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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Inert Surstain C_(18)(250 mm×4.6 mm,5μm)色谱柱;以0.386%醋酸铵溶液加氨水调节p H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1.0 m L·min~(-1);柱温25℃;进样体积为20μL。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0.099 6~9.960μg·m L~(-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0.242 3~48.45μg·m L~(-1)范围内呈良好的线性关系(r=0.999 6),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 reverse phase HPLC method for the determination of Hypoxanthine and its related substances in didanosine enteric-coated capsules. Methods: Inert Surstain C 18 column (250 mm × 4.6 mm, 5 μm) was used. With 0.386% ammonium acetate solution and ammonia solution, pH 8.0 was used as mobile phase A and acetonitrile-methanol 50:50 as mobile phase Phase B, the gradient elution determination of didanosine enteric-coated capsules hypoxanthine; mobile phase A mobile phase B (90:10) as mobile phase of didanosine enteric-coated capsules didanosine The detection wavelength was 254 nm. The flow rate was 1.0 m L · min -1. The column temperature was 25 ℃. The injection volume was 20 μL. Results: Under the selected chromatographic conditions, the main components and their related substances could be completely separated. The impurity hypoxanthine showed a good linearity (r = 1.000) in the range of 0.099 6 ~ 9.960 μg · m L -1. , The average recovery (n = 9) was 100.7% (RSD = 1.9%), the relative correction factor of hypoxanthine and didanosine was 0.62, and the values of didanosine between 0.242 3 ~ 48.45 μg · m L ~ -1) (r = 0.999 6). The average recovery (n = 9) was 99.9% (RSD = 1.4%). Conclusion: This method is validated by methodology and can be used for the related substances examination and determination of didanosine enteric-coated capsules.