RESPONSE SURFACE METHODOLOGY ASSISTED ULTRAPERFORMANCE LIQUID CHROMATOGRAPHIC METHOD OPTIMIZATION FOR THE...

Area of Science:

• Analytical Chemistry
• Chromatography
• Method Development

Background:

• Simultaneous estimation of vitamin D and K vitamers presents analytical challenges.
• Optimization of liquid chromatography (LC) is crucial for accurate quantification.

Purpose of the Study:

• To develop and optimize an LC method for the simultaneous estimation of six vitamin D and K vitamers.
• To apply response surface methodology (RSM) for efficient method optimization.

Main Methods:

• Utilized response surface methodology (RSM) with a Box-Behnken design (BBD) for parameter optimization.
• Employed an Accucore C18 column with a specific mobile phase composition and flow rate.
• Analyzed data using multiple regression analysis and validated using Q-ToF/MS detection.

Main Results:

• Optimized LC parameters achieved excellent separation of six vitamin D and K vitamers.
• High coefficients of determination (R2 > 0.983) indicated a significant regression model.
• The method demonstrated specificity, sensitivity, linearity, accuracy, precision, and robustness.

Conclusions:

• The developed LC method, optimized via RSM, is suitable for simultaneous quantification of vitamin D and K vitamers.
• The approach provides a reliable and efficient analytical solution for pharmaceutical analysis.
• The study highlights the utility of RSM in optimizing complex chromatographic separations.