The effects of five extraction solvents and their mixtures on the yield of metabolites in crude and fractionated extracts of Annona muricata L. leaves were investigated by direct comparison. Extraction media were prepared using simplex centroid mixtures of ethanol, ethyl acetate, dichloromethane, acetone, and chloroform. The effects of the mobile phase solvent strength and the analysis wavelength on the chromatographic separation were also investigated. Solvent mixtures rather than pure solvents were found to be the most efficient extractors for the different fractions. The results indicated that the mobile phase composed of methanol/acetonitrile/water (26:27:47 v/v/v) was most suitable for the basic fraction analysis at 254 nm, whereas the mobile phase composed of methanol/acetonitrile/water (35:35:30 v/v/v) was the most adequate for the organic fraction analysis at 254 nm. The results indicated that the chromatographic profiles and number of peaks were affected by the mobile phase strength and analysis wavelength.