Organic matrix components of modern oil paintings are complex and difficult to identify, and their analysis and identification is one of the difficulties in today’s scientific and technological research on the conservation of cultural relics. The use of micro-destructive analysis methods to investigate the structure, materials and techniques for complex oil painting matrices can elucidate the nature and condition of specific layers of painting. These physicochemical findings are crucial for conservation professionals to make restoration decisions as to the correct handling and concern the future state of restored painting, and thus have become a focus of attention for many researchers. In this paper, FT-IR, Py-GC/MS and THM-Py-GC/MS micro-destructive techniques were used to analyze the scientific and technological characteristics of an oil painting, Autumn on the Dnieper River, in the collection of the National Museum of China. It was found that the combination of these micro-destructive analysis techniques could effectively identify the binding media used for the oil painting and the type of additives present therein to maximally, accurately and integrally extract the information contained in the sample. The results show that the organic cementing materials for the oil painting are polyvinyl acetate materials and linseed oil, while dibutyl phthalate was used as a plasticizer in the polyvinyl acetate materials. The absence of varnish resin-like components in the pyrolysis products suggests that the glossy layer on the lower part of the painting is not the layer of varnish that the author intended to put on. In addition, beeswax was added to some areas of the painting as a modifying material that serves to increase pigment consistency and plasticity, and also to diminish gloss.