Fluorescent immunoliposomal nanovesicles for rapid multi-well immuno-biosensing of histamine in fish samples

Scombroid poisoning in fish-based and other food products has raised concerns due to toxicity outbreaks and incidences associated with histamine, thus measuring the amount of histamine toxic molecule is considered crucial quality indicator of food safety and human health. In this study, liposome-based measurement of histamine was performed via rupturing mechanism of sulforhodamine B dye encapsulated anti-histamine antibody conjugated liposomal nanovesicles. The immunosensing ability of immuno-liposomal format was assessed by monitoring the fluorescence at excitation/emission wavelength of 550/585 nm. Immuno-liposomal format assays were considered, one based on single wash procedure (Method 1), which had a detection limit of 10 ppb and quantification limit 15-80 ppb. While Method 2 based on one-by-one wash procedure had a detection limit of 2-3 ppb and quantification limit 8.5 ppb-200 ppm that required 2 h 30 min to perform. In view of better quantification limit, Method 2 was chosen for further tests required to validate its applicability in real samples. The feasibility of Method 2 was reconfirmed in fresh mackerel fish, and canned fish (tuna and salmon) with a similar detection limits but with low amplified fluorescence signals and sufficient levels of histamine recovery from fresh mackerel (73.50-99.98%), canned tuna (79.08-103.74%) and salmon (74.56-99.02%). The specificity and method accuracy were expressed as % CV in the range 5.34%-8.48%. Overall, the developed multi-well sensing system (Method 2) showed satisfactory specificity, cost effectiveness, rapidity, and stability for monitoring histamine toxicity as a practical food diagnostic device.