3,5-Dihydroxybenzoic Acid

Applications:

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Mixed-Mode Separation of Dihydroxybenzoic Acids

Primesep B offers better selectivity over traditional, reversed-phase C18 columns for separating regioisomers of aromatic dihydroxybenzoic acids (2,3-dihydroxybenzoic, 2,4-dihydroxybenzoic, 2,5-dihydroxybenzoic 3,4-dihydroxybenzoic, 3,5-dihydroxybenzoic acids) Primesep B combines reversed-phase and anion-exchange mechanism with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA).

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2,3-Dihydroxybenzoic Acid
2,4-Dihydroxybenzoic Acid
2,5-Dihydroxybenzoic Acid
3,4-Dihydroxybenzoic Acid
3,5-Dihydroxybenzoic Acid
Carboxylic Acids

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HPLC Separation of Dihydroxybenzoic Acid

Separation of hydrophilic isomers is very difficult when only one mechanism of retention, like reversed-phase, is available. Isomers have very similar hydrophobic properties. In case of mixed-mode chromatography, small difference in hydrophobic and ionic properties allows to separate isomers. Isomers of dihydroxybenzoic acids are separated on a Primesep D column with good selectivity and peak shape. This method can be used for separation of other hydrophilic acidic compounds as well as hydrophobic basic compound.

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2,3-Dihydroxybenzoic Acid
2,4-Dihydroxybenzoic Acid
2,5-Dihydroxybenzoic Acid
3,4-Dihydroxybenzoic Acid
3,5-Dihydroxybenzoic Acid

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HPLC Separation of Organic Acids in HILIC and Anion-Exclusion Mode on Primesep S2 Column

Organic acids were separated on a HILIC/cation-exchange column in HILIC/anion-exclusion mode. This column can be used for analysis of polar compounds in HILIC mode. If compounds are ionizable, additional mode of interaction can be added (cation-exchange or anion-exclusion).

Condition

Column	Primesep S2, 4.6x150 mm, 5 µm, 100A
Mobile Phase	MeCN/H2O - 85/15%
Buffer	AmAc pH 5.0 15 mM
Flow Rate	1.0 ml/min
Detection	ELSD, 50C UV 250 nm

 

Description

Class of Compounds	Nucleosides,  Hydrophilic, Ionizable
Analyzing Compounds	Toluenesulfonic acid, Naphthalenedisulfonic acid, 3.5 DHBA, Ascorbic acid,

 

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1,5-Naphthalenedisulfonic Acid
3,5-Dihydroxybenzoic Acid
Ascorbic Acid
Organic Acids
p-Toluenesulfonic Acid (PTSA)

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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, 3.2x100 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, 3.2x100 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

 

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2,4-Dihydroxybenzoic Acid
2,5-Dihydroxybenzoic Acid
3,5-Dihydroxybenzoic Acid

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