Objective Through comparing the drinking water, deionized water, pure water, double steamed water, purified water and sterilized water for injection and so on the various character water in vitro hemolysis test of hemolysis degree, we hope to provide the better positive control. Methods The hemocytolysis of various character water was observed by means of the routine test tube method in vivo (macroscopic observation methods) and modified hemolytic test method in vitro (spectrophotometry). Results Hemolytic crisis occur between the six character water, the hemolysis degree of sterilized water for injection is the largest, drinking water is minimum, the difference of several other character water is small, macroscopic observation method and spectrophotometry results are consistent. Conclusions In this test conditions, several character water to the rabbit red blood cells of hemolysis degree is sterilization water for injection > purified water > double steamed water > pure water >deionized water > drinking water, we recommended sterilized water for injection for hemolysis test positive control.