This study developed and optimized a methodology based on controlled centrifugation for the segmented extraction of extracellular fluid in Apis mellifera bees. Three critical variables were analyzed: relative centrifugal force, centrifugation time, and the number of individuals processed, ensuring the reproducibility and efficiency of the procedure. The results demonstrated significant differences in the volume of fluid recovered from different body segments, with the abdomen yielding the highest volumes, followed by the thorax and the head. UV-Vis spectroscopic characterization revealed distinct optical features for the samples, identifying specific absorbance peaks unique to each segment. Furthermore, biochemical analysis using Benedict's reagent confirmed the presence of reducing sugars, with head samples displaying the most intense coloration. These findings underscore the importance of segment-specific analysis to gain deeper insights into the physiology and metabolism of bees. The proposed methodology offers a novel and robust tool for physiological, metabolic, and ecotoxicological studies, facilitating the assessment of environmental and contaminant impacts on pollinator health.
