Quality by design-driven RP-HPLC method for simultaneous determination of α-mangostin and cardamonin: validation, process capability, and multidimensional sustainability assessment.
Abstract
α-Mangostin (AMG) and cardamonin (CDN) are bioactive phytoconstituents with complementary pharmacological properties and potential utility in melanoma therapy; however, a reliable simultaneous chromatographic procedure for their quantitative determination across pharmaceutical and biological matrices remains limited. The present study aimed to develop and validate an Analytical Quality by Design (AQbD)-assisted RP-HPLC method for simultaneous determination of AMG and CDN and to demonstrate its applicability to co-loaded bovine serum albumin (BSA) nanoparticles and human plasma. Risk assessment and Failure Mode and Effects Analysis were initially employed to identify influential chromatographic variables, followed by Taguchi L8 screening and Box-Behnken response-surface optimization. Acetonitrile content, flow rate, and column temperature were identified as critical method parameters. The optimized procedure employed a Phenomenex Kinetex C18 column, an isocratic mobile phase of acetonitrile-0.1% aqueous formic acid (40:60, v/v), a flow rate of 1.0 mL min-1, column temperature of 30 °C, and PDA detection at 273 nm. The method exhibited excellent linearity over 2-12 μg mL-1, with R2 values >0.999 for both analytes. LOD/LOQ values were 0.467/1.406 μg mL-1 for AMG and 0.587/1.768 μg mL-1 for CDN, respectively. Precision results showed %RSD values below 2%, while recoveries ranged from 98.62 ± 0.87% to 99.87 ± 0.24%. The method was successfully applied to optimized AMG-CDN co-loaded BSA nanoparticles, which exhibited a particle size of 75.62 ± 0.19 nm, zeta potential of -30.42 mV, and entrapment efficiencies of 90.25 ± 0.15% for AMG and 84.62 ± 0.18% for CDN. Recovery from the nanoparticle matrix ranged from 97.86 ± 0.26% to 98.12 ± 0.15%, while plasma recovery ranged from 97.14 ± 0.24% to 98.63 ± 0.47%. Multidimensional sustainability assessment demonstrated favorable environmental and practical profiles, with AGREE, AGREEprep, Analytical Eco-Scale, CaFRI, CACI, ComplexMOGAPI, and BAGI scores of 0.70, 0.68, 73, 77, 81, 89, and 75.0, respectively. The developed procedure therefore provides a robust, sustainable, and versatile analytical platform for simultaneous quantification of AMG and CDN in bulk materials, BSA nanoparticles, and human plasma, supporting further development of AMG-CDN combination therapy for melanoma management.
Abstract as supplied by the indexing source. © the authors / publisher.