TY - JOUR A1 - Grandke, Julia A1 - Oberleitner, Lidia A1 - Resch-Genger, Ute A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Quality assurance in immunoassay performance - comparison of different enzyme immunoassays for the determination of caffeine in consumer products N2 - Enzyme immunoassays with optical detection are amongst the most widely used bioanalytical tools. We defined seven parameters for the quality assessment of immunoassays that were addressed in a systematic study of direct and indirect immunoassays, using the enzymes horseradish peroxidase (HRP) and alkaline phosphatase (AP), the chromogenic substrates 3,3',5,5'-tetramethylbenzidine (TMB) and para-nitrophenyl phosphate, and the fluorescent substrates 3-(4-hydroxyphenyl)propionic acid and 4-methylumbelliferyl phosphate. The same monoclonal antibody against caffeine was used throughout the study. The four quality parameters regarding the standard curve were the test midpoint (sensitivity), the measurement range, the relative dynamic range of the signal, and the goodness of fit of the adjusted four-parameter logistic function. All HRP immunoassays showed a higher sensitivity compared to the AP assays. On the basis of all four criteria, it was established that the direct assay format is superior to the indirect format, the immunoassay using HRP TMB fulfilling all requirements best. In a second step, caffeine concentrations in 24 beverage and cosmetics samples were determined and three more quality parameters were assessed with this application. The direct HRP TMB assay showed one of the best intra- and inter-plate precisions and the best accuracy, defined by the correlation of results with those from the chosen reference method liquid chromatography tandem mass spectrometry (LC-MS/MS). Considering all criteria, HRP TMB seems to be the enzyme substrate system of choice preferably used in the direct assay format. KW - Enzyme immunoassay KW - Caffeine KW - Enzyme substrate KW - Competitive assay KW - Assay format KW - Direct KW - Indirect PY - 2013 U6 - https://doi.org/10.1007/s00216-012-6596-0 SN - 1618-2642 SN - 1618-2650 VL - 405 IS - 5 SP - 1601 EP - 1611 PB - Springer CY - Berlin AN - OPUS4-27639 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Grandke, Julia A1 - Oberleitner, Lidia A1 - Resch-Genger, Ute A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Quality assurance in immunoassay performance - carbamazepine immunoassay format evaluation and application on surface and waste water N2 - Carbamazepine (CBZ) is one of the most frequently detected pharmaceuticals in water samples. For the determination of this anthropogenic marker, various immunoassay formats were tested and evaluated in order to identify the most suitable one. For these direct competitive assays, the analyte was labelled with the enzyme horseradish peroxidase (HRP) or alkaline phosphatase (AP), and seven substrates with specific detection properties were used. The quality criteria for the standard curves were fulfilled by all HRP assays and the chemiluminescence AP format. Furthermore, intra- and inter-plate coefficients of variation as a measure of the achievable precision were determined for the samples. The application of the AP assays to surface water was unfeasible due to CBZ concentrations below the quantifiable concentration range. Surface as well as waste water samples could be analyzed with the HRP assays. Here, the HRP assay employing the chromogenic substrate 3,3',5,5'-tetramethylbenzidine yielded the best results. PY - 2013 U6 - https://doi.org/10.1039/c3ay40702k SN - 1759-9660 SN - 1759-9679 VL - 5 IS - 15 SP - 3754 EP - 3760 PB - RSC Publ. CY - Cambridge AN - OPUS4-28860 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -