Separation of Antibiotics in Mixed-mode chromatography

HPLC-Applikation • Datum: 2015/05/11
Separation of Antibiotics in Mixed-mode chromatography Chromatogramm

A complex mixture of sulphonamide, macrolide, tetracycline and fluoroquinolone antibiotics were separated in one run using mixed-mode chromatography with LC/MS -compatible conditions. All compounds are separated based on reversed-phase and/or ion-exchange mechanism. Method can be used for analysis of various classes of antibiotics and related impurities in different sample matrices (blood, urine, soil, waste water).

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

Methodenparameter

ColumnObelisc R, 2.1×150 mm, 5 µm, 100 Å
Mobile PhaseGradient MeCN – 0-25%, 6 min, 25-70% 14 min
BufferGradient Formic Acid – 0.05%-0.3%, 10 min, 14 min hold
Flow Rate1.0 mL/min
DetectionUV, 270 nm