Access the UV-Vis Spectrum SIELC Library

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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. The spectra presented here are measured with an acidic mobile phase that has a pH of 3 or lower.

Aromatic hydrocarbons are hydrophobic compounds which are well retained on any reverse column. Retention time is adjusted by the amount of ACN. Change in buffer concentration or buffer pH does not affect retention time. Method on Obelisc mixed-mode column shows retention and separation of PAHs by reverse phase mechanism. Change of pH is changing conformation and ionization of stationary phase on Obelisc R column, making it more or less hydrophobic. This changes interaction on the column and selectivity of separation. This method can be used for analysis of hydrophobic compounds and isomers by reverse phase mechanism, when fine tuning is required to achieve desired degree of separation.