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A new approach for evaluating carryover and its influence on quantitation in high‐performance liquid chromatography and tandem mass spectrometry assay
Author(s) -
Zeng Wei,
Musson Donald G.,
Fisher Alison L.,
Wang Amy Qiu
Publication year - 2006
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2353
Subject(s) - chemistry , chromatography , analyte , bioanalysis , sample (material) , relative standard deviation , sample preparation , liquid chromatography–mass spectrometry , analytical chemistry (journal) , detection limit , high performance liquid chromatography , standard deviation , mass spectrometry , statistics , mathematics
In a high‐performance liquid chromatography (HPLC)‐based analytical method, carryover denotes one type of systematic error that is derived from a preceding sample and introduced into the next sample. For typical bioanalytical method development, a significant amount of time and resources are spent on reducing carryover for some analytes. In this paper, the statistical characteristics of carryover were analyzed based on the experimental results. The relative carryover (RC), defined as the peak area ratio of a blank sample to the preceding sample, was constant for the analyte and independent of the concentration of the preceding sample. The influence of carryover on the quantitation of the next injected sample or the ‘following’ sample was proportional to the concentration ratio of two consecutive samples and the relative carryover. Based on these experiments and analyses, the influence of carryover on the quantitation of unknown samples in an HPLC assay can be evaluated by the estimated carryover influence (ECI), which is the product of the relative carryover and the concentration ratio. This new approach provides a quantitative estimation for the influence of carryover on the quantitation of the unknown sample, and removes the limit put on the dynamic range of the assay by the current criterion of carryover. In general, if the relative standard deviation (RSD) of a validated bioanalytical method is less than 10%, the carryover will not have a significant effect on the accuracy of the assay when the estimated carryover influence is less than 5%. Copyright © 2006 John Wiley & Sons, Ltd.