Generic approach in a gradient elution HPLC method development that enables troubleshooting free method transfer

• Published in: Journal of pharmaceutical and biomedical analysis 2022-01, Vol.207, p.114367-114367, Article 114367
• Main Authors: Milenković, Milan, Rašević, Marija, Otašević, Biljana, Zečević, Mira, Malenović, Anđelija, Protić, Ana
• Format: Article
• Language: English
• Subjects: (U)HPLC; Design of experiments; Dwell volume; Gradient elution method; Method optimization; Method transfer
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2021.114367

•Integration of the dwell volumes into the gradient methods optimization phase.•The intertwining of development and transfer of gradient method for quality control.•Dabigatran etexilate mesylate and nine impurities as fair chromatographic challenge.•Three (U)HPLC instruments with great difference in dwell volume were tested.•The validation studies confirmed adequacy of the method and proved its utility. [Display omitted] Nowadays, method development is strongly focused on reducing time needed for method development and execution. This subject specially concerns gradient elution methods regarding the usual need for troubleshooting assistance with uncertain outcome during the method transfer from one laboratory to another. One of the main reasons for this situation is the dwell volume difference between HPLC systems. Therefore, the aim of this study was to propose a novel method development methodology that would integrate the dwell volumes differences in the optimization process. The proposed approach could be quite useful in industry that has insight in HPLC instruments planned to be used during the method life cycle. It was tested on the model mixture consisting of dabigatran etexilate mesylate and its nine impurities by use of experimental design methodology. Three different (U)HPLC instruments with high dwell volume differences were selected to challenge the methodology. Plan of experiments was defined with Plackett-Burman design for screening phase and D-optimal design for optimization phase. Initial and final amount of organic modifier, time of the gradient elution and pH value of the aqueous phase were selected as variables significant for the gradient programme profile and included in the optimization stage along with dwell volume values. The separation criteria s between critical peak pairs was selected as output for method optimization while indirect modelling together with Monte Carlo simulations enabled selection of optimal and robust chromatographic conditions. They included 24% (v/v) of initial amount of acetonitrile, 54% (v/v) of the final amount of acetonitrile, 15 min of gradient elution run time and pH value equal to 4.9. The proposed method was successfully validated, met all validation criteria and thus proved its utility.