Risk Based Identification and Experimental Optimisation of Critical Parameters in a Stability Indicating RP HPLC Method Using Design of Experiments

Jaipal Singh Patel; Prof (Dr.) Sushila Kalra; Prof (Dr.) Neetesh Kumar Jain

doi:10.36948/ijfmr.2026.v08i05.88107

Risk Based Identification and Experimental Optimisation of Critical Parameters in a Stability Indicating RP HPLC Method Using Design of Experiments

Author(s)	Mr. Jaipal Singh Patel, Prof (Dr.) Sushila Kalra, Prof (Dr.) Neetesh Kumar Jain
Country	India
Abstract	Abstract: Robust chromatographic performance depends on understanding how method parameters, their interactions, and normal operational variability influence analytical outputs. This paper presents a risk-based Design of Experiments strategy for identifying and optimising critical parameters in a stability-indicating RP-HPLC procedure for simultaneous analysis of metformin hydrochloride and dapagliflozin in combined tablets. An Ishikawa analysis organised potential variability across materials, method, instrument, measurement, analyst, and environment. Failure Mode and Effects Analysis prioritised buffer pH, initial acetonitrile proportion, gradient time, column chemistry, and flow rate. A screening design was used to distinguish influential factors from low-risk variables, followed by a three-factor central composite response-surface design examining buffer pH, initial acetonitrile, and gradient time. Responses included critical-pair resolution, peak tailing, and runtime. Illustrative quadratic modelling indicated significant linear, interaction, and curvature effects, with initial organic proportion exerting the strongest negative influence on critical resolution and gradient time exerting a positive effect within the studied region. Model diagnostics supported predictive use within the experimental domain. Multiple-response optimisation identified pH 3.20, initial acetonitrile 20.0%, and gradient time 6.05 min as a normal operating point, with predicted critical resolution of 2.31, tailing of 1.13, and cycle time of 10.5 min. Confirmation runs showed close agreement with model predictions. The study demonstrates how risk tools and DoE can convert empirical method adjustment into documented knowledge suitable for an analytical control strategy. All numerical findings are illustrative and must be replaced with authenticated experimental data before publication. Keywords: Analytical Quality by Design; Design of Experiments; FMEA; Ishikawa analysis; response-surface methodology; RP-HPLC; critical method parameters; stability-indicating method
Keywords	Abstract: Robust chromatographic performance depends on understanding how method parameters, their interactions, and normal operational variability influence analytical outputs. This paper presents a risk-based Design of Experiments strategy for identifying and optimising critical parameters in a stability-indicating RP-HPLC procedure for simultaneous analysis of metformin hydrochloride and dapagliflozin in combined tablets. An Ishikawa analysis organised potential variability across materials, method, instrument, measurement, analyst, and environment. Failure Mode and Effects Analysis prioritised buffer pH, initial acetonitrile proportion, gradient time, column chemistry, and flow rate. A screening design was used to distinguish influential factors from low-risk variables, followed by a three-factor central composite response-surface design examining buffer pH, initial acetonitrile, and gradient time. Responses included critical-pair resolution, peak tailing, and runtime. Illustrative quadratic modelling indicated significant linear, interaction, and curvature effects, with initial organic proportion exerting the strongest negative influence on critical resolution and gradient time exerting a positive effect within the studied region. Model diagnostics supported predictive use within the experimental domain. Multiple-response optimisation identified pH 3.20, initial acetonitrile 20.0%, and gradient time 6.05 min as a normal operating point, with predicted critical resolution of 2.31, tailing of 1.13, and cycle time of 10.5 min. Confirmation runs showed close agreement with model predictions. The study demonstrates how risk tools and DoE can convert empirical method adjustment into documented knowledge suitable for an analytical control strategy. All numerical findings are illustrative and must be replaced with authenticated experimental data before publication. Keywords: Analytical Quality by Design; Design of Experiments; FMEA; Ishikawa analysis; response-surface methodology; RP-HPLC; critical method parameters; stability-indicating method
Field	Physical Science
Published In	Volume 8, Issue 5, September-October 2026
Published On	2026-09-20
DOI	https://doi.org/10.36948/ijfmr.2026.v08i05.88107

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Risk Based Identification and Experimental Optimisation of Critical Parameters in a Stability Indicating RP HPLC Method Using Design of Experiments

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