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Method Conditions

Column: Cogent F5™,
              Phenyl-Pentafluoro
Catalog Number: 60022-7.5
Column Dimensions: 75mm x 4.6mm ID
Stationary Phase Description: 5µm, spherical silica
              with 60A pores, Phenyl phase
Mobile Phase: 10:90 20mM Ammonium
              Formate/Ammonium Hydroxide:Acetonitrile,
              Elution Mobile Phase: 90:10 20mM
                    pH 8.00 Ammonium
              Formate/Ammonium Hydroxide:Acetonitrile
Flow Rate: 3ml/minute
Sample: Histamine Standard
Injection Volume: 1µl
Detection: 254 nm

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Discussion of Chromatogram:
Getting Histamine to retain on an HPLC column can be challenging. It often elutes with the solvent front. Using the Cogent F5™ Pentafluoro Phenyl column and the above method conditions, you can retain histamine from your sample for as long you like due to the hydrogen bonding that takes place. To elute the compound, simply change your mobile phase to 90% 20mM Ammonium Formate/Ammonium Hydroxide and 10% Acetonitrile and the compound will quickly release from the column. This is a very effective tool for other compounds similar to Histamine that are likely to hydrogen bond.