Accurate analysis of a blood stains which found at the crime scene is extremely important in reconstructing the crime. Because the bloodstain is evidence that is closely related to the occurrence of the crime. Currently, chemical analysis of bloodstain has been actively studied from many perspectives. However, it has problems to apply the results of the research to the field. One of the problems is the unknown amount of initial blood volume of bloodstain. For example, index materials of bloodstain age change based on initial quantities. Because of unknown amount of initial blood volume, we couldn’t apply the research results to the field. In order to solve these problems, we have found an internal standard metabolite that maintains a consistent amount of initial blood volume even after the passage of time. In this study, the sample for making bloodstain was collected from six healthy men and women in their 20s who did not take any medicine at least since a week ago. Using the HPLC-QTOF, metabolic profiling of bloodstain was performed according to the bloodstain age. We filtered metabolites that revealed on all samples. Among them, we selected metabolites with p-value <0.05. Then 36 metabolites were obtained by filtering which of selected metabolites had changed from the day 0 to the day 21 of less than 2-fold. The CV values were calculated from the abundance of all samples. Six metabolites with less than 5% CV was obtained. The candidate metabolites for internal standard found in this study will serve as a reference point for changes of the index materials and help prior study results of bloodstain analysis to be applied in the field. It can also be provided as a more objective clue.