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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.

Toluidine isomers and aminobyphenyl isomers are organic compounds each with a substituted amine group. Primesep 200 was used to separate these isomers. Primesep 200 separates isomers by interacting with polar molecules, retaining basic compounds, and additionally by reverse phase mechanism.

Two isomers of aminobiphenyl were successfully retained and separated in cation-exchange mode on a Primesep S2 column. Primesep S2 column is a HILIC/cation-exchange column that can be operated in HILIC, cation-exchange, and anion-exclusion modes. Method and column are compatible with ELSD/LC/MS and UV and can be used for routine analysis of polar molecules.

Aminobiphenyls consist of isomers that differ only in the location of a primary amine group on an aromatic ring. 2-, 3-, and 4-aminobiphenyl are separated on a Primesep D with good peak shape and a short retention time by a mixture of reversed-phase and ion-exclusion interactions. Peak order can be reversed for 2- and 3-aminobiphenyls by using ammonium formate buffer without pH adjustment. The HPLC separations use a mobile phase of water, acetonitrile (MeCN, ACN), ammonium formate buffer or acetic acid (HOAc) with UV detection at 250 nm.