Facultad de Farmacia, Universidad San Pablo-CEU, Urbanización Montepríncipe, Boadilla del Monte, 28668 Madrid, Spain.
J Pharm Biomed Anal. 2010 Oct 10;53(2):122-9. doi: 10.1016/j.jpba.2010.01.036. Epub 2010 Jan 25.
In this study, a screening on reversed-phase stationary phases (including C(8), C(18), CN, PEG and amide) was carried out in order to obtain an efficient HPLC method for the determination of sertraline and three of its more closely related synthetical and non-chiral impurities, without using ion-pair reagents. The best results in terms of both retention time and resolution of the target analytes were obtained with a Zorbax Bonus-RP column, which contains a polar amide group embedded in a C(14) alkyl chain. Once the most suitable stationary phase was chosen, the HPLC method was optimized by using a factorial design, evaluating three quantitative factors (column temperature, buffer pH and buffer concentration) in order to find the best conditions which maximize the resolution between impurities A and B (positional isomers) and minimize the total run time. The final HPLC conditions were set by means of a second experimental design, which allowed optimizing the effects of the buffer pH and the proportion of methanol in the mobile phase. The optimal conditions for simultaneously determining sertraline and its impurities, being baseline separated in less than 10 min, were finally obtained with Zorbax Bonus-RP column (150 mmx4.6mm, 5 microm), under isocratic conditions with phosphate buffer (pH 2.8; 10mM)-methanol (63:37, v/v) at 50 degrees C, at the flow-rate of 1.0 mL/min. UV detection was set at 220 nm. This method was successfully validated following ICH guidelines and it proved to be reliable for the determination of sertraline and related impurities in tablets as pharmaceutical forms.
在本研究中，进行了反相固定相（包括 C(8)、C(18)、CN、PEG 和酰胺）的筛选，以获得一种无需使用离子对试剂即可高效测定舍曲林及其三种更密切相关的合成和非手性杂质的 HPLC 方法。在保留时间和目标分析物的分辨率方面，Zorbax Bonus-RP 柱获得了最佳结果，该柱含有嵌入 C(14)烷基链中的极性酰胺基团。一旦选择了最合适的固定相，就通过析因设计优化了 HPLC 方法，评估了三个定量因素（柱温、缓冲液 pH 和缓冲液浓度），以找到最佳条件，最大限度地提高杂质 A 和 B（位置异构体）之间的分辨率，并最小化总运行时间。通过第二次实验设计确定最终的 HPLC 条件，该设计允许优化缓冲液 pH 和流动相中甲醇比例的影响。最终，通过 Zorbax Bonus-RP 柱（150mmx4.6mm，5μm）在等度条件下，使用磷酸盐缓冲液（pH 2.8；10mM）-甲醇（63:37，v/v），在 50°C 下，以 1.0mL/min 的流速，获得了同时测定舍曲林及其杂质的最佳条件，在不到 10 分钟内实现基线分离。UV 检测设置在 220nm。该方法按照 ICH 指南成功验证，证明其可用于测定片剂等药物制剂中的舍曲林和相关杂质。
