Eco-friendly HPTLC method for Trifluridine and Tipiracil determination: Quality-by-design meets green analytical chemistry
Amruta Balekundri, Eknath D. Ahire, Rakesh U. Shelke, Dinesh Rishipathak, Sanjay J. Kshirsagar
Abstract
• An eco-friendly, green chemistry approach was employed using the AGREE scale calculator to ensure sustainability. • The Quality by Design (QbD) methodology, utilizing Analytical Quality by Design (AQbD) tools, was applied to develop a high-performance thin-layer chromatography (HPTLC) method with robust quality attributes. • The method was optimized using Central Composite Design (CCD) under Response Surface Methodology (RSM) for chromatographic parameters. • The validation of the developed HPTLC method was conducted as per ICH Q2 (R1) guidelines, confirming excellent linearity, low limits of detection (LOD) and quantification (LOQ), and high precision (%RSD < 2%). • The environmental impact of the method was evaluated using the ComplexGAPI, AGREE, Eco-Scale, and BAGI tools, yielding high green metrics and confirming minimal environmental impact. Colorectal cancer (CRC) is a leading cause of cancer-related mortality worldwide, necessitating efficient treatment strategies and accurate analytical methods. This study presents a green analytical approach using a Quality by Design (QbD)-assisted high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantification of trifluridine (TRI) and tipiracil (TIP) in pharmaceutical formulations. Chromatographic parameters were optimized using a Central Composite Design (CCD) under Response Surface Methodology (RSM), with solvent volume and chamber saturation time identified as critical factors. The optimized method yielded reliable Rf values of 0.64 for TIP and 0.91 for TRI. Validation per ICH Q2 (R1) guidelines confirmed excellent linearity (R² = 0.9944 for TIP and R² = 0.9988 for TRI), low detection limits (0.0011 µg/mL for TIP and 0.0022 µg/mL for TRI), and high precision (intra-day %RSD <0.74, inter-day %RSD <0.92). Robustness testing demonstrated minimal variability in Rf values (%RSD <0.28). Environmental sustainability was assessed using ComplexGAPI, AGREE, and BAGI tools. The developed method achieved an AGREE score of 0.81, an Eco-Scale score of 86, and a BAGI score of 80, highlighting its eco-friendliness and practical applicability. This study demonstrates an efficient, precise, and environmentally sustainable analytical method for TRI and TIP quantification, aligning with green chemistry principles and ensuring minimal environmental impact.