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For optimal results in HPLC analysis, it is recommended to measure absorbance at a wavelength that matches the absorption maximum of the compound(s) being analyzed. The UV spectrum shown can assist in selecting an appropriate wavelength for your analysis. Please note that certain mobile phases and buffers may block wavelengths below 230 nm, rendering absorbance measurement at these wavelengths ineffective. If detection below 230 nm is required, it is recommended to use acetonitrile and water as low UV-transparent mobile phases, with phosphoric acid and its salts, sulfuric acid, and TFA as buffers.
For some compounds, the UV-Vis Spectrum is affected by the pH of the mobile phase. This analysis was done with the mobile phase made of 20% acetonitrile and 80% water.

Hydrophilic and hydrophobic quaternary amines, along with sodium ion, were separated by mixed-mode chromatography on a Primesep 200 column. Mechanism of retention for sodium and tetramethylammonium ions is cation exchange, while the tetrabutylammonium ion is retained by combination of reversed-phase and cation-exchange mechanisms. All three compounds are not UV-active and monitoring is done by ELSD/CAD.

Primesep B separates quaternary amines, such as t-butylamine with symmetrical peak shape by a combination of reversed-phase and ion-exclusion mechanisms. The embedded basic functional group on the stationary phase shields the underlying silanols to prevent peak tailing. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with evaporative light scattering detection (ELSD).