HPLC Separation of Three Isomers of Aminobenzoic Acid on Coresep 100 Mixed-Mode Column

HPLC Separation of Three Isomers of Aminobenzoic Acid on Coresep 100 Mixed-Mode Column chromatogram
Conditions of Experiment
Column:
Coresep 100
Separation Modes:
reversed-phase and cation-exchange
Column Dimenstions: 3.0 x 100 mm
Mobile Phase: 20% ACN with 0.1% H3PO4
Detection: UV 235 nm
Sample: 0.3 mg/ml
Injection: 1 uL
Flow rate: 0.6 mL/min
Analytes
1. 4-Aminobenzoic acid
2. 3-Aminobenzoic acid
3. 2-Aminobenzoic acid

Class of compounds: Amino acid, Aromatic compound, Isomer, Pyridine
Nature of compounds: Hydrophilic, Polar, Zwitterionic

Application description

Aminobenzoic acid are organic zwitterionic compounds with the same empirical formula and properties which are very similar to each other. Aminobenzoic acids are used as building blocks in organic and pharmaceutical chemistry. They also are used in azo dyes production as well as crosslinking agents. Three isomers of aminobenzoic acid (2-aminobenzoic acid, 3-aminobenzoic acid and 4-aminobenzoic acid) were separated on Coresep 100 column by reversed-phase and cation-exchange mechanisms. Mixed-mode chromatography explores a small difference in hydrophobic and ionic properties of molecules to enhance resolution between isomers and other close related compounds. Retention time is controlled by amount of ACN, buffer concentration and buffer pH. Method is fast, robust, reproducible, and can be used for analysis of isomers of aminobenzoic acid in various sample matrices. Various mobile phases can be used with Coresep 100 column to accommodate different detection techniques (UV, MS, CAD, ELSD detection). Isomers of other compounds can be retained and separated if they are hydrophobic and basic in nature. Core-shell mixed-mode columns provide a great combination of unique selectivity, speed, and efficiency.

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