Development and validation of an HPLC-PDA method for the analysis of phycocyanobilin in <i>Arthrospira maxima</i>

An high-performance liquid chromatography (HPLC)photodiode array (PDA) method was developed and validated for quantifying phycocyanobilin (PCB) in Arthrospira maxima extracts.While previous methods for PCB analysis lacked comprehensive validation and struggled with coelution issues, this approach demonstrated high specificity, accuracy, sensitivity, and reproducibility.Effective separation was achieved, with a retention time of 18.7 min.The calibration curve exhibited excellent linearity (r 2 = 1.0000) across a concentration range of 3.125-50 g/mL, with a limit of detection and limit of quantification of 0.22 and 0.67 g/ mL, respectively.Recovery rates ranged from 97.75 to 103.36%, with relative standard deviations (RSD) between 0.50 and 2.14%, confirming the accuracy and reliability of the method.The intra-and inter-day precisions yielded mean contents of 0.122 (RSD: 1.61%) and 0.128 mg/g (RSD: 0.71%), respectively.Notably, this study is the first to validate an HPLC-PDA method for direct quantification of PCB in A. maxima with full validation parameters.By offering a standard for PCB analysis, this study contributes to quality control in research and industrial processes, paving the way for advancements in pharmacology and biotechnology.