Dwell Time, a Critical Factor on Precision in N-in-One Assays Utilizing UPLC-MS/MS - April, 2014

UPLC typically results narrow peaks of less than 0.1 minute in width. As an accurate integration of a chromatography peak requires a minimum of 10 to 15 data points throughout the peak, a short dwell time becomes unavoidable in MS/MS analysis. The situation is more acute when multiple analytes are analyzed simultaneously (n-in-one assays). In this poster abstract, we describe an example of a five-in-one assay we developed. Due to the small peak width of the analytes (i.e., ~ 0.06 minute), the  maximum dwell time allowed for each analyte/internal standard (IS) trace was only 30 milliseconds (ms). The impact of short dwell time on assay performance was evaluated.

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