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Methodological approaches for histamine quantification using derivatization by chloroethylnitrosourea and ELISA measurement. Part II: Optimisation of the derivatization step |
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| Authors: | M Claeys-Bruno O Vandenabeele-Trambouze M Sergent M Geffard D Bodet M Dobrijevic A Commeyras R Phan Tan Luu |
| Affiliation: | aLaboratoire de Méthodologie de la Recherche Expérimentale, UMR 6171, Université Paul Cézanne, Aix-Marseille III, Faculté des sciences et techniques, Avenue Escadrille Normandie Niemen, 13397 Marseille cedex 20, France bOrganisation Moléculaire, Evolution et Matériaux Fluorés, UMR-CNRS 5073, Université Montpellier II, 2 Place E. Bataillon, 34195 Montpellier Cedex 5, France cENSCPB, PIOM, EPHE, 16, avenue Pey-Berland-33607 PESSAC Cedex, France dLaboratoire d'astrodynamique, d'astrophysique et d'aéronomie de Bordeaux, UMR-CNRS 5804, OASU, 2 rue de l'observatoire, BP 89, 33270 Floirac, France |
| Abstract: | In a previous paper (part I), new strategy was used for raising antibodies against hapten (< 300 Da) and the quantification of these hapten by ELISA using derivatization by chloroethyl nitrosourea (CENU). After raising antibodies against histamine, they were characterized and used for ELISA measurements. Optimal detection conditions were determined for the histamine quantification by ELISA method. The present study investigates the derivatization step of the histamine by chloroethylnitrosourea (CENU). Five factors (2 qualitative: nature of the solvent and nature of the antibodies and 3 quantitative: pH, % of solvent and time of derivatization) have been considered. Optimal reaction conditions were established by calculation of a validated model. |
| Keywords: | Experimental design Box Behnken Antibodies Derivatization Histamine quantification |
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