Background: Stereotypical beliefs and gender biases play a limiting and discouraging role in the development of women's entrepreneurship, which can have a significant impact on women's willingness to become entrepreneurs. Methods: This research aims to identify and analyze the gender-based limitations affecting the entrepreneurial opportunities of female entrepreneurs using a mixed-methods approach. This research is applied in terms of its objective. In the qualitative phase, 14 experts were selected using a purposeful sampling method. Data collection was conducted through library studies and semi-structured interviews. In this section, the data were analyzed in three stages: open, axial, and selective coding, which resulted in the identification of 51 indicators and 12 components. The research questionnaire was designed based on these indicators. In the quantitative section, the statistical population was all female entrepreneurs in the pharmaceutical companies under study, which totaled 425 people. Using Cochran's formula and a simple random sampling method, 170 people were selected as the sample size. Data collection was carried out using a researcher-made questionnaire, whose reliability was confirmed with a Cronbach's alpha coefficient of 0.89 and validity using expert opinion. Data analysis was performed using SPSS23 and PLS3 Smart software. Results: Based on the research findings, 12 components of social, cultural, economic, educational, legal, behavioral, personality, individual, commercialization, knowledge enhancement, entrepreneurial awareness, and market recognition limitations were identified under the title of dimensions and components of gender-based limitations affecting women's entrepreneurial opportunities. The results also showed that social limitations (β=-0/578), cultural limitations (β=-0/429), economic limitations (β=-0/408), educational limitations (β=-0/346), legal limitations (β=-0/297), behavioral limitations (β=-0/215), personality limitations (β=-0/189), and individual limitations (β=-0/138) have a negative and significant impact on entrepreneurial opportunities