Creatine, the precursor of creatine (guanidinoacetic acid), and the final metabolite of creatine (creatinine) were separated using a polymer-based amino column, Asahipak NH2P-40 2D, under HILIC mode. Hydrophilic compounds were retained at a high concentration of acetonitrile under HILIC mode condition. This is advantageous for the high sensitivity analysis by ESI-MS/MS.
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Sample : 5 μL
10 ng/mL each
(in H₂O/CH₃CN=1/3)
1. Creatinine
2. Creatine
3. Guanidinoacetic acid

Column       : Shodex Asahipak NH2P-40 2D (2.0 mm I.D. x 150 mm)
Eluent       : 20 mM HCOONH4 aq./CH3CN=30/70
Flow rate    : 0.2 mL/min
Detector     : ESI-MS/MS (MRM Positive)
Column temp. : 30 °C