## 5 Challenges and Opportunities
Yet the need for matrix-specific optimization can limit reproducibility across food types. Importantly, derivatization-based HPLC methods are less suitable for high-fat and animal-based matrices, where recovery variability tends to be higher, making LC-MS/MS the preferred option despite its cost and operational constraints. International guidelines such as the QuPPE method have been instrumental in standardizing the analysis of polar pesticides in food products. Nonetheless, their broader applicability to animal-derived or processed foods remains insufficient. As evidenced in multiple studies, protocols developed for products such as infant formula, eggs, and milk require significant modifications and validation to achieve consistent results. Moreover, the lack of harmonized calibration strategies contributes to the limited transferability and comparability of results across laboratories and regulatory contexts. Many researchers agree that the complexity of food matrices and the variety of samples make it difficult to apply these guidelines universally. To address these limitations, future efforts should focus on developing derivatization-free methods that are less sensitive to matrix variation. The integration of internal standards and matrix-matched calibration protocols would significantly enhance interlaboratory comparability and method robustness. Method validation must be extended to include a broader array of animal-derived and processed foods to more accurately reflect real-world dietary exposure. Streamlining sample preparation workflows is also crucial for facilitating high-throughput applications in routine testing laboratories, particularly in low-resource settings. Additionally, high-resolution mass spectrometry and non-targeted screening approaches should be employed to identify coformulants or transformation products that may contribute to a cumulative concentration of GLYP and AMPA in food matrices. Finally, greater international cooperation will be crucial in developing harmonized protocols that ensure global consistency in GLYP and AMPA monitoring. Taken together, while substantial progress has been achieved in the analytical determination of GLYP and AMPA, existing methodologies remain limited by challenges related to analyte chemistry, matrix variability, and operational demands. A coordinated interdisciplinary approach is necessary to refine and validate methods that are both sensitive and broadly applicable, thereby supporting effective food safety surveillance and regulatory compliance worldwide. Conceptualization, L.M.A.-E. and Z.V. ; methodology, A.D.G.-C., L.M.A.-E., I.V.-C., F.M.-E., J.M.R.-G., J.M.S.-J., C.A.V.-C., I.B.-L., R.I.A.-G. and Z.V. ; investigation, A.D.G.-C., L.M.A.-E., I.V.-C., F.M.-E., J.M.R.-G., J.M.S.-J., C.A.V.-C., I.B.-L., R.I.A.-G. and Z.V. ; writing--original draft preparation, A.D.G.-C., L.M.A.-E., I.V.-C., F.M.-E., J.M.R.-G., J.M.S.-J., C.A.V.-C., I.B.-L., R.I.A.-G. and Z.V. ; writing--review and editing, A.D.G.-C., L.M.A.-E., I.V.-C., F.M.-E. and Z.V. ; project administration, Z.V. ; funding acquisition, Z.V. All authors have read and agreed to the published version of the manuscript. **Funding:** This research was funded by Secretaria de Ciencia, Humanidades, Tecnologia e Innovacion (SECIHTI), project number 322722. **Acknowledgments:** The authors would like to thank Secretaria de Ciencia, Humanidades, Tecnologia e Innovacion (SECIHTI), Mexico, for financial support. [PERSON] (CVU: 1028213) gratefully acknowledge the financial support for the scholarship number 4025668 from SECIHTI-Mexico for Ph.D. studies in Science in Microbiology and Molecular Biotechnology program from the Centro Universitario de Ciencias Exactas e Ingenierias (CUCEI) of University of Guadalajara. **Conflicts of Interest:** The authors declare no conflicts of interest.