Polydivinylbenzene (pDVB) microspheres with the particle size of 14.2 mm were prepared with divinylbenzene as monomer by membrane emulsification and suspension polymerization at 75℃ for 20 h. The prepared pDVB microspheres had uniform particle size (Rspan=0.71), mean pore size of 65 nm and specific surface area of 160 m2/g. The pDVB microspheres were packed into f4.6 mm′250 mm stainless steel column by slurry packing method, and tested in separation of peptide drug by high performance liquid chromatography. Afterword, a very fast separation process of octreotide from synthetic peptide crude mixtures on pDVB reversed phase was carried out. Fourier-transformed infrared resonance (FT-IR) spectroscopy, and reversed-phase HPLC combined with electrospray ionisation mass spectrometry (ESI-MS) were used for analysis of resultant peptides. Under the optimized conditions, the octreotide was purified from 42.89% to 99.99%.