If you are looking for optimized HPLC method to analyze Trometamol (Tris, Tris(hydroxymethyl)aminomethane, Tromethamine, and or THAM) check our HPLC Applications library

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.

HPLC Method for Separation of  Trometamol (Tris) and Sodium on Primesep 100 by SIELC Technologies.

Separation type: Liquid Chromatography Mixed-mode

Trometamol, also known as Tris or Tromethamine, is a buffering agent (or Good’s buffer) frequently used in biochemical and molecular biological applicaitons. Tris is often mixed with Sodium Chloride as a buffer, and the Sodium ion and Tris will often co-elute, deliving inaccurate quantitation results. Tris and Sodium ion can be retained, separated, and analyzed on a Primesep 100 mixed-mode stationary phase column using several different isocratic analytical methods with a simple mobile phase of water, Acetonitrile (MeCN), and a Formic acid (FA) buffer. By simply changing the relative concentrations of MeCN and Water, the retention times and order of Tris and Sodium can be significantly manipulated to deliver high quality separations. This analysis method is compatible with evaporative detection methods such as Evaporative Light Scattering Detection (ELSD), Charged Aerosol Detection (CAD), and Electrospray Ionization (ESI) for Mass Spectrometry (LCMS).

High Performance Liquid Chromatography (HPLC) Method for Separation of  Trometamol (Tris) and Sodium

Condition

Description

TRIS or tris(hydroxymethyl)aminomethane, is a hydrophilic basic compounds widely used in chemistry, biochemistry, and molecular biology. TRIS pKa (8.1) makes it an effective buffer in pH range of 7-9.2. TRIS is not retained on reversed-phase column and is not UV active, making it hard to be analyzed by HPLC. TRIS was analyzed by mixed-mode HPLC on a Primesep 100 column. Retention time of compound is controlled by the amount and pH of the buffer. No ion-pairing reagent is required for HPLC retention of hydrophilic basic molecules like TRIS. TRIS can be monitored by ELSD/CAD with trifluoroacetic acid in the mobile phase, or by LC/MS with ammonium formate or acetate in the mobile phase.

High Performance Liquid Chromatography (HPLC) Method for Analyses of Trometamol (Tris, Tris(hydroxymethyl)aminomethane, Tromethamine, and or THAM)