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Real-time monitoring of culture medium- and species-specific transformation of zinc oxide nanoparticles in bacterial systems using coupled CyElFFF-ICPMS

Weichen Zhao,Zhangguo Wu,Meseret Amde,Hongwei Liu,Yan Li,Quanlong Wang,Yongguang Yin,Maoyong Song,Zhiqiang Tan,Yukui Rui,Qinghua Zhang,Bruce K. Gale,Guibin Jiang

Zinc oxide nanoparticles (ZnO NPs) have emerged as promising nanoagrochemicals due to their high utility and efficiency. For the first time, we employed an online-coupled cyclical electrical field-flow fractionation and inductively coupled plasma mass spectrometry (CyElFFF-ICPMS) system for rapid (<10 min) simultaneous characterization and quantification of ZnO NPs and Zn(II) ions in aqueous matrices, achieving a detection limit of 0.03 μg/mL and 92 % recovery. CyElFFF-ICPMS analysis demonstrated that ZnO NPs remain well-dispersed in ultrapure water but formed surface coronas in yeast mannitol broth prior to precipitation, demonstrating culture medium-specific stability profiles. Crucially, we observed divergent transformation mechanisms between beneficial bacteria. Gram-positive bacteria ( Bacillus subtilis ) stabilized ZnO NPs via biomolecular corona formation, while Gram-negative bacteria ( Sinorhizobium fredii ) promoted extracellular ZnO NPs aggregation and dissolution via protein-rich extracellular polymeric substance secretion, as validated by 3D excitation-emission matrix fluorescence spectroscopy. Intracellular transformation diverged markedly after 6 h: B. subtilis reduced internalized Zn(II) to ZnO nanocomposites through activated the detoxification mechanism, contrasting with S. fredii 's extensive intracellular dissolution and extracellular precipitation. These findings demonstrate that bacterial species-specific interactions critically govern the stability, transformation, and intracellular fate of ZnO NPs, providing essential mechanistic insights imperative for rational design of nanoagrochemicals and their environmental risk assessment.