Separation of Antibiotics in Mixed-mode chromatography
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).
Condition
Column
Obelisc R, 2.1x150 mm, 5 µm, 100A
Mobile Phase
Gradient MeCN - 0-25%, 6 min, 25-70% 14 min
Buffer
Gradient Formic Acid - 0.05%-0.3%, 10 min, 14 min hold
Analysis of Antibiotic Dapsone and Related Impurities in Mixed-Mode Chromatography
Separation type: Liquid Chromatography Mixed-mode
Dapsone is an antibiotic used to treat leprosy and acne. Obelisc R was used to analyze dapsone and its impurities. Obelisc R is a tri-modal reversed-phase, cation- and anion-exchange column. Method is compatible with LC/MS and prep chromatography.
Condition
Column
Obelisc R, 4.6x150 mm, 5 µm, 100A
Mobile Phase
Gradient MeCN - 0-25%, 6 min, 25-70% 14 min
Buffer
Gradient Formic Acid - 0.05%-0.3%, 10 min, 14 min hold
View on hplc.cloudAntibiotics are widely used for treatment and prevention of bacterial infections. Various antibiotics, particularly those with a sulfanilamide structure, were separated in HPLC using mixed-mode columns with varying strengths of ion-pairing groups. Primesep 200 has weak acidic ion-exchange pairing groups while Newcrom A has strong acidic ion-exchange groups. In addition, methanol can be used in the mobile phase on Newcrom A column. The antibiotics were resolved on both columns with a gradient mobile phase consisting of acetonitrile (ACN) or methanol (MeOH), water and sulfuric acid (H2SO4) buffer. UV detection at 275nm.