Seed color is an important trait affecting flour yield and quality in wheat. Seed color also is either tightly linked to or pleiotropically controls seed dormancy in wheat, because most of the red-seeded wheats are tolerant to pre-harvest sprouting in comparison to white-seeded wheats. Recently, metabolomics approaches have recently been used to assess the natural variance in metabolite content between individual plants, an approach with great potential for the improvement of the compositional quality of crops. Basically, in the study here, the simultaneous proteomic and metablomic approaches are being investigated to identify the expressed proteins of genes and specific metabolism responsible for the expression of red and white colors of seed. Red seed “Jinpum” and white seed “Kumkang” cultivars were used in this study to identify the storage proteins use of 2-DE, MALDI-TOF/MS. Here we optimize tissue extraction methods compatible with high-throughput, reproducible nuclear magnetic resonance (NMR) spectroscopy based metabolomics. It appears that the proteins expressed were different each other according to two different cultivars from the seeds of hexaploid wheat. Some selected protein spots were identified as follows: B3-hordein, Gamma-hordein-3,bifunctionalalpha amylase/subtilisin Inhibitor. To monitor metabolic profile, wheat grain was ground in liquid nitrogen, ensuring a homogeneous mix of the tissue, solution samples extracted from seed grains of two wheat cultivars were conducted to measurement of metabolite using 1H-1D NMR method. Representative 1H-1D NMR spectra showing the metabolic fingerprints of wheat grain extracted and presented in Fig. The different peaks, observed at 3.4 and 4.3 ppm, were detected and difference in each two cultivars. The metabolic fingerprint of each two wheat cultivars by 1H-1D NMR were analysed using partial least squares (PLS) in mutivariate analysis to confirm metabolic profiling between different cultivars and to screen chemical shift spectrum corresponding to metabolite specifically abundant in each cultivars. Profiling using 1H-1D NMR was applied to measure of abundance of major metabolite. In total metabolites were compared between “Jinpum” and “Kumkang” cultivars. Therefore, NMR based on the metabolic-phenotyping should be mostly applicable to systematic exploration of plant genetic resources as well as to metabolite based on the breeding program involved in crops productivity.