HPLC Separation of Vanilmandelic, Hydroxyindoleacetic, and Homovanillic Acids

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Condition 1

Column Newcrom B , 3.2x50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O - 20/80%
Buffer H3PO4 - 0.1%
Flow Rate 0.5 ml/min
Detection UV, 275 nm
Vanilmandelic, hydroxyindoleacetic and homovanillic acids are metabolites of various neurotransmitters that are used in urine tests to measure the levels of those neurotransmitters in the body. They can be baseline separated on a reverse-phase Primesep D HPLC column by varying the buffers and pH of the mobile phase consisting of water and acetonitrile (MeCN, ACN). UV detection at 230nm.

Condition 2

Column Primesep D, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN
Buffer TFA,  AmFm
Flow Rate 1.0 ml/min
Detection UV, 230 nm


Class of Compounds  Acid, Monocarboxylic acid,  Hydrophilic, Ionizable
Analyzing Compounds Vanilmandelic, Hydroxyindoleacetic, Homovanillic Acid
Application Analytes:
5-Hydroxyindole-3-Acetic Acid
Homovanillic Acid
Vanilmandelic Acid