Abstract

This paper described an organic solvent-free, rapid, simple, and space-saving method of sample preparation followed by HPLC coupled photo-diode array (PDA) detector for simultaneous quantification of cyromazine (CYR) and its decy-cropropylated metabolite, melamine (MEL), in milk. The HPLC-PDA was performed on an Inertsil? HILIC column with an isocratic aqueous mobile phase. Analytes were extracted from the sample using water, and purified by Mono-Spin?-C18, a centrifugal monolithic silica spin mini-columns, and quantified within 20 min. The method, performed under 100% aqueous conditions, obtained average recoveries for CYR and MEL in the range of 93.2% - 99.1% with relative standard deviations ≤ 2.8%. The quantitation limits were 8.5 ng/mL for CYR and 10 ng/mL for MEL, respectively. No organic solvents were used at any stage of the analysis.

Highlights

  • This paper described an organic solvent-free, rapid, simple, and space-saving method of sample preparation followed by HPLC coupled photo-diode array (PDA) detector for simultaneous quantification of cyromazine (CYR) and its decycropropylated metabolite, melamine (MEL), in milk

  • The HPLC-PDA was performed on an Inertsil® HILIC column with an isocratic aqueous mobile phase

  • Cyromazine (CYR) is a triazine insect growth regulator used as an insecticide

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Summary

Introduction

Cyromazine (CYR) is a triazine insect growth regulator used as an insecticide. In veterinary medicine, CYR is used as an ectoparasiticide and is added to animal feed to prevent fly from the manure, so as to improve the hygiene control of animal housing environments. The optimal harmonized method for the routine monitoring chemicals such as melamine in foods must be simple, small scale, economical in time and cost, and must cause no harm to the environment and humans. Previous techniques based on high-performance liquid chromatography (HPLC) for simultaneous determination of CYR and MEL in animal-derived foods [12,13,14,15,16] has crucial drawbacks: 1) all of methods consume organic solvents in the instrumental analyses as well as for extraction and de-proteinization in sample preparation. This article describes an ultrasafe, idiotproof, and inexpensive method to strictly monitor CYR and MEL residues in milk using a 100% aqueous solution in sample preparation and HPLC separation without organic solvent consumption

Reagents
Apparatus
HPLC Operating Conditions
Preparation of Stock Standards and Working Mixed Solutions
Preparation of Calibration Standards and Quality Control Samples
Sample Preparation
Method Validation
Optimum HPLC Conditions
Cost and Time Performance
Application to Residue Monitoring in Commercial Samples
Conclusion

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