Abstract
Melatonin is a bioactive compound that is present in fermented beverages, such as wine and beer, at concentrations ranging from picograms to nanograms per mL of product. The purpose of this study was to optimize a novel fluorescent bioassay for detecting melatonin based on a cell line that contains the human melatonin receptor 1B gene and to compare these results with LC-MS/MS as a reference method. Conditions that could affect cell growth and detection (cell number per well, stimulation time, presence or absence of fetal bovine serum and adhesion of cells) were tested in the TANGO® cell line. Food matrices (wine and grape must) could not be directly used for the cell line due to low response. Therefore, for the determination of melatonin in food samples, an extraction procedure was required before conducting the assay. We demonstrated an improvement in melatonin determination by the cell-based bioassay due to increased sensitivity and specificity and improved quantification in complex matrices. Therefore, this method is a good alternative to determine melatonin content in some food samples, especially for those containing very low melatonin levels.
Highlights
® presence or absence of fetal bovine serum and adhesion of cells) were tested in the TANGO cell line
We aimed to develop and optimize a new method for the detection of Mel in food samples using a BLA cell-based bioassay
A validated method, liquid chromatography (LC)-mass spectrometry (MS)/MS, was used as a reference method to determine whether this methodology could be a good alternative to analyse Mel in food samples
Summary
® presence or absence of fetal bovine serum and adhesion of cells) were tested in the TANGO cell line. We demonstrated an improvement in melatonin determination by the cell-based bioassay due to increased sensitivity and specificity and improved quantification in complex matrices. This method is a good alternative to determine melatonin content in some food samples, especially for those containing very low melatonin levels. Mel has been found in many fermented beverages, such as beer or wine, at concentrations ranging from picograms to nanograms per mL of product[8,9,10]. During the wine-making process, the concentration of Mel reaches its maximum between the first and second day of fermentation[11,12], highlighting the role of yeast in its production. Despite the occurrence of Mel in fermented beverages is low, these concentrations have been described as contributing sufficiently to the dietetic intake to exhibit measurable effects[13]
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