HPLC Separation of Surfactants

Surfactants are molecules that contain both hydrophilic and hydrophobic groups, usually in the form of a hydrophilic head and a hydrophobic tail. Surfactants are used in detergents where they can form micelles around hydrophobic dirt molecules and wash them away. Triton X-100 is a surfactant with a hydrophilic polyethylene oxide chain that can be separated on a Primesep D reverse-phase HPLC column based on the number of oxide units in the chain. The mobile phase is water, acetonitrile (MeCN, ACN) and sulfuric acid as buffer. UV detection at 210nm.


Column Primesep D,  4.6x50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer H2SO4
Flow Rate 1.0 ml/min
Detection UV 210nm


Class of Compounds Surfactant,  Hydrophobic, Ionizable
Analyzing Compounds Triton X-100, Cetylpyridinium Chloride
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Application Analytes:
Benzalkonium Chloride
Cetylpyridinium Chloride
Triton X100

Application Detection:
UV Detection