Intestinal coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses for the industry. It compromises the nutrition absorption, leading to weight loss and elevated mortality rates. Furthermore, the stress caused by the infection can compromise the immune system, making poultry more susceptible to secondary infections and reducing overall productivity. As a result, simple analytical techniques are critical for determining anticoccidial drugs. A new, sensitive, and environmentally friendly HPLC method was developed for determining amprolium (AMP), sulfaquinoxaline (SUL), diaveridine (DIV), and vitamin K3 (VIT K3) in their formulations for the first time. Stability tests were performed under diverse stress conditions to verify the safety and efficiency of the formulation throughout its designated shelf time. These investigations ascertain the influence of various environmental conditions on a drug's chemical stability and physical characteristics. A Supelcosil C18 column was used as the stationary phase, and 0.05 M KH