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HPLC/UHPLC Technical Tip

Level: Basic

Basic Analytes and the Use of High pH Stable Stationary Phases

The development of hybrid materials has greatly extended the high pH stability of HPLC sorbents. This new development allowed users to use high pH to deprotonate basic compounds, increasing their hydrophobicity (and hence retention) while also removing the possibility of ion exchange.

This approach is particularly useful when considering polar basic compounds as the combination of good peak shape with enhanced retention is useful, especially in LC-MS/MS where elution under higher organic mobile phase conditions has been shown to be beneficial for peak response. Columns such as Geminiâ„¢ and Kinetexâ„¢ EVO can be used in the pH range 1-12.

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Lidocaine (peak 3) run at pH 12 on a Twin technology, high pH stable material.

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.

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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

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