The invention discloses a three-wavelength vein blood oxygen concentration measuring method, a measuring device is composed of a blood oxygen signal, a blood oxygen probe, a signal amplifier, an analog-digital converter and a main controller, the blood oxygen probe collects a blood sample signal, carries out photoelectric conversion, converts a light intensity signal into a frequency signal, carries out signal amplification through a signal amplification circuit, performs ADC conversion, converts the frequency signal into a digital signal, transmits the digital signal to the main controller, the digital signal is processed by the main controller to obtain an electric signal in direct proportion to the light intensity, and the obtained electric signal is substituted into a three-wavelength calculation formula obtained by new derivation to obtain the corresponding venous blood oxygen concentration. According to the three-wavelength vein blood oxygen concentration measuring method, wavelength light sources on absorption points such as reduced hemoglobin and oxyhemoglobin serve as reference light sources, the three-wavelength vein blood oxygen concentration measuring method is obtained through derivation, the anti-interference capacity is higher, measurement is accurate, no time interval is generated in the measuring period, and real-time, continuous and effective parameters can be provided.