In recent years, a supplementation of livestock animals, including poultry, with β-Hydroxy-β-methylbutyrate (HMB) has gained attention for its effects on protein and fat metabolism. This study investigates the effects of HMB in the laying hen diet on egg quality, focusing on amino acid and fatty acid composition. Laying hens were supplemented with 0.02% HMB, with performance parameters and egg components analyzed. HMB supplementation led to increased albumen weight, influencing egg weight while also reducing feed intake per egg without affecting laying rate, yolk indices, fat, or cholesterol content. Notably, the study revealed significant changes in egg amino acid and fatty acid profiles due to HMB supplementation. Various amino acids, including glycine, serine, and isoleucine, were altered in the yolk, impacting nutritional value and potential health benefits. Regarding fatty acids, the study observed changes in both saturated as well as n-6 and n-3 fatty acids, affecting the overall lipid profile of egg yolks. However, the shifts in fatty acid composition could have implications for cardiovascular health due to altered ratios of n-6/n-3 fatty acids. Further research is required to comprehensively understand the implications of these findings for consumer-oriented egg quality and health benefits.