Abstract

Development of simple, fast and non-destructive technique such as fluorescence based method for the quantification of milk allergens in various dairy products is a highly rewarding task. In this contribution, a red emitting fluorogenic dye, quinaldine red (QR) is reported for the detection and quantification of a milk allergen, beta lactoglobulin (β-LG) in milk and whey matrices, utilizing its high selectivity and sensitivity towards β-LG. Detail spectroscopic investigation reveals that binding of QR to the hydrophobic calyx site of β-LG protein substantially reduces the torsional agility and propensity of TICT state formation of QR, rendering the dye highly fluorescent in nature. This enables estimation of β-LG with LOD 52.1(±0.9) nM in buffer solution and 0.21(±0.01) μM in 5 % bovine milk matrix respectively. Additionally, high selectivity and sensitivity, excellent repeatability, quick response, and emission in the biologically favorable red spectral region make QR based fluorometric quantification of β-LG a highly attractive choice. Finally, the estimated β-LG concentrations in milk and whey matrices from fluorometric titration and densitometry methods are found to match excellently with each other, suggesting potential of QR as an efficient turn-on fluorescent probe for the quantification of β-LG (milk allergen) in various dairy products.

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