Development and validation of a fast monoclonal based disequilibrium enzyme-linked immunosorbent assay for the detection of triphenylmethane dyes and their metabolites in fish

Malachite Green (MG), Crystal Violet (CV) and Brilliant Green (BC) are antibacterial, antifungal and antiparasitic agents that have been used for treatment and prevention of diseases in fish. These dyes are metabolized into reduced leuco forms (LMG, LCV, LBG) that can be present in fish muscles for a long period. Due to the carcinogenic properties they are banned for use in fish for human consumption in many countries including the European Union and the United States. HPLC and LC-MS techniques are generally used for the detection of these compounds and their metabolites in fish. This study presents the development of a fast enzyme-linked immunosorbent assay (ELISA) method as an alternative for screening purposes. A first monoclonal cell line producing antibodies to MG was generated using a hybridoma technique. The antibody had good cross-reactivates with related chromatic forms of triphenylmethane dyes such as CV, BC, Methyl Green, Methyl Violet and Victoria Blue R. The monoclonal antibody (mAb) was used to develop a fast (20 min) disequilibrium ELISA screening method for the detection of triphenylmethanes in fish. By introducing an oxidation step with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) during sample extraction the assay was also used to detect the presence of the reduced metabolites of triphenylmethanes. The detection capability of the assay was 1 ng g(-1) for MG, LMG, CV, LCV and BC which was below the minimum required performance limit (MRPL) for the detection method of total MG (sum of MG and LMG) set by the Commission Decision 2004/25/EC (2 ng g(-1)). The mean recoveries for fish samples spiked at 0.5 MRPL and MRPL levels with MG and LMG were between 74.9 and 117.0% and inter- and intra-assay coefficients of variation between 4.7 and 25.7%. The validated method allows the analysis of a batch of 20 samples in two to three hours. Additionally, this procedure is substantially faster than other ELISA methods developed for MG/LMG thus far. The stable and efficient monoclonal cell line obtained is an unlimited source of sensitive and specific antibody to MG and other triphenylmethanes. (C) 2011 Elsevier B.V. All rights reserved.

Paul Celan - poetique de la rencontre en quete de l'etre humain

This article deals with Paul Celans poetics of the encounter as a reflection on the anthropological question.

La concentration. Le mouvement du discours poetique dans le poeme EIN WURFHOLZ de Paul Celan

This article demonstrates how poetic discourse signifies by its movement of language at the example of a poem by Paul Celan.

Les voix et Paul Celan

This article pursues the role of voices in Celan's poetry and poetics.

Le travail du sens de l'Homme. La poetique de Paul Celan

This article presents the poetics of Paul Celan with regard to the constitution of the subject in poetry and the anthropological dimension of poetics.

Vom Innen und vom Aussen. Zum Raum in der Poetik Paul Celans

This article deals with the conception of space in the poetics of Paul Celan in relation to French theory (Kristeva, Blanchot, Foucault). It analyses in detail one poem to show how Celan constructs his concept of space in the poetic form, relating it to Martin Buber dialogical thinking.

Die Konzentration. Zur Bewegung des dichterischen Sprechens in der Poetik Paul Celans

This article demonstrates the mouvement of poetic speech in Celan's poetics at the exemple of one poem, thus stressing the significance of the poetic form.

The 'stance' in the poetics of Paul Celan

This article deals with the ethics of Paul Celan's poetics, presenting and analysing his notion of 'Stehen'.

Zur Poetik Paul Celans: Gedicht und Mensch - Die Arbeit am Sinn

This publication summarises my PhD on Paul Celans poetics, focussing on the constitution of meaning and establishing it as a poetological anthropology.