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Purpose: This study aimed to analyze the factors influencing the well-being of middle-aged non-shift female workers using health-determinant models. Methods: This study analyzed data from the fifth Korean Working Conditions Survey (KWCS), involving 5,449 participants. A hierarchical multiple regression analysis was employed to examine the relationships between various factors and well-being. Results: The analysis identified key factors affecting well-being in middle-aged female non-shift workers, including supervisor support, presenteeism, satisfaction with the working environment, autonomy in working hours, support from coworkers, workplace discrimination, occupation, work-life balance, sleep problems, workplace size, weekly work hours, and sickness absence. This study confirms that the well-being of middle-aged non-shift female workers is influenced by factors at the individual, social, and community levels as well as by conditions related to love. Conclusion: To enhance the well-being of middle-aged female non-shift workers, it is essential to reinforce positive factors such as support from coworkers and superiors. Additionally, addressing and mitigating negatively influencing factors such as workplace discrimination and sleep problems is crucial in promoting well-being. By implementing measures to improve these aspects, organizations and policymakers can contribute to a healthier and more supportive work environment for middle-aged, non-shift female workers.