Food Dye HPLC-DAD Method Analytical Validation: Precision and Reliability Studies, Optimized Using the Box–Behnken Experimental Design

Abstract

This study presents the validation of a high-performance liquid chromatography with diode array detection method for the simultaneous quantification of 11 food dyes. The method was developed using the Box–Behnken Design to maximize the efficiency of separation and improve the robustness of the analysis. Validation parameters for the method included linearity, accuracy, limit of detection|limits of detection, limit of quantification|limits of quantification, limit of repeatability-reproducibility, and recovery of samples by the internationally recognized methods. The method obtained excellent linearity (R2 > 0.998), low LODs, and high recovery (>90%). The limits of repeatability and limit of reproducibility had low relative standard deviations, indicating the reliability of the method. The validated method was tested on commercially available food products. Results were also compared with spectrophotometric analysis, demonstrating good consistency. This validated method provides a reliable and sensitive approach for routine quality control and regulatory monitoring of dyes in food matrices, supporting consumer safety and compliance with international food standards.