In the process of practical production, it is important to accurately analyze the proportion of mixed samples with high speed, which plays a great role for quality control and formulation design in food and agricultural processing. Traditional solution is to build statistical model with a large number of representative samples, which is both labor-intensive and time-consuming. In this paper, the proportion of alcohol and acids mixed samples, and their dilute solution mixed samples(used carbon tetrachloride (CCl4) which has no near-infrared absorption characteristics as the solvent medium)，as well as sheet tobacco leaf mixed samples are respectively analyzed by using near infrared spectroscopy, SG smooth and non-negative coefficients regression, which verifies the feasibility of analyzing the proportion of the mixed samples. The results show that, the analytic proportion of transmission spectra of alcohol and acids according to non-negative coefficients regression is closer to actual molar proportion with result error less than 4%. The result of the dilute solution is much better with error less than 4%. The analytic proportion of diffuse reflectance spectra of sheet tobacco leaf according to non-negative coefficients regression is highly accurate with error less than 10%. In the meantime, it has a highly consistency between actual spectra and analytic spectra of mixed samples; and the result of F-test and T-test shows that there is no significant difference between them and the confidence level is 0.01. It has the reliability of analytical proportion in theory. With the spectral data of several pure samples, the proportion of mixed samples can be thus analyzed, which has a good application prospect for quality control and formulation design in food and agricultural processing.