TY - JOUR A1 - Grandke, Julia A1 - Resch-Genger, Ute A1 - Bremser, Wolfram A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Quality assurance in immunoassay performance-temperature effects N2 - Temperature is one factor that influences the performance of immunoassays. Most commonly all incubation steps of the immunoassay are performed at ambient temperature. To systematically study the influence of temperature deviations on assay parameters like signal variation coefficients, limit of detection and measurement range, direct competitive enzyme immunoassays for the determination of two anthropogenic markers, caffeine and carbamazepine, were run at different temperatures above and below room temperature during individual assay steps. The temperature during the competition step had the greatest impact on the assay sensitivity yielding changes in test midpoints by a factor of 10 to 15. For carbamazepine, the test midpoints varied from 36 ng L-1, 108 ng L-1 to 378 ng L-1 employing 4 °C, 21 °C, or 37 °C as incubation temperature, respectively. The edge effect can be minimized resulting in a good plate homogeneity with small variation coefficients, when the assay is performed at ambient temperature. KW - Immunoassay KW - Temperature effects PY - 2012 U6 - https://doi.org/10.1039/c2ay05918e SN - 1759-9660 SN - 1759-9679 IS - 4 SP - 901 EP - 905 PB - RSC Publ. CY - Cambridge AN - OPUS4-25740 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 - 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 - TY - JOUR A1 - Oberleitner, Lidia A1 - Grandke, Julia A1 - Mallwitz, F. A1 - Resch-Genger, Ute A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Fluorescence polarization immunoassays for the quantification of caffeine in beverages N2 - Homogeneous fluorescence polarization immunoassays (FPIAs) were developed and compared for the determination of caffeine in beverages and cosmetics. FPIAs were performed in cuvettes in a spectrometer for kinetic FP measurements as well as in microtiter plates (MTPs) on a multimode reader. Both FPIAs showed measurement ranges in the µg/L range and were performed within 2 and 20 min, respectively. For the application on real samples, high coefficients of variations (CVs) were observed for the performance in MTPs; the CVs for FPIAs in cuvettes were below 4%. The correlations between this method and reference methods were satisfying. The sensitivity was sufficient for all tested samples including decaffeinated coffee without preconcentration steps. The FPIA in cuvettes allows a fast, precise, and automated quantitative analysis of caffeine in consumer products, whereas FPIAs in MTPs are suitable for semiquantitative high-throughput screenings. Moreover, specific quality criteria for heterogeneous assays were applied to homogeneous immunoassays. KW - caffeine KW - Decaffeinated coffee KW - Fluorescence polarization immunoassay KW - Homogeneous immunoassay KW - Quality assurance criteria PY - 2014 U6 - https://doi.org/10.1021/jf4053226 SN - 0021-8561 SN - 1520-5118 VL - 62 IS - 11 SP - 2337 EP - 2343 PB - American Chemical Society CY - Columbus, Ohio AN - OPUS4-30613 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -