Aquaphotomics Monitoring of Lettuce Freshness during Cold Storage

Fresh-cut leafy vegetables are one of the most perishable products because they readily deteriorate in quality even during cold storage and have a relatively short shelf life. Since these products are in high demand, methods for rigorous quality control and estimation of freshness that are rapid and non-destructive would be highly desirable. The objective of the present research was to develop a rapid, non-destructive near-infrared spectroscopy (NIRS)-based method for the evaluation of changes during cold storage of lettuce using an aquaphotomics approach to monitor the water molecular structure in lettuce leaves. The reference measurements showed that after 6 days of dark, cold storage, the weight and water activity of lettuce leaves decreased and β-carotene decreased, while chlorophylls slightly increased. Aquaphotomics characterization showed large differences in the lettuce leaves’ spectra depending on their growth zone. Difference spectra, principal component analysis (PCA) and linear discriminant analysis (LDA) confirmed the differences in the inner and outer leaves and revealed that spectra change as a function of storage time. Partial least squares regression (PLSR) allowed the prediction of the time spent in storage with a coefficient of determination of R(2) = 0.80 and standard error of RMSE = 0.77 days for inner, and R(2) = 0.86 and RMSE = 0.66 days for outer leaves, respectively. The following water absorbance bands were found to provide the most information in the spectra: 1348, 1360, 1373, 1385, 1391, 1410, 1416, 1422, 1441, 1447, 1453, 1466, 1472, 1490, 1503, 1515, 1521, 1534 and 1571 nm. They were further used as water matrix coordinates (WAMACs) to define the water spectral patterns (WASPs) of lettuce leaves. The WASPs of leaves served to succinctly describe the state of lettuces during storage. The changes in WASPs during storage reveled moisture loss, damage to cell walls and expulsion of intracellular water, as well as loss of free and weakly hydrogen-bonded water, all leading to a loss of juiciness. The WASPs also showed that damage stimulated the defense mechanisms and production of vitamin C. The leaves at the end of the storage period were characterized by water strongly bound to collapsed structural elements of leaf tissues, mainly cellulose, leading to a loss of firmness that was more pronounced in the outer leaves. All of this information was reflected in the changes of absorbance in the identified WAMACs, showing that the water molecular structure of lettuce leaves accurately reflects the state of the lettuce during storage and that WASPs can be used as a multidimensional biomarker to monitor changes during storage.