HPLC Separation of β-Lactam Antibiotics on Primesep 100 Column

Separation type: Liquid Chromatography Mixed-mode

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High Performance Liquid Chromatography (HPLC) Method for Analysis of Amoxicillin and Cephalexin. Both amoxicillin and cephalexin are beta-lactam antibiotics. Beta-lactam antibiotics contain a beta-lactam ring in their molecular structure. As a group, these drugs are active against many gram-positive, gram-negative and anaerobic organisms. Both compounds can be retained and baseline separated with an isocratic method in 30/70 Acetonitrile (ACN) and water mobile phase with a Sulfuric acid (H2SO4) buffer. UV detection at 250 nm.


Column Primesep 100, 4.6x100 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer TFA, H2SO4
Flow Rate 1.0 ml/min
Detection UV, 250 nm


Class of Compounds Drug, Antibiotics, Hydrophobic, Ionizable
Analyzing Compounds Cephalexin,  Amoxicillin
Application Analytes:
Amoxicillin hydrate (1:3)