Introduction: Respect for patients’ rights and observance of professional ethics standards improves medical services that will have an essential role in promoting patients’ health. Objective: This study aimed to determine the relationship between awareness of patient’s rights charter and professional ethics with patient’s rights charter observance. Materials and Methods: This analytical correlational study was performed on 285 staff members (196 nurses and 85 midwives) working in 2 hospitals affiliated to Alborz University of Medical Sciences, Iran, and 2 social security hospitals. They were selected using a stratified random sampling method. The study data were collected using the sociodemographic characteristics questionnaire, awareness and observance of the patient’s rights, and professional ethics charter questionnaires. Descriptive statistics and inferential statistics, including the independent t-test, Pearson correlation, Chi-square tests, and multivariate linear regression, were used for data analysis. Results: The Mean±SD ages of nurses and midwives were 36.2±8.3 and 37.5±7.9 years, respectively. The results also showed an association of professional ethics and awareness of patient’s rights charter with patient’s rights charter observance in both nurses (P=0.0001) and midwives (P=0.0001). Based on multivariate linear regression analysis, there were relationships between gender (B=-0.3; 95%CI; -2.0, 2.0; P=0.000), work experience (B=-0.6; 95%CI; -4.0, 3.0; P=0.000), marital and employment status (B=-3.2; 95%CI; -6.0, -0.0; P=0.045), in nurses and between job position (B=9.5; 95%CI; 5.4, 13.5; P=0.000), interest to work (B=-7.9; 95%CI; -11.5, -4.3; P=0.000), and income status (B=6.3; 95%CI; 3.3, 9.2; P=0.000), in midwives with Observance of the Charter of Patient Rights. Conclusion: It is possible to improve compliance with the charter of patient’s rights and its dimensions by strengthening the knowledge of nurses and midwives in this area and their professional ethics, which results in patient satisfaction in this area.