Mosquitoes (Anopheles and Culex), the vector of many diseases, are highly resistant to insecticides in many developing countries. It would be of great relevance to search for alternatives to control mosquito-borne diseases. This study aimed to run phytochemical screening for Calotropis procera Ait flower parts, thin layer chromatography (TLC) test, in addition to test their potentialities against Anopheles and Culex larvae. Fresh flowers of C. procera were collected from the gardens of the main campus, University of Gezira and were separated into four main parts: petals, corona, gynostigium and both styles and ovary. The dried plant materials were used to run the phytochemical, TLC and the toxicity tests, in the Faculty of Engineering and Technology, University of Gezira. Samples of Anopheles and Culex larvae were collected from the breeding sites of some villages near to Wad Medani City. The results showed that, petal and corona parts contain tannins, flavonioids, glycosides and steroids, while the other parts lack tannins and glycosides. Similar number of spots were detected using TLC test among flower parts, but with different Rf values. At 1.2 g/L, corona and gynostigium parts resulted in 90%-100% mortalities on Anopheles and on Culex larvae, while Ovary and style part showed low toxicity (10% - 30%). It was clear that, Culex larvae were relatively susceptible to C. procera flower parts more than Anopheles larvae. Similar studies should be run for all Sudanese aromatic and medicinal plants in order to establish local database.