Rheum tanguticum (R. tanguticum) has been widely used for the treatment of inflammatory diseases in clinical. However, limited research exist on the quality evaluation of various R. tanguticum locations, which has certain drawbacks. In this study, Fourier-transform infrared spectroscopy (FTIR) and high-performance liquid chromatography (HPLC) were used to comparative study on the chemical contents of R. tanguticum, to clarify the relationship between the chemical contents and the spatial distribution of R. tanguticum. First of all, the FTIR spectra of 18 batches of R. tanguticum were examined. Following the cluster analysis, the FTIR spectra of various production locations differed. To some extent, establishing the double index analysis sequence of common and variation peaks may differentiate distinct production locations of medicinal materials. The HPLC fingerprint of R. tanguticum was constructed to further explore the link between components and their origin. PCA of common peaks of 18 batches of R. tanguticum indicated that R. tanguticum grown in Gannan and Qinghai had a tendency to separate t[2], however this trend was not noticeable. Then, OPLS-DA model was established, and the key differential components of R. tanguticum produced in Gannan and Qinghai were discovered to be R16, R37, R46, and R47 (Aloe emodin) (VIP ≥ 1 and P < 0.05). At last, Pearson's test was used to examine the relationship between longitude, latitude, altitude, and composition. Longitude was significantly positively correlated with R28 and R30 (P < 0.05), and a very significantly positively correlated with R35, R36, R37, R46, and R47 (P < 0.01). Latitude was significantly negatively correlated with R34, R35, and R40 (P < 0.05), and extremely significantly negatively correlated with R28, R30, R36, R37, R46, and R47 (P < 0.01). Altitude was significantly positive correlation with R36 and R37 (P < 0.01). The results of our study can provide insights into R. tanguticum quality control and aid in establishing a natural medication traceability system.