Work settings are the focal determining factor for the motivation of workers. As the public health care segment in Kenya suffers from problems of professionalization with human resource, the private healthcare sector is expanding fast with the emphasis on management of human resource lying at the centre of the success of private health facilities. The purpose of the study was to establish the role of human resource management practices on the healthcare workers motivation in Machakos County. The specific objectives of the study were to establish the effect of recruitment on motivation of healthcare workers, determine the effect of training on motivation of healthcare workers, examine the effect of performance management on healthcare workers’ motivation and assess the effect of reward management on motivation of healthcare workers in Machakos County. The study adopted a descriptive research design in which the target population was 1179 healthcare employees in Machakos County. Disproportionate stratified sampling and simple random sampling was used to sample 299 respondents. The questionnaire was the selected tool for data collection. Trained research assistants were utilized to oversee the issuance of questionnaires to the participants of this study. Quantitative data collected was analysed by the use of descriptive statistics. The study also used inferential statistics namely correlation and regression analysis to test the relationship between the dependent and the independent variables. The findings are presented in figures and tables. The findings revealed that recruitment, training, and reward management positively influenced healthcare workers motivation. There was however a negative relationship between performance management and employee motivation. The study recommended that first, healthcare facilities should enhance the use recruitment in order to attract the best talents. Secondly, they should embrace the use of training to enhance employee skills and thirdly, embrace performance management with the goal of enhancing employee motivation. Finally, that the health care organizations should use more of rewards to motivate their employees.