Vertical stratification in marine sediment profiles indicates physical and chemical sedimentary processes and, thus, is the first step in sedimentary research and in studying their relationship with global climate change. Traditional technologies for studying vertical stratification have low efficiency; thus, new technologies are highly needed. Recently, visible and near-infrared spectroscopy (VNIR) has been explored to rapidly determine sediment parameters, such as clay content, particle size, total carbon (TC), total nitrogen (TN), and so on. Here, we explored vertical stratification in a sediment column in the South China Sea using VNIR. The sediment column was 160 cm and divided into 160 samples by 1 cm intervals. All samples were classified into three layers by depth, that is, 0–50 cm (the upper layer), 50–100 cm (the middle layer), and 100–160 cm (the bottom layer). Concentrations of TC and TN in each sample were measured by Elementa Vario EL III. Visible and near-infrared reflectance spectra of each sample were collected by Agilent Cary 5000. A global model and several classification models for vertical stratification in sediments were established by a Support Vector Machine (SVM) after the characteristic spectra were identified using Competitive Adaptive Reweighted Sampling. In the classification models, K-means clustering and Density Peak Clustering (DPC) were employed as the unsupervised clustering algorithms. The results showed that the stratification was successful by VNIR, especially when using the combination of unsupervised clustering and machine learning algorithms. The correct classification rate (CCR) was much higher in the classification models than in the global model. And the classification models had a higher CCR using K-means combined with SVM (94.8%) and using DPC combined with SVM (96.0%). The higher CCR might be derived from the chemical classification. Indeed, similar results were also found in the chemical stratification. This study provided a theoretical basis for the rapid and synchronous measurement of chemical and physical parameters in sediment profiles by VNIR.