Abstract

The study utilizes an analytical quality-by-design (AQbD) approach to establish a robust and efficient HPTLC method for plumbagin quantification in plumbago species. The highest-risk critical method parameters (CMP; concentration of plumbago and composition of mobile phase) and most appropriate critical analytical attributes (CAA; Peak area and retention factor [Rf]) were employed in the AQbD methodology. These CMP were tuned for higher plumbagin recovery using the central composite design (CCD) experiment. The created approach was validated by the requirements outlined in the International Conference on Harmonization (ICH) Q2 (R2). Every plate involved in the proposed method validation has been subjected to the passing requirements of the system suitability test (SST). The low LOD (16.93 ng/mL) and LOQ (49.68 ng/mL) values and the minimal divergence in intra- and inter-day studies validate the developed method’s high precision and sensitivity. Further experiments employing the proposed method to determine the amount of plumbagin from the two distinct plant sources harvested under two different climate conditions determined the highest plumbagin content in P. indica, which is in line with the results. To summarize, the AQbD method may be used in other plant systems to optimize various factors and get a greater concentration of the desired plant metabolite.

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