Development and Validation of New Ultra-High-Performance Liquid Chromatography-Hybrid Quadrupole-Orbitrap Mass Spectrometry Method for Determination of Melatonin in Fruits

Abstract

A new reliable analytical method based on ultra-high-performance liquid chromatography coupled with hybrid quadrupole-Orbitrap mass spectrometry was developed for the analysis of melatonin (MEL) in food products. In-house validation was performed using deuterated melatonin (MEL-d4) as an internal standard to ensure method selectivity and accuracy and to evaluate the efficiency of a robust ethyl acetate extraction technique used for sample preparation. The analysis of 18 tart cherry varieties and 28 tomato varieties was performed at optimized conditions. The method was linear (R2> 0.99) over the concentration range of 5-200 pg/g. A very low limit of quantification (10 pg/g) was provided for both analyzed matrices. The determined average recoveries (102 and 110%) and the values of intraday repeatability (6.30 and 10.9%) for cherry and tomato matrices, respectively, indicated a good accuracy and precision. The elaborated procedure proved the absence of MEL in any of tart cherries (<10 pg/g), whereas the concentration levels in tomatoes were found to be in the range of <10-149 pg/g, where the highest concentrations were determined in "Cherry," "Cherry Red" and "Rome" tomatoes grown in the Netherlands.