A number of 17 sulfonamides (SNs) determination in porcine tissues using two new materials including Enhanced Matrix Removal for Lipid (EMR-L) and Oasis PRiME hydrophilic-lipophilic balance (HLB), and the conventional liquid-liquid extraction with n-hexane (LLE) sample preparation methods were evaluated and compared. Samples were extracted uniformly with acidified acetonitrile and cleaned up by the three sample preparation methods, and then, analytes were further separated by ultrahigh-performance liquid chromatography (UHPLC) and detected by a triple quadrupole mass spectrometry (MS/MS) or a quadrupole-time-of-flight tandem mass spectrometry (Q-TOF/MS). The results showed that the matrix effects from the EMR-L and HLB were significantly lower than that from LLE, suggesting that these two new materials are superior to n-hexane in the precipitation of proteins and the adsorption of lipids. Moreover, the recoveries of 17 SNs were quantified by the matrix-matched calibration curve at spiked level of 5, 10, and 20 μg/kg, and 97.0% of the results satisfied method validation requirements. The samples cleaned up by EMR-L and HLB achieved the highest average recovery in liver and kidney with high moisture content, and muscle which is high in fat, respectively. In addition, Q-TOF/MS could play a good role in aided verification based on the result of repeated validation test. In summary, either combination of approaches could be used to achieve monitoring purposes; it is still worthwhile to adopt a specific sample preparation method and MS detector for the quantification in a specific matrix.