Development and validation of a new derivatization-based quantitative analysis of multi-components by single marker...

Simultaneous quantitative analysis of multiple biogenic amines (BAs) in food is challenging due to the high cost of reference standards, weak ultraviolet absorption, and complex matrix interference. To address these limitations, this study developed a strategy integrating dansyl chloride (DNS-Cl) derivatization with the quantitative analysis of multi-components by single marker (QAMS) approach to enable simultaneous quantification of nine BAs in fermented wine using HPLC-UV. A key methodological innovation of this work is the first-time adoption of a weighted scoring system for the systematic selection of the optimal internal standard (IS) for QAMS. This system comprehensively evaluates the stability of the relative correction factor (RCF), retention time suitability, and response intensity, moving beyond traditional empirical choices. Through this rigorous evaluation, tyramine (Tyr) was selected as the optimal internal standard. The established RCFs proved reproducible and stable across different instruments and operational conditions (RSD < 10%). Method validation demonstrated good linearity (R² > 0.9985), precision (RSD < 2.28%), and accuracy (recovery: 93.92%-107.43%). Bland-Altman analysis showed that 100% of data points lay within the 95% confidence interval, demonstrating strong agreement with conventional external standard methods. In conclusion, the developed derivatization-based QAMS method is accurate, reliable, and cost-effective for the simultaneous quantification of nine biogenic amines in fermented wine, offering a practical alternative to traditional approaches. This method provides a low-cost analytical tool for quality control of BAs in fermented foods and expands the applicability of QAMS to compounds with weak UV absorption.