Chromatographic Retention of Polar Compounds: What are the options?

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Diane Diehl;Eric S. Grumbach;Jeffrey R. Mazzeo;Uwe D. Neue [Waters]
Pittcon 2003
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Content Subtype:
Amoxicillin; Albuteral; Allantoin; Amitriptyline; Adenine; Bamethan;Thiourea; 5-Fluorocytosine; Guanosine-5''- monophosphate; Thymine; Uracil; 5-Fluorouracil; Nortriptyline;
Atlantis dC18 5 µm Steel 4.6 mm 150 mm Atlantis HILIC Silica 5 µm Steel 4.6 mm 150 mm Atlantis dC18 3 µm Steel 2.1 mm 50 mm
Many lead compounds under research in the pharmaceutical industry today are polar compounds that are difficult to retain on standard reversed-phase columns. This makes HPLC method development more demanding than it was in the past. What options are available to retain very polar compounds, and what are their relative advantages and disadvantages? In this presentation, we will discuss the different techniques that are available for the retention of polar compounds. These include reversed-phase techniques as well as techniques that can be summarized as hydrophilic interaction chromatography (HILIC). Among reversed-phase chromatography, we will discuss special phases that do not exhibit hydrophobic collapse in water as eluent and phases with embedded polar groups. Another option is the increase in retention that is possible through pH changes, if the analytes are ionizable. Among HILIC techniques, several options are available as well, ranging from pure HILIC to the use of the ion-exchange properties of HILIC phases. In all these comparisons, we are using mobile phases that are compatible with high-sensitivity MS detection.

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