HPLC separation of Dihydroxybenzoic acid in Hydrogen-Bonding mode on SHARC 1 HPLC column

 

 

Application Notes: Dihydroxybenzoic acids are polar acidic compounds containing two hydroxyl groups as well as a carboxylic acid. Isomers of dihydroxybenzoic acids are very polar with varying degrees of acidity. While the isomers can be analyzed by mixed-mode chromatography on Primesep D column, the presence of hydroxyls and carboxylic acid fragments also make these compounds good candidates for using the new hydrogen bonding columns for analysis. Since all these compounds have the same amount of hydroxyls they are separated based on accessibility of this hydroxyl group. Retention time can be adjusted by the amount of ACN, MeOh and concentration of additives (formic acid, ammonium formate, triethylamine, etc.). Our method is fully compatible with LC/MS and prep chromatography and can be used for other hydroxy and carboxylic acids containing compounds..

Application Columns: SHARC 1, 3.2×100 mm, 5 um, 100A. To learn more about SHARC 1 columns click here. To order this column click here. To see more chromatographic separations check our web site.

Application Compounds: 2,6-dihydroxybenzoic acid, 2,4-dihydroxybenzoic acid, 3,5-dihydroxybenzoic acid, 2,5-dihydroxybenzoic acid

Detection Technique: UV, LC/MS

Condition

Column Sharc 1, 3.2×100 mm, 5 µm, 100A
Mobile Phase MeCN/MeOH
Buffer AmFm, Formic acid
Flow Rate 1.0 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Vitamin, Supplements
Analyzing Compounds 2,6-dihydroxybenzoic acid, 2,4-dihydroxybenzoic acid, 3,5-dihydroxybenzoic acid, 2,5-dihydroxybenzoic acid

 

Application Column

SHARC 1

Column Diameter: 3.2 mm
Column Length: 100 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
2,4-Dihydroxybenzoic Acid
2,5-Dihydroxybenzoic Acid
3,5-Dihydroxybenzoic Acid

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.