Online Multimodal HILIC–ESI-HRMS/ICP-MS/MS Speciation Reveals Remodeling of Sulfur- and Phosphorus-Containing Metabolites Associated with Gold Nanoparticle Exposure in Transgenic ( cp 4 EPSPS Gene) Soybean Callus

Abstract A multimodal HILIC–ESI-HRMS/ICP-MS/MS workflow profiled sulfur- and phosphorus-containing metabolites in transgenic soybean (Glycine max) calli exposed to 100 μg L–1 gold nanoparticles (Au NPs). Calli were cultured on proliferation medium for 30 days, with or without Au NPs. Thirty-eight analytically supported P- or S-containing features corresponded to 28 putative annotations. Exploratory analysis showed lower relative signals for reduced sulfur metabolites in Au NP-treated calli: methionine (log2FC = −5.20) and l-homomethionine (log2FC = −4.10). Oxidized sulfur metabolites (methionine sulfoxide, cysteine sulfinic and sulfonic acids, cystine, and sulfolactate) were detected only in the Au NP-treated profile. At the same time, thiamine showed a higher relative signal (log2FC = +2.13) in the treated sample. Phosphorylcholine showed a lower relative signal (log2FC = −6.21), whereas LPC(16:0) showed a higher relative signal (log2FC = +1.82) in the Au NP-treated profile. These patterns are consistent with differences in sulfur redox, thiamine-associated, nucleotide-sugar, and phospholipid metabolism.