Workplace stress is a growing occupational health concern, adversely affecting mental well-being, productivity, and organisational outcomes. Evidence from Indian professional populations remains limited, particularly at the regional level. A cross-sectional study was conducted among 430 professionals working in various organisations in Pune (Maharashtra) between September and October 2024. Participants were selected using convenience sampling. Workplace stress was assessed using the standardised Workplace Stress Scale (American Institute of Stress). Data were analysed using SPSS version 18. Descriptive statistics, chi-square tests, and binary logistic regression were applied. The results showed that mean workplace stress score was 23.4 ± 7.73, indicating a moderate stress level. Overall, 45.8 percent of participants reported moderate stress, 32.3 percent mild stress, and 21.9 percent severe stress. Significant associations were observed between stress levels and age, gender, profession, years of service, and working hours (p < 0.05). Regression analysis showed that professionals working >10 hours/day (OR = 2.41; 95% CI: 1.52–3.81) and healthcare professionals (OR = 1.98; 95% CI: 1.26–3.12) had significantly higher odds of moderate-to-severe stress. To conclude, workplace stress is highly prevalent among professionals in Pune, particularly among healthcare workers and those with longer working hours. Organisational interventions focusing on workload management, mental health support, and policy reforms are urgently required.