was 0.62.The liner range of didanosine was 0.2423-48.45 μg·mL^-1(r=0.9996),and the average recovery was 99.9% with RSD of 1.4% (n=9).Conclusion:The established method can be used for the content determination of related substances in didanosine entric-coated capsules.

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