Analytical quality by design-based RP-HPLC method for dobutamine quantification: development, optimization, and validation

The study presents method development, optimization, and validation of RP-HPLC technique to measure dobutamine, a cardiotonic agent utilized in the treatment of acute heart failure and cardiogenic shock. An Analytical Quality by Design was used to optimize chromatographic parameters, such as composition of the mobile phase, flow rate, and temperature of the column, applied Central Composite Design. The resulting optimized procedure showed superior system suitability, consisting of tailing factor 1.0, number of plates 12036 and similarity factor of 98.9 percent, which was very high in terms of resolution and reproducibility. The six repeated injections used to measure the system precision were 2106, 310.67 in terms of mean peak area and the low %RSD was 0.3 which indicated a high reproducibility. According to forced degradation studies, dobutamine was found to be stable when exposed to acidic conditions, basic conditions and peroxide conditions and also under thermal conditions; though these findings were accompanied by a high photolytic degradation (9%) and thus need to be stored under light protection. The technique also showed linearity over the broad concentration range (50 %, 150 %) with an R2 of 0.99996 and at three different concentrations of 50 %, 100 %, and 150 %, the recovery studies have shown accurate results with low %RSD values (0.2, 0.4). The robustness assurance ensured that there is minimum change in USP tailing, plate counts and % similarity factor with different chromatographic conditions hence robustness of this methodology. The standardized AQbD-informed RP-HPLC method exhibits great precision, accuracy, and reliability in the analysis of dobutamine during the pharmaceutical quality control with the ICH compliance, contributing to the safety of patients by the guarantee of consistent therapeutic efficacy.