In the present work, a freshness indicator was developed by immobilizing extracted betacyanins from prickly pear fruit (PFB) onto cellulose filter paper to visually determine fish spoilage. Various properties of the prepared indicator, including color stability, moisture uptake, sensivity to hydrogen sulfide (H2S) gas and pH changes, and chemical analysis were studied. The PFB-based indicator exhibited good color stability at various temperatures and times, with no significan color changes observed during storage at 25, 4, and −18 °C for 8 days. The moisture absorption of the indicator based on PFB was lower than that of cellulose paper, which is crucial for colorimetric indicators. Basic nitrogen and sulfur compounds are the main volatile compounds produced during fish spoilage. The results of the present study showed that both total volatile basic nitrogen (TVB-N) and H2S gas produced during fish spoilage contributed to the color changes of betacyanin-based colorimetric indicators. The prepared freshness indicators were pH-sensitive, with significant color changes due to TVB-N and H2S production during fish storage. A significant correlation was observed between fish quality (pH, TVB-N and total viable count) and the visible color changes of the indicators. The color of the designed indicator shifted from dark pink to pale pink/white during fish storage. These prepared freshness indicators were inexpensive, easy to use, and non-toxic, making them suitable for assessing the quality and shelf life of fish fillets.
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