| Summary: | Fermentation is a technology that enhances biologically active ingredients, improves the absorption rate and induces the generation of new functional ingredients by the catalytic action of enzyme systems possessed by microorganisms. In this study, changes in the content of five kinds of bioactive compounds (deacetylasperulosidic acid, asperulosidic acid, scopolin, asperuloside and scopoletin) of <i>Morinda citrifolia</i> L. were confirmed by fermentation, and a high-performance liquid chromatography-photodiode array (HPLC-PDA) analysis method for measuring analytes was developed and validated. HPLC method for the determination of five bioactive compounds in <i>Morinda citrifolia</i> L. extracts (MCE) was validated in terms of sensitivity, linearity, selectivity, limit of detection (LOD) and quantification (LOQ), precision and accuracy. The coefficient of determination of the calibration curve for bioactive compounds (1.56–100 μg/mL) showed linearity (R<sup>2</sup> ≥ 0.9999). LOD and LOQ were in the range 0.04–0.97 and 0.13–2.95 μg/mL, respectively. The range of intra- and intraday accuracies values (recovery) were 97.5–121.9% and 98.8–118.1%, respectively, and precision value (RSDs) of the bioactive compounds were <4%. In addition, changes in the content of five bioactive compounds in MCE by fermentation were confirmed. These results indicate that the developed fermentation and analysis method could be applied in the development of potential functional food ingredients.
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