Using Australian grain-fed beef as the subject of study, four thawing methods, i.e., low-frequency electric field thawing, room temperature thawing, running water thawing and refrigerator thawing were applied to frozen beef. The impact of these methods on pH, color difference, water-holding capacity, texture properties, total volatile basic nitrogen(TVB-N) mass fraction, free amino acids, and soluble protein content was examined. The integrity of the muscle microstructure was observed using scanning electron microscopy to compare the effects of the four thawing methods on beef quality. The results showed that, among the four methods, low-frequency electric field thawing resulted in a pH closest to that of fresh meat(6.00). The L* was the highest at 48.46, while centrifugal loss and drip loss were the lowest, at 21.53% and 1.93%, respectively. Hardness, springiness, and chewiness were the highest, recorded at 8 784.61 g, 0.86, and 2 704.82 g, respectively. The TVB-N mass fraction was the lowest at 5.13 mg/100 g, and the soluble protein mass fraction was the highest at 74.67 mg/g. The proportion of sweet amino acids was the highest, while the proportion of bitter amino acids was the lowest. Scanning electron microscopy results revealed that the original muscle structure of beef thawed by low-frequency electric field remained intact. In conclusion, low-frequency electric field thawing can reduce quality loss during the thawing process and maintain the fresh quality and original flavor of beef.