Characterization of Impurities in the Fungicide Flutriafol Using UNIFI and UPLC-ToF-MSE

Significance of the topic

The comprehensive characterization of impurities in technical grade agrochemicals is essential for product safety, regulatory compliance, and optimization of synthetic routes. Identifying and quantifying trace contaminants supports risk assessment and ensures that active ingredients meet stringent registration thresholds, typically 0.1% of total formulation.

Study objectives and overview

This application note describes the use of Waters UNIFI Screening Platform combined with UPLC MSE to profile structurally related impurities in the fungicide flutriafol. The goals were to detect minor components above 0.1%, elucidate their structures, and demonstrate a streamlined workflow for impurity analysis in pesticide development.

Methodology and instrumentation

• Sample Preparation 1 mg/mL flutriafol stock in methanol
• Chromatography ACQUITY UPLC I Class System, CORTECS C18 column 2.1 x 100 mm 1.6 μm, 0.1% formic acid in water and acetonitrile gradient, 0.6 mL/min, 50 °C, 1 μL injection
• PDA Detection ACQUITY UPLC PDA, 210–400 nm, 20 Hz sampling
• Mass Spectrometry Xevo G2 XS QTof, ESI plus, MS E resolution mode, low CE 4 eV, high CE ramp 15–30 eV, scan range 50–950 m/z, 0.15 s scan time
• Data Processing UNIFI software with Scientific Library, component summary, extractable ion chromatograms, structural elucidation tools, ChemSpider search, fragment match, common fragment search

Main results and discussion

Chromatographic separation revealed three minor peaks plus the primary flutriafol peak, each exceeding the regulatory threshold. High accuracy mass and MSE fragmentation data enabled:

• Components 1 and 2 were assigned as flutriafol isomers sharing the elemental composition C16H13F2N3O and common diagnostic fragments at m/z 70.0399 and 109.0450.
• Component 3 displayed m/z 314.1303, elemental composition C17H16FN3O2, and shared core fragments. Structural elucidation combined elemental composition analysis, MS MS fragment matching, knowledge of synthetic intermediates, and comparison to the active ingredient.

Benefits and practical applications

This integrated UPLC MSE and UNIFI approach dramatically simplifies impurity screening by capturing precursor and product ion spectra in a single injection. Automated component extraction and library driven workflows enhance confidence in assignments, accelerate synthetic route optimization, and support regulatory submissions.

Future trends and potential applications

Advances in high resolution time of flight detectors and artificial intelligence driven spectral interpretation will further improve sensitivity and structure elucidation. Integration of orthogonal techniques such as NMR microflow detection and ion mobility separation promises deeper insight into complex impurity profiles.

Conclusion

The study demonstrated a rapid, sensitive, and information rich method for profiling flutriafol impurities. The UNIFI Screening Platform with UPLC MSE enabled confident identification of isomeric byproducts and a previously unknown component. Adoption of this workflow can streamline agrochemical development, ensure product safety, and accelerate registration processes.

References

1. Commission Regulation EU 544 2011 on conditions for active substance approval in crop protection products
2. An overview of the principles of MSE The engine that drives MS performance Waters White Paper p n 720004036en October 2011
3. Chang M, Kim TH, Kim HD Stereoselective synthesis of flutriafol Tetrahedron Asymmetry 19 1503 1507 2008