The volatilization loss of manganese during the vacuum smelting process is one of the key factors that determines the manufacturing cost and quality of manganese steel. In this study, the laboratory experiments and thermodynamic calculations were performed to investigate volatilization behavior of manganese from molten steels with different alloying methods in vacuum process. Based on the thermodynamic analysis, with the increase of manganese content, the partial vapor pressure of the manganese component increased, resulting in manganese being easily volatilized from molten steel. The carbon content in the steel shows an evident influence on partial vapor pressure of manganese component, and a higher carbon content in steel leads to a lower partial vapor pressure of manganese, but it not influenced by the silicon content. Compared with the alloying method of high carbon ferromanganese, the volatilization loss of manganese in the alloying method of silicon manganese presents faster decay, agreeing well with the thermodynamic analysis. Besides, the volatile fraction generated in the alloying method of high-carbon ferromanganese is composed of a large amount of MnO nanorods with a lateral length approximately 500 nm and a small number of Mn3O4/Mn nanoparticles with a diameter less than 500 nm. Additionally, the volatile fraction generated in the alloying method of silicon manganese shows Mn3O4 nanoparticles as the main phase. It can be inferred that the existence of the manganese oxide phase is attributed to the high chemical activity of nanoscale particles within air.