N-Nitrocarcosine is very polar compound with LogP of -0.5. It is a nitro derivative of glycine which is slightly acidic. HPLC method was developed on Obelisc N HILIC mixed-mode HPLC column. Nitrosarcosine is retained by combination of hydrophilic interaction chromatography and anion-exchange interaction. Compound exists in two forms which can be separated by adjusting pH of the mobile phase. Method is LC/MS compatible and uses ammonium formate as mobile phase modifier.