Separation of Model Compounds in Reversed-Phase and Mixed-Mode

Separation type: Liquid Chromatography Mixed-mode








 
Many compounds are difficult, if not impossible, to separate on reverse-phase columns in HPLC. Other compounds cannot be separated on ion-exchange columns. That’s where the mixed-mode columns come in. By using a stationary phase with both hydrophobic and ion-exchange properties, allows the chromatographer to have additional controls over separation conditions. Here, we demonstrate the separation of compounds that can’t be achieved on a C18 column. By using both an organic gradient and buffer gradient of ammonium formate (AmFm), we can separate structurally similar compounds that can’t be separated on a reverse-phase column alone.




 

Condition

Column Primesep 100, 3,2×50 mm, 2,7 µm, 100A
Mobile Phase Gradient  MeCN – 10-60%, 5 min
Buffer Gradient AmFm pH 3.5- 30 – 70 mM, 5 min
Flow Rate 1.2 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug,  Basic, Hydrophilic, Hydrophobic, Ionizable.
Analyzing Compounds Adenosine, 3,4-Difluroaniline, 4-Amino-2-chloropyridine, 5-Aminoindole, 4-Amino-3-chloropyridine, 2-Amino 5-methylthiadiazole, 4-Ethylaniline

 

Application Column

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.

Select options
Application Analytes:
2-Amino-5-Methylthiadiazole
2-Amino-5-methyl-thiazole
3,4-Difluoroaniline
4-Amino-2-Chloropyridine
4-Amino-3-Chloropyridine
4-Ethylaniline
5-Aminoindole
Adenosine
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.