Development and validation of a HPLC method for the determination of sertraline and three non-chiral related impurities
ABSTRACT 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.
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ABSTRACT: In this paper, a novel, precise, specific, accurate and rapid reversed-phase high performance liquid chromatographic method was developed, optimized and validated for determining sumatriptan succinate in niosomes with the best chromatographic peak resolution, reduced run time and low cost of analysis. The formulation has been previously optimized in terms of composition and preparation technique to obtain a high drug encapsulation efficiency and adequate vesicle size distribution. This method showed the best resolution by using Spherisorb(®) OSD2 C18 column (250mm×4.6mm, 5μm) using phosphate buffer (0.05M):acetonitrile (80:20, v/v; pH adjusted to 6.0) as a mobile phase at a flow rate of 1mL/min and wavelength of 214nm. The main objective of this research was to demonstrate the robustness of the reversed-phase HPLC method development by applying the Taguchi robust methodology. The signal-to-noise ratio (S/N) was employed as a quality measurement. This tool permits to establish the influence of some selected factors (acetonitrile:phosphate ratio, pH buffer, oven temperature and flow rate) on two responses (peak areas and retention time). On the basis of the results obtained, we can conclude that this analytical method was robust for all the factors studies, as exception of the flow rate, where the higher quality was obtained for the fewer values (0.8mL/min). Therefore, this parameter must be carefully controlled when this method was employed, to avoid any modification in the peak areas overall.Journal of Pharmaceutical and Biomedical Analysis 09/2012; 72:251-260. DOI:10.1016/j.jpba.2012.08.030 · 2.83 Impact Factor
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ABSTRACT: HPLC method development and validation play important role in the discovery, development and manufacture of agro chemicals , pharmaceutical products. This article mainly focuses on the optimization of HPLC conditions and other important perspectives during method development and validation. Various critical steps related to analytical method development and validation is discussed. A sequence of events required for method development and analytical validation are described. The steps involved in developing a stability-indicating HPLC method influences the analysis of degradation products/impurities in stability study and its validation demonstrate the suitability for its intended purpose.High Performance Liquid Chromatography (HPLC) is one of the most widely used analytical techniques in industry. It is used to separate and analyse compounds through the mass-transfer of analytes between stationary and mobile phases1-3. The technique is employed in a broad range of activities, such as the analysis of foods, drugs and agrochemicals. The technique of HPLC utilises a liquid mobile phase to separate the components of a mixture. The components themselves are first dissolved in a solvent and then forced to flow (via the mobile phase) through a column (stationary phase) under highpressure. The mixture is resolved into its components within the column and the amount of resolution is dependent upon the interaction between the solute components and the column stationary phase (immobile packing within the column) and liquid phase. The interaction of the solute with the mobile and stationary phases can be manipulated through different choices of both solvent and stationary phases