HPLC Method for Separation of Chlorhexidine and Cetylpyridinium Chloride on Primesep B Column

HPLC Separation of Chlorhexidine and Cetylpyridinium chloride on Primesep B Column_1222 Chlorhexidine gluconate, or simply chlorhexidine, is a biguanide used as an antiseptic and disinfectant. It is a component of mouthwash rinses that has been shown to reduce plaque, gingivitis and oral bacteria. It’s also used as a topical agent for skin disinfection. Cetylpyridinium chloride is another type of antiseptic used in mouthwash rinses. Both compounds are cationic. They can be separated using HPLC on SIELC’s reverse-phase (RP) mixed-mode Primesep B column with the mobile phase of acetonitrile (ACN) and water with formic acid buffer and UV detected at 270nm.


Column Primesep B,  3.2x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer Formic Acid - 0.5%
Flow Rate 0.5 ml/min
Detection UV 270 nm


Class of Compounds Surfactant,  Hydrophobic, Ionizable
Analyzing Compounds Chlorhexidine, Cetylpyridinium Chloride

Application Analytes:
Cetylpyridinium Chloride