The purpose of this in vitro study was to investigate the efficacy of micro-Raman spectroscopy on detecting mineral content change during the demineralization and de/remineralization cycling process. The enamel samples (n=55) were randomly divided into three groups and separately treated with demineralization solution (n=20), de/remineralization cycling solution (n=30), and distilled water (n=5). Micro-Raman spectroscopy, microhardness (MHS), and the released calcium ions concentration were performed before and after treatment, respectively. A one-way analysis of variance (ANOVA) with a post hoc Tukey test was used to analyze the results. The Spearman correlation coefficients among the parameters of Raman relative intensity decrease (RRID%), the percentage of MHS loss (PML), and the released calcium ions concentration were also analyzed. In demineralization group, RRID%, PML, and released calcium ions concentration were highly correlated with each other (r=0.979, p<0.001; r=0.984, p<0.001; and r=0.983, p<0.001, respectively). While for the de/remineralization cycling group, there also existed a high correlation between RRID% and PML (r=0.987, p<0.001). In conclusion, micro-Raman spectroscopy could effectively monitor the mineral content change, and its efficacy was validated by the measurement of released calcium ions concentration and MHS.