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

Chlorhexidine gluconate (commonly known as chlorhexidine) is a biguanide antiseptic widely used in mouthwash formulations to reduce plaque, gingivitis, and oral bacteria. It is also employed as a topical disinfectant for skin. Cetylpyridinium chloride, another cationic antiseptic, is also frequently included in oral rinses for its antimicrobial properties.

Both chlorhexidine and cetylpyridinium chloride, being cationic compounds, can be effectively separated and analyzed using HPLC on  Zodiac HST B reverse-phase mixed-mode column. The analysis utilizes a mobile phase composed of acetonitrile (ACN), water, and formic acid buffer. Detection is performed via UV absorbance at 270 nm, providing clear resolution of both analytes.