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.
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 |
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 options2-Amino-5-methyl-thiazole
3,4-Difluoroaniline
4-Amino-2-Chloropyridine
4-Amino-3-Chloropyridine
4-Ethylaniline
5-Aminoindole
Adenosine