Surfactants are frequently found in pharmaceutical and biopharmaceutical drug applications, as well as common household products. Surfactants can be polar, non-polar, or amphoteric. This structural diversity of the surfactants and complexity of the sample matrix can make the separation and identification of surfactants by HPLC challenging. Historically, this has been achieved using Reversed Phase Chromatography supplemented with Ion Exchange Chromatography. When performed individually, these techniques can be used to characterize the surfactant profile.We have evaluated multiple RP-HPLC columns and an amine functionalized HILIC column for the separation of polar and non-polar surfactants in a single injection using 2-D LC. The use of either mode individually was unable to retain both concurrently. The effect of carbon content, porosity, and particle size on the separation and selectivity was also studied. When used in 2-D LC, the orthogonal relationship between RP-HPLC and HILIC provided the broad range selectivity and retention of both polar and non-polar surfactants. A non-porous IEX column was also investigated for the separation of surfactants.Here, we show the effectiveness of the use of online 2-D LC using Reversed Phase Chromatography in the 1st dimension followed by HILIC in 2nd dimension in the separation of anionic, cationic, and hydrotropic surfactants. The 2-D RP-HPLC � HILIC separation of surfactants using MS compatible mobile phase conditions yielded excellent peak shapes and reproducible retention time and peak area. The results shown here is expected to be useful in the analysis of surfactants present in common household products and drugs.

Use of reversed phase-HPLC and HILIC in the 2D LC separation of cationic, anionic, and hydrotropic surfactants

248th National Meeting (2014)

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