This study was carried out to evaluate the effects of salt and phosphate on pH and protein extraction of beef muscle homogenate and their effects on separation of salt-soluble proteins by high performance liquid chromatography (HPLC). Beef muscle homogenate was prepared with 12 different levels of salt(4 levels of NaC1;0, 1, 3 or 5%) and phosphate (3 levels of sodium tripolyphosphate; 0, 0.1 or 0.2%). As the percent salt or phosphate increased, the pH and salt-soluble protein solubility increased linearly. Four protein peaks with approximate retention times (RT) of 5.8, 7.7, 10.5 and 13.3 min were observed in salt-soluble protein fraction by HPLC, and these proteins were calculated to approximate molecular weights of 1.5million, 500,000, 52,000 and 15,000 daltons, respectively. Salt or phosphate addition decreased linearly area of protein peak with 5.8 min RT (actomyosin likely) and increased linearly area of protein peak with 7.7 min RT (myosin likely). Among 12 salt and phosphate combinations, 3% salt-0.2% phosphate and 5% salt-0.2% phosphate showed relatively higher pH, salt-soluble protein solubility and protein peak area of 7.7 min RT, and their results were comparable.