Introduction and aim of the work: We still need a complete understanding of the biochemical changes that occur during starvation and the impact of Zamzam water. The objective of this study is to investigate the biochemical impacts of Zamzam water on rats, as well as to compare the effects of Zamzam water and regular water on rats under conditions of severe starvation. Materials and Methods: We divided the rats into three groups: the normal control group, the starved group that drank tap water, and the starved group that drank Zamzam water. The biochemical analysis was carried out for Rats [total bilirubin, protein, urea, creatine, phosphorus, magnesium (Mg), iron (Fe), low-density lipoprotein (LDL), and high-density lipoprotein (HDL)]. Results: Rats-starved groups showed a significant reduction in body weight without a significant difference between the normal water and Zamzam groups. Biochemical analysis of the blood samples revealed lower levels of amylase, LDL, phosphorus, and total protein in the starved rats compared to the control group. On the other hand, the levels of creatinine and GOT were higher. On the other hand, there is no significant change in the other biochemical analyses between the control group and the starved groups. However, when comparing the two starved groups with each other, the experiments revealed that there are more increases in GPT, GOT, and HDL in the Zamzam group than in the tap water group, but in general, there are no significant changes in the other biochemical tests between the Zamzam water and tap water groups. Independent T-sample tests were conducted to compare means between groups, while a one-way ANOVA test was employed to assess differences among the three groups. The results revealed significant decreases in total protein levels in both starved groups compared to the control group, indicating the catabolic effects of prolonged fasting on protein metabolism. However, no significant differences were observed in most biochemical parameters between the starved groups with normal water and Zamzam water, except for amylase, creatinine, and magnesium levels. The one-way ANOVA test showed significant differences in total protein, amylase, creatinine, and magnesium levels among the three groups. These findings highlight the complex interactions between nutritional status, hydration, and metabolic pathways in response to starvation. Further research is warranted to elucidate the underlying mechanisms and clinical implications of these observations