An apparatus for non-invasively estimating a biological component is provided. The apparatus may include: a sensor configured to measure a calibration spectrum for a first duration and measure a biological component estimation spectrum for a second duration, based on light returning from an object; and a processor configured to remove a signal of a biological component from the calibration spectrum to obtain a background spectrum for the first duration, and estimate the biological component, based on the background spectrum and the biological component estimation spectrum, for the second duration in response to a command for measuring the biological component.