Selecting Buffer pH in Reversed-Phase HPLC

8 Tel: 1-800-255-8324 Fax: 847-948-1078 Specialists in Chromatography Selecting the proper Buffer pH isnecessary to reproducibly separateionizable compounds by reversed-phaseliquid chromatography. Selecting animproper pH for ionizable analytes oftenleads to asymmetric peaks that are broad,tail, split, or shoulder. Sharp, symmetricalpeaks are necessary in quantitativeanalysis in order to achieve low detectionlimits, low relative standard deviations(RSD) between injections, andreproducible retention times. We willdiscuss when buffering is needed, how toselect Buffer pH, and how to choosethe proper Buffer system for that is Buffering Needed?In Reversed-Phase chromatography,mobile phase pH values are usually between2 and Buffers are needed whenthe analyte is ionizable under Reversed-Phase conditions or the sample solution isoutside this pH range. Analytes ionizableunder Reversed-Phase conditions oftenhave amine or acid functional groups withpKa s between 1 and 11.

8 “Specialists in Chromatography” Tel: 1-800-255-8324 • Fax: 847-948-1078 Selecting the proper buffer pH is necessary to reproducibly separate ionizable compounds by reversed-phase liquid chromatography. Selecting an

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  Phases, Selecting, Buffer, Reserved, Selecting buffer ph in reversed phase

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