Determination of Sulfonamide Residues in Chicken Muscle by Agilent Bond Elut QuEChERS AOAC Kit and HPLC-FLD

Significance of the Topic

Sulfonamides are widely used antimicrobial agents in veterinary medicine for disease control and growth promotion in poultry. Residues in edible tissues pose potential health risks, including allergic reactions and the development of antibiotic-resistant bacteria, necessitating reliable monitoring methods.

Objectives and Overview of the Study

This study aimed to develop and validate a multi-residue method for nine sulfonamides in chicken muscle using a QuEChERS sample preparation protocol combined with precolumn fluorescamine derivatization and HPLC-FLD analysis.

Methodology and Sample Preparation

A buffered QuEChERS extraction (AOAC 2007.01) was performed on homogenized chicken muscle, followed by dispersive SPE cleanup. Derivatization of primary amine groups was achieved using a fluorescamine reagent, enhancing fluorescence detection of sulfonamides.

Instrumental Setup

• Agilent 1200 Series HPLC system with binary pump and fluorescence detector (Ex 405 nm, Em 495 nm).
• Agilent ZORBAX Eclipse Plus C18 column (4.6 × 75 mm, 3.5 μm).
• Agilent Bond Elut QuEChERS AOAC Buffered Extraction kit (p/n 5982-5755) and Dispersive SPE kit (p/n 5982-5158).
• Fluorescamine derivatization reagent (0.02% w/v in acetone).

Main Results and Discussion

• Complete derivatization within 60 minutes and separation via methanol–acetate buffer gradient (pH 4.5).
• Limits of detection ranged from 0.02 to 0.39 ng/g and LOQs from 0.25 to 1.30 ng/g, all below regulatory MRLs.
• Recoveries between 76.8% and 95.2% with RSDs of 1.5% to 4.7% across fortification levels (50–150 ng/g).

Benefits and Practical Applications of the Method

The method offers rapid, cost-effective, and robust multi-residue analysis suitable for routine monitoring of sulfonamide residues in poultry products, ensuring compliance with food safety regulations.

Future Trends and Potential Applications

• Integration with tandem mass spectrometry to improve selectivity and lower detection limits.
• Automation and high-throughput adaptations of QuEChERS workflows.
• Expansion to other antibiotic classes and diverse food matrices.
• Development of portable, in-field screening platforms.

Conclusion

A validated HPLC-FLD method with QuEChERS sample preparation and fluorescamine derivatization provides reliable detection of nine sulfonamides in chicken muscle, with performance metrics meeting or exceeding regulatory requirements.

References

1. A. Posyniak, J. Zmudzki, K. Mitrowska, J. Chromatogr. A 1087 (2005) 259–264.
2. I. Pecorelli, R. Bibi, L. Fioroni, R. Galarini, J. Chromatogr. A 1032 (2004) 23–29.
3. A. R. Long, L. C. Hsieh, M. S. Malborough, C. R. Short, S. A. Barker, J. Agric. Food Chem. 38 (1990) 423–426.
4. D. Kim, J. Choi, J. S. Kim, D. W. Lee, Bull. Korean Chem. Soc. 23 (2003).
5. V. Gamba, C. Terzano, L. Fioroni, S. Moretti, G. Dusi, R. Galarini, Anal. Chim. Acta 637 (2009) 18–23.
6. S. Wang, H. Y. Zhang, L. Wang, Z. J. Duan, Food Addit. Contam. 23 (2006) 362–384.
7. X. Wang, K. Li, D. Shi, N. Xiong, X. Jin, J. Yi, D. Bi, J. Agric. Food Chem. 55 (2007) 2077.
8. L. Zhao, J. Stevens, Analysis of Pesticide Residues in Spinach Using Agilent Bond Elut QuEChERS AOAC Kits by LC/MS/MS, Agilent Technologies publication, 5990-4248EN.
9. G. Stoev, A. Michailova, J. Chromatogr. A 871 (2000) 37–42.
10. N. Takeda, Y. Akiyama, J. Chromatogr. B 558 (1991) 175–180.