[en] This research studies the optimal conditions for extraction, separation and determination of Tylosin & Spiramycin compounds in poultry samples, using High Performance Liquid Chromatography technique with Diode Array Detector (HPLC-DAD).The best recovery ratios for Tylosin and Spiramycin were 90.0, 93.0% respectively at 0.10ppm, and 91.90, 92.90% respectively at 1.0ppm; using an extraction solvent of Mcllvaine-EDTA at pH:2.6. The study revealed an excellent recovery ratios of 96.50, 97.50% respectively for Tylosin and Spiramycin at 0.10ppm and 98.10, 100.50% respectively at 1.0ppm, at the extraction by ultrasonic water bath with a percentage relative standard deviation less than 1.1% during half an hour. On the other hand, Cartridges solid phase extraction Oasis HLB. (Hydrophilic Lipophilic Balance Copolymer) (500mg, 5ml) showed also an excellent response for the recovery of Tylosin and Spiramycin: 96.50, 97.50% respectively at 0.10ppm and 100.10, 100.40% respectively at 1.0ppm in comparison with other cartridges. The results showed that the recovery ratios of Tylosin and Spiramycin were not affected by increasing the volume of methanol used in elution more than 3.5ml. The study showed that the holding time needed for the quantitative determination of both compounds from six poultry samples was about four hours, which considered as a good time for the application of this method in the control laboratories (Food Section). The calibration curves of Tylosin and Spiramycin were found to have an excellent linearity and correlation coefficients; R2= 0.99990, 0.99997 respectively within a range of concentrations between 0.080-1.20ppm. (author)