HPLC Separation of Antibiotics

HPLC.cloud View on hplc.cloud Antibiotics 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.

Condition 1

Column Primesep 200, 4.6x100 mm, 5 µm, 100A
Mobile Phase MeCN Gradient
Buffer H2SO4 - 0.5%
Flow Rate 1.0 ml/min
Detection UV 275 nm

Condition 2

Column Newcrom A, 3.2x100 mm, 5 µm, 100A
Mobile Phase MeOH Gradient
Buffer H2SO4 Gradient
Flow Rate  0.5 ml/min
Detection UV 275 nm


Class of Compounds Drugs, Antibiotics
Analyzing Compounds Sulfanilamide, Sulfadiazine, Sulfaguanidine, Dapsone, Sulfaquinoxaline

Application Analytes: