Application of a heterogeneous immunoassay for the quality control testing of release-active forms of diclofenac.
We report on a specially designed diclofenac-ELISA for the determination of diclofenac in the presence of release-active forms of diclofenac in lactose dissolved in water solutions according to a predefined schedule in single-blind experiments. In accordance with the objective of this project, a number of experiments were conducted to determine the optimal ELISA conditions for detecting potential modulatory effects of release-active forms of diclofenac depending on their ability to affect the binding of diclofenac to anti-diclofenac antibodies. As a feature, the diclofenac antibodies were previously incubated with manufactured pharmaceutical samples containing release-active forms of diclofenac or placebo. For comparison of the sample types, measured in ELISA optical densities were chosen. For statistic analysis, Student's two-sample t-test and single-factor ANOVA were applied. The extremely low concentrations of diclofenac of 0.01, 0.05 and 0.1 ng mL(-1) seem most appropriate for routine assay performance. The source of diclofenac used for standard solution preparation is not important but it could be important as the source of diclofenac for release active form of diclofenac preparation. As an outcome, the ELISA appeared to be suitable for the detection of the modifying effects of release-active forms of diclofenac toward the pharmaceutical substance in vitro.
Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, 81377 München, Germany. Electronic address: michael.pschenitza@mytum.de.
Michael
M
Pschenitza
OOO "NPF" Materia Medica Holding, 3rd Samotyochny per., 9, Moscow 127473, Russia. Electronic address: GavrilovaES@materiamedica.ru.
Elena S
ES
Gavrilova
OOO "NPF" Materia Medica Holding, 3rd Samotyochny per., 9, Moscow 127473, Russia. Electronic address: TarasovSA@materiamedica.ru.
Sergey А
SА
Tarasov
Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, 81377 München, Germany. Electronic address: dietmar.knopp@ch.tum.de.
Dietmar
D
Knopp
Institute of Hydrochemistry and Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, 81377 München, Germany. Electronic address: reinhard.niessner@ch.tum.de.
Reinhard
R
Niessner
OOO "NPF" Materia Medica Holding, 3rd Samotyochny per., 9, Moscow 127473, Russia. Electronic address: Nauka@materiamedica.ru.
Oleg I
OI
Epstein
D000906:Antibodies / Q000378:metabolism; D004008:Diclofenac / Q000276:immunology / Q000378:metabolism*; D065546:Drug Liberation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007785:Lactose / Q000378:metabolism; D011485:Protein Binding; D011786:Quality Control; D012015:Reference Standards; D014867:Water / Q000378:metabolism
D000906:Antibodies; D014867:Water; D004008:Diclofenac; D007785:Lactose
Diclofenac derivatives; ELISA; Quality control; Release activity