Enhancing Pesticide Residue Quantification in Honey through Integrated Chemo-physical and Chemometric Separation using LC-MS Spectrometry: Comparison with Multiple Reaction Monitoring

Honey, a naturally occurring product generated by bees, holds considerable nutritional and ethnomedical significance. Its constitution, primarily comprising carbohydrates and water, demonstrates variability rooted in the sources of nectar. The widespread consumption of honey on a global scale has precipitated the imperative for rigorous quality control and vigilance against contamination within commercial transactions. The utilization of pesticides in agricultural practices, pivotal for augmenting yields, can engender the presence of pesticide residues in foodstuffs, thereby necessitating the establishment of Maximum Residue Limits (MRLs). The main objective of the present study is to monitor residues of some major pesticides in honey samples collected from important areas of honey production in the country. Divergences in MRLs across different nations have engendered contentious discussions in international trade circles. The comprehensive analysis of pesticide residues within honey demands the application of sophisticated methodologies owing to its intricate matrix [1]. Modern analytical techniques such as GC-MS/MS and UHPLC-MS/MS, coupled with proficient extraction and purification protocols, offer stringent detection thresholds for the analysis of multiple pesticide residues, congruent with established MRLs. Among these methods, LC-MS/MS in Multiple Reaction Monitoring (MRM) mode emerges as a benchmark approach for the analysis of intricate samples [2]. This mode enables the selective isolation of target analytes while excluding background noise, delivering enhanced sensitivity and reproducibility. Alternatively, LC-MS operated in full scan mode, reliant solely on a single-quadrupole mass spectrometer, has the potential to circumvent challenges like peak overlapping. Nevertheless, full scan LC-MS is marked by a lack of selectivity, thereby complicating the resolution and precise quantification of targeted compounds, particularly when compared to MRM. The phenomenon of overlapping peaks and the presence of interfering substances are commonplace in the quantitative analysis of complex samples like biological fluids. To surmount these intricacies, a shrewd approach capitalizing on three-way LC-MS data and second-order calibration was devised. This strategy achieves quantitative precision comparable to MRM, leveraging the concept of mathematical separation in lieu of the conventional optimization of physical separations. This innovative strategy bestows amplified selectivity and sensitivity upon the analytical process. Its practicality is underscored by its efficiency, cost-effectiveness, and eco-friendliness, thus emerging as a viable alternative for the precise determination of target analytes within intricate systems. In a specific project context, the analysis of 38 pesticides within honey samples was accomplished through the utilization of LC-MS data and advanced chemometric techniques. The inadequacies of the chromatographic approach in addressing peak separation intricacies were compensated through the employment of chemometrical separation. The integration of fchemometrical and physico-chemical separation techniques led to a discernible augmentation in the selectivity of the analytical methodology.