Thummala Veera Raghava Raju, Seshadri Raja Kumar, Tharlapu Satya Sankarsana Jagan Mohan, Ivaturi Mrutyunjaya Rao, Nittala Someswara Rao
Analytical Research and Development, Integrated Product Development, Dr. Reddy's Laboratories Ltd., Bachupally, Hyderabad-500 072, India.
School of Chemistry, Andhra University, Visakhapatnam-530003, A.P., India.
Sci Pharm. 2014 Jan 16;82(2):307-26. doi: 10.3797/scipharm.1310-17. Print 2014 Apr-Jun.
Statistical experimental design was used to optimize the chromatographic separations of two pharmaceutical compounds from their respective potential impurities. A fractional factorial design was utilized to study the effects of pH, organic solvent in mobile phases A&B, and flow rate on the resolution of Rabeprazole and Rabeprazole Sulfone, which had closely eluting peaks. A desirability function applied to the optimized conditions predicted the peak resolution between 2.2 and 2.7 for the Rabeprazole & Rabeprazole Sulfone impurity. The chromatographic method employed an Acquity UPLC, BEH C18 column (100 × 2.1 mm i.d., 1.7 μm particle size) with the mobile phase consisting of a phosphate buffer, pH 6.5, and acetonitrile in a gradient program. The flow rate and injection volumes were 0.45 mL/min & 5 μl, respectively, and detection was done at 254 nm. The chromatographic method was validated for linearity, accuracy, precision, specificity, and ruggedness according to ICH guidelines. The results clearly showed that the quality by design concept could be effectively applied to optimize a UPLC chromatographic method with fewer trials and error-free experimentation.
采用统计实验设计来优化两种药物化合物与其各自潜在杂质的色谱分离。利用析因设计研究pH值、流动相A和B中的有机溶剂以及流速对雷贝拉唑和雷贝拉唑砜分辨率的影响，这两种物质的洗脱峰很接近。应用合意函数于优化条件，预测雷贝拉唑与雷贝拉唑砜杂质之间的峰分辨率在2.2至2.7之间。该色谱方法采用Acquity UPLC、BEH C18柱（内径100×2.1 mm，粒径1.7μm），流动相由pH 6.5的磷酸盐缓冲液和乙腈组成，采用梯度洗脱程序。流速和进样体积分别为0.45 mL/min和5μl，检测波长为254 nm。根据ICH指南对该色谱方法的线性、准确度、精密度、特异性和耐用性进行了验证。结果清楚地表明，设计质量理念可以有效地应用于以较少的试验和无错误实验来优化UPLC色谱方法。
