Objective To determine the incidence of vitamin B1 deficiency in critically ill patients, to compare vitamin B1 levels between septic and non-septic patients, and to explore the relationship between vitamin B1 levels and lactate levels. Methods Using a retrospective study method, critically ill patients admitted to the Department of Intensive Care of Nanjing Drum Tower Hospital from February 2022 to November 2022 were included in the study, and the patients were divided into sepsis and non-sepsis groups according to the admission diagnosis, and the differences in the vitamin B1 levels of the patients between the two groups were analyzed, as well as the correlation between the vitamin B1 levels and the lactic acid levels. Results There was a significant difference in serum vitamin B1 levels between the sepsis patients and the non-sepsis patients [(1.6±0.3)ng/mL vs. (2.1±0.2)ng/mL, P=0. 009]. For all patients, there was no correlation between vitamin B1 levels and lactate levels. But when the patient was in a hyperlactate state (lactate level ≥2 mmol/L), vitamin B1 levels were significantly negatively correlated with lactate levels (r=–0. 229, P=0. 004). Conclusions Vitamin B1 deficiency is prevalent in critically ill patients and is strongly correlated with whether or not the patient is septic. Vitamin B1 levels are significantly and negatively correlated with lactate levels when the patient's lactate level is ≥2 mmol/L.