Techniques for colorimetric based test strip analysis and reader system are provided. In one aspect, a method of test strip analysis includes: illuminating a test strip wetted with a sample with select spectrums of light, wherein the test strip includes test pads that are configured to change color in the presence of an analyte in the sample; obtaining at least one digital image of the test strip; and analyzing color intensity from the at least one digital image against calibration curves to determine an analyte concentration in the sample with correction for one or more interference substances in the sample that affect the color intensity. A calibration method and a reader device are also provided.