Analytical quantitation and confirmation of cyano metabolite by high-resolution mass spectrometry: Residue depletion study in shrimp treated with nitrofurazone.

Authors: ESCOBAR LOAIZA, HERNANDO - Us Food & Drug Administration (FDA); TAYLOR, KENNETH - Us Food & Drug Administration (FDA); VEACH, BRIAN - Us Food & Drug Administration (FDA); ANDERSEN, WENDY - Us Food & Drug Administration (FDA); Singh, Anuradha; HERATH, KITHSIRI - Us Food & Drug Administration (FDA)

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/30/2025
Publication Date: 1/6/2026
Citation: Escobar Loaiza, H., Taylor, K.M., Veach, B.T., Andersen, W.C., Singh, A., Herath, K.B. 2026. Analytical quantitation and confirmation of cyano metabolite by high-resolution mass spectrometry. Residue depletion study in shrimp treated with nitrofurazone.. Journal of Agricultural and Food Chemistry. 74:2342-2351. https://doi.org/10.1021/acs.jafc.5c12391?urlappend=%3Fref%3DPDF&jav=VoR&rel=cite-as.
DOI: https://doi.org/10.1021/acs.jafc.5c12391?urlappend=%3Fref%3DPDF&jav=VoR&rel=cite-as

Interpretive Summary: Historically, nitrofurazone was used as a feed additive in farm animals, including chickens, turkeys, cattle, fish, shrimp and pigs, to prevent bacterial diseases and promote growth. But its use was banned in the U.S. and most U.S. trading partners due to toxicity concerns. Regulatory agencies monitor illegal use of nitrofurazone by measuring a marker compound called semicarbazide (SEM) in animal-derived food products. Because SEM is produced naturally, it has produced many false positive results. Therefore, another metabolite of nitrofurazone, a cyano metabolite (COBS), was tested to find if it could be used as an alternative marker for nitrofurazone. A mass spectrometry method was developed and validated to measure COBS in shrimp. The developed method could be applied in other animal food products for nitrofurazone surveillance.

Technical Abstract: A high-resolution mass spectrometry (HRMS) method was developed and optimized for quantitative and confirmatory analysis of the nitrofurazone (NFZ) cyano metabolite in shrimp. Extraction with 80% acetonitrile: buffer, salting out, and solid phase cartridge clean-up resulted in 87-112% recovery, <10% precision, and good sensitivity with 0.02 ppb limit of quantitation. Current monitoring of unauthorized NFZ use in aquaculture relies upon semicarbazide (SEM) metabolite detection; however, the natural occurrence of SEM in some crustaceans has raised interest for an alternative biomarker. The developed method was used to investigate the suitability of cyano metabolite as an alternate biomarker for unauthorized use of NFZ. In NFZ-incurred shrimp, the cyano metabolite depleted rapidly, with no detection after 6 days, while SEM was detected for at least 20 days. The short-lived stability of cyano metabolite limits its potential for nitrofurazone surveillance in shrimp. However, the method may be adequate for NFZ surveillance in other food animal matrices.