The applicability of non-destructive near infrared reflectance spectroscopic (NIRS) method was tested to determine the protein and oil contents of intact soybean [Glycine max (L.) Merr.] seeds. A total of 198 soybean calibration samples and 101 validation samples were used for NIRS equation development and validation, respectively. In the developed non-destructive NIRS equation for analysis of protein and oil contents, the most accurate equation was obtained at 2, 8, 6, 1(2nd derivative, 8 nm gap, 6 points smoothing, and 1 point second smoothing) and 2, 1, 20, 10 math treatment conditions with Standard Normal Variate and Detrend (SNVD) scatter correction method and entire spectrum (400-2500 nm) by using Modified Partial Least Squares (MPLS) regression, respectively. Validation of these non-destructive NIRS equations showed very low bias (protein: 0.060%, oil: -0.017%) and standard error of prediction (SEP, protein: 0.568 %, oil : 0.451 %) as well as high coefficient of determination (R2 , protein: 0.927, oil: 0.906). Therefore, these non-destructive NIRS equations can be applicable and reliable for determination of protein and oil content of intact soybean seeds, and non-destructive NIRS method could be used as a mass screening technique for selection of high protein and oil soybean in breeding programs