Electrophoretically Mediated Microanalysis (EMMA) assays involving small molecules suffer from limited sensitivity, particularly when UV absorption is the mode of detection employed.  We explore here the use of electrophoretic stacking to improve the sensitivity of an in-line assay for creatinine. One accepted clinical technique for quantifying creatinine is the Jaffe reaction between alkaline picrate and creatinine resulting in formation of a red Jaffe product.  EMMA is attractive for the Jaffe chemistry as it is faster than the traditional methodology and generates less waste.  Under certain reaction conditions, the doubly anionic Jaffe product formed with the in-line reaction will undergo a transient isotacophoretic stacking as it leaves the reaction zone.  If the assay can be performed such that the timing of detection corresponds with this stacking, the method sensitivity can be significantly increased.   Characterization of what governs this particular phenomenon with both experiment and computer simulation will be presented.

Understanding transient isotacophoresis of an in-line generated product using EMMA

244th National Meeting (2012)

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