In modern education, mobile applications offer flexibility for learning anytime and anywhere, yet biochemistry lacks mobile-based virtual laboratories. This study aimed to develop a 2D Mobile-Based Virtual Laboratory Simulation (2D-MViLS) for blood glucose level measurement and evaluate its impact on pharmacy students' learning experience. In stage 1, storyboards were developed and expanded to show the overall flow and simulation in interactive 2D form using Unity for Android devices. In stage 2, usability testing was carried out. In Stage 3, reflective essays from first-year pharmacy students were analyzed using both qualitative and quantitative methods. Three key themes were identified: students' feelings, their understanding of the learning outcomes, and their reflections on the need to enhance their current knowledge and skills. Thematic analysis revealed a positive shift in students' feelings after using the 2D-MViLS. Quantitatively, 44% of students demonstrated a highly positive change, moving from pessimistic to optimistic feelings, 17% showed moderate positive change, and 51% expressed optimism before and after the simulation. Quiz marks were consistently high across all groups, with minimal variation regardless of the number of learning outcomes mentioned. Additionally, 73.9% of students identified areas for knowledge and/or skill enhancement, highlighting their engagement with the simulation. In conclusion, students' reflections reveal the positive impact of 2D-MViLS on their learning experience, indicating that implementing mobile-based virtual laboratories could significantly enhance the learning experience in biochemistry education and potentially lead to improved teaching methods in pharmacy and related disciplines.