A sensitive method for the simultaneous analysis of five estrogens in sewage sludge was developed. The extraction and purification steps were optimized and the matrix effects were evaluated. The chromatographic gradient was optimized to limit matrix effects and the analysis step was performed by LC-MS/MS. The method consists of an ASE® extraction with a solvent mixture water/methanol 80/20v/v at 100°C followed by two consecutive purifications on Oasis HLB® and florisil cartridges. A thorough validation of the developed method was performed. Recoveries determined at two different spiking levels ranged between 86% and 126% depending on the molecule. Repeatability was evaluated on five replicates of the same sludge sample spiked at two different levels and measuring native estrogens in triplicates of 12 sludge samples. Relative standard deviations obtained a range of between 2% and 27%. Reproducibility was also studied by analyzing the same sludge on four different days: the relative standard deviation ranged between 14% and 20% for E1, βE2 and E3. For αE2, poor reproducibility (68%) was observed but it was linked to the very low quantity of αE2 present in the sludge sample and not to the method performance. The specificity of the method was evaluated on various sludge samples spiked at different spiking levels showing that performances of the proposed method were not modified by matrix effects. Finally, sensitivity of the method was evaluated taking into account both instrumental sensitivity and matrices; the estimated limits of quantification were around 1 ng/g for E1, between 2 and 4 ng/g for αE2, βE2, and E3 and around 5 ng/g for EE2.