HPLC Separation of Isomers of Amino Salicylic Acid


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Condition 1

Column Newcrom A, 4.6×50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 20/80%
Buffer H3PO4 – 0.5%
Flow Rate 0.5 ml/min
Detection UV 275 nm

 


4-Aminosalicylic acid (PAS) is an antibiotic used in treatment of tuberculosis. It is a polar amino acid with limited retention on traditional C18 (reversed-phase) columns. Other isomers of aminosalicylic acid exist, but the main impurity in PAS is 5-Aminosalicylic acid, which also serves as anti-inflammatory drug. Both compounds are isomers with similar empirical structure and properties. These two isomers were separated on a Primesep 100 column with UV, ELSD and LC/MS compatible mobile phase. Method can be used a generic approach for separation of isomers of basic and zwitter ionic compounds. Isomers are retained and separated based on reversed-phase and cation-exchange properties. Retention time is controlled by the amount of acetonitrile, buffer concentration and buffer pH. Buffer pH is affecting ionization of these two compounds and thus serves as a powerful tool to adjust selectivity of separation.

Condition 2

Column Newcrom A, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 50/50%
Buffer AmFm pH 3.0- 40 mM
Flow Rate 1.0 ml/min
Detection UV 256 nm,  MS-compatible mobile phase

Description

Class of Compounds Acids
Analyzing Compounds 5-aminosalicylic acid, 4-aminosalicylic acid

Application Column

Newcrom A

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

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Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
4-Aminosalicylic Acid
5-Aminosalicylic Acid (Mesalamine)
Mesalamine hydrochloride

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.