New EU legislation in force New European legislation for reference point of action (RPA) for nitrofuran metabolites will be in force from 28 November 2022 (EU 2019/1871). For the known metabolites SEM, AHD, AMOZ and AOZ a RPA of 0.5 µg/kg is set. This legislation is also applicable for DNSH, the metabolite of Nifursol.

Nifursol is a nitrofuran banned as a feed additive in the European Union and other countries. Nifursol is metabolised to 3,5-dinitrosalicylic acid hydrazide (DNSH) in living organisms. DNSH is a marker for the detection of illegal use of nifursol in animal husbandry.

ELISA test available

R-Biopharm Nederland launches an ELISA test for monitoring the presence of DNSH in meat and seafood: the EuroProxima 5091 DNSH ELISA. This test was developed and validated in accordance with Commission Implementing Regulation 2021/808

Sample preparation and procedure

A sample preparation method based on salting-out assisted liquid-liquid extraction was developed and optimized. The developed method is simple and does not require an evaporation step. The competitive ELISA is based on a polyconal antibody raised against a DNSH hapten-conjugate. The total procedure takes 60 minutes.

Validation of the ELISA test

The R-Biopharm test was validated with incurred samples, obtained in a feeding study with nifursol. The samples were analysed by the ELISA and by a UPLC-LC/MS method. A good correlation was obtained.

Here you can find more information about the DNSH test.