In this paper, polyethylene glycol modified (PEGylated) magnetic carbon nanotubes were developed as solid-phase extraction nanosorbents for the determination of methylprednisolone in rat plasma. The procedure mainly involved two steps including preparation of PEGylated magnetic nanosorbents and bioanalysis. Monodisperse magnetites (Fe3O4) anchored onto multi-walled carbon nanotubes (MWCNTs) were synthesized by a facile solvothermal synthesis method. The obtained MWCNTs-Fe3O4 nanomaterials were further non-covalently functionalized by a surfactant phospholipids-polyethylene glycol (DSPE-PEG). Owing to dispersibility and high enrichment ability, water-soluble PEGylated MWCNTs-Fe3O4 nanomaterials can provide more efficient way for the extraction of methylprednisolone than only MWCNTs-Fe3O4 used. The methylprednisolone could be easily extracted via π-π stacking interactions with PEGylated MWCNTs-Fe3O4. The captured methylprednisolone/nanosorbents were isolated from the matrix by placing a magnet, and desorbed by the elution solvent composed of acetonitrile. Extraction conditions such as amount of nanosorbents added, adsorption time, desorption solvent, and desorption time were investigated and optimized. The method recoveries were obtained from 88.2% to 92.9%. Limits of quantification and limits of detection of 0.01 and 0.005μg/mL were acquired, respectively. The precision ranged from 4.2% to 7.8% for within-day measurement, and for between-day variation was in the range of 5.5-9.0%. Moreover, the analytical performance obtained by PEGylated magnetic MWCNTs was compared with that of magnetic MWCNTs. The results indicated that the approach based on PEGylated magnetic MWCNTs was useful for the analysis of methylprednisolone in the complex plasma.