biostudies-literatureUnknown20(1)Kothapalli PSRM Institute of Science & TechnologyTrigonelline is an alkaloidal plant derived bioactive compound reported for its various pharmacological activities. Recent advances in the utilization of this compound in drug development have highlighted the importance of establishing a reproducible and efficient analytical method for estimating trigonelline (TRG) in both its pure form and formulated nano systems. Current research aims to develop and validate a green RP-HPLC method for estimating TRG by integrating Analytical Quality by Design (AQbD) with the principles of Green Analytical Chemistry. Optimization of the chromatographic conditions was done by employing a rotatable central composite design with amount of mobile phase and its flow rate selected as critical variables and tailing factor (T<sub>f</sub>), retention time (R<sub>tBMC chemistry46https://www.ebi.ac.uk/biostudies/studies/S-EPMC12949508biostudies-literatureGreen RP-HPLC method for estimation of trigonelline: AQbD based development, validation, and application to nanoformulations.PMC12949508Kothapalli PVasanthan MfalseGreen RP-HPLC method for estimation of trigonelline: AQbD based development, validation, and application to nanoformulations.Trigonelline is an alkaloidal plant derived bioactive compound reported for its various pharmacological activities. Recent advances in the utilization of this compound in drug development have highlighted the importance of establishing a reproducible and efficient analytical method for estimating trigonelline (TRG) in both its pure form and formulated nano systems. Current research aims to develop and validate a green RP-HPLC method for estimating TRG by integrating Analytical Quality by Design (AQbD) with the principles of Green Analytical Chemistry. Optimization of the chromatographic conditions was done by employing a rotatable central composite design with amount of mobile phase and its flow rate selected as critical variables and tailing factor (T<sub>f</sub>), retention time (R<sub>t2026-01-01T00:00:00Z2026 Feb2026-07-16T23:15:19.6Z2026-07-12T03:11:06.649ZS-EPMC129495084165260210.1186/s13065-026-01734-z