The states of protons within food items are highly related to their physical attributes. In this study, the effect of cooking methods including boiling, steaming, roasting and frying on proton dynamics, physicochemical parameters and microstructure of Spanish mackerel was assessed by low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance imaging (MRI) techniques. The treatment of cooking resulted in a significant reduction of proton mobility and declined freedom of protons. The state changes of protons can be monitored easily in an intuitive and non-destructive manner during various cooking process. The treatments of boiling, steaming, roasting and frying resulted in different cooking loss and similar water-holding capability. A significant increase of total carbonyl content and thiobarbituric acid reactive substances was found, while a decrease of the values for free thiols and surface hydrophobicity was observed. The analysis of circular dichroism spectroscopy and cryo-scanning electron microscopy showed significant structural change. The correlation coefficients of R cal 2 and R cv 2 from partial least squares (PLS) regression models were more than 0.980, suggesting good correlation between LF-NMR data and hardness, resilience, springiness, chewiness, gumminess, and adhesiveness. Good recoveries and a relatively small coefficient of variation (CV) were obtained from the PLS regression models, indicating good reliability and accuracy in predicting texture parameters for mackerel samples.Foods 2020, 9, 364 2 of 16 and texture characteristics. However, it is a challenge to characterize the water dynamics, protein denaturation and lipid oxidation during various thermal processes of Spanish mackerel. Therefore, it is vital to determine the effect of different cooking methods on protein and lipid state changes in Spanish mackerel. The states of protons from water, proteins, lipid and polysaccharide are highly related to fish muscle destruction and protein denaturation during thermal processes. Most of the protons coming from the water within the fish meat can be reflected through monitoring relaxation parameters by radio frequency with the low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance imaging (MRI) techniques [3]. The protons are chemically exchanged back and forth from water to macromolecules in fish meat, so that their transverse component of the magnetization vector can be measured by the spin-to-spin relaxation time (T 2 ). Different food samples have different relaxation time T 2 , and therefore the parameters of T 2 can reflect the change of physical attributes [4]. In addition, the MRI can provide visualized internal information of the food items during processing and storage [5]. At present, the non-destructive LF-NMR and MRI technology has exhibited prominent superiority in assessing food proton change during various food-processing methods, such as rehydration of sea cucumbers [6], drying of surf calm and chicken [7], cooking change of striped snakehead fish, and evalua...