Forensic investigations utilizing bloodstains analysis have experienced rapid growth. Samples collected from crime scenes may undergo exposure to various conditions before analysis. Several factors, including blood aging, environmental conditions like temperature and humidity, and pH levels, can impact the appearance and preservation of bloodstains. This study sought to assess the impact of different temperature gradients on bloodstains pattern, examining temperature variations (4°C, 37°C, and 50°C) and aging durations (0, 24, 48, and 72 hrs). Using human and cat blood samples, macroscopic changes were observed, followed by phenolphthalein and Takayama tests for blood identification. Results reveal that stains exposed to higher temperatures and longer durations exhibited darker coloration, dryness, and cracking, while those at 4°C remained relatively unchanged, except for increased gelatinous consistency over time. Both phenolphthalein and Takayama tests successfully identified blood in stains across temperature and time variations, with variations in color intensity and crystal morphology observed, particularly at higher temperatures and longer durations. This research underscores the importance of considering environmental factors when analyzing bloodstains for forensic purposes.