This study evaluated 173 university students' different understanding levels on their conceptions of the particulate nature of the matter. To develop an assessing tool with high validity and reliability to evaluate macroscopic, symbolic and microscopic conception of students' understanding levels for the particulate nature of the matter in chemistry equilibrium. This study applied SPSS statistic method to analyze students' understanding levels. There were two dynamic results to show in the following. Firstly, there were 3 reviewers to present substantial inter-rater reliability which Kendall's coefficient of concordance was 0.61 with significant level p<.05. Secondly, 61.8% university students' understanding levels present low level 1 and level 2. It would be expected that the struggles of the improve chemistry instruction strategy for their concept understanding of the particulate nature of the matter in the future.