A series of analogs of endomorphin-2 (EM-2) with phenylglycine (Phg) in position 3 or 4 were synthesized. In electrospray ionization Fourier transform ion cyclotron resonance (ESI-FT-ICR) MS/MS spectra of these compounds, some b, y, a, and internal ions were observed and slight mass differences between the calculated and observed results are obtained. Their sequences were derived successfully. However, the MS/MS patterns of these analogs with D Phg and L Phg were very similar. It is hard to distinguish them by MS/MS spectra. Moreover, if the third position was substituted by phenylglycine (l or d), a rearrangement could occur in MS/MS experiment to lose proline residue.