Abstract: The numbing sensation serves as one of the important indicators for evaluating the quality of Zanthoxylum plants. In this study, pericarp samples from 113 Zanthoxylum accessions of different geographical origins were subjected to qualitative and quantitative analyses of their primary numbing substances—hydroxyl-ε-sanshool, hydroxyl-α-sanshool, hydroxyl-γ-sanshool, and hydroxyl-β-sanshool—using high-performance liquid chromatography (HPLC). In addition, cluster analysis and principal component analysis (PCA) were employed to decipher the compositional characteristics and content variations of numbing compounds across different origins. The research results show that there are significant differences in the composition patterns and contents of pungent substances among different geographical sources and varieties of Zanthoxylum. Horizontally, longitude is extremely significantly negatively correlated with the content of hydroxy-γ-sanshool, and a larger longitude leads to a smaller content of hydroxy-γ- sanshool, which is lower than the content of hydroxy-ε-sanshool it contains, thus changing the composition characteristics of the pungent substances of Zanthoxylum. Vertically, altitude is extremely significantly positively correlated with the total content of pungent substances. In areas with higher altitudes, the content of pungent substances in Zanthoxylum is also higher. Hydroxy-α-sanshool had the highest proportion in the total numbing substances, serving as the primary contributor to the characteristic numbing sensation. Notably, Hanyuanhuajiao exhibited distinct differentiation due to their significantly elevated levels of hydroxyl-γ-sanshool. This study systematically elucidates the compositional profiles and quantitative variations of numbing substances in Zanthoxylum plants, providing a scientific basis for the selection of superior cultivars and enriching the foundational research framework on the chemical constituents responsible for its characteristic numbing sensation.