Ultra-Fast HPLC Analysis of Aromatic Hydrophilic Acids on Coresep SB Column in Reversed-Phase and Anion-Exchange Modes
Application description
Homovanillic and vanilmandelic acid are major catecholamine pathway metabolites. VMA and HVA can be found in urine along with other metabolites. Homovanillic and vanilmandelic acids were separated by combination of weak reversed-phase and anion-exchange mechanisms on the core-shell column. This method can be used for analysis of hydrophilic acidic compounds with LC. MS compatible conditions. Both compounds elute with a good peak shape. Retention time is controlled by amount of acetonitrile, buffer concentration, and buffer pH. Stationary phase does not change its ionization state with the change of buffer pH, but ionization state of acidic compounds is greatly affected by buffer pH.
Conditions of Experiment
Column: | Coresep SB |
Separation Modes: | reversed-phase and anion-exchange |
Column Dimenstions: | 3.2 x 100 mm, 2.7 um, 90A |
Mobile Phase: | 50% ACN with 50 mM sodium phosphate (monobasic) pH 3 |
Detection: | UV 230 nm |
Sample: | 0.3 mg/ml |
Injection: | 1 uL |
Flow rate: | 0.8 ml/mim |
Analytes
Class of compounds: | Aromatic acid, Catecholamine, Neurotransmitter, Organic acid |
Nature of compounds: | Acidic, Hydrophilic, Polar |
Compounds: | Homovanillic acid, Vanilmandelic acid |