The design of experiments (DoE) is one of the quality-by-design tools valued in analytical method development, not only for cost reduction and time effectiveness, but also for enabling analytical method control and understanding via a systematic workflow, leading to analytical methods with built-in quality. This work aimed at using DoE to enhance method understanding for a developed UHPLC enantioseparation of terbutaline (TER), a model chiral drug, and to define quality assurance parameters associated with using chiral mobile phase additives (CMPA). Within a response surface methodology workflow, the effect of different factors on both chiral resolution and retention was screened and optimized using Plackett-Burman and central composite designs, respectively, followed by multivariate mathematical modeling. This study was able to delimit method robustness and elucidate enantiorecognition mechanisms involved in interactions of TER with the chiral modifiers. Among many CMPAs, successful TER enantioresolution was achieved using hydroxypropyl β-cyclodextrin (HP-β-CD) added to the mobile phase as 5.4 mM HP-β-CD in 52.25 mM ammonium acetate. Yet, limited method robustness was observed upon switching between the different tested CMPA, concluding that quality can only be assured with specific minimal pre-run conditioning time with the CMPA, namely 16-column volume (60 min at 0.1 mL/min). For enantiorecognition understanding, computational molecular modeling revealed hydrogen bonding as the main binding interaction, in addition to dipole-dipole inside the CD cavity for the R enantiomer, while the S enantiomer was less interactive.

Chemometrics, which applies mathematical and statistical methods to aid in experimental design, data analysis, and process optimization, is gaining more importance. It plays an important role in enhancing the sustainability of analytical methods when integrated with green analytical chemistry. Applying chemometrics to liquid chromatographic methods can speed up the analytical method, saving time, energy, and solvent consumption. This would be reflected by a low energy footprint and environmental impact. This review discusses the application of chemometric methods like multivariant data analysis, design of experiments, retention time prediction, and peak deconvolution in green liquid chromatographic analysis of pharmaceuticals. The review highlights the impact of chemometric approaches on the global push for sustainable practices. The integration of chemometrics into sustainable chromatographic analysis will continue to revolutionize the field leading to better performance of analytical methods.

Chemometrics, which applies mathematical and statistical methods to aid experimental design, data analysis, and process optimization, is gaining more importance. It plays an important role in enhancing the sustainability of analytical methods when integrated with green analytical chemistry. Applying chemometrics to liquid chromatographic methods can speed up the analytical method, saving time, energy, and solvent consumption. This would be reflected by a low energy footprint and environmental impact. This review briefly discusses the application of chemometric methods like multivariant data analysis, design of experiment, retention time prediction, and peak deconvolution in green liquid chromatographic analysis of pharmaceuticals. The integration of chemometrics in sustainable chromatographic analysis will continue to revolutionize the field and further whitening analytical methods with better performance.