For security purposes
FOR SECURITY PURPOSES - because Internet Explorer is no longer supported by Microsoft, we suggest that you interact with our secure site through one of our supported browsers - Google Chrome, Firefox, or MS Edge. If you continue to use this website with Internet Explorer you do so at your own risk and you may encounter problems.


HPLC/UHPLC Technical Tip

Level: Basic

Overcoming Peak Tailing of Basic Analytes: Silica Type A Stationary Phases RP


Silica-based stationary phases are currently the most frequently used phases in liquid chromatography. Traditional C18 columns often suffered from poor peak shape and tailing of basic analytes at low pH, and limited stability due to gradual silane hydrolysis at high pH. Through cross-linking and end-capping technologies, silica‘s undesired secondary interactions and limited stability have been significantly improved.

The most recent development introduced for analysis of basic analytes at low pH levels is the production of C18 columns with a residual positive charge on the silica support. This combination allows for 100% aqueous conditions to be used, enhancing retention of basic compounds. It also minimizes (ionic) secondary interactions between positively charged bases and silanol groups due to charge repulsion at the silica surface. The result is that basic compounds can be retained and eluted with good peak shape under acidic conditions, which are generally favored. Figure 1 shows an example of enhanced peak shape of chlorpheniramine, a basic antihistamine, using a Kinetexâ„¢ PS C18 column with this technology.

tn-1255-chromatogram-1

Figure 1. Improved reversed-phase retention and peak shape of chlorpheniramine with the Kinetex PS C18.

tech-tips-app22721

Figure 2. Analysis of nicotine and other metabolites with the Kinetex EVO C18 column.

Basic Compounds by LC/MS using a High pH Mobile Phase

We investigate the effectiveness of using high pH mobile phases for the quantitation of basic compounds in ESI+ LC/MS/MS by comparing limits of detection (LOD), limits of quantitation (LOQ), linearity ranges, precision, and accuracy observed in high and low pH mobile phases.

gemini-application-note-4241


Using pH-LC to Control Selectivity of Acidic and Basic Compounds by HPLC

We investigate the effectiveness of using pH-LC techniques in the separation of a mixture of acidic (naproxen), basic (amitriptyline), and neutral (toluene) compounds using a Gemini 5 um NX-C18 column

gemini-application-note-5847