Many studies have examined the impact of factors affecting accident severity in rural areas; however, little attention has been paid to different lighting conditions (LCs), and less to the detailed categories and precise determining of twilight. In this paper, solar altitude angle (SAA), as a basis for differentiating and categorizing LCs, is proposed to investigate explanatory variables in much greater detail. For each LC, namely, dark, twilight, dark lit (dark with street lights) and daylight, separate random parameter models are developed to investigate the impacts of some factors on crash injury severity data of 2017 and 2018 in two lane rural roads of Texas. The model estimation results indicated that different LCs have various contributing factors, indeed, to each injury severity, further stressing the significance of investigating crashes based on SAA. The key differences include crash location, marked lane, grade direction, no passing zone, shoulder width, weekday and collision type. The important findings were that developing artificial lighting at intersections and LED raised pavement markers on two lane rural roads could lead to enhanced road safety under dark LCs. Furthermore, increasing shoulder width in straight segments of two lane rural roads is important for decreasing severe injury in daylight conditions.