NMR-based metabolomics is a useful and reliable approach for comprehensively evaluating the metabolic products in a wide range of biological systems such as animals, plants, and microbial ecosystems, and was also used for comprehensive analysis of the metabolites in complex microbial ecosystems in our previous studies1. 1D 1H-NMR spectra are common method that is widely used for the NMR-based metabolomic approach, however, some overlapped signals such as saccharide region were major drawback for annotation and assignment of each signal and for subsequent multivariate statistical analysis. Here we described an integrated metabolomics approach using solution-state 1H- and 13C-NMR spectra in combination with HSQC, HSQC-TOCSY and J-RES NMR spectroscopy with time course variations to evaluate the degradation process of biological complex mixtures from plants in complex microbial ecosystem. As the 13C-labeled plant samples, we prepared stem of soybeans, stem of tomato, and leaf of Japanese mustard spinach. The freeze-dried and milled samples were incubated with agricultural field soil at 30 °C for 50 days. From 1H-NMR spectra, metabolic dynamics of produced organic acids in each sample were elucidated although some saccharides couldn’t be evaluated because of the signal overlapping. To compensate for the deficiencies, 13C-DEPT and HSQC NMR spectroscopy were performed and the obtained data were combined with 1H-NMR spectral data. We successfully visualized the time course differences of metabolic pathway from sugars to amino acids and organic acids in each complex microbial ecosystem by using this integrated approach. In this meeting, we will discuss a relationship between each metabolic pathway and microbiota.