Provided is a method for measuring a target polypeptide in a biological sample abundantly containing impurities by mass spectrometry. A method for measuring a target polypeptide in a biological sample includes: a first reaction step of bringing a liquid containing a biological sample into contact with a first antibody-immobilizing carrier to bind the target polypeptide in the biological sample with the first carrier; a first washing step of washing the first carrier; a first eluting step of eluting the target polypeptide from the first carrier by using an acidic solution to obtain a first eluate; a neutralizing step of obtaining a first purified solution by adding a neutral buffer to the first eluate; a second reaction step of bringing the first purified solution into contact with a second antibody-immobilizing carrier, to bind the target polypeptide in the first purified solution with the second carrier; a second washing step of washing the second carrier; a second eluting step of eluting the target polypeptide from the second carrier by using an acidic solution to obtain a second purified solution; and a step of detecting the target polypeptide in the second purified solution by mass spectrometry.