Premium
A novel approach for in‐process monitoring and managing cross‐contamination in a high‐throughput high‐performance liquid chromatography assay with tandem mass spectrometric detection
Author(s) -
Chang Min S.,
Kim Elaine J.,
ElShourbagy Tawakol A.
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.2578
Subject(s) - contamination , chemistry , bioanalysis , chromatography , analyte , sample (material) , contamination control , throughput , sample preparation , process engineering , detection limit , calibration , standard solution , quality (philosophy) , biochemical engineering , computer science , statistics , mathematics , ecology , telecommunications , engineering , wireless , biology , philosophy , epistemology
Cross‐contamination among wells of a high‐throughput, high‐density assay is a risk that cannot be detected or controlled by the performance of calibration standards and quality control samples. In the current practice, carryover and cross‐contamination is detected only when analytes are detected in blank, zero, placebo, pre‐dose samples, in a low standard or low quality control sample. There is no mechanism that allows bioanalytical scientists to determine if cross‐contamination has occurred among other samples. As a result, erroneous results can be released to clients even though a batch meets the acceptance criteria. We tested a new approach that quantifies the cross‐contamination of each sample and allows the scientist to make quality decisions with documentation. The approach will also detect carryover in over 90% of the wells. Briefly, two additional analytes were added as contamination markers. The markers were added to a multi‐well plate alternatively creating a pattern of a checkerboard. The spiked multi‐well plate was then used to perform the assay. If both markers were detected in a well, the sample was considered contaminated. The amount of the unexpected marker detected in a well measures the degree of contamination and may be used to make deactivation decisions. Depending on the relative impact of the contamination, a scientist can choose to tolerate the bias, reject the sample, reject the batch or raise the lower limit of quantitation for the batch. A guideline for rejection decisions is presented for discussion. Copyright © 2006 John Wiley & Sons, Ltd.