Simultaneous determination of five metabolites of nitrofurans including the metabolite nifursol in shrimp and fish by UPLC- MS/MS: in-house method validation according to commission implementing regulation (EU) 2021/808

Abstract

For the simultaneous identification and quantification of five nitrofurans metabolites in farmed shrimp and fish, 3-amino-2-oxazolidinone (AOZ), 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), 1-aminohydantoine (AHD), semicarbazide (SEM), and 3,5-dinitrosalicylic acid hydrazide (DNSH), an accurate, precise, and specific method was developed. The mixture of water and methanol (60/40; v/v) was found to be the final optimised solvent for injection. The analytical run duration was 7 min, and the mobile phase included 2 mM methanol and ammonium formate. The new reference point for action (RPA) of 0.50 µg kg⁻¹ as per EC/1871/2019 was taken into consideration and evaluated for the performance characteristics as per the CIR (EC)/2021/808 criteria. Specificity, relative retention time (≤0.25%) relative ion ratio (≤40%), linearity (0.25 to 2.0 µg kg⁻¹), trueness (between 82.8 and 118.1%), repeatability (RSD_r ≤14%), within lab reproducibility (RSD_wr ≤16.9%), CCα (0.32–0.36 μg kg⁻¹), ruggedness and relative matrix effect (≤14.26%) achieved acceptable values.

Keywords:

• Nitrofuran metabolites
• nifursol metabolite
• aquaculture products
• UPLC-MS/MS
• method validation

Acknowledgments

The authors gratefully acknowledge the support provided by the Export Inspection Council, New Delhi, India, and the Export Inspection Agency-Kochi in completing the work. We would like to express our sincere gratitude to Dr. Padmakar Waugh and Mr. Pawan, Water India Pvt., Ltd., India for the technical support provided during the work. We would like to thank an anonymous reviewer for providing feedback on the article. This work did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Correction Statement

This article has been republished with minor changes. These changes do not impact the academic content of the article.