ABSTRACT Vital signs have been widely used to assess the disease severity of patients, but there is still a lack of research on their circadian rhythms. The objective is to explore the circadian rhythms of vital signs in critically ill patients and establish an in-hospital mortality prediction model. Study patients from the recorded eICU Collaborative Research Database were included in the present analyses. The circadian rhythms of vital signs are analyzed in critically ill patients using the cosinor method. Logistic regression was used to screen independent predictors and establish a prediction model for in-hospital mortality by multivariate logistic regression analysis and to show in the nomogram. Internal validation is used to evaluate the prediction model by bootstrapping with 1000 resamples. A total of 29,448 patients were included in the current analyses. The Mesor, Amplitude, and Peak time of vital signs, such as heart rate (HR), temperature, respiration rate (RR), pulse oximetry-derived oxygen saturation (SpO2), and blood pressure (BP), were significant differences between survivors and non-survivors . Logistic regression analysis showed that Mesor, Amplitude, and Peak time of HR, RR, and SpO2 were independent predictors for in-hospital mortality in critically ill patients. The area under the curve (AUC) and c-index of the prediction model for the Medical intensive care unit (MICU) and Surgical intensive care unit (SICU) were 0.807 and 0.801, respectively. The Hosmer-Lemeshow test P-values were 0.076 and 0.085, respectively, demonstrating a good fit for the prediction model. The calibration curve and decision curve analysis (DCA) also demonstrated its accuracy and applicability. Internal validation assesses the consistency of the results. There were significant differences in the circadian rhythms of vital signs between survivors and non-survivors in critically ill patients. The prediction model established by the Mesor, Amplitude, and Peak time of HR, RR, and SpO2 combined with the Acute Physiology and Chronic Health Evaluation (APACHE) IV score has good predictive performance for in-hospital mortality and may eventually support clinical decision-making. Abbreviations: ICU: Intensive care unit; MICU: Medical intensive care unit; SICU: Surgical intensive care unit; HR: Heart rate; RR: Respiration rate; SpO2: Pulse oximetry-derived oxygen saturation; BP: Blood pressure; SBP: Systolic blood pressure; DBP: Diastolic blood pressure; APACHE: Acute Physiology and Chronic Health Evaluation; bpm: beats per min; BMI: Body mass index; OR: Odd ratio; CI: Confidential interval; IQR: Interquartile range; SD: Standard deviation; ROC: Receiver operating characteristic; AUC: area under the curve; DCA: Decision curve analysis; IRB: Institutional review board.